KR20110049321A - A targetable fluorescent probe for natural ros of living cells , preparation of the same, detection probe and detection method of ros using the same - Google Patents

Abstract

PURPOSE: A thiazolidine derivative compound for detecting intracellular active oxygen fluorescence is provided to ensure high detection sensitivity. CONSTITUTION: A thiazolidine derivative compound for detecting intracellular active oxygen fluorescence is denoted by chemical formula 1. The compound of chemical formula 1 is prepared by reacting coumarine aldehyde compound of chemical formula 2 and an anline derivative of chemical formula 3 containing ortho substituent. A sensor for selective detection of active oxygen contains the thiazolidine derivative compound. The active oxygen is detected by dissolving the thiazolidine derivative compound in a solvent and contacting to cells for detecting the active oxygen. The solvent is water. The cell is a mitochondria.

Description

Thiazolidine derivative compounds for detecting active oxygen fluorescence in a cell, a method for preparing the same, and a method for detecting active oxygen fluorescence in a cell including the same, and a method for detecting the same, detection probe and detection method of ROS using the same}

The present invention relates to a thiazolidine derivative compound for detecting active oxygen fluorescence in a cell, a method for preparing the same, a sensor for detecting active oxygen in a cell, and a detection method including the same.

Free radicals in cells are closely related to Alzheimer's, Parkinson's disease, and cancer. Higher free radicals in cells promote disease and aging. In particular, mitochondria in which aerobic respiration occurs have a lot of free radicals. If the amount of free radicals can be detected and quantified, it will be helpful for early diagnosis of modern diseases.

Recently, effective methods having good selectivity and sensitivity to free radicals based on phosphors have been studied, but there are no studies showing high sensitivity to free radicals in cells. Therefore, research results of the degree to which the change of the phosphors can be seen only when intentionally added active oxygen such as hydrogen peroxide and stimulation from the outside are reported.

Accordingly, the present invention has high sensitivity, and thus light emission occurs without external active oxygen being introduced into the cell to detect active oxygen such as hydrogen peroxide (H ₂ O ₂ ), superoxide (O ₂ ⁻ ), hydroxy radical (HO ^. ), And the like ^. It is an object of the present invention to provide a thiazolidine derivative compound for detecting active oxygen fluorescence in a cell and a method of preparing the same.

In another aspect, the present invention is hydrogen peroxide (H ₂ O _2), superoxide (O ₂ ^-) in the number of free radicals mitochondrial living using the fluorescence probe compound can detect active oxygen, such as, hydroxy radical ^(HO.) Another object is to provide a free radical fluorescence detection sensor and a detection method in a cell.

The present invention provides a thiazolidine derivative compound for detecting active oxygen fluorescence in a cell, which is represented by the following Chemical Formula 1 in order to achieve the above object.

<Formula 1>

In Formula 1, X is selected from S, O, and NH, and R ₁ , R _2, and R ₃ are each independently hydrogen, a linear or nonlinear C1 to C20 alkyl group, or a linear or nonlinear C1 to C20 alkyl group. Is selected from a ketone group having, a ester group having a linear or nonlinear C1 to C20 alkyl group, a substituted or unsubstituted aryl group of C5 to C24, or an arylalkyl group of C6 to C24, each of R ₄ and R ₅ is independently linear Or a nonlinear C1 to C20 alkyl group, a ketone group having a linear or nonlinear C1 to C20 alkyl group, an ester group having a linear or nonlinear C1 to C20 alkyl group, a substituted or unsubstituted aryl group or C6 to C24 To C24 arylalkyl group.)

In another aspect, the present invention is a thiazoli represented by Chemical Formula 1 by reacting the cumin aldehyde compound represented by Chemical Formula 2 with the imine compound represented by Chemical Formula 3 having an ortho substituent in order to achieve the above object. Provided is a method for preparing a thiazolidine derivative compound for detecting active oxygen fluorescence in a cell, the method comprising preparing a dean derivative compound.

<Scheme 1>

(X in Scheme 1 is selected from S, O, NH, R ₁ , R ₂ and R ₃ are each independently hydrogen, linear or nonlinear C1 to C20 alkyl group, linear or nonlinear C1 to C20 alkyl group Is selected from a ketone group having, a ester group having a linear or nonlinear C1 to C20 alkyl group, a substituted or unsubstituted aryl group of C5 to C24, or an arylalkyl group of C6 to C24, each of R ₄ and R ₅ is independently linear Or a nonlinear C1 to C20 alkyl group, a ketone group having a linear or nonlinear C1 to C20 alkyl group, an ester group having a linear or nonlinear C1 to C20 alkyl group, a substituted or unsubstituted aryl group or C6 to C24 To C24 arylalkyl group.)

In another aspect, the present invention provides a sensor for detecting active oxygen fluorescence in the cell comprising a thiazolidine derivative compound represented by the formula (1) to achieve the above object.

In another aspect, the present invention is to dissolve the thiazolidine derivative compound represented by the formula (1) in a solvent in order to achieve the above object and to contact the solution with cells for detecting active oxygen to detect the active oxygen through fluorescence change It provides a method for detecting active oxygen fluorescence in a cell, characterized in that.

The thiazolidine derivative compound represented by Chemical Formula 1 according to the present invention may detect active oxygen such as intracellular hydrogen peroxide (H ₂ O ₂ ), superoxide (O ₂ ⁻ ), hydroxy radical (HO ^. ), And the like ^. As a novel fluorescent precursor compound, the light emission selectively occurs only in mitochondria rich in active oxygen without entering cellular tissues such as other nuclei, and the light emission can be observed under a microscope even at a concentration of about 10 uM. At about 0.1 uM, the fluorescence intensity shows a linear dependence on the concentration, so that the fluorescence change can be observed even in the presence of very low concentration of active oxygen in the cell. Therefore, it can be usefully used for active oxygen fluorescence detection sensor in cells.

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

Thiazolidine derivative compounds for detecting active oxygen fluorescence in cells according to the present invention are represented by the following formula (1).

<Formula 1>

In Formula 1, X is selected from S, O, and NH, and R ₁ , R _2, and R ₃ are each independently hydrogen, a linear or nonlinear C1 to C20 alkyl group, or a linear or nonlinear C1 to C20 alkyl group. Is selected from a ketone group having, a ester group having a linear or nonlinear C1 to C20 alkyl group, a substituted or unsubstituted aryl group of C5 to C24, or an arylalkyl group of C6 to C24, each of R ₄ and R ₅ is independently linear Or a nonlinear C1 to C20 alkyl group, a ketone group having a linear or nonlinear C1 to C20 alkyl group, an ester group having a linear or nonlinear C1 to C20 alkyl group, a substituted or unsubstituted aryl group or C6 to C24 To C24 arylalkyl group.)

The thiazolidine derivative compound represented by Formula 1 is a compound of Formula 4, wherein X is S, R ₁ , R _2, and R ₃ are hydrogen, and R ₄ and R ₅ are cyclic phenyl, X is O, R ₁ , R ₂ and R ₃ are hydrogen and R ₄ and R ₅ are cyclic phenyl or a compound of formula 5 or X is S, R ₁ , R ₂ and R ₃ are hydrogen and R ₄ and R ₅ are It is preferable to select from the compound represented by following formula (6) which is cyclic phenyl.

<Formula 4>

<Formula 5>

<Formula 6>

The present invention provides a method for more effectively preparing a thiazolidine derivative compound represented by Chemical Formula 1.

More specifically, the thiazolidine derivative compound represented by Chemical Formula 1 according to the present invention is represented by Chemical Formula 3 having an ortho substituent and a cumin aldehyde compound represented by Chemical Formula 2 as shown in Scheme 1 below. It is prepared by reacting an imine compound.

<Scheme 1>

(X in Scheme 1 is selected from S, O, NH, R ₁ , R ₂ and R ₃ are each independently hydrogen, linear or nonlinear C1 to C20 alkyl group, linear or nonlinear C1 to C20 alkyl group Is selected from a ketone group having, a ester group having a linear or nonlinear C1 to C20 alkyl group, a substituted or unsubstituted aryl group of C5 to C24, or an arylalkyl group of C6 to C24, each of R ₄ and R ₅ is independently linear Or a nonlinear C1 to C20 alkyl group, a ketone group having a linear or nonlinear C1 to C20 alkyl group, an ester group having a linear or nonlinear C1 to C20 alkyl group, a substituted or unsubstituted aryl group or C6 to C24 To C24 arylalkyl group.)

The thiazolidine derivative compound represented by Chemical Formula 1 in Scheme 1 may be easily reacted at a ratio of 1 to 2 mol of the amine compound represented by Chemical Formula 3 having an ortho substituent with respect to 1 mol of the cumin aldehyde compound represented by Chemical Formula 2 It may be prepared a thiazolidine derivative compound represented by the formula (1).

The reaction may be carried out in the presence of an organic solvent, and organic solvents, including dichloromethane, chloroform, tetrahydrofuran, methanol and ethanol, may be used generally used in the field of organic synthesis.

After the reaction is completed, the reaction solution is diluted with water, the organic solvent layer is fractionated using an organic solvent, the solvent is evaporated, and then separated by column chromatography to obtain a thiazolidine derivative compound represented by Chemical Formula 1.

The thiazolidine derivative compound represented by Chemical Formula 1 according to the present invention prepared by the above method may be utilized in various fields as a fluorescent precursor.

In particular, the thiazolidine derivative compound represented by Chemical Formula 1 is useful for a sensor for selectively detecting active oxygen such as hydrogen peroxide (H ₂ O ₂ ), superoxide (O ₂ ⁻ ), hydroxy radical (HO ^. Can be used. Preferably, the thiazolidine derivative compound represented by Formula 1 may be usefully used for an active oxygen fluorescence detection sensor that selectively detects naturally occurring active oxygen in the mitochondria. Accordingly, the present invention provides a sensor for detecting active oxygen fluorescence in a cell comprising a thiazolidine derivative compound represented by Chemical Formula 1.

Hereinafter, a detection method for selectively detecting active oxygen in the intracellular mitochondria will be described in detail.

First, the thiazolidine compound represented by Chemical Formula 1 is dissolved in a solvent. The solvent may be one containing water.

In this case, the thiazolidine compound concentration represented by Chemical Formula 1 is not necessarily limited, but in order to visually check the fluorescence change of the phosphor, it is preferable to use 10 μM or more. However, even when the concentration of the thiazolidine compound is less than 10 μM, since the fluorescence change can be easily confirmed by a microscope or a UV lamp, the concentration of the thiazolidine compound represented by Chemical Formula 1 may be variously changed according to the application environment. .

The solution containing the thiazolidine compound represented by Formula 1 according to the present invention and the sample to detect the presence of active oxygen are contacted. Contact here includes adding a sample to a solution or adding a solution to a sample.

After addition of the sample, the presence of free radicals is detected in the cell through color change. In this case, when the concentration of the active oxygen is 1 μM or more, the fluorescence change can be visually confirmed. If the concentration of the active oxygen is less than 1μM fluorescence change can be easily confirmed with a microscope or UV lamp.

Hydrogen peroxide (H ₂ O ₂₎ in the sample, superoxide (O ₂ ^-), hydroxyl radical ^(. HO) If the active oxygen contains, as the aromatic reaction by the dehydrogenation reaction proceeds as shown in the following scheme 2 fluorescence changes in Is generated. At this time, the presence of active oxygen can be confirmed by changing the fluorescence.

<Scheme 2>

In Scheme 2, X and R ₁ to R ₅ are the same as defined in Chemical Formula 1.

As shown in Scheme 2, the thiazolidine derivative compound represented by Chemical Formula 1 according to the present invention is dehydrogenated by 1 equivalent of active oxygen, and becomes an aromatic compound, thereby emitting light. Such luminescence is very selectively observed only by active oxygen. In particular, fluorescence changes are prominent in mitochondria rich in free radicals in cellular tissues.

As a specific example, the compound of Formula 4 is dehydrogenated by active oxygen to produce the compound of Formula 7, as shown in Scheme 3 below, and the compound of Formula 7 emits strong light.

<Scheme 3>

In particular, after the addition of hydrogen peroxide to the aqueous solution containing the compound represented by the formula (4) and looking at the UV absorption spectrum according to the unit time, as shown in Figure 1 the absorption at about 399 nm is rapidly reduced absorption at about 458 nm It can be seen that the increase rapidly. Therefore, when hydrogen peroxide is contained in the sample, the color is changed by the change in the absorption spectrum, and the increase in fluorescence makes it easy to check whether or not active oxygen such as hydrogen peroxide is contained.

Hereinafter, the present invention will be described in more detail with reference to the following examples, which are only presented to aid the understanding of the present invention, but the present invention is not limited thereto.

Synthesis Example 1

63.6 mg (0.25 mmol) of 7-diethylamino-3-formylcumarin and 2-aminobenzenethiol (0.25 mmol) are dissolved in 1 mL of ethanol, respectively. Slowly add 2-aminobenzenethiol while stirring the 7-diethylamino-3-formylcumarin solution. The reaction solution is stirred at room temperature for 3 hours. If a yellowish precipitate occurs, filter it out, rinse with ethanol two or three times and dry. Yield: 85%. The ¹ H and ¹³ C-NMR and MS measurement results of the product is as follows, as a result it was confirmed that the structural formula is a compound represented by the formula (4).

1 H NMR (DMSO-d6, 300 MHz): 7.76 (s, 1H), 7.48 (d, J = 9.0 Hz, 1H), 7.00 (d, J = 7.2 Hz, 1H), 6.92 (t, J = 7.8 Hz , 1H), 6.86 (d, J = 2.7 Hz, 1H), 6.74 (d, J = 7.8 Hz, 1H), 6.70 (dd, J = 9.0 Hz, J = 2.1 Hz, 1H), 6.63 (t, J = 7.2 Hz, 1H), 6.56 (d, J = 2.1 Hz, 1H), 6.17 (d, J = 2.7 Hz, 1H), 3.46 (q, J = 6.9 Hz, 4H), 1.14 (t, J = 6.9 Hz, 6H)

13C NMR (DMSO-d6, 75 MHz): 161.09, 155.73, 150.88, 147.66, 137.90, 130.04, 125.76, 125.60, 122.19, 121.84, 119.53, 110.04, 109.64, 107.74, 96.81, 63.89, 44.50, 12.75 (18 carbon peaks )

HRMS (FAB +, m-NBA): m / z calculated for C 20 H 21 N 2 O 2 S: 353.1324, observed: 353.1319

Synthesis Example 2

63.6 mg (0.25 mmol) of 7-diethylamino-3-formylcumarin and 2-aminophenol (0.25 mmol) are dissolved in 1 mL of ethanol, respectively. Slowly add 2-aminobenzenethiol while stirring the 7-diethylamino-3-formylcumarin solution. The reaction solution is stirred at room temperature for 3 hours. If you have a scarlet pink filter, filter it out, rinse with ethanol two or three times and dry. Yield 74%. ¹ H-NMR measurement results of the product is as follows, as a result it was confirmed that the structural formula is a compound represented by the formula (5).

1 H NMR (DMSO-d 6, 300 MHz): 8.96 (s, 1H), 8.82 (s, 1H), 8.66 (s, 1H), 7.52 (d, J = 9.0 Hz, 1H), 7.19 (d, J = 7.8 Hz, 1H), 7.07 (t, J = 7.5 Hz, 1H), 6.90 (d, J = 7.8 Hz, 1H), 6.82 (m, 2H), 6.52 (s, 1H), 3.53 (q, J = 6.9 Hz, 4H), 1.18 (t, J = 6.9 Hz, 6H)

&Lt; Synthesis Example 3 &

Dissolve 63.6 mg (0.25 mmol) of 7-diethylamino-3-formylcumarin and diaminobenzene (0.25 mmol) in 1 mL of ethanol, respectively. Slowly add 2-aminobenzenethiol while stirring the 7-diethylamino-3-formylcumarin solution. The reaction solution is stirred at room temperature for 3 hours. If you have a red precipitate, filter it out, rinse with ethanol two or three times and dry. Yield: 78%. ¹ H-NMR measurement results of the product is as follows, as a result it was confirmed that the structural formula is a compound represented by the formula (6).

1 H NMR (DMSO-d6, 300 MHz): 8.75 (s, 1H), 8.56 (s, 1H), 7.62 (d, J = 9.0 Hz, 1H), 7.06 (d, J = 7.8 Hz, 1H), 6.96 (t, J = 7.5 Hz, 1H), 6.82 (d, J = 9.0 Hz, 1H), 6.72 (d, J = 7.8 Hz, 1H), 6.62 (s, 1H), 6.56 (t, J = 7.5 Hz , 1H), 5.29 (s, 2H), 3.50 (q, J = 6.9 Hz, 4H), 1.18 (t, J = 6.9 Hz, 6H)

&Lt; Example 1 >

Confirmation of Fluorescence Change by the Compound Represented by Formula 4

30 uM of the compound represented by Formula 4 prepared in Synthesis Example 1 was dissolved in water (HEPES buffer pH 7.4) and isopropanol (3: 2, v / v), and at least 1 equivalent of hydrogen peroxide (H ₂ O ₂ ) was added thereto. After the change in the UV absorption spectrum with time was measured, the results are shown in FIG.

As shown in FIG. 1, when the hydrogen peroxide is added to an aqueous solution containing a thiazolidine compound represented by Chemical Formula 4 according to the present invention, the UV absorption spectrum according to unit time decreases rapidly at 399 nm. It can be seen that the absorption at 458 nm increases rapidly. Therefore, when hydrogen peroxide is contained in the sample, it can be seen that the presence of hydrogen peroxide can be easily confirmed through the change in color represented by the change in the absorption spectrum.

<Example 2>

Fluorescence Change Measurement by Compound of Formula 4

Change in fluorescence over time after dissolving 10 uM of the compound represented by Formula 4 in water (HEPES buffer pH 7.4) and isopropanol (3: 2, v / v) and adding 500 equivalents of hydrogen peroxide (H ₂ O ₂ ) Was measured and the result is shown in FIG. In addition, the fluorescence intensity of the compound represented by Formula 7 is also shown.

As shown in FIG. 2, the fluorescence intensity at 510 nm increases gradually over time and reaches its saturation point after 6 hours. At this time, the fluorescence pattern shows the same shape as the fluorescence pattern of Chemical Formula 7, a compound of Chemical Formula 4 This hydrogen implies that the reaction proceeded to the compound represented by the formula (7).

<Example 3>

Measurement of Fluorescence Changes of Compounds of Formula 4 According to Concentration of Hydrogen Peroxide

Dissolve 3 uM of the compound represented by the formula (4) in water (HEPES buffer pH 7.4) and isopropanol (3: 2, v / v) and fluorescein while changing the amount of hydrogen peroxide (H ₂ O ₂ ) from 0.1 uM to 10 uM The change was observed and the result is shown in FIG.

As shown in FIG. 3, it can be observed that the fluorescence intensity increases linearly with the concentration of hydrogen peroxide. In addition, it can be seen that the quantitative analysis of hydrogen peroxide according to the fluorescence intensity is possible by using the linear relationship of the fluorescence intensity.

<Example 4>

Measurement of Fluorescence Changes of Compounds of Formula 4 According to pH

After 500 equivalents of hydrogen peroxide (H ₂ O ₂ ) was added to 10 uM of the compound represented by Formula 4, the change in fluorescence intensity at 510 nm according to pH was measured (triangle display) and the results are shown in FIG. 4. As a control, the change in fluorescence intensity at 510 nm according to pH in the state without adding hydrogen peroxide was measured (circle) and the results are shown together in FIG. 4.

As shown in FIG. 4, the control group that does not include hydrogen peroxide has little change between pH 7-9, but it can be seen that the fluorescence intensity is greatly changed when hydrogen peroxide is included. This shows that the compound represented by Formula 4 can be usefully applied to a living body.

Example 5

Measurement of Fluorescence Intensity of Cells with Time and Concentration of Compound of Formula 4

After dropping the compound represented by the formula (4) to a predetermined concentration in living 293T cells, the fluorescent photographs of the cells according to the incubation time were taken and the results are shown in FIG. 5. At this time, the concentration of the compound represented by the formula (4) and the incubation time are as follows.

(a) concentration 1.0 uM, 30 min incubation, (c) concentration 1.0 uM, 1 hour incubation, (e) concentration 3.0 uM, 30 min incubation, (g) concentration 3.0 uM, 1 hour incubation, (i) concentration 10 uM, 30 minute incubation (k) concentration 10 uM, 1 hour incubation, (m) concentration 30 uM, 30 minute incubation, (o) concentration 30 uM, 1 hour incubation.

In Fig. 5, (b), (d), (f), (h), (j), (l), (n) and (p) are respectively (a), (c), (e), (g) ), (i), (k), (m) and the general picture of (o) is shown.

As shown in FIG. 5, even when the incubation time is about 30 minutes and the compound of Formula 4 is about 10 uM, fluorescence is expressed in the cells.

<Example 6>

Cytotoxicity Testing of Compounds of Formula 4

After dropping 30 uM of the compound represented by Formula 4 into MCF-7 cells, which are breast cancer cells, and 293T cells, which are human kidney embryonic cells, the cells were cultured for 1 hour, followed by fluorescence photographing of the cells, and the results are shown in FIG. 6.

In Figure 6 (a) to (d) is a culture of MCF-7 cells of breast cancer cells, (e) to (h) is a culture of 239T cells of human kidney embryonic cells, specifically as follows.

(a) no compound of formula 4, (c) 30 uM of compound of formula 4, incubated for 1 hour, (e) no compound of formula 4, (g) 30 uM of compound of formula 4, incubated for 1 hour.

 In FIG. 5, (b), (d), (f) and (h) show general photographs of (a), (c), (e) and (g), respectively.

As shown in FIG. 6, it was confirmed that there is almost no cytotoxicity, since a certain number of individuals was shown regardless of whether the compound of Formula 4 was treated.

<Example 7>

Measurement of Cell Tissue Selectivity of Compounds of Formula 4

In order to confirm whether the compound represented by Formula 4 selectively penetrated into and expressed in the tissue, expression location tracking experiment was performed using MitoTracker (MitoTracker ^ㄾ Red FM, Invitrogen, Eugene, OR), a mitochondrial tracer. The expression tracking experiment was performed by treating the compound of Formula 4 and MitoTracker Red for 30 minutes on MCF-7 cells, and image image using a confocal laser scanning microscope (LSM 710, Carl Zeiss MicroImaging GmbH, Jena, Germany) Photographed and shown in FIG. 7.

Figure 7 (A) is a result of observing where the formula 4 is present in the cell using a green filter, (B) is a result of observing the position of MitoTracker Red in the same cell sample using a red filter. (C ) Is an image obtained by combining the image using the green filter with the image using the red filter.

In FIG. 7C, a yellow portion is a region formed by overlapping the red of MitoTracker Red and the green of Formula 4. That is, it shows the part generated when MitoTracker Red and Formula 4 are in the same position. Looking at this video image, it can be observed that most of the yellow area coincides with the area of MitoTracker. That is, it can be seen that the compound represented by the formula (4) in most of the cells is selectively located in the mitochondria in the cell in which MitoTracker is located.

1 is a graph showing the change in UV absorption spectrum with unit time after the addition of hydrogen peroxide to the aqueous solution containing the compound of formula (4).

FIG. 2 is a graph showing the fluorescence change over time after adding hydrogen peroxide to an aqueous solution containing the compound of Formula 4 and the fluorescence of the compound represented by Formula 7. FIG.

3 is a fluorescence intensity measured after adding to the aqueous solution containing the compound of formula 4 while changing the concentration of hydrogen peroxide.

Figure 4 is a graph measuring the change in fluorescence according to the pH of the compound of formula 4 with hydrogen peroxide (triangular display) and without hydrogen peroxide (circle display).

FIG. 5 is a fluorescence photograph of culture time of live 293T cells according to the concentration of the compound of Formula 4.

FIG. 6 shows fluorescence photographs of cytotoxicity experiments on MCF-7 and 293T cells of the compound of Formula 4. FIG.

7 is a confocal microscopy cell photo showing mitochondrial selectivity of MCF-7 cells.

Claims (7)

Thiazolidine derivative compounds for detecting active oxygen fluorescence in cells, which are represented by the following Chemical Formula 1. <Formula 1>

In Formula 1, X is selected from S, O, and NH, and R ₁ , R _2, and R ₃ are each independently hydrogen, a linear or nonlinear C1 to C20 alkyl group, or a linear or nonlinear C1 to C20 alkyl group. Is selected from a ketone group having, a ester group having a linear or nonlinear C1 to C20 alkyl group, a substituted or unsubstituted aryl group of C5 to C24, or an arylalkyl group of C6 to C24, each of R ₄ and R ₅ is independently linear Or a nonlinear C1 to C20 alkyl group, a ketone group having a linear or nonlinear C1 to C20 alkyl group, an ester group having a linear or nonlinear C1 to C20 alkyl group, a substituted or unsubstituted aryl group or C6 to C24 To C24 arylalkyl group.) The thiazolidine derivative compound represented by Chemical Formula 1 is represented by Chemical Formula 4, wherein X is S, R ₁ , R _2, and R ₃ are hydrogen, and R ₄ and R ₅ are phenyl cyclic. A compound represented by the following formula (5) wherein X is O, R ₁ , R ₂ and R ₃ are hydrogen and R ₄ and R ₅ are phenyl cyclic or X is S and R ₁ , R ₂ and R ₃ A thiazolidine derivative compound for detecting active oxygen fluorescence in a cell, wherein is hydrogen and R ₄ and R ₅ are phenyl in a ring form. <Formula 4>

<Formula 5>

<Formula 6>

As shown in Scheme 1 below, the reaction of the cumin aldehyde compound represented by the formula (2) and the aniline derivative represented by the following formula (3) having an ortho substituent reacts to prepare the compound represented by the formula (1). Method for preparing a thiazolidine derivative compound for detecting oxygen fluorescence. <Scheme 1>

(X in Scheme 1 is selected from S, O, NH, R ₁ , R ₂ and R ₃ are each independently hydrogen, linear or nonlinear C1 to C20 alkyl group, linear or nonlinear C1 to C20 alkyl group Is selected from a ketone group having, a ester group having a linear or nonlinear C1 to C20 alkyl group, a substituted or unsubstituted aryl group of C5 to C24, or an arylalkyl group of C6 to C24, each of R ₄ and R ₅ is independently linear Or a nonlinear C1 to C20 alkyl group, a ketone group having a linear or nonlinear C1 to C20 alkyl group, an ester group having a linear or nonlinear C1 to C20 alkyl group, a substituted or unsubstituted aryl group or C6 to C24 To C24 arylalkyl group.) An active oxygen selective detection sensor containing a thiazolidine derivative compound for detecting active oxygen fluorescence in a cell of claim 1 or 2. A thiazolidine derivative compound for detecting active oxygen fluorescence in a cell of claim 1 or 2 is dissolved in a solvent, and contacted with a cell for detecting active oxygen, thereby detecting active oxygen through fluorescence change. Active oxygen fluorescence detection method. The method of claim 5, wherein the solvent comprises water. The method of claim 5, wherein the cell is a live mitochondria.

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