KR100930969B1 - Acridine derivatives, preparation method thereof and selective detection method of mercury ion and / or cadmium ion using the same - Google Patents

Abstract

The present invention relates to an acridine derivative, a method for preparing the same, and a method for selectively detecting mercury ions and / or cadmium ions using the same, and more specifically, an acridine derivative and a 4,5-bis (bromomethyl) acrylic. Reacting by adding an azacrown compound to a solution of a dine and a base in a solvent; Stirring the reaction product of the above step at room temperature and then filtering and evaporating the solvent; And a method for preparing an acridine derivative, which comprises performing column chromatography on the result of evaporating the solvent, and a method for selectively detecting mercury ions and / or cadmium ions existing in an aqueous solution environment or a living body using the same. to provide.

In the acridine derivative according to the present invention, a large chelate amplification fluorescence effect was observed at pH 7.4 only for the addition of mercury ions and cadmium ions, and conventionally, while heavy metal ions were mainly detected in organic solvents, The compound can not only detect mercury ions and cadmium ions in aqueous solution, but can also be permeated into cells to detect intracellular mercury ions and cadmium ions. It can be usefully used.

Mercury ions, cadmium ions, acridine, chelate amplified fluorescence

Description

Acridine derivatives, preparation method etc. and selective detection method of mercury ion and / or cadmium ion using the same

The present invention provides an acridine derivative, more specifically an acridine derivative that effectively recognizes mercury ions and cadmium ions at PH 7.4, ie, azacrown or azacia crown ligands immobilized on the acridine phosphor and thus in aqueous solution or in vivo. Acridine derivatives exhibiting chelate enhanced fluorescence (CHEF) effects on mercury ions and cadmium ions, methods for their preparation, and selective detection of mercury ions and / or cadmium ions in aqueous environments or in vivo using them It is about a method.

The design and research of new sensors for key substances and ions in vivo has been actively conducted. Recent understanding and research on supramolecule chemistry has shown great potential for the design of host compounds that can selectively bind ions or other types of guest compounds. In addition, it has been of great help in the development of fluorescent chemosensors, which can easily observe selective binding with guest compounds using fluorescence changes.

Fluorescence is a photochemical phenomenon that occurs when photons with a specific light wave (excitation wavelength) collide with an indicator molecule and electrons are excited to a high energy level as a result of the collision. Among the various analytical methods, fluorescence has the advantage of being able to observe signals even at 10 ^-9 M concentrations due to their excellent sensitivity. Recently, studies on fluorescence chemical sensors for cations, anions and neutral organic molecules using these properties have been published.

High selective detection of metal ions in physiological conditions is very important in the development of fluorescence chemical sensors for environmental and biological applications [ Fluorescent Chemosensors for Ion and Molecular Recognition , Czarnik, AW Ed., American Chemical Society: Washington, DC, 1993.]. In particular, mercury ions and cadmium ions are biologically important heavy metals because they have a detrimental effect on human health. Emissions by oceans and volcanoes [Renzoni, A; Zino, F .; Franchi, E. Environ . Res . 1998 , 77 , 68.], gold mining [Malm, O. Environ . Res . 1998 , 77 , 73.], or mercury contamination by solid waste incineration has been a major issue because of extreme toxicity to the immune system, genes and nervous system. On the other hand, chronic exposure to cadmium can cause kidney dysfunction may increase the incidence of certain cancers in the [Cadmium in the Human Environment : Toxicity and Carcinogenicity ; Nordberg , GF, Herber, RFM, Alessio, L., Eds .; Oxford University Press: Oxford, 1992]. As such, mercury and cadmium are very dangerous poisonous pollutants, but unfortunately they are present in our environment. Mercury and cadmium in the environment accumulate throughout the food chain. The higher the level of accumulation, the higher the level of contaminants that accumulate in humans through food intake. Mercury and cadmium are ionic forms that dissolve well in water through several pathways (Hg ^{2 +} , Cd ^{2 +} ) and accumulates in fish and other foods. Therefore, naturally, the accumulation of mercury and cadmium occurs in humans who eat them. Therefore, monitoring the mercury and cadmium content in foods can be one of the important activities to protect human health.

Therefore, great attention is being paid to the development of a new fluorescence chemical sensor having sufficient selectivity for the metal ion. Many fluorescence chemical sensors have been reported to selectively detect mercury ions and cadmium ions, but mercury ions [KS Kim et al . , Org . Lett . 2007 , 9 , 907; X. Qian et al . , Org . Lett . 2006 , 8 , 3721; S.-K. Chang et al . , Org . Lett . 2006 , 8 , 3413; J.-G. Xu et al . , Org . Lett . 2006 , 8 , 859; Chang, S.-K et al. Org . Lett . 2006 , 8 , 371; J. Tae et al . , J. Am . Chem . Soc . 2005 , 127 , 16760; CJ Chang et al . , J. Am . Chem . Soc . 2005 , 127 , 10124; L. Jia et al . , J. Am . Chem . Soc . 2004 , 126 , 2272; DC Magri et al., Tetrahedron 2005 , 61 , 8551] and cadmium ions [X. Peng, et al . , J. Am . Chem . Soc . 2007 , 129 , 1500; PB Savage et al . , Org . Lett . 2005 , 7 , 1105; S.-K. Chang et al., Tetrahedron Lett . 2005 , 46 , 125; R. Parkesh et al . , Org . Lett . 2003 , 5 , 4065; J. Yoon et al . , Bull . Korean Chem. Soc . 2003 , 24 , 1032; J. Soto et al . , J. Chem . Soc ., Dalton Trans . 2002 , 1769; J. Yoon et al . , Org . Lett . 2001 , 3 , 3455 are rare examples of fluorescence chemical sensors that selectively increase fluorescence.

Accordingly, the present inventors can detect mercury ions and cadmium ions in aqueous solution or cells by introducing acridine derivatives, azacrown or azacia crown, while studying compounds selective for mercury ions and cadmium ions. Confirmed and completed the present invention.

It is an object of the present invention to provide an acridine derivative.

Another object of the present invention is to provide a method for preparing an acridine derivative.

Still another object of the present invention is to provide a method for selectively detecting mercury ions and / or cadmium ions existing in an aqueous solution environment or a living body using an acridine derivative.

In order to achieve the above object, the present invention provides an acridine derivative.

The present invention also provides a method for preparing an acridine derivative.

Furthermore, the present invention provides a method for the selective detection of mercury ions and / or cadmium ions present in an aqueous solution environment or a living body using an acridine derivative.

In the acridine derivative according to the present invention, a large chelate amplification fluorescence effect was observed at pH 7.4 only for the addition of mercury ions and cadmium ions, and conventionally, while heavy metal ions were mainly detected in organic solvents, The compound can not only detect mercury ions and cadmium ions in aqueous solution, but can also be permeated into cells to detect intracellular mercury ions and cadmium ions. It can be usefully used.

The present invention provides an acridine derivative represented by the following formula (1).

In the above formula, X is O or S.

The acridine derivative of Formula 1 according to the present invention is preferably an acridine-azacrown derivative of Formula 2 or an acridine-azacia crown derivative of Formula 3 below.

In addition, the present invention as shown in Scheme 1,

Reacting 4,5-bis (bromomethyl) acridine of Formula 4 by adding azacrown compound of Formula 5 to a solution of a base dissolved in a solvent (step 1);

Stirring the reaction product of step 1 at room temperature and then filtering and evaporating the solvent (step 2); And

It provides a method for producing an acridine derivative comprising the step (step 3) of purifying by performing column chromatography on the reaction product of step 2.

(X is O or S in the above scheme.)

Hereinafter, the present invention will be described in more detail step by step.

Step 1 according to the present invention is a step of reacting 4,5-bis (bromomethyl) acridine of Formula 4 by adding azacrown compound of Formula 5 to a solution of a base dissolved in a solvent.

The acridine derivative according to the present invention preferably uses 4,5-bis (bromomethyl) acridine as a starting material as shown in Scheme 1, and the base of step 1 is preferably potassium carbonate, Triethylamine, sodium carbonate, etc. can be used, More preferably, potassium carbonate can be used.

As the solvent of step 1, chloroform, dichloromethene, acetonitrile, or the like may be used, and chloroform may be preferably used.

In addition, the azacrown compound of step 1 may use 4,13-diaza-18-crown-6-ether or 1,10-diaza-4,7,14,17-tetrasiacyclooctadecane.

Step 2 according to the present invention is a step in which the reaction product of step 1 is stirred at room temperature, filtered, and then the solvent is evaporated.

The stirring may preferably be carried out for 24 hours and may be evaporated under vacuum.

Step 3 according to the present invention is a step of purifying by performing column chromatography on the reaction product of step 2.

As the separator of the column chromatography, methyl chloride and methanol may be mixed in a ratio of 99: 1.

In another aspect, the present invention provides a method for the selective detection of mercury ions and / or cadmium ions present in the aqueous solution environment or living body using the acridine derivative.

The detection of the mercury ions and / or cadmium ions may be carried out by exciting a complex of the acridine derivative and the mercury ions and / or cadmium ions to confirm chelate amplification fluorescence (CHEF) effects. Fluorescence imaging can detect mercury ions and / or cadmium ions present in vivo or in cells. In this case, the excitation wavelength for chelate amplification is preferably 356 nm.

The imaging can be carried out using an azacrown-acridine or azaciacrown-acridine and a complex of mercury ions and / or cadmium ions according to the invention using an image restoration microscope, for example a DeltaVision microscope. Can be.

Hereinafter, the present invention will be described in more detail with reference to Examples and drawings. However, the following examples are merely to illustrate the invention, but the content of the present invention is not limited by the following examples.

< Example  1> Azacrown - Acridine  Produce

To a reaction mixture of 4,5-bis (bromomethyl) acridine (500 mg, 1.36 mmol) and potassium carbonate (751 mg, 5.4 mmol) 4,13-diaza-18-crown 6-ether (360 mg) , 1.36 mmol) was added.

The reaction mixture to which the azacrown was added was stirred at room temperature for 24 hours and then filtered, and the solvent was evaporated under vacuum. The crude product was purified using alumina column chromatography using methylchloride and methanol as separators to give azacrown-acridine (540 mg, 84%) as a pale yellow oil.

¹ H-NMR (250 MHz, CDCl ₃ ) 8.71 (s, 1H), 7.87 (t, 4H, J = 7.7 Hz), 7.51 (t, 2H, J = 7.2 Hz), 4.71 (s, 4H), 3.28 -3.71 (m, 16H), 3.09 (t, 8H, J = 6.1 Hz);

¹³ C-NMR (62.5 MHz, CDCl ₃ ) 147.1, 139.5, 135.9, 130.7, 127.1, 126.3, 125.4, 70.4, 69.6, 55.1, 54.9;

MS (FAB) m / z = 466.2707 (M + H) &lt; + &gt;, calc. for C ₂₇ H ₃₆ N ₃ O ₄ = 466.2706.

< Example  2> Azacia Crown - Acridine  Produce

To a reaction mixture of 4,5-bis (bromomethyl) acridine (500 mg, 1.36 mmol) and potassium carbonate (751 mg, 5.4 mmol) 1,10-diaza-4,7,14,17-tetra Ciacyclooctadecane (440 mg, 1.36 mmol) was added.

The reaction mixture to which the azacia crown was added was stirred at room temperature for 24 hours and then filtered, and the solvent was evaporated under vacuum. The crude product was purified using alumina column chromatography using methylchloride and methanol as separators to give azaciacrown-acridine (530 mg, 70%) as a pale yellow oil.

¹ H-NMR (250 MHz, CDCl ₃ ) 8.69 (s, 1H), 7.89 (dd, 2H, J = 8.5 Hz & 1.2 Hz), 7.69 (d, 2H, J = 6.8 Hz), 7.45 (dd, 2H , J = 8.3 Hz & 6.8 Hz), 4.57 (s, 4H), 3.03 (t, 8H, J = 6.9 Hz), 2.74 (m, 8H), 2.46 (m, 8H);

¹³ C-NMR (62.5 MHz, CDCl ₃ ) 147.2, 136.6, 134.7, 131.3, 127.9, 126.5, 125.3, 55.9, 53.2, 32.5, 29.4;

MS (FAB) m / z = 530.1786 (M + H) &lt; + &gt;, calc. for C ₂₇ H ₃₆ N ₃ S ₄ = 530.1792.

<Experimental Example 1> aza crown-observed fluorescence change following the binding of acridine and heavy metal ions

1-1. Azacrown - Acridine and  Fluorescence titration experiment for the binding of heavy metal ions

Experiments include Ag ⁺ , Ca ^{2 +} , Cd ^{2 +} , Co ^{2 +} , Cs ⁺ , Cu ^{2 +} , Hg ^{2 +} , K ⁺ , Li ⁺ , Mg ^{2 +} , Mn ^{2 +} , Na ⁺ , Ni ²⁺ and Zn ^{2 +} was used as the heavy metal ions.

Fluorescent metal ion titration solution was prepared using demineralized water distilled from a stock solution (1 mM) of metal perchlorate salts in duplicate and prepared azacrown in Example 1 A stock solution of acridine (0.01 mM) was prepared using dimethylsulfoxide (DMSO).

For the test solution, 40 μl of the probe stock solution was added to the test tube, and an appropriate amount of each metal stock solution (0 to 400 μl, for example, 4 μl for 1 equivalent of metal and 40 μl for 10 equivalents of metal) was added thereto, followed by 100 mM. Prepared by diluting the solution to 4 mL with HEPES buffer (pH 7.4). All fluorescence changes were measured at excitation wavelength (excitation, 356 nm). The slit width of excitation and emission was 5 nm and the results are shown in FIG. 1.

As shown in FIG. 1, azacrown-acridine according to the present invention exhibited a unique change in emission spectrum for the addition of mercury ions and cadmium ions at pH 7.4. That is, the addition of mercury ions showed a large chelate amplification fluorescence effect, and the addition of cadmium ions showed a relatively smaller chelate amplification fluorescence effect than that of hydrogen ions. On the other hand, no significant change was seen for the addition of 100 equivalents of other heavy metal ions. The reason why the azacrown-acridine exhibits a chelate amplification fluorescence effect selectively only on hydrogen ions and cadmium ions is determined by the nitrogen of the azacrown ligand and acridine. Therefore, it can be seen that the azacrown-acridine according to the present invention binds selectively to mercury ions and cadmium ions.

1-2. Fluorescence titration experiment of azacrown-acridin for concentration-specific mercury ions

In order to determine the binding properties between the azacrown-acridine and mercury ions, the following experiment was performed.

A stock solution of mercury ions (10 mM) was prepared using deionized water distilled off in duplicate. In addition, the stock solution of azacrown-acridine prepared in Example 1 (0.01 mM) was prepared using dimethyl sulfoxide (DMSO).

12 μl of the test solution was added to the test tube so that the final concentration of the probe stock solution was 3 μM, and 1.5 to 60 μM of the aliquot of the mercury ion storage solution was added to the solution, followed by 100 mM HEPES buffer (pH 7.4). Prepared by diluting the solution to 4 ml. All fluorescence changes were measured at excitation wavelength (excitation, 356 nm). The slit width of excitation and emission was 5 nm and the results are shown in FIG. 2.

As shown in Fig. 2, the azacrown-acridine according to the present invention has a total fluorescence increase of 10 times or more as the addition amount of mercury ions increases from 0.5 equivalents to 20 equivalents when the final concentration is 3 μM at pH 7.4. Indicated. Through titration experiments with mercury ions, the binding constant of the compound of the present invention and the mercury ion complex was found to be 1.18 × 10 ⁵ M ⁻¹ .

1-3. Fluorescence titration experiment of azacrown-acridin for concentration-specific cadmium ions

In order to determine the binding properties between the azacrown-acridine and cadmium ions, the following experiment was performed.

A stock solution of cadmium ions (10 mM) was prepared using deionized water distilled off in duplicate. In addition, the stock solution of azacrown-acridine prepared in Example 1 (0.01 mM) was prepared using dimethyl sulfoxide (DMSO). The stock solutions were used on the day of preparation.

The test solution contained 12 μl of the probe stock solution (the stock solution of azacrown-acridine of the present invention) into the test tube so that the final concentration was 3 μΜ, and 5 to 200 equivalents of aliquots of the cadmium ion stock solution were added. After 15 to 600 μM was added, the solution was diluted with 4 mL of 100 mM HEPES buffer (pH 7.4). All fluorescence changes were measured at excitation wavelength (excitation, 356 nm). The slit width of excitation and emission was 5 nm and the results are shown in FIG. 3.

As shown in Figure 3, the azacrown-acridine according to the present invention, when the final concentration is 3 μM at pH 7.4, when the addition amount of cadmium ion increases from 5 equivalents to 200 equivalents, the overall increase in fluorescence is more than 5 times Indicated. Through titration experiments with cadmium ion, the binding constant of the compound of the present invention and the cadmium ion complex was found to be 4.48 × 10 ³ M ^-1 .

Experimental Example 2 Observation of Fluorescence According to Azacia Crown-Acridin and Heavy Metal Ions

2-1. Azacia Crown - Acridine and  Fluorescence titration experiment for the binding of heavy metal ions

Experiments include Ag ⁺ , Ca ^{2 +} , Cd ^{2 +} , Co ^{2 +} , Cs ⁺ , Cu ^{2 +} , Hg ^{2 +} , K ⁺ , Li ⁺ , Mg ^{2 +} , Mn ^{2 +} , Na ⁺ , Ni ²⁺ and Zn ^{2 +} was used as the heavy metal ions.

Fluorescent metal ion titration solution was prepared using demineralized water distilled off a stock solution (1 mM) of metal perchlorate salts. In addition, the stock solution of azacia crown-acridine prepared in Example 2 (0.01 mM) was prepared using dimethyl sulfoxide (DMSO).

For the test solution, 40 μl of the probe stock solution was added to the test tube, and an appropriate amount of each metal stock solution (0 to 400 μl, for example, 4 μl for 1 equivalent of metal and 40 μl for 10 equivalents of metal) was added, followed by 100 mM. Prepared by diluting the solution to 4 mL with HEPES buffer (pH 7.4). All fluorescence changes were measured at excitation wavelength (excitation, 356 nm). The slit width of excitation and emission was 5 nm and the results are shown in FIG. 4.

As shown in Figure 4, the emission spectrum of azaciacrown-acridine according to the present invention showed a unique change for the addition of mercury ions and cadmium ions at pH 7.4. The addition of mercury ions showed a small chelate amplification fluorescence effect, but showed a large selectivity, whereas the addition of cadmium ions showed a better chelate amplification fluorescence effect than a mercury ion. On the other hand, no significant change was shown for the addition of 100 equivalents of other anions. The reason why azacia crown-acridine exhibits chelate amplification fluorescence effect selectively only for hydrogen ions and cadmium ions is determined by the nitrogen of azacia crown ligand and acridine. Therefore, it can be seen that the azacia crown-acridine according to the present invention binds selectively to mercury ions and cadmium ions.

2-2. Fluorescence Titration Experiment of Azaciacrown-Acridin for Concentrations of Mercury Ions

In order to determine the binding properties between the azacia crown-acridine and mercury ions, the following experiment was performed.

A stock solution of mercury ions (10 mM) was prepared using demineralized water distilled twice. In addition, the stock solution of azacia crown-acridine prepared in Example 2 (0.01 mM) was prepared using dimethyl sulfoxide (DMSO).

The test solution contains 12 μl of the final concentration of the probe storage solution into the test tube, add 0.6-6 μM of the aliquot of the mercury ion storage solution to 0.2 to 2 equivalents, and then add 100 mM HEPES buffer (pH 7.4). ) To dilute the solution to 4 ml. All fluorescence changes were measured at excitation wavelength (356 nm). The slit width of excitation and emission was measured at 5 nm and the results are shown in FIG. 5.

As shown in Fig. 5, the azacrown-acridine according to the present invention has a total fluorescence increase of 4 times or more as the addition amount of mercury ions increases from 0.2 equivalents to 2 equivalents when the final concentration is 3 μM at pH 7.4. Indicated. Through titration experiments with mercury ions, the binding constant of the compound of the present invention and the mercury ion complex was found to be more than 10 ⁸ it was found to be very selective to mercury ions.

2-3. Fluorescence Titration Experiments of Azacia Crown-Acridines on Cadmium Ions by Concentration

In order to determine the binding properties between the azacia crown-acridine and cadmium ions, the following experiment was performed.

A stock solution of cadmium ions (10 mM) was prepared using deionized water distilled off in duplicate. In addition, the stock solution of azacia crown-acridine prepared in Example 2 (0.01 mM) was prepared using dimethyl sulfoxide (DMSO).

12 μl of the test solution was added to the test tube so that the final concentration of the probe storage solution was 3 μM, and 3 to 900 μM of the aliquot of the cadmium ion storage solution was added to 1 to 300 equivalents, followed by 100 mM HEPES buffer (pH 7.4). Prepared by diluting the solution to 4 ml. All fluorescence changes were measured at the excitation wavelength (356 nm). The excitation and emission slit width was 5 nm and the results are shown in FIG. 6.

As shown in Figure 6, azacrown-acridine according to the present invention, when the final concentration is 3 μM at pH 7.4, a total fluorescence increase of 6 times or more as the amount of addition of cadmium ions increases from 1 equivalent to 300 equivalents Indicated. Through titration experiments with cadmium ions, the binding constant of the compound of the present invention and the cadmium ion complex was found to be 3.28 × 10 ⁴ M ^-1 .

analysis

One. Azacrown - Acridine and  Analysis of Fluorescence Titration Results of Complexes of Heavy Metal Ions

In addition to the complex of azacrown-acridine with mercury and cadmium ions, the binding of other heavy metal ions showed only a slight chelate amplification fluorescence effect, and no significant changes were observed. For azacrown-acridine and mercury ion complexes, when the final concentration was 3 μM at pH 7.4, a total fluorescence increase of more than 10-fold could be observed as the amount of mercury ions added increased from 0.5 equivalents to 20 equivalents.

In addition, for the complex of azacrown-acridine and cadmium ion, when the final concentration was 3 μM at pH 7.4, a total fluorescence increase of 10 times or more was observed as the amount of cadmium ion added increased from 5 equivalents to 200 equivalents. .

From the above results, it can be seen that azacrown-acridine according to the present invention binds selectively to mercury ions and cadmium ions.

2. Azacia Crown - Acridine and  Analysis of Fluorescence Titration Results of Complexes of Heavy Metal Ions

As shown in 1, only a slight chelate amplification fluorescence effect was observed for the binding of other heavy metal ions in addition to the complex of azacia crown-acridine, mercury ions, and cadmium ions, and no significant change was observed.

For azaciacrown-acridine and mercury ion complexes, when the final concentration was 3 μM at pH 7.4, a four-fold increase in total fluorescence was observed as the addition of mercury ions increased from 0.5 equivalents to 20 equivalents. .

In addition, for the complex of azacia crown-acridine and cadmium ion, when the final concentration is 3 μM at pH 7.4, a total fluorescence increase of 6 times or more can be observed as the amount of cadmium ion added increases from 1 equivalent to 200 equivalents. there was.

From the above results, it can be seen that the azacia crown-acridine according to the present invention binds selectively to mercury ions and cadmium ions.

< Experimental Example  3> NMR Using Azacia Crown - Acridine and  Experiment to measure the formation of complex of mercury ions

The azaciacrown-acridine is dissolved in 1 ml of DMSO- d ₆ at a concentration of 3 mM and mercury ions are prepared using DMSO- d ₆ at a concentration of 0.25 M. ¹ H NMR of the solution in which azacia crown-acridine was dissolved and a solution in which 0.4 equivalent, 1 equivalent and 2 equivalents of mercury ions were added to azacia crown-acridine were measured, and the results are shown in FIG. 7. The peaks near the aromatics shifted to the left when mercury ions were added as compared to the DMSO- d ₆ solution in which only azaciacrown-acridine (3 mM) was dissolved. For example, in the case of the solution in which only the azacia crown-acridin was dissolved, the H-9 peak could be observed at 9.09 ppm (see FIG. 7 (a)), and the solution was added at 9.49 ppm when 2 equivalents of mercury ions were added. The H-9 peak could be observed (see FIG. 7D). The above results are believed to indicate that azaciacrown-acridine is present in combination with mercury ions.

< Experimental Example  4> Azacrown - Acridine  And Azacia Crown - Acridine and  Experimental Measurement of Intracellular Operation of Complexes of Mercury Ions and Cadmium Ions

SK-N-SH cells were cultured in culture solution (MEM containing 10% FBS, 2 mM L-glutamine, 100 units / ml penicillin, 100 μg / ml streptomycin) in a thermostat. Cell line HaCaT human keratinocytes and cell line HCT116 human colon cancer cells were also cultured in RPMI 1640 containing 100 units / ml penicillin, 100 μg / ml streptomycin, and 10% fetal calf serum. The cells were incubated on a cover glass of 24-can plates at 104 cell density per cell. After 24 hours, 50-200 μM of mercuric chloride or cadmium chloride was added to the cells, and the cells were cultured in a medium at 37 ° C. and 5% carbon dioxide, washed three times with PBS (phosphate-buffered saline), and residual mercury ions or cadmium ions. Was removed. After washing with PBS, the treated cells were placed in 50 μM of the azacrown-acridine or azacia crown-acridine and incubated in medium at 37 ° C. for 1 hour. After washing three times with PBS, the residual compound was removed, the cells were fixed for 4 minutes with 4% paraformaldehyde and rinsed again with PBS, and then the cover glass was attached to the slide glass. Fluorescence images of the cells were observed under a DeltaVision microscope. All images were measured under fixed conditions such as exposure time (0.1 sec), brightness, contrast, and the like, and the results are shown in FIG. 8.

(A) and (c) of FIG. 8 show the results of observing azacrown-acridine and azaciacrown-acridine alone with a DeltaVision microscope, respectively, and no fluorescence image was observed. FIG. 8 (b) Is a complex of azacrown-acridine and mercury ions observed in HaCaT human keratinocyte line, and (d) a complex of azacia crown-acridin and cadmium ion in SK-N-SH human neuroblast line. (E) and (f) show the optical and fluorescence images of azaciacrown-acridine and cadmium ions in the HCT-116 human colorectal cancer cell line, respectively. It was observed that the dean and the azaciacrown-acridine exhibit chelate amplification fluorescence effects in combination with mercury ions and / or cadmium ions.

1 is aza from pH 7.4 (100mM HEPES buffer) crown-acridine (3 μM) in 100 equivalent mercury ions, cadmium ions and other metal ions (Ag ^+, the Ca ^{2 +,} Cd ^{2 +,} Co ^{2 +,} Cs ^+, and Cu ^{+ 2,} Hg ^{+ 2,} K ^+, Li ^+, Mg ^{+ 2,} Mn ^{+ 2,} Na ^+, Ni ^{+ 2} and Zn ^{+ 2)} a graph showing a change in fluorescence when the a,

FIG. 2 is a graph showing the fluorescence titration when the concentration of mercury ions (0.5 to 20 equivalents) was added to azacrown-acridine (3 μM) at pH 7.4 (100 mM HEPES buffer).

Figure 3 is a graph showing the fluorescence titration when the concentration of cadmium (5-200 equivalents) was added to azacrown-acridine (3 μM) at pH 7.4 (100 mM HEPES buffer),

Figure 4 pH 7.4 (100mM HEPES buffer) aza cyano crown-acridine (3 μM) 100 equivalents of mercury ions, cadmium ions and other metal ions (Ag ^+, of the Ca ^{2 +,} Cd ^{2 +,} Co ^{2 +} , and ^{Cs +, Cu 2+, Hg 2} +, K +, Li +, Mg 2 +, Mn 2 +, Na +, Ni 2 + , and Zn ^{+ 2)} a graph showing a change in fluorescence when the a,

Figure 5 is a graph showing the fluorescence titration when the concentration of mercury (5-200 equivalents) was added to azacia crown-acridine (3 μM) at pH 7.4 (100 mM HEPES buffer),

FIG. 6 is a diagram showing the fluorescence titration when different concentrations of cadmium (1 to 300 equivalents) were added to azaciacrown-acridine (3 μM) at pH 7.4 (100 mM HEPES buffer).

7 shows mercury ions ((b): 0.4 equivalents, (c): 1 equivalents, (d): 2 equivalents) in azacrown-acridine (3 mM, (a)) and dimethyl sulfoxide (DMSO). Is a graph showing the ¹ H NMR spectra of the azacrown-acridine (3 mM) complex,

FIG. 8 shows fluorescence images (b) in a HaCaT human keratinocyte line of azacrown-acridine (50 μM, (a)), a complex of mercury ions (50 μM) and azacrown-acridine, azacia crown Fluorescence images (d), cadmium ions in SK-N-SH human neuroblast cell line of the complex between acridine (50 μM, (c)), cadmium ion (50 μM) and azaciacrown-acridine 50 μM) and HCT-116 of the complex of azaciacrown-acridine HCT-116 in human colorectal cancer cell line (e) and HCT-116 of the complex of cadmium ions (50 μM) and azaciacrown-acridine It is a photograph showing the fluorescence image (f) in the human colorectal cancer cell line.

Claims (9)

Acridine derivative represented by the following formula (1). [Formula 1]

Wherein X is O or S The acridine derivative according to claim 1, wherein the acridine derivative of Formula 1 is an azacrown-acridine derivative of Formula 2 or an azacia crown-acridine derivative of Formula 3. [Formula 2]

[Formula 3]

As shown in Scheme 1 below, Reacting by adding azacrown compound of formula 5 to a solution of 4,5-bis (bromomethyl) acridine of formula 4 and a base selected from the group consisting of potassium carbonate, triethylamine and sodium carbonate in a solvent (Step 1); Stirring the reaction product of step 1 at room temperature and then filtering and evaporating the solvent (step 2); And A method for preparing the acridine derivative according to claim 1, comprising the step (step 3) of purifying by performing column chromatography on the reaction product of step 2. Scheme 1

(Wherein X is O or S) delete  The method of claim 3, wherein the solvent of step 1 is selected from the group consisting of chloroform, dichloromethane and acetonitrile. The compound of claim 3, wherein the azacrown compound of formula 5 of step 1 is 4,13-diaza-18-crown-6-ether or 1,10-diaza-4,7,14,17-tetrasiacycloocta A method for producing an acridine derivative, characterized in that the dican. Method for selective detection of mercury ions and / or cadmium ions present in an aqueous solution environment or a living body using the acridine derivative of claim 1. 8. The method of claim 7, wherein the detection of mercury ions and / or cadmium ions is carried out by exciting a complex of the acridine derivative of claim 1 and the mercury and / or cadmium ions to confirm chelate amplification fluorescence (CHEF) effects. Characterized in that the selective detection of mercury ions and / or cadmium ions. The method of claim 8, wherein the mercury ions and / or cadmium ions present in the cell are detected by imaging the amplification fluorescence effect of the complex.

Images