Lysosome hypochlorous acid fluorescence probe as well as preparation method and application thereof

A fluorescent probe and hypochlorous acid technology, which is applied in the field of small molecule probes, can solve the problems of few reports of two-photon hypochlorous acid fluorescent probes, and achieve the effects of high sensitivity, wide concentration range and strong specificity

Inactive Publication Date: 2017-11-07
UNIV OF JINAN
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

But so far, there are very few reports on tw

Method used

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  • Lysosome hypochlorous acid fluorescence probe as well as preparation method and application thereof
  • Lysosome hypochlorous acid fluorescence probe as well as preparation method and application thereof
  • Lysosome hypochlorous acid fluorescence probe as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] Example 1 Synthesis of fluorescent probe TPS-Lyso-HClO

[0043] (1) Synthesis of compound 4-bromo-N-(2-morpholinoethyl)naphthalimide:

[0044]In a 100 mL round bottom flask, add 1 mmol of 4-bromo-1,8-naphthalic anhydride, 1.2 mmol of aminoethylmorpholine, and 5 mL of ethanol, heat to reflux for 3 h, cool to room temperature, and depressurize After filtering, the filter cake was washed with ethanol for 2-3 times, and dried under vacuum to obtain 4-bromo-N-(2-morpholinoethyl)naphthalimide as a light gray solid. Yield: 81%.

[0045] 1 H NMR (400 MHz, DMSO- d 6 ) δ 8.57 (dd, J 1 = 7.2 Hz, J 2 = 0.8 Hz, 1H), 8.55 (dd, J 1 = 8.4 Hz, J 2 = 0.8 Hz, 1H), 8.34 (d, J = 7.6 Hz, 1H), 8.22 (d, J =7.6 Hz, 1H), 8.00 (dd, J 1 = 8.4 Hz, J 2 = 7.2 Hz, 1H), 4.18 (t, J = 6.8 Hz, 2H), 3.53 (t, J = 4.8 Hz, 4H), 2.57 (t, J = 7.2 Hz, 2H), 2.47 (s, 4H); 13C NMR (100 MHz, DMSO- d 6 ): 163.30, 163.25, 133.09, 132.07, 131.82, 131.44, 130.20, 129.63, 129.27, 128.68, 123.11, 122.33, 66.6...

Embodiment 2

[0058] Example 2 The selectivity of TPS-Lyso-HClO to different molecules and ions

[0059] The TPS-Lyso-HClO hypochlorous acid fluorescent probe prepared in Example 1 was dissolved in ethanol to prepare a 1 mM stock solution.

[0060] Prepare 5 mL of various conventional ions and active oxygen in phosphate buffer (10 mmol / LPBS, pH=5.5 and pH=7.4) with a concentration of 40 mM, respectively.

[0061] Add 25 μL probe mother solution, 75 μL ethanol and 100 mmol / L analyte solution to the test tube, and dilute to 10 mL with 10 mmol / L PBS with pH=5.5 and pH=7.4 respectively, shake well and perform fluorescence detection (λ ex =390 nm; pH 7.4, λ em =446 nm and λ em =585 nm; pH 5.5, λ em =490 nm and λ em =585 nm), the ratio of the short-wave fluorescence intensity to the long-wave fluorescence intensity is the ordinate (I 446 / I 585 or I 490 / I 585 ), to establish the fluorescence intensity ratio map of different molecules and ions, such as image 3 As shown, 1-12 are probes...

Embodiment 3

[0062] Example 3 TPS-Lyso-HClO Fluorescence Intensity Variations with Different Concentrations of Hypochlorous Acid

[0063] Prepare 10 mL of sodium hypochlorite mother liquor with a concentration of 100 mmol / L for later use (sodium hypochlorite dissociates into ClO in aqueous solution - ).

[0064] Dilute the probe stock solution prepared in Example 1 to a fluorescent probe working solution with a concentration of 2.5 μmol / L, react with 15 hypochlorous acid (0-250 μmol / L) at equal concentrations, and perform fluorescence detection (λ ex =390 nm; pH 7.4, λ em =446 nm and λ em=585 nm; pH 5.5, λ em =490 nm and λ em =585 nm), calculate the fluorescence intensity in each system, take the fluorescence intensity as the ordinate and the scanning wavelength as the abscissa as Figure 4 In a, b, and d, the normalized fluorescence intensity is taken as the ordinate, and the scanning wavelength is taken as the abscissa. Figure 4 In c, where a and b are when the pH is 7.4, and c a...

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Abstract

The invention provides a lysosome hypochlorous acid fluorescence probe which has a chemical name of 8,8,9-trimethyl-5-(2-morpholinoethyl)-9,10-dihydrobenzo[de] pyrrole[2,3-9]isoquinoline-4,6(5H,8H) diketone, called TPS-Lyso-HClO for short. The lysosome hypochlorous acid fluorescence probe is capable of detecting the content of hypochlorous acid in solution and lysosome: the excitation wavelength of the solution is detected to be 380nm; when the pH value is 7.0-7.4, the emission wavelength is 446nm; when the pH value is 5-5.5, the emission wavelength is 490nm; the two-photon cell fluorescence imaging excitation wavelength is 780nm, and the emission bands refer to 480-540nm and 560-620nm; and the single photon cell fluorescence imaging excitation wavelength is 405nm, and the emission bands refer to 480-540nm and 560-620nm. The lysosome hypochlorous acid fluorescence probe disclosed by the invention is high in specificity of a hypochloric acid reaction and is capable of resisting against multiple interferents; and the fluorescence probe has lysosome targeting property and is high in sensitivity, low in detection limit and wide in detected hypochloric acid concentration range. The lysosome hypochlorous acid fluorescence probe disclosed by the invention is a simple, rapid and sensitive hypochloric acid molecular specificity detection reagent and has wide application prospects in the field of bio-molecular detection.

Description

technical field [0001] The invention relates to a photosensitive fluorescent probe for rapidly detecting hypochlorous acid and its preparation and application in detecting lysosome hypochlorous acid, belonging to the field of small molecule probes. Background technique [0002] Hypochlorous acid (HClO) is a kind of active oxygen, as a highly effective bactericide, it plays an important role in the immune system of life. Endogenous hypochlorous acid in cells is mainly produced by the myeloperoxidase system in leukocytes (such as monocytes, eosinophils, neutrophils, etc.). Cellular immune response can produce hypochlorous acid, but once the concentration of hypochlorous acid in cells is abnormal, it will cause various diseases including rheumatoid arthritis, cardiovascular disease and cancer. In intracellular lysosomes, as a kind of relatively important organelles, hypochlorous acid can maintain the balance of lysosome oxidation-reduction, and play a very important role in th...

Claims

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Application Information

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IPC IPC(8): C07D471/06C09K11/06G01N21/64
CPCC07D471/06C09K11/06C09K2211/1029C09K2211/1033G01N21/6428
Inventor 林伟英唐永和马燕燕
Owner UNIV OF JINAN
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