No-wash aggregation-inducible cell membrane targeting staining reagent based on purine skeleton and its preparation method and application

An aggregation-induced, cell membrane technology, applied in the preparation of test samples, styryl dyes, chemical instruments and methods, etc., can solve the problems of low accuracy of imaging results, complicated operation process, inability to connect sensing, etc., to avoid Interference from background light, economical availability of raw materials, and low overall cost

Active Publication Date: 2020-02-07
SICHUAN UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] The purpose of the present invention is to provide a purine skeleton-based no-wash aggregation-inducible cell membrane targeting staining reagent and its preparation method and application to solve the problem of the existing cell membrane phospholipid molecular layer targeting dyes due to the need for multiple washings. The process is complicated, time-consuming, the accuracy of imaging results is low, and the problem of inability to connect to sensing

Method used

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  • No-wash aggregation-inducible cell membrane targeting staining reagent based on purine skeleton and its preparation method and application
  • No-wash aggregation-inducible cell membrane targeting staining reagent based on purine skeleton and its preparation method and application
  • No-wash aggregation-inducible cell membrane targeting staining reagent based on purine skeleton and its preparation method and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0067] The preparation method of the purine skeleton-based aggregation-inducible cell membrane targeting staining reagent of this embodiment comprises the following steps:

[0068] (1) Synthesis of the first intermediate: 2,6-dichloro-9-n-propyl-9-hydrogen-purine

[0069] The synthetic route is as follows:

[0070]

[0071] 2,6-Dichloropurine (1.0 mmol), 1-bromopropane (1.5 mol) and potassium carbonate (3.0 mmol) were mixed and stirred in DMSO (5 mL) for 6 hours, then 100 mL of water was added. The organic layer was separated, and the aqueous layer was extracted with ethyl acetate (30 mL×3). The organic extracts were washed with brine and treated with Na 2 SO 4 dry. After the solvent was removed and distilled under reduced pressure, it was purified by 200-300 mesh silica gel column chromatography. Elution with petroleum ether / ethyl acetate (3:2) gave the first intermediate as a white solid with a yield of 57%. The eluent is ethyl acetate / petroleum ether=2:3 (V / V). Fi...

Embodiment 2

[0090] This example is basically the same as Example 1, except that the third intermediate R is changed 3 The substituent, its synthetic route is as follows:

[0091]

[0092]After adding the third intermediate (381 mg, 1 mmol) and 1,4-lutidine-1-iodide (235 mg, 1 mmol) in ethanol (10 mL), piperidine (0.05 mL) was dropped into the stirring liquid. Then the mixture was stirred at room temperature for about 12 hours. After the completion of the reaction was monitored by thin-layer chromatography, the solvent was removed by rotary evaporation, and then dissolved with saturated potassium hexafluorophosphate acetone solution (10 mL). After stirring at room temperature for 2 hours, the acetone was distilled off under reduced pressure, and then the crude product was obtained by filtration. The crude product was purified by neutral alumina column chromatography. Elution with methanol / dichloromethane=20 / 1 (V:V) gave a yellow solid with a yield of 37%.

[0093] 1 H NMR (400MHz, D...

Embodiment 3

[0096] This example is basically the same as Example 1, except that the picoline salt in step (4) is changed to 4-methyl-1-(3-(trimethylammonium) propyl)pyridinium-1-ammonium bromide , its synthetic route is as follows:

[0097]

[0098] A dark brown solid was obtained in 33% yield. 1 H NMR (400MHz, DMSO-d 6 )δ9.23-9.20(d,1H),9.09-9.03(m,3H),8.71-8.69(s,1H),8.62-8.57(d,2H),8.37-8.33(d,2H),8.20- 8.14(d,1H),8.02-7.98(d,2H),7.74-7.67(m,2H),7.47-7.42(t,1H),7.34-7.28(t,1H),6.95-6.93(d,1H ),4.65-4.60(t,2H),4.40-4.34(t,2H),3.11-3.7(s,9H),1.99-1.92(m,2H),0.89-0.83(t,3H). 13 C NMR (101MHz, DMSO-d 6 )δ157.08,154.10,153.50,149.06,146.25,145.04,140.83,139.78,137.37,135.68,130.69,129.15,129.00,128.93,124.83,124.58,124.36,123.26,121.57,121.29,116.71,109.99,108.61,62.25,57.24 ,52.91,45.50,24.47,23.06,11.51.HRMS(ESI):m / z:Calcd for C 35 h 39 f 6 N 7 P + :702.2903; [M-PF 6 ] + Found: 702.2902.

[0099] The hydrogen spectrum, carbon spectrum and high-resolution mass spectrum of t...

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Abstract

Disclosed in the present invention is a purine skeleton-based no-wash, aggregation-induced plasma membrane targeted staining reagent, and a preparation method and an application thereof, the present invention using a purine skeleton as a base for a plasma membrane stain, obtaining, by means of reasonable design regulating a fat-soluble end and a hydrophilic end, a purine skeleton-based plasma membrane targeted staining reagent, the reagent being capable of both rapid targeted staining and remaining on a plasma membrane for a relatively long time, benefiting long-term detection. In addition, given that the staining reagent has characteristics of an aggregation-induced compound, emitting light weakly or not emitting light in solution, and exhibiting strong fluorescence when not in solution, causing the stain to further have specific no-wash performance. The preparation method of the present invention has high yield and mild reaction conditions, and the prepared staining reagent has a large Stokes shift and high targeting.

Description

technical field [0001] The invention relates to the field of biochemistry, in particular to the technical field of biomembrane targeted staining, in particular to a purine skeleton-based wash-free aggregation-inducible cell membrane targeted staining reagent and its preparation method and application. Background technique [0002] The cell membrane (also known as the plasma membrane or cytoplasmic membrane), consisting of a phospholipid bilayer with intercalated proteins, is a biological membrane that separates the interior of a cell from its external environment and protects the cell from its environment. important components of cells. It has been shown to be involved in various cellular processes and biological functions, such as cell migration, cell spreading, phagocytosis, endocytosis, exocytosis, and selective permeability of substances. Cell membrane abnormalities are important markers of poor cell status and various diseases. Therefore, the development of highly sel...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07D473/34C07D473/00C09B23/14C09K11/06G01N1/30G01N21/64
CPCC07D473/00C07D473/34C09B23/14C09K11/06G01N1/30G01N21/64
Inventor 李坤石磊余孝其刘艳红于抗抗
Owner SICHUAN UNIV
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