KR101528256B1 - Preparation method of BODIPY based dual fluorogenic probe for tandem superoxide and Hg2+ chemosensing - Google Patents

Abstract

The present invention relates to a bipyridyl derivative compound which is a fluorescent molecule that simultaneously detects mercury metal ions and active oxygen. More specifically, the present invention relates to a fluorescent material which exhibits fluorescence characteristics when bound to a mercury metal ion and an active oxygen including an oxygen element, a sulfur element, a selenium element or a tellurium element and an aryl group for metal bonding in a BODIPY-based fluorescent base, And a method for producing the same.

Description

[0001] The present invention relates to a chemical sensor compound for simultaneously chemically detecting mercury ions and active oxygen, and a preparation method thereof,

The present invention relates to a chemical sensor compound which is a BODIPY-based fluorescent substance which simultaneously chemically detects mercury ions and active oxygen, and a method for producing the same.

Reactive oxygen species (ROS) in the body are known to have strong oxidizing power during the physiological action. In particular, active oxygen is known to be an important causative agent in aging, various cancers, and neurodegenerative diseases. However, the timing, location and cause of active oxygen are not well known. Development of a system that can detect the generation and distribution of active oxygen is a key aid in the diagnosis and treatment of disease.

Mercury ions are used in many industries such as barometers, thermometers, batteries, mercury lamps, and dental amalgam. As many people live with the potential or direct exposure to mercury. Mercury can be absorbed through the skin or mucous membranes, and mercury vapor can be absorbed through the respiration. When absorbed in the body, it is not easily released and acts as a toxic ion to infiltrate the cerebral blood vessels, affecting the central nervous system, causing diseases in digestive organs and kidneys, and especially diseases such as Minamata disease. That is, in the case of a person or animal exposed to a mercury environment, it should be possible to perform mercury analysis immediately if necessary.

Thus, mercury and active oxygen are known to be very dangerous substances that cause aging or toxicity in vivo. Therefore, it is necessary to develop a method for easily detecting and diagnosing mercury and active oxygen present in the body cheaply and cheaply.

Although the detection method of active oxygen and mercury can be detected by an electrochemical method, a biological method, and a chemical method using expensive analytical equipment, researches on a detection method by a chemical method have been actively conducted, and many technologies have been developed . Chemical detection methods have high sensitivity and selectivity and can be detected in an economical way. In particular, chemical detection technology using coloring or fluorescence using a compound is known as a powerful technique. Many researches have been carried out because of high selectivity for analytes, sensitivity, economy, ease of manufacture, and ease of detection.

However, existing chemical compounds for chemical detection are usually a single compound, and only one kind of analyte can be analyzed. Therefore, in order to detect various kinds of compounds, it is necessary to use several chemical sensor compounds for each function separately.

Mercury is also generally known as a substance that quenches fluorescence. Sensing can be done by quenching method, but coloring and fluorescence emission is more effective than quenching. However, existing detection methods have high detection limits and can not detect mercury and active oxygen at the same time.

The present invention provides a chemical sensor compound and a method for producing the same, which can easily detect mercury and active oxygen simultaneously with high sensitivity using one compound.

It is another object of the present invention to provide a chemical sensor compound which is capable of detecting mercury ions and active oxygen, exhibiting color development and fluorescence even at a very low concentration, and which can be easily produced at low cost, .

The present invention provides a chemical sensor compound selected from compounds represented by the following general formulas (1) and (2).

[Chemical Formula 1]

(2)

In the formulas (1) and (2)

E ₁ , E ₂ and E ₃ are each independently O, S, Se or Te,

R ₁ , R ₂ , R ₃ , R ₄ , R ₅ and R ₆ are each independently hydrogen or an alkyl group having 1 to 20 carbon atoms or an aryl group having 6 to 30 carbon atoms, which may contain a hetero atom;

X is a halogen element.

The chemical sensor compound exhibits a change in fluorescence property when it is combined with a mercury metal ion or active oxygen, or exhibits absorption and color development, and is characterized in that a mercury metal ion, an active oxygen atom (ROS) or a mercury metal ion and an active oxygen atom (ROS) at the same time.

In such a chemical sensor compound, it is preferable that R ₁ and R ₂ are each independently a 2-pyridyl group.

In the chemical sensor compound, it is preferable that each of E ₁ , E _2, and E ₃ is independently S.

The present invention also relates to a process for the preparation of a compound of formula

Reacting an aldehyde compound having a substituent represented by the following formula (3) or (4) with 2,4-dimethylpyrrole and p-chloriranyl

A method for producing the chemical sensor compound described above.

(3)

[Chemical Formula 4]

In the formulas (3) and (4)

E ₁ , E ₂ and E ₃ are each independently O, S, Se or Te,

R ₁ and R ₂ are each independently hydrogen or an alkyl group having 1 to 20 carbon atoms or an aryl group having 1 to 20 carbon atoms, which may contain a hetero atom,

X is a halogen element.

The present invention relates to a chemical sensor compound capable of simultaneous detection of mercury and active oxygen in aqueous solution or in a living body such as a cell by high sensitivity and selectivity through fluorescence or coloring when reacting with mercury and active oxygen simultaneously There is an effect to provide.

Fig. 1 shows the fluorescence properties of active oxygen (ROS) and mercury ion of the compound 1-1 of Example 1. Fig.
Fig. 2 shows fluorescence characteristics for the active oxygen (ROS) of the compound 2-1 of Example 2. Fig.
Fig. 3 shows the crystal structure of the compound 2-1 of Example 2 (a: viewed from the side, and b: viewed from below).
Figure 4 is the first embodiment of the metal ion of the compound 1-1 ^{(Cd 2 +, Hg 2 +} , Fe 2 +, Ca 2 +, Cu 2 +, Co 2 +, Mn 2+, Mg 2 +, Pb 2 + , Zn ^{2 +} , Ag ⁺ ).
FIG. 5 shows the results of reversible reaction of Compound 1-1 of Example 1 with various amino acids. FIG.
6 shows the results of Job plot analysis for confirming the binding ratio of the compound 1-1 of Example 1 with mercury.
FIG. 7 shows Stern-Volmer plots of various concentrations of Hg ^{2 +} ions and probes with the concentration of KO ₂ fixed to the compound 1-1 of Example 1. FIG.
8 shows the reactivity of the compound 1-1 of Example 1 in neuroblasticoma cells (a: when Compound 1-1 was treated, b: Compound 1-1, Hg ^{2 +} , and KO ₂ were treated simultaneously ).
9 shows a double PET passage mechanism acting upon the reaction of Compound 1-1 of Example 1 with O ₂ ^- , Hg ^{2 +} .
10 shows the 'AND' molecular logic circuit of the compound 1-1 of Example 1. FIG.

While the invention is susceptible to various modifications and alternative forms, specific embodiments thereof are shown by way of example in the drawings and will herein be described in detail. It should be understood, however, that the invention is not intended to be limited to the particular forms disclosed, but includes all modifications, equivalents, and alternatives falling within the spirit and scope of the invention.

Hereinafter, the present invention will be described in detail.

According to one embodiment of the present invention, there is provided a chemical sensor compound selected from compounds represented by Chemical Formulas 1 and 2 below.

[Chemical Formula 1]

(2)

In the formulas (1) and (2)

E ₁ , E ₂ and E ₃ are each independently O, S, Se or Te,

R ₁ , R ₂ , R ₃ , R ₄ , R ₅ and R ₆ are each independently hydrogen or an alkyl group having 1 to 20 carbon atoms or an aryl group having 6 to 30 carbon atoms, which may contain a hetero atom;

X is a halogen element.

The present invention relates to a chemical sensor compound which is a fluorescent substance that simultaneously chemically detects mercury ions and active oxygen, and a method for producing the same. More particularly, the present invention relates to a method for preparing a fluorophosphate-based fluorescent base, which is capable of coordinating an oxygen element, a sulfur element, a selenium element or a tellurium element with an aryl group (preferably a nitrogen atom-containing functional group) Thereby providing a chemical sensor compound that changes color and fluorescence when mercury metal binds. The present invention also provides a chemical sensor compound capable of simultaneously detecting mercury and active oxygen by changing coloring and fluorescence by binding oxygen atom, sulfur atom, selenium atom or tellurium contained in the compound to active oxygen, and a method for producing the same. do. Such a method of the present invention relates to a technique for detecting a chemical method of active oxygen and mercury outside the body.

A fluorescent molecule is a substance that strongly emits a wavelength of a specific visible light due to a characteristic peculiar to a molecule when light is externally applied. In the present invention, a boron-dipyrromethene (BODIPY) molecule is used as a basic fluorescent material. BODIPY is a stable fluorescent material with two pyrrole molecules attached and bonded by boron difluoride. In addition, the compound has stability that is not easily denatured, and it is a substance that detects and detects changes in fluorescence by reacting various external substances by adding various functional groups. In particular, fluorescence is a method that is easy to detect because it emits a specific fluorescence even at a trace concentration of micromolar or nanomolar.

In view of this point, the present invention relates to a fluorescent molecule compound having mercury ion, active oxygen, or a selectivity capable of simultaneously detecting mercury ions, active oxygen, or the like based on a body fluid compound exhibiting fluorescence properties, Or a derivative thereof.

In the compounds of formula (1) or (2) of the present invention, it is preferable that each of E ₁ , E ₂ and E ₃ is independently S. Each of R ₁ and R ₂ independently represents a 2-pyridyl group, R ₃ - R ₆ is preferably methyl, and X is bromine.

Accordingly, the chemical sensor compound of the present invention may include a compound represented by the following general formula (1-1) or (2-1).

[Formula 1-1]

[Formula 2-1]

The chemical sensor compound of the present invention generates a strong fluorescence signal even at a low concentration of several nanometers to a few nanometers to a subnumolar concentration, so that a compound capable of detecting mercury and active oxygen can be provided. Therefore, the chemical sensor compound of the present invention is characterized in that it has the function of simultaneously detecting mercury metal ions, active oxygen species (ROS), or mercury metal ions and active oxygen species (ROS).

In addition, the present invention aims to provide a fluorescent detection compound that operates in a turn-on manner instead of a turn-off method in the case of mercury. For this purpose, the development of specific functional groups capable of binding metal ions is of utmost importance. In addition, for the detection of active oxygen, a functional group reactive with active oxygen is required.

Accordingly, the present invention is based on the finding that the use of boron dipyrrole (BODIPY) as a catalyst for the synthesis of sulfur-containing sulphides and thienyl functional groups, or the use of compounds substituted with oxygen, selenium or tellurium instead of sulfur, Which is a derivative thereof. Since the chemical sensor compound has a functional group capable of reacting with mercury metal ion and active oxygen, it can detect mercury and active oxygen in the conventional atmosphere economically and can help diagnosis and treatment of disease.

The chemical sensing of the present invention is to design a highly sensitive molecule in the electron donor and electron acceptor duplex by connecting the 3-thienyl sulfur element side to the aryl sulfide group. In addition, the present invention can perform chemical sensing using oxygen, selenium or tellurium instead of the sulfur element.

When E ₁ , E ₂ and E ₃ are respectively sulfur atoms in the formula 1 or 2 of the present invention, the chemical sensitizing compound is a compound in which the sulfide group, which is a functional group in the compound, reacts with the ROS substance to form sulfoxide or R ₂ S -OOOO ^- materials. In the chemical sensor compound of Chemical Formula 1 or Chemical Formula 2, when a compound in which E ₁ , E ₂ , and E ₃ are each substituted with an oxygen element instead of the sulfur element is used, the ether group that is a functional group in the compound reacts with the ROS substance, There is a change phenomenon. Therefore, the present invention can detect ROS materials using this method.

In the chemical sensor compound of Chemical Formula (1) or Chemical Formula (2), when a compound in which E ₁ , E ₂ , and E ₃ are each substituted with a selenium element instead of the sulfur element is used, the selenide group, which is a functional group in the chemical sensor compound, And the ROS material can be detected using the phenomenon that the material is converted into selenium oxide material.

Further, in the chemical sensor compound of Chemical Formula (1) or Chemical Formula (2), when a compound in which E ₁ , E ₂ , and E ₃ are each substituted with a tellurium element in place of the sulfur element is used, the telluride group that is a functional group in the compound reacts with the ROS substance ROS materials can be detected using a phenomenon that is converted to an oxide material.

Further, the present invention relates to a method for producing a silver halide photographic light-sensitive material by using a functional group in which two sulfide groups and one thienyl group are contained in a BODIPY-based compound represented by the general formula (1) or (2) And mercury can be detected using the phenomenon that the characteristic of the light absorption and generation changes. Further, by using a compound substituted with an oxygen element, a selenium element, or a tellurium element in place of the sulfur element in the same manner, mercury ions are detected using a phenomenon in which the characteristics of light absorption and color development are changed when mercury metal ions are bonded .

In this case, the ROS substance may be detected by using a compound substituted with an oxygen element instead of the sulfur element, and a phenomenon that a functional ether group in the compound reacts with an ROS substance to convert to an oxide substance. Also, by using a compound substituted with a selenium element or a tellurium element instead of the sulfur element, a selenide group or a telluride group, which is a functional group in the compound, is converted into a selenium oxide or tellurium oxide material by reacting with the ROS substance ROS material can be detected.

The present invention also relates to a method for producing a compound of formula (I) or (II) wherein a mercury metal ion and a ROS substance are combined with a compound in a cell using a functional group in which two sulfide groups and one thienyl group are contained in a BODIPY- , Mercury ions and ROS can be detected at the same time using the phenomenon that the characteristic of fluorescence changes.

In addition, the present invention relates to a method for preparing a compound of formula (I) or (II) by using a compound substituted with an oxygen element, a selenium element or a tellurium element instead of a sulfur element, Mercury ions and ROS can be detected at the same time.

In addition, the present invention relates to a method for producing a silver halide photographic light-sensitive material, which comprises using a functional group in which two sulfide groups and one thienyl group are contained in a BODIPY-based compound represented by Chemical Formula 1 or 2, , Cysteine, histidine, arginine, and lysine may be used to remove mercury ions from the compound to regenerate the compound.

In addition, the present invention relates to a method for producing a cysteine, a histidine, or a cysteine compound, which uses a compound substituted with an oxygen element, a selenium element or a tellurium element instead of a sulfur element in the structure of the formula (1) , Arginine, and lysine to remove mercury ions from a compound to thereby regenerate the compound.

In addition, when the chemical sensor compound of the present invention is treated with an amino acid such as cysteine or histidine, the color development and decolorization reaction of mercury ions can be reversibly confirmed. In particular, the compound represented by the formula (1) may exhibit a hypsochromic shifting phenomenon when histidine, arginine, lysine or cysteine is present in the presence of mercury ions. In addition, when applied to neuron neurons, it reacts with superoxide and mercury to generate fluorescence of a specific wavelength, and it is confirmed that diagnosis is possible.

As a result of screening analysis of the above formula (1) or (2) with various types of ROS, the sulfide groups of the compounds 1 and 2 are changed to sulfoxide and sulfone groups when they react with ROS.

Based on this point, the chemical sensor compound of Formula 1 or 2 of the present invention exhibits an 'AND' logic gate when mercury ion and ROS ion are present.

That is, the present invention relates to a method for preparing a compound of formula (I) or (II) using a BODIPY-based compound having two sulfide groups and one thienyl group in a compound of formula ND ' molecular logic circuit using the generated reaction can be provided.

In the following, the reaction mechanism will be described more specifically by taking, as an example, a sulfur element among the chemical sensor compounds of the present invention.

In the chemical sensor compound of Chemical Formula (1) or (2) of the present invention, the 2-pyridine sulfur conjugate can be changed due to the oxidation of ROS and can act as a ligand under coordination of metal ions. In addition, the chemical sensor compound of the present invention is designed to have a stable sensing structure by strengthening the aromatic ring molecular structure. Therefore, it is preferable that the frame of the molecule of the chemical sensor compound of the present invention uses a BODIPY frame.

In the formula (1) or (2) of the present invention, the Aryl-sulfide group and the 3-thienyl group can serve as electron donors and acceptors of the dimer. Sulfur ions of 2-pyridyl-S-arm can detect both ROS and transition metal ions simultaneously, and detectability is greatly enhanced by the structural rigidity of the meso group when reacting with ROS and transition metal ions .

Accordingly, the chemical sensing of the present invention can simultaneously sense metal and ROS by the sulfur element of Aryl-sulfide group and 3-thienyl group.

As a specific example of the fluorescence intensity of the chemical sensor compound of the present invention, when the mercury ion and the superoxide substance KO ₂ are simultaneously reacted, the mercury ion increases by 25 times. In addition, there is a weak increase in fluorescence intensity when the chemical sensor compound works separately with mercury ion and superoxide.

In addition, the chemical sensor compound of the present invention can detect mercury ions through a discoloration reaction that shifts from yellow to red by 25 nm.

In the case of the present invention, the mercury ion and the active oxygen can be chemically sensed by substituting the oxygen element, the selenium element or the celium element instead of the sulfur element with the similar principle.

According to another embodiment of the present invention, a step of reacting an aldehyde compound having a substituent represented by the following formula (3) or (4) with 2,4-dimethylpyrrole and p-chloranil under a solvent, a base and a Lewis acid catalyst A process for producing a chemical sensor compound according to claim 1, comprising the steps of:

(3)

[Chemical Formula 4]

In the formulas (3) and (4)

E ₁ , E ₂ and E ₃ are each independently O, S, Se or Te,

R ₁ and R ₂ are each independently hydrogen or an alkyl group having 1 to 20 carbon atoms or an aryl group having 6 to 30 carbon atoms, which may contain a hetero atom,

X is a halogen element.

In Formula 3 or 4, R ₁ and R ₂ are each independently an aryl group having 6 to 30 carbon atoms and having a nitrogen atom. Most preferably, each of R ₁ and R ₂ may independently be a 2-pyridyl group.

The chemical sensor compound of the present invention is based on the production method of the body compound and can be easily synthesized by a cheap and easy method.

The method of preparing a chemical sensor compound of the present invention also includes a method of synthesizing an aldehyde substance or a carboxyl compound for synthesizing a BODIPY compound in which two sulfide groups and one thienyl group are contained in a compound.

Accordingly, the present invention first prepares an aldehyde compound having a substituent represented by the formula (3) or (4) used for meso position in order to synthesize the bipyridyl derivative compound of the formula (1) or (2). This is to stabilize the molecular structure of the aromatic ring and to provide stable chemical detection.

In a preferable example, the conjugation form of BODIPY in the synthesis of the chemical sensor compound of the present invention is a halogen reaction of 3-thiophene carboxaldehyde in a DMF solvent It is important to synthesize 2,5-dibromo-3-thiophene carboxaldehyde (2,5-bromo-3-thiophenecarboxaldehyde). This process can be carried out by the following Reaction Scheme 1.

[Reaction Scheme 1]

Wherein E ₁ , E ₂ , E ₃ , R ₁ , R ₂ , X are as defined above,

In addition, the aldehyde group of the compound may include a disulfide group of the formula (3) and an aldehyde compound converted to a monosulfide group of the formula (4) by copper coupling.

The aldehyde compound having a substituent represented by the general formula (3) or (4) thus obtained can be produced as a chemical sensor compound represented by the general formula (1) or (2) as a BODIPY derivative compound by bonding with 2,4-dimethylpyrrole have. At this time, the reaction is carried out in the presence of a solvent, a base, and a Lewis acid catalyst, and may be carried out according to a method of producing a bipiper using p-chloranil. According to this method, the chemical sensor compound of Formula 1 or 2 can be produced at a yield of 42 to 46%.

On the other hand, in the method of the present invention, the Lewis acid catalyst can be usually used for the organic synthesis, but it is preferable to use trifluoroacetic acid (TFA). In the above reaction, at least one acid selected from the group consisting of acetic acid and hydrochloric acid may be further used. The 2,4-dimethylpyrrole compound may be reacted at a molar ratio of 2 to 4 with respect to 1 mole of the aldehyde compound having a substituent represented by the following general formula (3) or (4). Furthermore, the base is not particularly limited, since a material well known in organic synthesis can be used. Examples of the base include triethylamine, piperidine, N, N -isopropylethylamine and the like, and N, N -isopropylethylamine is more preferably used.

Further, the reaction for preparing the aldehyde compound of formula (3) or (4) may be carried out at a temperature of 50 ° C to 100 ° C for 1 hour to 24 hours.

The process for preparing a bipyridyl derivative of formula (1) or (2) may be carried out at a temperature of 0 ° C to 10 ° C for 6 hours to 24 hours.

The chemical sensor compound obtained according to this method can be usefully used in a method for detecting mercury metal ion and active oxygen material.

Also, in the case of the present invention, it is possible to provide a compound containing a functional group such as a sulfonyl group in the E2 or E3 group by oxidizing a chemical sensor compound selected from the compounds represented by the above-mentioned formulas (1) and (2). The oxidizing agent may be any ordinary oxidizing agent used in organic synthesis, and is not particularly limited.

The compound obtained through the oxidation may include the following formula (5) or (6).

[Chemical Formula 5]

[Chemical Formula 6]

Best Mode for Carrying Out the Invention Hereinafter, the function and effect of the present invention will be described in more detail through a specific embodiment of the present invention. It is to be understood, however, that these embodiments are merely illustrative of the invention and are not intended to limit the scope of the invention.

< Example >

Experimental General : All reagents used in the experiments were purchased commercially from companies such as Aldrich, Acros and Junsei. The preparation of BODIPY followed the method of the following reference: (a) C. Brkner, V. Karunaratne, SJ Rettig, D. Dolphin, Can J. J. Chem. 1996, 74, RW Wagner, JS Lindsey, Pure Appl . Chem . 1996, 68, 1373. (c) CH Lee, JS Lindsey, Tetrahedron 1994, 50, 11427.)

¹ H NMR and ¹³ C NMR were performed on a Bruker Avance 400 MHz spectrometer and TMS was used as an internal standard. ¹ H NMR and ¹³ C NMR are internally calibrated by reliable protio impurity and carbon resonance of CDCl ₃ . High-resolution hybrid tandem LC-MS / MS (manufacturer: Bruker Daltonik (Germany)) in KAIST was measured with the help of instrument operator to obtain mass information of the compound.

Absorption and emission spectroscopy (Absorbance and Emission spectroscopy ) : The BODIPY probe was dissolved in acetonitrile to a concentration of 10 ^-6 M. The absorbance was measured using a CARY 300 Bio UV-Vis spectrometer and a fluorescence spectrometer RF-53011 PC spectrometer. The excitation wavelength of the fluorescence used the wavelength having the maximum absorbance. To measure the quantum yield (φ _F ), fluorescein was dissolved in 0.1 N NaOH (φ _F = 0.92) as the calibration solution and used.

&Lt; Preparation Examples 1 and 2 &

Synthesis of Compounds 3 and 4

References (AK Verma, J. Singh, R. Chaudhary, Tetrahedron Letters 2007, 48, 7199). First, CuI (0.5 mol%) and benzotriazole (1.0 mol%) were dissolved in 2 ~ 3 mL of DMSO. An aryl halide (here, 2,5-dibromo-3-thiophene carbaldehyde) (1.0 g, 3.7 mmol) was added thereto and stirred for 10 minutes. Then 2-pyridyl-thiol (2-pyridyl-thiol) ( 0.87g, 7.77 mmol) and Cs ₂ CO ₃ (1.69 g, 5.18 mmol) was added thereto, followed by stirring for 10 to 12 hours while maintaining the temperature at 90 to 95 ° C. The reaction procedure was monitored by TLC. After the reaction was completed, 30 ml of ethyl acetate was added and the layers were washed three times with 10 ml of water each. The solution was dehydrated with Na ₂ SO ₄ and the solvent was removed by rotary evaporation. The viscous material was subjected to column chromatography (dichloromethane / methanol) to obtain compounds 3 and 4

Compound 3 :

Yield = 0.55 g, 44.9%. _{^{1 H-NMR (CDCl 3,}} δ 7.24, 400 MHz): δ 10.0 (s, 1H 6), 8.40 (m, 1H 11), 8.36 (m, 1H 16), 7.66 (s, 1H 4), 7.53 ( _{m, 1H 9), 7.48 (} m, 1H 14), 7.19 (dd, 3 J H -H = 8.1 Hz, 4 J H H = 1.2 Hz, 1H 8), 7.07 (m, 1H 10), 7.03 (dd _{^{, 3 J HH = 8.0 Hz,}} 4 J H -H = 1.2 Hz, 1H 13), 7.0 (m, 1H 16).

_{^{13 C (CDCl 3, δ 77.0}} , 400 MHz) δ 184.0 (C 6), 158.7 (C 12), 156.3 (C 7), 149.4 (C 11), 149.3 (C 16), 145.9 (C 2), 141.9 _{(C 3), 137.1 (C} 9), 137.9 (C 14), 134.5 (C 4), 132.7 (C 5), 122.1 (C 8), 121.5 (C 10), 120.9 (C 13), 120.6 (C ₁₅ ).

ESIMS (positive mode, CHCl ₃ + CH ₃ OH) = [M + Na] ⁺ = 352.9853 (cal.), 352.9850 (exp.); _{[M + Na + CH 3 OH} ] + = 385.0115 (cal.), 385.0103 (exp.).

Compound 4 :

Yield = 0.150 g, 13.5%. _{^{1 H-NMR (CDCl 3,}} δ 7.24, 400 MHz): δ 9.98 (s, 1H 6), 8.44 (m, 1H 11), 7.58 (m, 1H 9), 7.51 (s, 1H 4), 7.15 ( ^{_{dd, 3 J H -H = 8.1}} Hz, 4 J H H = 1.0 Hz, 1H 8), 7.12 (m, 1H 10).

_{^{13 C (CDCl 3, δ 77.0}} , 400 MHz) = 183.7 (C 6), 157.1 (C 7), 149.8 (C 11), 143.8 (C 2), 141.4 (C 3), 137.3 (C 9), 129.3 (C ₄ ), 121.7 (C ₅ ), 121.5 (C ₈ ), 116.9 (C ₁₀ ).

ESI-MS (positive mode, CHCl ₃ + CH ₃ OH) = [M + Na] ⁺ = 321.8972 (cal.), 321.8963 (exp.); [M + H] &lt; ⁺ &gt; = 299.9152 (cal.), 299.9090 (exp.); _{[M + Na + CH 3 OH} + CH 3 + H] + = 299.9547 (cal.), 369.9223 (exp.).

&Lt; Example 1 >

Synthesis of Compound 1-1

Compound 3 (0.250 g, 0.7565 mmol), 2,4-dimethylpyrrole (0.16 mL, 1.51 mmol), TFA (trifluoroacetic acid) (0.01 mL, 0.076 mmol), p- (p-Chloranil) (0.205 g , 0.832 mmol), N, N - diisopropylethylamine (N, N -diisop ℃ ropylethylamine) (1.32 mL, 7.57 mmol) and BF ₃ and _{Et 2 O (1.03 mL, 7.57} mmol ) Was used to produce a chemical sensor compound 1 by reacting by a method of making BODIPY.

That is, a 250 mL two-neck round bottomed flask was filled with argon gas, and the compound 3, 2,4-dimethylpyrrole and TFA were added to dichloromethane maintained at 0 占 폚. After about 1 hour, N, N-diisopropylethylamine ( N, N- isopropylethylamine) was added to neutralize the pH to 7, p-chloranil was added while maintaining the temperature at 0 ° C , And stirred for about 3 hours. Again, N, N-di-isopropylethylamine was added to neutralize the pH to 7, BF ₃ Â OEt ₂ was added, and the mixture was stirred at room temperature for 2 hours. After completion of the reaction, the reaction product was purified by silica column chromatography to obtain Compound 1-1.

Yield = 0.20 g, 41.5%. _{^{1 H-NMR (CDCl 3,}} δ 7.24, 400 MHz): δ 8.38 (m, 2H 15 + 20), 7.49 (m, 2H 13 +18), 7.19 (d, 3 J H H = 8.4 Hz, 1H 12 _{), 7.13 (s, 1H 10} ), 7.12 (d, 3 J H -H = 7.6 Hz, 1H 18), 7.03 (m, 2H 14 +19), 6.0 (s, 2H 2), 2.5 (s, 6H ₂₁ ), 1.7 (s, 6H, ₂₂ ).

_{^{13 C (CDCl 3, δ 77.0}} , 400 MHz) = 160.0 (C 16), 156.0 (C 11), 155.4 (C 1), 149.5 (C 15 +20), 142.7 (C 3), 137.7 (C 5) _{, 136.9 (C 13), 136.7} (C 18), 135.9 (C 10), 134.4 (C 9), 134.0 (C 6), 133.4 (C 7), 131.3 (C 4), 123.4 (C 2), 121.5 (C ₁₂ ), 121.3 (C ₁₄ ), 120.7 (C ₁₇ ), 120.6 (C ₁₉ ), 14.6 (C ₂₁ ), 13.9 (C ₂₂ ).

¹¹ B-NMR (CDCl ₃ , BF ₃ Â OEt ₂ , δ 0.00): 5.41 (t, 32.86 Hz). ESI-MS (positive mode, CHCl ₃ + CH ₃ OH) = [M + Na] ⁺ = 571.1044 (cal.), 571.1062 (exp.).

&Lt; Example 2 >

Synthesis of Compound 2-1

Compound 4 (0.10 g, 0.33 mmol) , 2,4- dimethyl-pyrrol (2,4-dimethylpyrrole) (0.07 mL , 0.67 mmol), TFA (0.003 mL, 0.033 mmol), Chloranil (0.090 g, 0.366 mmol), N , N- i-isopropylethylamine (0.58 mL, 3.3 mmol) and BF ₃揃 Et ₂ O (0.45 mL, 3.3 mmol) were used in place of N, N -diisopropylethylamine The chemical sensor compound 2-1 was synthesized based on the BODIPY compound in the same manner.

Yield = 0.080 g, 46.3%. _{^{1 H-NMR (CDCl 3,}} δ 7.24, 400 MHz): δ 8.38 (dd, 3 J HH = 4.6 Hz, 4 J H -H = 1.2 Hz, 1H 15), 7.46 (m, 1H 13), 7.14 ( ^{_{d, 3 J H -H = 8.0}} Hz, 2H 12), 7.03 (m, 1H 14), 6.90 (s, 1H 10), 5.93 (s, 2H 2), 2.51 (s, 6H 16), 1.64 (s , 6H ₁₇ ).

_{^{13 C-NMR (CDCl 3,}} δ 77.0, 400 MHz) = 156.1 (C 1), 156.7 (C 11), 149.5 (C 15), 142.8 (C 4), 138.5 (C 9), 136.7 (C 13) _{, 133.6 (C 7), 131.3} (C 3), 129.7 (C 10), 129.2 (C 6), 123.0 (C 12), 121.4 (C 2), 121.3 (C 14), 116.9 (C 5), 14.6 (C ₁₆ ), 13.8 (C ₁₇ ).

¹¹ B-NMR (CDCl ₃ , BF ₃ O EE ₂ , δ 0.00): 5.34 (t, 32.0 Hz). ESI-MS (. Exp) ( positive mode, CHCl 3 + CH 3 OH) = [M + Na + 2H] + = 542.0319 (cal.), 542.0147.

&Lt; Examples 3 and 4 >

Synthesis of Compound 5 and Compound 6

Compound 2-1 (0.040 g, 0.0772 mmol) was dissolved in 20 mL of dichloromethane and placed in an ice bath to cool. M- CPBA (meta-chloroperoxybenzoic acid, 0.033 g, 0.193 mmol) was slowly added thereto, followed by reacting for 2 hours while stirring the solution. After 2 hours, the eluent (dichloromethane / methanol = 99/1) was confirmed by TLC, and R _f New spots were identified in ~ 0.25 (for 5) and 0.40 (for 6). Separation and purification were conducted using column chromatography to obtain compounds 5 and 6. [

Compound 5 :

Yield = 0.020 g, 48.8%. _{^{1 H-NMR (CDCl 3,}} δ 7.24, 400 MHz): δ 8.49 (dd, 3 J HH = 4.36 Hz, 4 J H -H = 1.1 Hz, 1H 19), 7.96 (d, 3 J H -H = _{7.9 Hz 1H 16), 7.88 (} ddd, 3 J HH = 7.7 Hz, 3 J H -H = 7.7 Hz, 4 J H -H = 1.8 Hz, 2H 17), 7.33 (ddd, 3 J H H = 7.4 Hz _{^{, 3 J HH = 4.6 Hz,}} 4 J H -H = 1.3 Hz, 1H 18), 6.93 (s, 1H 14), 6.02 / 5.98 (s, 1H 2, 1H 8), 2.54 (s, 6H 20/22 ), 1.71 / 1.61 (s, 6H 21/23).

¹³ C-NMR (CDCl ₃ ,? 77.0, 400 MHz) = 163.3, 157.2, 156.7, 149.9, 144.4, 143.2, 143.1, 140.2, 138.1, 131.4, 131.1, 130.6, 130.5, 125.5, 122.1, 121.5, , 14.7, 13.9, 13.7.

¹¹ BNMR (CDCl ₃ , BF ₃ Â OEt ₂ , δ 0.00): 5.40 (t, 31.3 Hz). ESI-MS (positive mode, CHCl ₃ + CH ₃ OH) = [M + Na + 2H] ⁺ = 558.0268 (cal.), 542.0072 (exp.).

Compound 6 :

Yield = 0.008 g, 18.6%. _{^{1 H-NMR (CDCl 3,}} δ 7.24, 400 MHz): δ 8.54 (m, 1H 15), 7.78 (d, 3 J H -H = 7.9 Hz, 1H 12), 7.64 (ddd, 3 J H -H ^{_{= 7.7 Hz, 3 J H H}} = 7.7 Hz, 4 J HH = 1.7 Hz, 2H 13), 7.33 (ddd, 3 J H -H = 7.7 Hz, 3 J H -H = 4.7 Hz, 4 J H -H _{= 1.2 Hz, 1H 14),} 6.98 (s, 1H 10), 5.80 (s, 2H 2), 2.50 (s, 6H 16), 1.38 (s, 6H 17).

¹³ C-NMR (CDCl ₃ ,? 77.0, 400 MHz) = 156.8, 155.9, 150.1, 142.6, 138.9, 138.5, 137.7, 132.4, 130.5, 127.4, 124.2, 122.0, 121.5, 14.6, 13.5.

¹¹ B-NMR (CDCl ₃ , BF ₃ Â OEt ₂ , δ 0.00): 5.23 (t, 33.6 Hz). ESI-MS (positive mode, CHCl ₃ + CH ₃ OH) = [M + Na + 2H] ⁺ = 574.0217 (cal.), 542.0019 (exp.).

<Experimental Example>

1. Fluorescence Experiment

For the compound 1-1 of Example 1 and the compound 2-1 of Example 2, mercury metal ion and active oxygen material detection experiments were conducted. Fig. 1 shows the fluorescence characteristics of mercury ion and active oxygen for the compound 1-1 of Example 1. Fig. The fluorescence properties for the active oxygen of Compound 2-1 of Example 2 are shown in Fig.

At this time, the value of φ _F was 0.019 and 0.026 in water and acetonitrile (? 30:? 70%) solvent, respectively. When the S-2-Py group is substituted with Br, the signal changes. In the absence of the aryl 8-substituent group, strong fluorescence appears because of the typical PET quenching effect.

1, in the case of compound 1-1, when mercury ions were reacted, there was a small fluorescence increase, but no fluorescence was observed after the oxidation. Also, intermediates such as S _sulfide -OOK were not separated and compounds 5 and 6 could be separated.

Also, it can be confirmed that the fluorescence turns on while titrating KO ₂ . This probe is sulfoxide or sulfone and R ₂ S-OO ^- means that it has changed to the compound and the like.

That is, the ROS material of m-CPBA, KO ₂ , H ₂ O ₂ , NaOCl, tBuOOH, · OH, and OtBu was reacted with 1-1 and 2-1 in MeCN / H ₂ O (70:30). As a result, KO ₂ showed a strong change (FIG. 2). From the above experiment, it was found that the compound 1-1 and 2-1 reacted with KO ₂ to form a sulfoxide group or a sulfone group. In addition, when the compound 1-1 and 2-1 were reacted with KO ₂ , the fluorescence was found to be 150% stronger, and m -CPBA was slightly increased by 20%.

At this time, Compound 2-1 was used to synthesize mono-oxidized, di-oxidized sulfur atom-containing molecules by using KO ₂ in acetonitrile-H ₂ O solvent. However, the compound was decomposed and could not be synthesized. And then synthesized using m -CPBA in a CH ₂ Cl ₂ solvent. When the Φ _F value of the resultant product was measured, it was 0.87 and 0.77, respectively. No fluorescence was observed when the Hg ^{2 +} ion was titrated using the newly synthesized oxidized probe.

In compound 2-1, N _py and S _sulfide groups showed reactivity of metal ion bond and ROS. This reaction was confirmed to be due to the quenching effect of PET and to have a 'turn on' effect (maximum wavelength 524 nm).

2. Decision data

The crystal structure was measured for Compound 2-1 of Example 2 by a conventional method, and the results are shown in Fig. In Fig. 3, a is a side view of compound 2-1 and b is a bottom view of compound 2-1.

Compound _{2-1 (CCDC 877969): C 22} H 19 BBrF 2 N 3 S 2, FW: 518.24.

B = 11.291 (2) Å, c = 27.486 (5) Å, β = 95.942 (8) °, V = 2327.7 (7) Å, Monoclinic, P2 (1) / n, Z = 4, a = 7.5410 ) A, T = 296 (2) K, N. reflections = 38634, No. independent reflections = 6972 [R (int) = 0.1663], R1 = 0.0671, wR2 = 0.1157, Abs. coeff. (mm ^-1 ) = 1.976, Largest diff. peak and hole = 0.793 and 0.553 e ^- / Å ³ .

Looking at the molecular geometry of FIG. 3, the structure of compound 2-1 has a single 2-Py-S group, which is in the second position of the thienyl group. This is confirmed by instrumental analysis such as NMR. In the compound 2-1, the C-Br group is located at the 5-position. The twist angle of the thienyl group and the BODIPY group in the molecular structure of the compound 2-1 is approximately 80 degrees. This shows that there is a steric effect or an electronic effect. The Py group had a twist angle of about 64 degrees with respect to the thienyl group, and no determination of the bonded state of the metal ion was obtained.

3. Metal ion selectivity test

In solution with respect to ^{Cd 2 +, Hg 2 +,} Fe 2 +, Ca 2 +, Cu 2 +, Co 2 +, Mn 2 +, Mg 2 +, Pb 2 +, Zn 2 + , and Ag ⁺ metal perchlorate compounds , The reactivity of the compound 1-1 and the compound 2-1 was confirmed. The results of Compound 1-1 are shown in Fig.

From the results of FIG. 4, in the case of the compound 1-1, a fluorescence wavelength shift occurred by reacting with mercury, thereby confirming discoloration of the solution.

4. Reversible reaction experiment with amino acid

Compounds 1-1 and 2-1 were examined for their ability to proceed with the reverse reaction by amino acid reaction. FIG. 5 shows the results of reversible reaction of Compound 1-1 of Example 1 with various amino acids. FIG.

In FIG. 5, only Hg ^{2 +} reacted among various metals, and the initial light yellow to strong orange color was discolored. Further, when the absorption wavelength was measured, it was confirmed that the? _Max changed from 506 nm to 532 nm. At the compound 1-1 concentration of 10 ^-6 M, sufficient color change was observed even with this degree of bathochromic shift for visual observation.

5. Bonding ratio test with mercury ion

Compounds 1-1 and 2-1 were each titrated with mercury ions and their binding ratios were measured. As a result, it was found that Compound 1-1 and mercury ion were bonded at a ratio of 1: 1, respectively, and confirmed by Job plot analysis. 6 shows the results of Job plot analysis for confirming the binding ratio of the compound 1-1 of Example 1 with mercury.

6. Amino acid treatment test

Amino acids were treated in the binding state of mercury ion and compound 1-1. That is, the compound 1-1 of Example 1 was bound to various concentrations of Hg ^{2 +} ions while the concentration of KO ₂ was fixed, and various kinds of amino acids were treated. FIG. 7 shows Stern-Volmer plots of various concentrations of Hg ions and probes with the concentration of KO ₂ fixed to the compound 1-1 of Example 1. FIG.

From the results of FIG. 7, it can be seen that the mercury is separated from the compound 1-1 and the reaction proceeds in a reversible reaction, and the color development again appears. At this time, the amino acids reacted with mercury were cysteine, histidine, arginine, and lysine, and they were found to be completely inverted at mercury binding sites. The fluorescence efficiency was recovered when 4 equivalents of amino acid were treated.

In addition, when the fluorescence was examined, the quenching effect was confirmed when the mercury was titrated, and the quenching effect was weak in the case of copper. Other ions did not affect fluorescence. The quenching effect of mercury-induced fluorescence is believed to be due to the induction of spin-orbit coupling between mercury and probe.

In addition, Stern-Volmer plots of Hg ^{2 +} ions and Compound 1-1 were obtained at various concentrations while the concentration of KO ₂ was fixed. Compound 1-1 exhibited saturating behavior at high concentrations. Artificial neural networking was attempted to optimize the relationship between optical yield and fluorescence of the reactants.

7. Reactivity of probes in neuroblastoma cells

With respect to the compound 1-1 of Example 1, the reactivity of probes in neuroblastoma cells was examined. 8 shows the reactivity of the compound 1-1 of Example 1 in neuroblasticoma cells (a: when Compound 1-1 was treated, b: Compound 1-1, Hg ^{2 +} , and KO ₂ were treated simultaneously ).

In the neuroblasticoma cells, fluorescence was observed in the probe and ROS in the absence of Hg ^{2 +} . However, cytotoxicity was very low.

8. Confirmation of PET passage mechanism of Compound 1-1

9 shows a double PET passage mechanism acting upon the reaction of Compound 1-1 of Example 1 with O ₂ ^- , Hg ^{2 +} .

In the case of the compound 1-1, it is considered that the bonding of the Hg ^{2 +} ion and the superoxide substance is bonded to the meso group of the compound 1-1. The reaction of one Hg ^{2 +} ion with one compound 1-1 molecule was confirmed by Job analysis. It is believed that the mercury ions are bonded to each other near the center, forming a _sulfide -OOK type material. When mercury ions bind to S _thi and N _py of S _sulfide -OOK molecule, it is considered that inducing PET inactivation by inducing the upper charge to the central part. Gearing occurs as the rotation of the aryl group of the enhanced steric group is weakened and appears to restore fluorescence. Oxidation of ROS by other than sulfur is considered to be difficult. This is because, when compound 1-1 is auto-oxidized to form a dimer, a strong red-shift must be observed and a better position for the turn-on phenomenon of mercury ions can not be established. Second, the formation of N _pyr -oxide by mercury ions is impossible. When mercury ions were added to excess of superoxide, the fluorescence remained turned on. When BODIPY is oxidized at positions 2 and 6 of the dyad acceptor, this is a reaction caused by electrophilic addition and is not suitable for describing stronger fluorescence. Moreover, such substituted materials are not suitable for describing strong fluorescence in the presence of mercury ions.

9. "AND" molecular logic circuit verification experiment

10 shows the 'AND' molecular logic circuit for the compound 1-1 of Example 1. FIG.

From the results of Figure 10, it was confirmed that the fluorescence strong regardless of the loading procedure of KO ₂ and Hg ^{2 +,} only one component was the less "turn on" the effect of the fluorescence can be when the Hg ^{2 +,} KO ₂ act as and , It is thought to be a useful result because the reaction occurs at a very low detection compound concentration. When both components interact with the compound, a change in geometry is observed in the donor atoms of [NS _thi N] and [S _sulfide S _sulfide ]. Initially, it acts as a support for the metal ion, but when added to the ROS material, it becomes a vertical structure. This is a multi-input system molecule, meaning "AND" logic gate configuration is possible. Regardless of the order in which they are input, it gives usability by the o- Py nitrogen group and contrasts with the previously reported p- Tol group.

Claims (7)

A chemical sensor compound selected from compounds represented by the following formulas (1) and (2).
[Chemical Formula 1]

(2)

In the formulas (1) and (2)
E ₁ , E ₂ and E ₃ are each independently O, S, Se or Te,
R ₁ , R ₂ , R ₃ , R ₄ , R ₅ and R ₆ are each independently hydrogen or an alkyl group having 1 to 20 carbon atoms or an aryl group having 6 to 30 carbon atoms, which may contain a hetero atom;
X is a halogen element.
The method according to claim 1,
A chemical sensor compound having the ability to simultaneously detect mercury metal ions, active oxygen species (ROS), or mercury metal ions and reactive oxygen species (ROS).
The method according to claim 1,
Wherein R &lt; ₁ &gt; and R &lt; ₂ &gt; are each independently a 2-pyridyl group.
2. A chemical sensor compound according to claim 1, wherein E ₁ , E ₂ and E ₃ are each independently S.
The method of claim 1, wherein the chemical sensor compound
A chemical sensor compound selected from compounds represented by the following formulas (1-1) and (2-1).
[Formula 1-1]

[Formula 2-1]

Under solvent, base and Lewis acid catalyst,
Reacting an aldehyde compound having a substituent represented by the following formula (3) or (4) with 2,4-dimethylpyrrole and p-chloriranyl
&Lt; RTI ID = 0.0 &gt; 1, &lt; / RTI &gt;
(3)

[Chemical Formula 4]

In the formulas (3) and (4)
E ₁ , E ₂ and E ₃ are each independently O, S, Se or Te,
R ₁ and R ₂ are each independently hydrogen or an alkyl group having 1 to 20 carbon atoms or an aryl group having 6 to 30 carbon atoms, which may contain a hetero atom,
X is a halogen element.
The method according to claim 6,
Wherein the 2,4-dimethylpyrrole compound is reacted at a molar ratio of 2 to 4 with respect to 1 mole of the aldehyde compound having a substituent represented by the general formula (3) or (4).

Images