KR101815791B1 - Bis(naphthalimide-piperazine) derivative, method for preparing the same and method for detecting strong acid using the same - Google Patents

Abstract

The present invention relates to a bis (naphthalimide-piperazine) derivative, a method for preparing the same, and a method for detecting strong acid using the same. According to the present invention, the bis (naphthalimide-piperazine) derivative reduces fluorescence by delivering nitrogen of piperazine to naphthalimide residues through a light-induced electron transfer reaction in neutral and alkaline pH levels and increases the fluorescence intensity by inhibiting the light-induced electron transfer reaction by a proton (H^+) in the acidic pH level, thereby detecting an acidic pH domain and strong acid. Further, the bis (naphthalimide-piperazine) derivative of the present invention has high selectivity and responsiveness to the proton and can detect the proton from alkaline and heavy metal ion or biomolecule. The bis (naphthalimide-piperazine) derivative of the present invention is highly stable in the strong acidic and alkaline states and can be used in monitoring pH changes without chemical degradation. Accordingly, the bis (naphthalimide-piperazine) derivative of the present invention can be utilized as a composition for detecting the strong acid, a pH sensor for detecting the strong acid, or a kit for detecting the strong acid in the processes in the chemistry, biology, and environment engineering fields.

Description

Bis (naphthalimide-piperazine) derivative, a process for producing the same, and a method for detecting a strong acid using the same (Bis (naphthalimide-piperazine) derivative,

The present invention relates to a bis (naphthalimide-piperazine) derivative, a process for producing the same, and a strong acid detection method using the same.

 The measurement of pH is very important in biological fluids, chemistry and the environment. For example, intracellular pH plays an important role in protein activity involved in proliferation, apoptosis, intracellular entry and other intracellular processes. Abnormal pH values affect cell function, growth, and cleavage, which are generally observed in pathogenic conditions. In addition, some industrial processes produce acids such as hydrochloric acid (HCl) and hydrofluoric acid (HF). Strong acid chemicals are fatal to the ecosystem. Therefore, these toxic species should be strictly regulated at the workplace and their emissions should be monitored. Therefore, it is necessary to develop an effective and quick sensor for the change of pH.

It is necessary to quickly and accurately analyze the concentration of various ions contained in a specific sample in various fields including clinical diagnosis. In this analysis, various kinds of chemical sensor materials using a substance having a specific ion selectivity are used. In general, a sensor senses an electrical property such as electricity, resistance, electric potential difference, and optical properties such as color and fluorescence. Among these, the change of color and the change of fluorescence can be easily discriminated by the naked eye, so it is one of the easy measurement methods without special equipment. The sensor that observes the change of fluorescence is called a fluorescence chemical sensor. Fluorescence chemistry sensors typically include photo-induced electron / energy transfer (PET), metal-ligand charge transfer (MLCT), intramolecular charge transfer (ICT), eximer / exiplex formation, imine isomerization, chelation-enhanced fluorescence (CHEF) and fluorescence resonance energy transfer (FRET) .

To date, several optical pH sensors have been developed for various chemical, biological and environmental processes. In particular, synthetic fluorescence probes are widely used for pH measurement with high sensitivity and simplicity. The most commonly used organic dyes are rhodamine, fluorescein, BODIPY, cyanine dyes and their derivatives.

Recently, naphthalimide has excellent photophysical properties such as absorption (350 to 450 nm) and emission (450 to 550 nm) wavelength in the visible region, high fluorescence quantum yield and large absorption coefficient, And is highly stable and potentially utilizable in environmental and biological sensing. However, only a few pH probes using naphthalimide have been reported so far. Also, as a pH sensing part, piperazine is widely used because it is bonded to various fluorophores due to the property of changing fluorescence intensity through electron transfer.

Japanese Patent No. 4638485

It is an object of the present invention to provide a composition for detecting a strong acid comprising a bis (naphthalimide-piperazine) derivative or a pharmaceutically acceptable salt thereof, a pH sensor for detecting a strong acid or a kit for detecting a strong acid.

It is still another object of the present invention to provide a method for producing a bis (naphthalimide-piperazine) derivative by reacting a bis (naphthalimide-piperazine) derivative with a sample containing a strong acid-derived proton (H ⁺ And measuring the at least one indicator selected from the group consisting of < RTI ID = 0.0 > a < / RTI >

In order to achieve the above object, the present invention provides a bis (naphthalimide-piperazine) derivative represented by the following formula (I) or a pharmaceutically acceptable salt thereof.

[Chemical Formula 1]

The present invention also provides a composition for detecting a strong acid comprising a bis (naphthalimide-piperazine) derivative represented by the following formula (1) or a pharmaceutically acceptable salt thereof, a pH sensor for detecting a strong acid or a kit for detecting a strong acid do.

[Chemical Formula 1]

The present invention also relates to a method for preparing a reaction product comprising the steps of adding a sample containing a strong acid-derived proton (H ⁺ ) to a bis (naphthalimide-piperazine) derivative represented by the following general formula (1) And measuring the at least one indicator selected from the group consisting of optical changes.

[Chemical Formula 1]

The bis (naphthalimide-piperazine) derivatives of the present invention transfer the nitrogen of piperazine to the naphthalimide moiety through a photoinduced electron transfer reaction at neutral and basic pH to reduce fluorescence, H ⁺ ), it is possible to detect the acidic pH region and the strong acid by increasing the fluorescence intensity. In addition, the derivatives have high selectivity and sensitivity to protons and can detect protons from alkali and heavy metal ions or biomolecules and are very stable in strong acid and base states and can be used to monitor pH changes without chemical degradation .

Therefore, the bis (naphthalimide-piperazine) derivative of the present invention can be utilized in chemical, biological and environmental engineering processes with a composition for detecting a strong acid, a pH sensor for detecting a strong acid, or a kit for detecting a strong acid.

1 schematically shows the pH sensing mechanism of bis (naphthalimide-piperazine) derivative 1. Fig.
Fig. 2 shows the synthesis method and chemical structure of bis (naphthalimide-piperazine) derivative 1.
(A) UV / Vis absorption of bis (naphthalimide-piperazine) derivative 1 and (b) absorption of bis (naphthalimide-piperazine) ) Fluorescence spectrum and (c) fluorescence intensity, FI (516 nm vs. pH).
4 (a) shows a plot of the fluorescence intensity (pH vs. log [(I _max -I) / (II _min )] of the bis (naphthalimide-piperazine) derivative 1 according to the pH value, (b) shows a fluorescence change photograph of the bis (naphthalimide-piperazine) derivative 1 according to the pH value.
To 5 is added to 50% (v / v) proton (H ⁺⁾ to pH 4.0 buffer containing perchlorate and 50% (v / v) CH ₃ CN of the metal ions in the pH 7.0 buffer containing CH ₃ CN (A) fluorescence reaction and (b) fluorescence photograph of the resulting bis (naphthalimide-piperazine) derivative 1 at 390 nm are shown here.
Figure 6 shows the reversible fluorescence change of the bis (naphthalimide-piperazine) derivative 1 between pH ~ 1 and pH 11 and the pH value was adjusted by adding hydrochloric acid (HCl) or sodium hydroxide (NaOH) all data was measured in an aqueous solution containing 50% (v / v) CH 3 CN to 390 nm here.

The inventors of the present invention synthesized bis (naphthalimide-piperazine) derivatives using two naphthalimides in order to improve the sensitivity, and these derivatives were synthesized by the method of the present invention using piperazine By providing a pH-sensitive fluorescent off-on signal by increasing the fluorescence intensity by inhibiting the photoinduced electron transfer reaction by the proton at acidic pH by reducing the fluorescence by transferring the nitrogen of the naphthalimide residue to the acid pH region and And the present inventors completed the present invention.

The present invention provides a bis (naphthalimide-piperazine) derivative represented by the following formula (I) or a pharmaceutically acceptable salt thereof.

[Chemical Formula 1]

The present invention also provides a composition for detecting a strong acid comprising a bis (naphthalimide-piperazine) derivative represented by the following formula (1) or a pharmaceutically acceptable salt thereof.

[Chemical Formula 1]

The present invention also provides a pH sensor for detecting a strong acid comprising a bis (naphthalimide-piperazine) derivative represented by the following formula (1) or a pharmaceutically acceptable salt thereof.

[Chemical Formula 1]

The present invention also provides a kit for detecting a strong acid comprising a bis (naphthalimide-piperazine) derivative represented by the following formula (1) or a pharmaceutically acceptable salt thereof.

[Chemical Formula 1]

The present invention also relates to a method for producing a biodegradable polymer comprising the steps of adding a sample containing a strong acid-derived proton (H ⁺ ) to a bis (naphthalimide-piperazine) derivative represented by the general formula (1) And measuring the at least one indicator selected from the group consisting of the change.

Preferably, the strong acid may be selected from the group consisting of hydrochloric acid, hydrofluoric acid, hydrochloric acid, and nitric acid, but is not limited thereto.

Preferably, the indicator can be measured using any one or more selected from the group consisting of a UV-Vis (Ultraviolet-visible) spectrophotometer, a fluorescence photometer, an electrospray ionization mass spectrometer and a confocal microscope, Or not.

Preferably, the coupling of the bis (naphthalimide-piperazine) derivative and the proton may be accomplished through a photoinduced electron transfer reaction.

Preferably, the sample may be, but not limited to, a sample containing a strong acid-derived proton used for industrial wastewater, a semiconductor process, an organic compound production process, an ink production process, a food additive production process, do.

Hereinafter, the present invention will be described in more detail with reference to Examples. It is to be understood by those skilled in the art that these embodiments are only for describing the present invention in more detail and that the scope of the present invention is not limited by these embodiments in accordance with the gist of the present invention .

Example  1: Preparation of reagent

All reagents and chemicals including metal ions were purchased from Sigma Aldrich (St. Louis, Mo., USA). All solvents were used as high performance liquid chromatography (HPLC) grade reagents, and triple-ionized water was used in the assay. ¹ H and ¹³ C NMR (nuclear magnetic resonance) spectra were measured with a Varian INOVA spectrometer (400 MHz), all chemical shifts are using tetramethylsilane (tetramethylsilane, TMS) as an internal standard were measured in ppm values. Electrospray ionization mass spectrometry (ESI-MS) analysis was performed using LC / MS-2020 Series (Shimadzu).

Example  2: UV / Vis  And fluorescence spectroscopy

A stock solution of bis (naphthalimide-piperazine) derivative 1 and a perchlorate of a metal ion was prepared using acetonitrile (CH ₃ CH). The various pH buffers are 50 mM potassium chloride (for pH 1-2), potassium hydrogen phthalate (for pH 3-5), potassium dihydrogen phosphate (pH 6 (For pH 9 to 10), sodium tetraborate (for pH 9 to 10), and sodium bicarbonate (for pH 11). The pH was adjusted by adding 0.1 M sodium hydroxide (NaOH) and 0.1 M hydrochloric acid (HCl) solution. UV / Vis absorption and fluorescence spectra were measured using S-3100 (Scinco) and RF-5301 PC (Shimadzu) spectrophotometer, respectively. The excitation was performed at 480 nm with a 3 nm excitation and emission slit width.

Example  3: Bis ( Naphthalimide - piperazine) derivatives

One. Bis ( Naphthalimide - piperazine) derivative 2 Synthesis

The bis (naphthalimide-piperazine) derivative 3 (200 mg, 0.60 mmol) and piperazine (518 mg, 6.0 mmol) were dissolved in 2-methoxyethanol (100 ml). The reaction mixture was stirred and refluxed under nitrogen atmosphere for one day. After removing the solvent under reduced pressure, the product was dissolved in CH ₂ Cl ₂ (100 ml) and water to give an organic layer. The CH ₂ Cl ₂ layer was dried over anhydrous magnesium sulfate (anhydrous MgSO ₄ ). After removal of the solvent, the product was purified by silica gel using CH ₂ Cl ₂ / methanol (v / v, 40: 3) to give the bis (naphthalimide- piperazine) derivative 2 (150 mg, 74% ).

ESI-MS m / z [M + H] ⁺ calc 338.19, obs 338.05. ¹ H NMR (400 MHz, CDCl ₃ )? 8.58 (dd, J = 8.00 Hz, 1H); 8.52 (dd, J = 8.00 Hz, 1H); 8.42 (dd, J = 8.00 Hz, 1H); 7.69 (t, J = 8.00 Hz, 1H); 7.22 (d, J = 8.00 Hz, 1H); 4.19 (t, J = 8.00 Hz, 2 H); 3.26 (s, 4 H); 3.23 (s, 4 H); 2.53 (s, 1H); 1.73 ~ 1.68 (m, 2H); 1.48-1.42 (m, 2H); 0.98 (t, J = 8.00 Hz, 3 H). ^{13 C} NMR (100 MHz, CDCl ₃ ): 164.4, 163.9, 156.1, 132.5, 130.9, 130.1, 129.8, 126.1, 125.6, 123.2, 116.8, 114.9, 54.1, 46.0, 40.0, 30.2, 20.4, 13.8 ppm.

2. Bis ( Naphthalimide - piperazine) derivative 1 Synthesis

Dichloro-p-xylene (30 mg, 0.15 mmol) and K ₂ CO ₃ (40 mg, 0.30 mmol) were added to a solution of bis (naphthalimide-piperazine) derivative 2 mmol) is dissolved in CH ₃ CN (100 ml). The reaction mixture was stirred and refluxed under a nitrogen atmosphere for 12 hours. After removing the solvent under reduced pressure, the product was dissolved in CH ₂ Cl ₂ (100 ml) and water to give an organic layer. The CH ₂ Cl ₂ layer was dried over anhydrous magnesium sulfate (anhydrous MgSO ₄ ). After removal of the solvent, the product was purified by silica gel using CH ₂ Cl ₂ / methanol (v / v, 20: 1) to give the bis (naphthalimide-piperazine) derivative 1 (160 mg, 70% ).

ESI-MS m / z [M + H] < ⁺ > calc 777.41, obs 777.35; [M + H + Na] ⁺ calc 800.40, 800.25 obs. ¹ H NMR (400 MHz, CDCl ₃ )? 8.58 (dd, J = 7.20 Hz, 2 H); 8.52 (dd, J = 8.01 Hz, 2 H); 8.41 (dd, J = 8.41 Hz, 2H); 7.68 (t, J = 7.81 Hz, 2H); 7.39 (s, 3 H); 7.22 (d, J = 8.01 Hz, 2 H); 4.17 (t, J = 7.4 Hz, 4 H); 3.70 (s, 4 H); 3.32 (s, 8 H); 2.82 (s, 8 H); 1.74 ~ 1.67 (m, 4 H); 1.47-1.42 (m, 4 H); 0.97 (t, J = 7.4 Hz, 6 H). ¹³ C NMR (100 MHz, CDCl ₃ ): 164.6, 164.2, 132.7, 131.2, 130.3, 130.0, 129.5, 126.3, 125.7, 123.5, 115.1, 62.9, 53.3, 40.2, 30.4, 29.8, 20.5, 14.0 ppm.

Experimental Example  One : Bis ( Naphthalimide -Piperazine) Derivative 1 and Fluorescence Spectrum

FIG. 1 is a schematic diagram showing a pH sensing mechanism of a bis (naphthalimide-piperazine) derivative 1, and FIG. 2 shows a synthesis method and chemical structure of a bis (naphthalimide-piperazine) derivative 1. The naphthalimide derivative 3 was synthesized by referring to a previously reported article (Anal Chem 81: 9993). Synthesis of bis (naphthalimide-piperazine) derivative 2 by the coagulation reaction of bis (naphthalimide-piperazine) derivative 3 and piperazine and synthesis of bis (naphthalimide-piperazine) derivative 1 Treated with diclo-p-xylene in the presence of potassium carbonate (K ₂ CO ₃ ). The chemical structure of the bis (naphthalimide-piperazine) derivatives 1 to 3 was confirmed by ¹ H NMR, ¹³ C NMR and ESI-MS spectroscopy.

To analyze the characteristics of bis (naphthalimide-piperazine) derivative 1 according to pH, absorption and fluorescence spectra of bis (naphthalimide-piperazine) derivative 1 were measured in various pH buffers. 3 (a) and 3 (b), the absorption band of bis (naphthalimide-piperazine) derivative 1 shifts from 400 nm to 380 nm as acidified from pH 11 to pH 1 , And it was confirmed that the fluorescence intensity was increased at 516 nm. Also, referring to FIG. 3 (c), it was confirmed that the fluorescence intensity was extremely weak in the range of pH 11 to 4.5, but the fluorescence intensity was increased about 22 times when the pH was lowered to 4.5 to 1.0. Thus, it can be seen that the fluorescent turn-on of the bis (naphthalimide-piperazine) derivative 1, which reacts with the acidic pH, inhibits the photoinduced electron transfer reaction through the proton of the piperazine moiety.

In previous studies, it was reported that naphthalimide is based on a photoinduced electron transfer reaction mechanism for pH sensing. The naphthalimide-piperazine derivative of the present invention confirmed a photoinduced transfer reaction involving fluorescence off-on signal change as an intracellular pH sensing probe. In the present invention analogs of the naphthalimide-piperazine deficient in the piperazine ring were prepared and the pH dependent fluorescence reaction was compared to the naphthalimide-piperazine. Analogs in which the piperazine ring was deficient showed little change in fluorescence at various pHs. Based on this, it can be seen that the pH-dependent fluorescence off-on change of the bis (naphthalimide-piperazine) derivative 1 is mediated by the photoinduced electron transfer reaction from the piperazine to the naphthalimide residue.

In addition, 4 (a) is a header Branson - using (pH) Henderson-Hasselbalch type mass action equation, (log [(I max - I) / (I - - I min)] = pK a) Hasselt Balkh equation The fluorescence intensity (516 nm vs. pH) log [(Imax-I) / (I-Imin)]. The pK _a value of the bis (naphthalimide-piperazine) derivative 1 was 2.88 ± 0.13, which can be usefully used for measuring acidic pH range, particularly pH 4.5 to 1.0 range. Also, referring to FIG. 4 (b), it was confirmed that the strong green fluorescence sharply increases in the range of pH 4.5 to 1.0. Thus, the bis (naphthalimide-piperazine) derivative 1 can be measured in the pH range of 4.5 to 1.0 and can be used to detect strong acidity such as hydrochloric acid and boric acid.

Experimental Example  2: For strong acid Bis ( Naphthalimide - < / RTI > piperazine) derivative 1

Experiments were conducted to confirm the selectivity of the bis (naphthalimide-piperazine) derivative 1 to protons (H ⁺ ) in alkali and heavy metal ions. As a result, referring to FIG. 5, it was confirmed that the bis (naphthalimide-piperazine) derivative 1 exhibited strong green fluorescence in the buffer of pH 4.0. However, in the buffer of ^{pH 7.0 Na +, K +,} Ag +, Ba 2 +, Ca 2+, Cu 2 +, Cd 2 +, Fe 3 +, Fe 2 +, Mg 2 +, Zn 2 +, Pb 2 + , Hg ^{2 +} , Ni ^{2 +} ions, and other alkali and heavy metal ions. Thus, it can be seen that the bis (naphthalimide-piperazine) derivative 1 exhibits selective fluorescence change for the proton.

Fluorescence changes of bis (naphthalimide-piperazine) derivatives 1 were also observed in the presence of biomolecules such as glutathione (GSH) and bovine serum albumin (BSA). As a result, it was confirmed that the bis (naphthalimide-piperazine) derivative 1 exhibits a weak fluorescence change to biomolecules. Thus, the bis (naphthalimide-piperazine) derivative 1 can selectively detect acidic pH regions and provide acid imaging without interfering with other competitive metal cations in various chemical, biological and environmental systems.

In addition, reversible fluorescence changes of bis (naphthalimide-piperazine) derivative 1 to pH diversity were performed using fluorescence spectroscopy. As a result, referring to FIG. 6, the reversibility of the bis (naphthalimide-piperazine) derivative 1 between pH 1 and pH 11 was found to be higher than that of the bis (naphthalimide- Lt; / RTI > derivative 1 in equilibrium mode. In addition, the bis (naphthalimide-piperazine) derivative 1 is very stable in relatively strong acidic and basic conditions and can be used to monitor pH changes in a variety of situations without chemical degradation.

While the present invention has been particularly shown and described with reference to exemplary embodiments thereof, it is clearly understood that the same is by way of illustration and example only and is not to be construed as limiting the scope of the invention. Accordingly, the actual scope of the present invention will be defined by the appended claims and their equivalents.

The scope of the present invention is defined by the appended claims, and all changes or modifications derived from the meaning and scope of the claims and their equivalents should be construed as being included within the scope of the present invention.

Claims (8)

delete A composition for detecting a strong acid comprising a bis (naphthalimide-piperazine) derivative represented by the following formula (1) or a pharmaceutically acceptable salt thereof:
[Chemical Formula 1]

A pH sensor for detecting a strong acid comprising a bis (naphthalimide-piperazine) derivative represented by the following formula (1) or a pharmaceutically acceptable salt thereof:
[Chemical Formula 1]

A kit for detecting a strong acid comprising a bis (naphthalimide-piperazine) derivative represented by the following formula (1) or a pharmaceutically acceptable salt thereof:
[Chemical Formula 1]

Adding a sample containing a strong acid-derived proton (H ⁺ ) to a bis (naphthalimide-piperazine) derivative represented by the following general formula (1); And
And measuring at least one indicator selected from the group consisting of fluorescence wavelength, fluorescence intensity, and optical change of the reacted reactant.
[Chemical Formula 1]

6. The method of claim 5, wherein the strong acid is selected from the group consisting of hydrochloric acid, hydrofluoric acid, hydrochloric acid, and nitric acid. 6. The method according to claim 5, wherein the indicator is measured using at least one selected from the group consisting of a UV-Vis (Ultraviolet-visible) spectrophotometer, a fluorescence photometer, an electrospray ionization mass spectrometer and a confocal microscope. / RTI > 6. The method of claim 5, wherein the coupling of the bis (naphthalimide-piperazine) derivative and the proton is performed through a photoinduced electron transfer reaction.

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