KR101845875B1 - Tripeptide derivatives, preparation method thereof, chemosensor having the same and detection method of aluminium ion and trivalent metal ion using the same - Google Patents

Abstract

The present invention relates to a tripeptide derivative, a preparation method for the same, a chemical sensor comprising the same, and a detection method for aluminum ions (Al^(3+)) and trivalent metal ions. The tripeptide derivative of the present invention, represented by chemical formula 1 can selectively, quantitatively, and simultaneously or at separate times detect aluminum ions (Al^(3+)), gallium ions (Ga^(3+)), indium ions (In^(3+)), and iron ions (Fe^(3+)) even in the presence of other metal ions that may act as interfering materials; and can easily penetrate into a living cell, and thus may be beneficially used as a fluorescence-sensitive chemical sensor capable of detection through changes in fluorescence intensity due to binding to aluminum ions (Al^(3+)), gallium ions (Ga^(3+)), indium ions (In^(3+)), and iron ions (Fe^(3+)) inside the living cell.

Description

TECHNICAL FIELD The present invention relates to a tripeptide derivative, a method for producing the tripeptide derivative, a chemical sensor including the tripeptide derivative, and a method for detecting an aluminum ion and a trivalent metal ion using the tripeptide derivative, a chemosensor having the same and detection method of aluminum ion and trivalent metal ion the same}

The invention is tripeptide derivatives, their preparation, chemical sensors, and using the same aluminum ion containing them (Al ^{+ 3)} and 3 relates to a method of detecting a metal ion (M ^3+).

The design and study of new sensors for the major substances and ions in vivo has been actively underway and has attracted much interest. Recently, supramolecule chemistry has shown great potential for the design of host compounds that can bind selectively to ions or various other guest compounds using various molecules such as amino acids, peptides, etc. Recently, The development of a fluorescent chemosensor capable of more easily detecting the change in fluorescence generated upon binding with a specific guest compound has been actively studied.

Fluorescence is a photochemical phenomenon that occurs when a photon with a specific wavelength (excitation wavelength) collides with an indicator molecule and excites the electron to a high energy level as a result of the collision. Among the various analytical methods, the fluorescence method has a great advantage of observing signals at a concentration of 10 ^-9 M due to its excellent sensitivity. Therefore, this analytical method has been widely used. In recent years, studies have been made on the application to fluorescent chemical sensors for the detection of cations, anions and neutral organic molecules by using such properties (Non-Patent Document 1).

Aluminum (Al) is the most abundant metal in the metal, and among all the elements, oxygen and silicon are the next most abundant elements. Also, it is widely used in many industrial fields such as automobile and computer manufacturing industries and medical fields. However, trivalent aluminum ions (Al ^{3 +} ) are known to be toxic in biochemical terms. High levels of aluminum ions in the body cause serious damage to the nervous system, including Alzheimer's and Parkinson's. In addition, high concentrations of aluminum ions cause soil acidification and damage to the aquatic environment. For example, aluminum in water inhibits plant growth because it inhibits phosphorus absorption. Therefore, the US Environmental Protection Agency (EPA) strictly regulates the amount of aluminum ions in drinking water and surface water. For this reason, the detection of low concentrations of aluminum ions in aqueous solutions is of great environmental importance. Although conventional methods using instruments such as an inductively coupled plasma mass spectrometer (ICP-MS) and an Atomic Absorption Spectrophotometer (AAS) are still used to identify low levels of aluminum ions, The methods are expensive and require a lot of time, and it is impossible to detect aluminum in real time.

Gallium ions (Ga ^{3 +} ), a trivalent ion in the same group as aluminum in the periodic table, are present in small amounts in water, vegetables, and fruits. Gallium compounds in the body can cause neck pain, dyspnea and chest pain. In addition, exposure to gaseous forms of gallium can lead to very severe conditions, such as pulmonary edema and partial paralysis. Indium (In) is also an element that is analogous to aluminum and gallium. It is known that the indium ion (In ^{3 +} ) in aqueous solution interferes with the metabolism and damages the heart, liver and lung.

Iron (III) in the trivalent ion is a compatible material in the enzyme-catalytic reaction because it has a rapid oxidation-reduction reaction rate and is used as a co-factor in the electron transport system. Thus, imbalance of iron ions in the body can cause many diseases. In the presence of excess trivalent iron, radicals can be generated to damage DNA and organ proteins, and deficiency of trivalent iron can lead to anemia, liver damage, kidney damage, diabetes and heart disease.

Therefore, it is very important to develop a detection agent capable of selectively detecting a trivalent metal ion (M ³⁺ ), and many studies have been made. However, a detection agent that simultaneously detects a trivalent metal ion (M ³⁺ ) It is very difficult to develop. In fact, chemical sensors that detect trivalent metal ions (M ³⁺ ) in addition to aluminum ions (Al ³⁺ ) are not well known.

Fluorescence is the most useful method for detecting low concentrations of analytes because the equipment required is small, inexpensive, and simple yet high in sensitivity. Thus, some kind of fluorescent chemical sensors for detecting the aluminum ions (Al ^{+ 3)} have been reported. Some reported fluorescence chemical sensors show a sensitive reaction to aluminum ions (Al ^{3 +} ), but most of them are not soluble in water and can only be used in organic solvents or solvents in which water and organic solvents are mixed (Patent Document 1) . Aluminum ion in water (Al ^{3 +)} and trivalent metal ions (M ³⁺⁾ is because it acts as a Lewis acid, an aluminum ion in a solvent mixture of water and an organic solvent (Al ³⁺⁾ and trivalent metal ions (M ^{3 +} ) In the solution causes the pH of the solution, which affects the fluorescence emission of the chemical sensor, to change, making accurate measurements impossible. In addition, the solubility and dissolution of the aluminum ion species (Al ^{3 +) 3} metal ion (M ^3+), including in the water is highly dependent on pH. For example, aluminum ions (Al ^{+ 3)} is only poorly soluble in water in an acidic pH, with increasing pH OH ^- complex ^{(Al (OH) 2+, Al} (OH) 2 +, Al (OH) 3, Al (OH) _{4 <} ^- >). In addition, most aluminum chemistry sensors commonly use a Schiff base (imine) as an important ligand to house aluminum. However, since the immigration is unstable in aqueous solution, it is necessary to develop a new fluorescence chemical sensor containing a new receptor for detecting various concentrations of aluminum in the actual sample solution.

On the other hand, studies for detecting various kinds of transition metals by synthesizing heavy metal fluorescence sensors using peptides have been actively conducted. Fluorescent peptide sensors are attracting a lot of attention in several ways: Fluorescence sensors including peptides can be synthesized relatively easily in a short time using a solid phase synthesis method, and the selectivity and sensitivity for a desired metal ion can be controlled by using various combinations of amino acids. In addition, since it melts well in an aqueous solution similar to physiological conditions, it is easy to use without using an organic solvent, so it is environment-friendly and has excellent ability to bind to metal ions. Further, the synthesized fluorescent peptide sensor can be easily bonded to a solid-phase device, and application such as removal of heavy metal adsorption can be applied. Recently, a method for detecting divalent metal ions using a synthesized fluorescent peptide sensor has also been studied (Patent Document 2). However, aluminum ion (Al ^{3 +} ) and trivalent metal ion (M ³⁺ ) recognition fluorescence sensors using a peptide as a receptor have not yet been attempted.

Thus, the inventors of aluminum ions (Al ^{+ 3)} and trivalent metal ions (M ³⁺⁾ is a compound derived from one, selective peptide was able to detect developing sensors that aluminum ion in the acidic aqueous solution (Al ^{3 +} ) And a trivalent metal ion (M ³⁺ ) can be selectively detected. Thus, the present invention has been completed.

Korean Patent No. 10-1105334 Korean Patent No. 10-1523311

A. P. de Silva., Et al., Chem. Rev. 97, (1997)

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

Another object of the present invention is to provide a method for producing the tripeptide derivative.

It is still another object of the present invention to provide a fluorescence-sensitive chemical sensor comprising the tripeptide derivative.

Another object of the invention is to provide a method of detecting the aluminum ion (Al ^3+), gallium ion (Ga ^{3 +),} indium ion (In ^{+ 3)} and iron ion (Fe ^{+ 3)} with the tripeptide derivative will be.

In order to achieve the above object,

The present invention provides a tripeptide derivative represented by the following formula (1), or an optical isomer thereof.

[Chemical Formula 1]

In Formula 1,

R is -NH ₂ or -OH; And

n is 1 to 4;

The present invention also relates to a process for producing a compound represented by the formula (1)

Reacting a compound supported on a solid phase represented by formula (2) with a compound represented by formula (3) to prepare a compound represented by formula (4) (step 1);

Reacting the compound of Formula 4 prepared in Step 1 with a compound of Formula 3 to prepare a compound of Formula 5 (Step 2);

Reacting the compound of Formula 5 prepared in Step 2 with a compound of Formula 6 to prepare a compound of Formula 7 (Step 3);

Reacting the compound of Formula 7 prepared in Step 3 with 5- (dimethylamino) naphthalene-1-sulfonyl chloride to prepare a compound of Formula 8 (Step 4); And

(Step 5) of removing the t-butyl protecting group from the compound of formula (8) prepared in step 4 and separating from the solid phase to obtain the tripeptide derivative represented by formula (1a). Of the present invention.

[Reaction Scheme 1]

In the above Reaction Scheme 1,

n is from 1 to 4;

은 고체상으로써, 아마이드가 연결된 메틸벤즈히드릴아민(MBHA) 수지, 왕(Wang) 수지, 폴리에틸렌글리콜-폴리스틸렌(PEG-PS) 수지, 실리카 나노 입자, 티타늄옥사이드 나노 입자 및 키토산으로 이루어진 군으로부터 선택되는 1종인 것을 특징으로 하고; 및

Is selected from the group consisting of methylbenzhydrylamine (MBHA) resin, Wang resin, polyethylene glycol-polystyrene (PEG-PS) resin, silica nanoparticles, titanium oxide nanoparticles and chitosan in the solid phase 1 < / RTI > And

Fmoc is a protecting group, fluorenylmethyloxycarbonyl chloride; And

Formula (1a) is a derivative of Formula (1).

Further, the present invention relates to a process for the preparation of

Reacting a compound supported on a solid phase represented by formula (2a) with a compound represented by formula (3) to prepare a compound represented by formula (4a) (step 1);

Reacting the compound of formula (4a) prepared in step 1 with a compound of formula (3) to prepare a compound of formula (5a) (step 2);

Reacting the compound of formula 5a prepared in step 2 with a compound of formula 6 to prepare a compound of formula 7a (step 3);

Reacting the compound of formula (7a) prepared in step 3 with 5- (dimethylamino) naphthalene-1-sulfonyl chloride to prepare a compound of formula (8a) (step 4); And

Removing the t-butyl protecting group from the compound of formula (8a) prepared in step 4 and separating from the solid phase to prepare a tripeptide derivative represented by formula (1b) (step 5). Of the present invention.

[Reaction Scheme 2]

In the above Reaction Scheme 2,

X is halogen;

n is from 1 to 4;

은 고체상으로써, 2-클로로트리틸클로라이드(2-Chlorotrityl chloride) 수지, 왕(Wang) 수지, 폴리에틸렌글리콜-폴리스틸렌(PEG-PS)수지, 실리카 나노 입자, 티타늄옥사이드 나노입자 및 키토산으로 이루어진 군으로부터 선택되는 1종인 것을 특징으로 하고;

Is selected from the group consisting of 2-chlorotrityl chloride resin, Wang resin, polyethylene glycol-polystyrene (PEG-PS) resin, silica nanoparticles, titanium oxide nanoparticles and chitosan as solid phase. And is characterized in that:

Fmoc is a protecting group, fluorenylmethyloxycarbonyl chloride; And

Formula 1b is a derivative of Formula 1 above.

Further, the first invention is selected from the group consisting of aluminum ions (Al ^3+), gallium ion (Ga ^{+ 3),} indium ion (In ^{+ 3)} and iron ion (Fe ^{+ 3)} containing the tripeptide derivative Provided is a fluorescence-sensitive chemical sensor capable of detecting an object to be detected or more.

Further, the above present invention is one member selected from the group consisting of the tripeptide derivative of aluminum ions (Al ^{+ 3),} gallium ion (Ga ^3+), indium ion (In ^{+ 3)} and iron ion (Fe ^{+ 3)} Into a sample containing the detection target (step 1); And

Aluminum ions (Al ^{3 +)} containing; measuring a fluorescence signal that is the reaction product of a tripeptide derivative of the first detection target in the general formula (1) or more species present in the sample of step 1 occurs (step 2) provides, gallium ion (Ga ^{+ 3),} indium ion method of detecting a (In ^{+ 3)} and iron ion (Fe ^3+).

Tripeptide derivative represented by the formula (I) according to the invention in the presence of other metal ions which may act as interfering substances also aluminum ions (Al ^{+ 3),} gallium ion (Ga ^3+), indium ion (In ^{+ 3)} and iron ions (Fe ^{+ 3)} optionally, can be quantitatively detected, and is easy to penetrate into the living cells of aluminum ions (Al ^{+ 3),} gallium ions in living cells (Ga ^3+), indium ion (in ^{3 +} ) And iron ions (Fe ^< ^{3 + &} gt ^; ).

1 is a schematic diagram showing the principle of luminescence of a chemical sensor according to the present invention.
2 is the aluminum ion (Al ^{3 +)} electrospray ionization mass spectrometry (ESI-MS) spectrum of the complex of Example 1 compound.
FIG. 3 is a fluorescence spectrum (A) showing the fluorescence change of the compound-metal ion complex of Example 1 according to the kind of the metal ion and a photograph (B) showing the fluorescence change.
4 is a fluorescence spectrum showing fluorescence change of the compound-metal ion complex of Example 1 according to the type of trivalent metal ion.
5 is a graph showing the fluorescence intensities of the 516 nm wavelength of the compound prepared in Example 1 in the fluorescence spectrum of each test solution for each type of metal ion, and the symbols shown in FIG. 5 are as shown in Table 1 below.
6 is aluminum ions (Al ^{+ 3)} in Example 1. The compounds according to the concentration gradient - a graph showing a fluorescence intensity change of the aluminum ion complex fluorescent spectrum picture showing (A) and the fluorescence changes (B).
FIG. 7 is a fluorescence spectrum showing the change in fluorescence intensity of the compound-gallium ion complex of Example 1 according to the change in gallium ion (Ga ^{3 +} ) concentration.
FIG. 8 is a fluorescence spectrum showing the change in fluorescence intensity of the compound-indium ion complex of Example 1 according to the change in the concentration of indium ion (In ^{3 +} ).
FIG. 9 is a fluorescence spectrum showing the change in fluorescence intensity of the compound of Example 1-iron ion complex according to the change of iron ion (Fe ^{3 +} ) concentration.
Figure 10 is one embodiment the compounds according to the change in concentration of aluminum ions (Al ^{+ 3)} - the absorption spectrum shows the absorbance change of the aluminum ion complex.
11 is a fluorescence spectrum showing the fluorescence intensity change of 516 nm of the compound of Example 1 according to the concentration change of aluminum ion (Al ^{3 +} ).
Figure 12 is a plot graph of the Job's aluminum ion Example 1 compound and the aluminum ion (Al ³⁺⁾ with the mole fraction of the (Al ^{3 +).}
FIG. 13A is a 1 H NMR chart of the compound of Example 1, and FIG. 13B is a 1 H NMR chart of a mixed sample of the compound of Example 1 and aluminum ions.
Figure 14 is one embodiment the compounds according to the pH change - a spectrum showing the fluorescence change of the aluminum ions (Al ^{+ 3)} complexes.

Hereinafter, the present invention will be described in detail.

The present invention provides a tripeptide derivative represented by the following formula (1), or an optical isomer thereof.

[Chemical Formula 1]

In Formula 1,

R is -NH ₂ or -OH; And

n is 1 to 4;

Preferably,

Wherein R is -NH ₂ or -OH; And

n is 1 or 2;

Most preferably, the tripeptide derivative of Formula 1, or an optical isomer thereof,

(S) -2- (5- (dimethylamino) naphthalene-1-sulfonamido) -3-hydroxypropanamide Butanamido) -5-oxopentanoic < / RTI >acid;

(S) -2- ((S) -2- (5- (dimethylamino) naphthalene-1-sulfonamido) -3-hydroxypropanamido) butane Amide Pentanedioic acid;

(S) -2- (5- (dimethylamino) naphthalene-1-sulfamoido) -3-hydroxypropanamide ) Propanamido) -4-oxobutanoic < / RTI >acid; And

(S) -2 - ((S) -3-carboxy-2 - ((S) -2- (5- (dimethylamino) naphthalene- 1 -sulfonamido) -3- hydroxypropanamido) It is an acid.

The present invention also relates to a process for producing a compound represented by the formula (1)

Reacting a compound supported on a solid phase represented by formula (2) with a compound represented by formula (3) to prepare a compound represented by formula (4) (step 1);

Reacting the compound of Formula 4 prepared in Step 1 with a compound of Formula 3 to prepare a compound of Formula 5 (Step 2);

Reacting the compound of Formula 5 prepared in Step 2 with a compound of Formula 6 to prepare a compound of Formula 7 (Step 3);

Reacting the compound of Formula 7 prepared in Step 3 with 5- (dimethylamino) naphthalene-1-sulfonyl chloride to prepare a compound of Formula 8 (Step 4); And

Removing the t-butyl protecting group from the compound of formula (8) prepared in step (4) and separating from the solid phase to prepare the tripeptide derivative represented by formula (1a) (step 5) A method for producing a derivative is provided.

[Reaction Scheme 1]

In the above Reaction Scheme 1,

n is from 1 to 4;

은 고체상으로써, 아마이드가 연결된 메틸벤즈히드릴아민(MBHA) 수지, 왕(Wang) 수지, 폴리에틸렌글리콜-폴리스틸렌(PEG-PS) 수지, 실리카 나노 입자, 티타늄옥사이드 나노 입자 및 키토산으로 이루어진 군으로부터 선택되는 1종인 것을 특징으로 하고; 및

Is selected from the group consisting of methylbenzhydrylamine (MBHA) resin, Wang resin, polyethylene glycol-polystyrene (PEG-PS) resin, silica nanoparticles, titanium oxide nanoparticles and chitosan in the solid phase 1 < / RTI > And

Fmoc is a protecting group, fluorenylmethyloxycarbonyl chloride; And

Formula (1a) is a derivative of Formula (1).

Hereinafter, the production method of Reaction Scheme 1 according to the present invention will be described in detail for each step.

In the preparation of Reaction Scheme 1 according to the present invention, Step 1 is a step of reacting a compound supported on a solid phase represented by Formula 2 with a compound of Formula 3 to prepare a compound of Formula 4, more specifically, 25% Of the compound of formula (2) is removed using a piperidine / dimethylformamide solution of the formula (3), and then the compound of formula (3) is coupled to prepare the compound of formula (4).

In this case, the coupling reaction is carried out with diisopropylethylamine or triethylamine in the presence of benzotriazol-1-yl-oxy-tris (dimethylamino) -phosphonium hexafluorophosphate (Py-BOP) Dihydroxy-7-aza-benzotriazole (HATU), hydroxybenzotriazole (HOBt), dicyclohexylcarbodiimide (DCC), diisopropylcarbodiimide - (3-dimethylaminopropyl) carbodiimide (EDC) or carbonyldiimidazole (CDI), preferably diisopropylcarbodiimide (DIC) and hydroxybenzotriazole Can be used together.

In addition, as the usable organic solvent, the reaction can be carried out using methanol, dimethylformamide (DMF), tetrahydrofuran (THF), dichloromethane (DCM) or toluene which does not adversely affect the reaction, Dimethylformamide (DMF) may be used.

Further, in step 1 of the present invention, the compound of formula (2) and the compound of formula (3) may be mixed in a molar ratio of 1: 2 to 10, preferably 1: 3 to 7 have.

When the mixing ratio of the compound of Formula 2 and the compound of Formula 3 is less than 1: 2, there is a problem that the efficiency of synthesis of the target compound is low due to the small amount of the compound of Formula 3 introduced, and the mixing ratio is 1:10 There is a problem that the amount of the compound of the general formula (3) which is unreacted and discarded after the reaction is large, which is uneconomical.

In the preparation of Reaction Scheme 1 according to the present invention, Step 2 is a step of reacting the compound of Chemical Formula 4 with the compound of Chemical Formula 4 once in Step 1 to prepare Compound 5, more specifically, 25% , The Fmoc protecting group at the terminal of the amino group of the compound of Formula 4 is removed using a piperidine / dimethylformamide solution of the compound of Formula 4, and then the Fmoc protecting group of Formula 3 and the glutamic acid protected by the t-butyl protecting group are bonded, .

In this case, the coupling reaction is carried out with diisopropylethylamine or triethylamine in the presence of benzotriazol-1-yl-oxy-tris (dimethylamino) -phosphonium hexafluorophosphate (Py-BOP) Dihydroxy-7-aza-benzotriazole (HATU), hydroxybenzotriazole (HOBt), dicyclohexylcarbodiimide (DCC), diisopropylcarbodiimide - (3-dimethylaminopropyl) carbodiimide (EDC) or carbonyldiimidazole (CDI), preferably diisopropylcarbodiimide (DIC) and hydroxybenzotriazole Can be used together.

In addition, as the usable organic solvent, the reaction can be carried out using methanol, dimethylformamide (DMF), tetrahydrofuran (THF), dichloromethane (DCM) or toluene which does not adversely affect the reaction, Dimethylformamide (DMF) may be used.

Further, in the step 2 of the present invention, the compound of the formula (4) and the compound of the formula (3) may be mixed in a molar ratio of 1: 2 to 10, preferably 1: 3 to 7 have. When the mixing ratio of the compound of formula (4) and the compound of formula (3) is less than 1: 2, there is a problem that the efficiency of synthesis of the target compound is low due to the small amount of the compound of formula (3) introduced. There is a problem that the amount of the compound of the general formula (3) which is unreacted and discarded after the reaction is large, which is uneconomical.

In the preparation of Reaction Scheme 1 according to the present invention, Step 3 is a step of reacting Compound (5) prepared in Step 2 with Compound (6) to prepare Compound (7) Dimethylformamide solution, the Fmoc protecting group at the end of the amino group of the compound of formula (5) is removed and the Fmoc protecting group of the formula (6) and the serine protected by the t-butyl protecting group are combined to give the compound of the formula .

In this case, the coupling reaction is carried out with diisopropylethylamine or triethylamine in the presence of benzotriazol-1-yl-oxy-tris (dimethylamino) -phosphonium hexafluorophosphate (Py-BOP) Dihydroxy-7-aza-benzotriazole (HATU), hydroxybenzotriazole (HOBt), dicyclohexylcarbodiimide (DCC), diisopropylcarbodiimide - (3-dimethylaminopropyl) carbodiimide (EDC) or carbonyldiimidazole (CDI), preferably diisopropylcarbodiimide (DIC) and hydroxybenzotriazole Can be used together.

The organic solvent that can be used in step 3 may be a reaction that does not adversely affect the reaction using methanol, dimethylformamide (DMF), tetrahydrofuran (THF), dichloromethane (DCM), or toluene And preferably dimethylformamide (DMF) can be used.

Further, in the step 3 according to the present invention, the compound of the formula (5) and the compound of the formula (6) may be mixed in a molar ratio of 1: 2 to 10, preferably 1: 3 to 7 have. When the mixing ratio of the compound of the general formula (5) and the compound of the general formula (6) is less than 1: 2, there is a problem that the synthesis efficiency of the desired compound is low due to the small amount of the compound of the general formula (6) 10, there is a problem in that the amount of the compound of the formula (6) which is unreacted and discarded after the reaction is large, which is uneconomical.

In the preparation of Reaction Scheme 1 according to the present invention, Step 4 is a step of reacting the compound of Formula 7 prepared in Step 3 with 5- (dimethylamino) naphthalene-1-sulfonyl chloride to prepare a compound of Formula 8 Step, more specifically, the Fmoc protecting group at the amino terminus of the compound of formula (7) is removed using 25% piperidine / dimethylformamide solution and reacted with 5- (dimethylamino) naphthalene-1-sulfonyl chloride To prepare a compound of formula (8).

At this time, as the usable organic solvent, the reaction can be carried out using methanol, dimethylformamide (DMF), tetrahydrofuran (THF), dichloromethane (DCM) or toluene which does not adversely affect the reaction, Dimethylformamide (DMF) may be used.

In addition, the reaction temperature is not particularly limited, but may be performed within a range of room temperature to reflux temperature of the solvent.

In the preparation of Reaction Scheme 1 according to the present invention, Step 5 is a step of removing the protecting groups of the compound of Formula 8 prepared in Step 4 and isolating the compound from the solid phase to prepare the tripeptide derivative represented by Formula 1a, More specifically, it is a step of removing the t-butyl protecting group of formula (8) using trifluoroacetic acid and tripropylsilane and separating from the solid phase to prepare the tripeptide derivative represented by formula (1a).

In this case, trifluoroacetic acid (TFA) and tripropylsilane used for removing the solid phase can be used as a reaction solvent by mixing with distilled water. Preferably, trifluoroacetic acid, tripropylsilane and distilled water are mixed with 95 : 2.5: 2.5 by volume ratio.

In addition, the reaction temperature is not particularly limited, but may be performed within a range of room temperature to reflux temperature of the solvent.

In addition,

As shown in Reaction Scheme 2 below,

Reacting a compound supported on a solid phase represented by formula (2a) with a compound represented by formula (3) to prepare a compound represented by formula (4a) (step 1);

Reacting the compound of formula (4a) prepared in step 1 with a compound of formula (3) to prepare a compound of formula (5a) (step 2);

Reacting the compound of formula 5a prepared in step 2 with a compound of formula 6 to prepare a compound of formula 7a (step 3);

Reacting the compound of formula (7a) prepared in step 3 with 5- (dimethylamino) naphthalene-1-sulfonyl chloride to prepare a compound of formula (8a) (step 4); And

Removing the t-butyl protecting group from the compound of formula (8a) prepared in step 4 and separating from the solid phase to prepare a tripeptide derivative represented by formula (1b) (step 5). A method for producing a derivative is provided.

[Reaction Scheme 2]

In the above Reaction Scheme 2,

X is halogen;

n is from 1 to 4;

은 고체상으로써, 2-클로로트리틸 클로라이드(2-Chlorotrityl chloride) 수지, 왕(Wang) 수지, 폴리에틸렌글리콜-폴리스틸렌(PEG-PS)수지, 실리카 나노 입자, 티타늄옥사이드 나노 입자 및 키토산으로 이루어진 군으로부터 선택되는 1종인 것을 특징으로 하고;

Is selected from the group consisting of 2-chlorotrityl chloride resin, Wang resin, polyethylene glycol-polystyrene (PEG-PS) resin, silica nanoparticles, titanium oxide nanoparticles and chitosan as solid phase. And is characterized in that:

Fmoc is a protecting group, fluorenylmethyloxycarbonyl chloride; And

Formula 1b is a derivative of Formula 1 above.

Hereinafter, the production method of Reaction Scheme 2 according to the present invention will be described in detail for each step.

In the preparation of Reaction Scheme 2 according to the present invention, Step 1 is a step of reacting a compound supported on a solid phase represented by Formula 2a with a compound of Formula 3 to prepare a compound of Formula 4a.

At this time, an organic base such as triethylamine, pyridine, diisopropylethylamine, 1,8-diazabicyclo [5.4.0] -7-undecene (DBU) or the like, sodium hydroxide, sodium carbonate, Potassium carbonate, cesium carbonate and the like can be used in an equivalent amount or in an excess amount, and triethylamine can be preferably used.

In addition, as the usable organic solvent in the step 1, the reaction can be carried out using methanol, dimethylformamide (DMF), tetrahydrofuran (THF), dichloromethane (DCM) or toluene which does not adversely affect the reaction And preferably dimethylformamide (DMF) can be used.

Further, in step 1, the compound of formula (2a) and the compound of formula (3) may be mixed in a molar ratio of 1: 2 to 10, preferably 1: 3 to 7. When the mixing ratio of the compound of the formula (2a) and the compound of the formula (3) is less than 1: 2, there is a problem that the synthesis efficiency of the desired compound is low due to the small amount of the compound of the formula (3) introduced. There is a problem that the amount of the compound of the general formula (3) which is unreacted and discarded after the reaction is large, which is uneconomical.

In the preparation of Reaction Scheme 2 according to the present invention, Step 2 is a step of reacting the compound supported on the solid phase represented by Formula 4a with the compound of Formula 3 once more to prepare the compound of Formula 5a, And then reacting the compound of formula (4) with a compound of formula (3) to prepare a compound of formula (5a).

In this case, the coupling reaction is carried out with diisopropylethylamine or triethylamine in the presence of benzotriazol-1-yl-oxy-tris (dimethylamino) -phosphonium hexafluorophosphate (Py-BOP) Dihydroxy-7-aza-benzotriazole (HATU), hydroxybenzotriazole (HOBt), dicyclohexylcarbodiimide (DCC), diisopropylcarbodiimide - (3-dimethylaminopropyl) carbodiimide (EDC) or carbonyldiimidazole (CDI), preferably diisopropylcarbodiimide (DIC) and hydroxybenzotriazole Can be used together.

In addition, as the usable organic solvent in the step 2, the reaction can be carried out using methanol, dimethylformamide (DMF), tetrahydrofuran (THF), dichloromethane (DCM) or toluene which does not adversely affect the reaction And preferably dimethylformamide (DMF) can be used.

Further, in step 2, the compound of formula (4a) and the compound of formula (3) may be mixed in a molar ratio of 1: 2 to 10, preferably 1: 3 to 7. When the mixing ratio of the compound of the formula (4a) and the compound of the formula (3) is less than 1: 2, there is a problem that the synthesis efficiency of the desired compound is low due to the small amount of the compound of the formula (3) introduced. There is a problem that the amount of the compound of the general formula (3) which is unreacted and discarded after the reaction is large, which is uneconomical.

In the preparation of Reaction Scheme 2 according to the present invention, Step 3 is a step of preparing a compound of Formula 7a by coupling a compound of Formula 6 with the compound of Formula 5a prepared in Step 2, (Fmoc) protecting group of the compound of Chemical Formula 5a is removed by using a piperidine / dimethylformamide solution of the compound of Chemical Formula 5a, and then the compound of Chemical Formula 6 is coupled to prepare the compound of Chemical Formula 7a.

In this case, the coupling reaction is carried out with diisopropylethylamine or triethylamine in the presence of benzotriazol-1-yl-oxy-tris (dimethylamino) -phosphonium hexafluorophosphate (Py-BOP) Dihydroxy-7-aza-benzotriazole (HATU), hydroxybenzotriazole (HOBt), dicyclohexylcarbodiimide (DCC), diisopropylcarbodiimide - (3-dimethylaminopropyl) carbodiimide (EDC) or carbonyldiimidazole (CDI), preferably diisopropylcarbodiimide (DIC) and hydroxybenzotriazole Can be used together.

The organic solvent that can be used in step 3 may be a reaction that does not adversely affect the reaction using methanol, dimethylformamide (DMF), tetrahydrofuran (THF), dichloromethane (DCM), or toluene And preferably dimethylformamide (DMF) can be used.

Further, in the step 3 according to the present invention, the compound of the formula (5a) and the compound of the formula (6) may be mixed in a molar ratio of 1: 2 to 10, preferably 1: 3 to 7 have. When the mixing ratio of the compound of the general formula (5a) and the compound of the general formula (6) is less than 1: 2, there is a problem that the amount of the compound of the general formula (6) 10, there is a problem in that the amount of the compound of the formula (6) which is unreacted and discarded after the reaction is large, which is uneconomical.

In the process for preparing Reaction Scheme 2 according to the present invention, Step 4 is a step of reacting the compound of Formula 7a prepared in Step 3 with 5- (dimethylamino) naphthalene-1-sulfonyl chloride to prepare a compound of Formula 8a As a step, more specifically, the Fmoc protecting group at the amino terminus of the compound of formula (7a) is removed using 25% piperidine / dimethylformamide solution and reacted with 5- (dimethylamino) naphthalene-1-sulfonyl chloride To give a compound of formula (8a).

At this time, as the usable organic solvent, the reaction can be carried out using methanol, dimethylformamide (DMF), tetrahydrofuran (THF), dichloromethane (DCM) or toluene which does not adversely affect the reaction, Dimethylformamide (DMF) may be used.

In addition, the reaction temperature is not particularly limited, but may be performed within a range of room temperature to reflux temperature of the solvent.

In the preparation of Reaction Scheme 2 according to the present invention, Step 5 is a step of removing the protecting groups of the compound of Formula 8a prepared in Step 4 and separating from the solid phase to prepare the tripeptide derivative represented by Formula 1b, More specifically, it is a step of removing the t-butyl protecting group of formula (8a) using trifluoroacetic acid and tripropylsilane and separating from the solid phase to prepare the tripeptide derivative represented by formula (1b).

In this case, trifluoroacetic acid (TFA) and tripropylsilane used for removing the solid phase can be used as a reaction solvent by mixing with distilled water. Preferably, trifluoroacetic acid, tripropylsilane and distilled water are mixed with 95 : 2.5: 2.5 by volume ratio.

In addition, the reaction temperature is not particularly limited, but may be performed within a range of room temperature to reflux temperature of the solvent.

The present invention relates to a group consisting of an aluminum ion (Al ^{3 +} ), a gallium ion (Ga ^{3 +} ), an indium ion (In ^{3 +} ) and an iron ion (Fe ^{3 +} ), which comprises a tripeptide derivative represented by the above formula And a fluorescence-sensitive chemical sensor which is capable of detecting at least one detection object selected from the group consisting of the fluorescence-

Furthermore, the present invention from the group consisting of a tripeptide derivative of the formula (1) to aluminum ion (Al ^{3 +),} gallium ion (Ga ^{3 +),} indium ion (In ^{3 +)} and iron ion (Fe ^{3 +)} Into a sample containing one or more detection targets to be selected (step 1); And

Aluminum ions (Al ^{3 +)} containing; measuring a fluorescence signal that is the reaction product of a tripeptide derivative of the first detection target in the general formula (1) or more species present in the sample of step 1 occurs (step 2) provides, gallium ion (Ga ^{+ 3),} indium ion method of detecting a (In ^{+ 3)} and iron ion (Fe ^3+).

Hereinafter, the detection method according to the present invention will be described in detail for each step.

In the detection method according to the present invention, the step 1 is the aluminum ion (Al ^{3 +),} gallium ion (Ga ^{3 +),} indium ion (In ^{3 +)} and iron ions to tripeptide derivatives represented by the formula (1) ( a step of input on the sample to determine the first detection subject the presence or absence or more member selected from the group consisting of Fe ^{3 +),} more particularly to a metal that can act as interfering substances ions represented by the general formula (1) in the dissolved sample with in the following the tripeptide derivatives, selected from the group consisting of aluminum ions (Al ^3+), gallium ion (Ga ^3+), indium ion (in ^{+ 3)} and iron ion (Fe ^{+ 3)} existing in the sample Is selectively reacted with a tripeptide derivative represented by the formula (I) to form a complex.

As in the detection method according to the present invention, the step 2 is the aluminum ion (Al ^{3 +),} gallium ion (Ga ^{3 +),} indium ion (In ^{3 +)} and iron ion (Fe ^{3 +)} formed in the step 1 (Al ^{3 +} ), gallium ion (Ga ^{3 +} ), indium ion (Ga ^{3 +} ), and the like are measured by fluorescence spectroscopy to measure the fluorescence signal emitted by the complex of the tripeptide derivative represented by formula (In ^{3 +} ) and iron ions (Fe ^{3 +} ). The tripeptide derivative according to the present invention can be detected in the presence of other metal ions which may act as an interfering substance FIG aluminum ions (Al ^{3 +),} gallium ion (Ga ^{3 +),} indium ion (in ^{3 +)} and iron ions indicates a large fluorescence intensity changes by a combination of the (Fe ^{3 +)} (experimental example 1, 3 , Figs. 4 and 5).

In addition, the tripeptide derivatives of the present invention are aluminum ions (Al ^{+ 3),} gallium ion (Ga ^3+), indium ion (In ^{+ 3)} and iron ions first detection of two or more selected from the group consisting of (Fe ^{+ 3)} And selectively detects the fluorescence signal and detects the fluorescence signal by the fluorescence signal difference.

Referring to simultaneous detection performance results of tripeptide derivative of the present invention, while the tripeptide derivatives of the present invention showing a large fluorescence intensity increase in the emission wavelength of 516 nm by a combination of the aluminum ions (Al ^{3 +),} iron (Fe ^{3 +} ) showed a decrease in fluorescence intensity at an emission wavelength of 550 nm. This aluminum ions (Al ^{3 +)} and iron ions by indicating a large fluorescence change in each of a different wavelength with respect to the tripeptide derivatives of aluminum ions (Al ^{3 +)} and iron ion (Fe ^{3 +)} of the invention (Fe ^{3 +)} (Al ^{3 +} ) and iron ions (Fe ³⁺ ) can be distinguished from each other due to the different fluorescence intensity change characteristics even when they exist simultaneously (Experimental Examples 2 and 3) 4).

Further, in the detection method of the present invention, the sample can be applied not only to an organic solvent but also to an aqueous solution or distilled water.

The aqueous solution may be in the form of an aqueous solution containing at least one organic solution selected from the group consisting of methanol, ethanol, dimethylformamide (DMF) and acetonitrile, and the range of the sample is not limited thereto.

At this time, it is preferable to use methanol, ethanol, dimethylformamide (DMF), acetonitrile or the like as the organic solution contained in the aqueous solution, but it is not limited thereto.

When the organic solvent is mixed with the aqueous solution, the organic solvents such as methanol, ethanol, dimethylformamide (DMF) or acetonitrile have high solubility in water, so that aluminum ions (Al ^{3 +} ), gallium ions (Ga ^{3 +} ), Indium ion (In ^{3 +} ), and iron ion (Fe ^{3 +} ), whereby the aluminum ion (Al ^{3 +),} gallium ion (Ga ^{+ 3),} it is possible to detect the indium ion (In ³⁺⁾ and iron ion (Fe ^3+).

In the detection method of the present invention, the sample may be a constant, sewage, wastewater, river, seawater, biological fluid, etc., but the sample is not limited thereto.

As described above, the tripeptide derivatives of the formula (I) according to the invention in the presence of other metal ions which may act as interfering substances also aluminum ions (Al ^3+), gallium ion (Ga ^{+ 3),} indium ion ( in ^{3 +)} and iron ion (Fe ^{3 +)} selectively, and can be quantitatively detected, is easy to aluminum ions (Al ^{3 +),} gallium ion (Ga ^{3 +)} in the body cells penetrate into the living cells of, It can be useful as a fluorescence-sensitive chemical sensor capable of detecting fluorescence intensity change by binding with indium ion (In ^{3 +} ) and iron ion (Fe ^{3 +} ).

Hereinafter, the present invention will be described in detail with reference to Examples and Experimental Examples.

However, the following Examples and Experimental Examples are merely illustrative of the present invention, and the present invention is not limited to the following Examples and Experimental Examples.

< Example  1> (S) -5-Amino-4 - ((S) -4- Carboxy -2 - ((S) -2- (5- ( Dimethylamino ) Naphthalene-1-sulfonamido) -3-hydroxypropanamido) butanamido) -5-oxopentanoic acid Eshi Manufacture of de

(S) -2- (5- (dimethylamino) naphthalene-1-sulfonamido) -5-amino-4 - ((S) ) -3-hydroxypropanamido) butanamido) -5-oxopentanone &lt; / RTI &gt;

Step 1: Preparation of compound of formula 10

First, Rink amide MBHA resin (235 mg, 0.1 mmol) in which the terminal amine of formula (9) was protected with Fmoc-group was added to a ribra tube in anhydrous dimethylformamide (3 mL ) And pre-swelled for about 15 minutes. Next, 25% piperidine / dimethylformamide mixed solution (3 mL) was added to the swollen resin and activated for 15 minutes to remove the amino terminal Fmoc protecting group of the dissolved resin. Then, Fmoc-L-glutamic acid (Glu) (OtBu) -OH (128 mg, 0.3 mmol), diisopropylcarbodiimide (DIC, 47 μL, 0.3 mmol) in the anhydrous dimethylformamide mmol) and hydroxybenzotriazole (HOBt, 40 mg, 0.3 mmol) were added and reacted with the linkamide methylbenzohydrillamine resin solution in which the amino terminal Fmoc protecting group was activated for 15 minutes in advance, for about 4 hours .

After the reaction, the reaction solution was filtered, and the filtered resin was washed several times with dimethylformamide and methanol, and then a kaiser test was conducted. When the Kaiser test was positive, the activated solution was added again, and when it was negative, the reaction was stopped to prepare the compound of formula (10).

Step 2: Preparation of compound of formula (11)

To the compound of the formula (10) obtained in the above step 1, 25% piperidine / dimethylformamide mixed solution (3 mL) was further added and the mixture was activated for 15 minutes to remove the Fmoc protecting group at the terminal of the amino group. Then, Fmoc-L-glutamic acid (Glu) (OtBu) -OH (128 mg, 0.3 mmol), diisopropylcarbodiimide (DIC, 47 μL, 0.3 mmol) in the anhydrous dimethylformamide mmol) and hydroxybenzotriazole (HOBt, 40 mg, 0.3 mmol) were added and reacted with the solution in which the amino terminal Fmoc protecting group, which had been activated for 15 minutes in advance, was removed for about 4 hours.

After the reaction, the reaction solution was filtered, and the filtered resin was washed several times with dimethylformamide and methanol, and then a kaiser test was conducted. When the Kaiser test was positive, the above-mentioned activated solution was added again, and when it was negative, the reaction was stopped to prepare the compound of formula (11) as the target compound.

Step 3: Preparation of the compound of formula (12)

To the compound of Formula 11 obtained in Step 2, 25% piperidine / dimethylformamide mixed solution (3 mL) was added and activated for 15 minutes to remove the Fmoc protecting group at the terminal of the amino group. Then, Fmoc-L-serine (Ser) (tBu) -OH (128 mg, 0.3 mmol), diisopropylcarbodiimide (DIC, 47 μL, 0.3 mmol) of the formula 6a in anhydrous dimethylformamide (3 mL) mmol) and hydroxybenzotriazole (HOBt, 40 mg, 0.3 mmol) were added and reacted with the solution in which the amino terminal Fmoc protecting group, which had been activated for 15 minutes in advance, was removed for about 4 hours.

After the reaction, the reaction solution was filtered, and the filtered resin was washed several times with dimethylformamide and methanol, and then a kaiser test was conducted. When the Kaiser test was positive, the activated solution was added again, and when it was negative, the reaction was stopped to prepare the compound of formula (12).

Step 4: Preparation of the compound of formula (13)

25% piperidine / dimethylformamide mixed solution (3 mL) was added to the compound of the formula (12) obtained in the step 3, and the mixture was activated for 15 minutes to remove the Fmoc protecting group at the terminal of the amino group. Then, 5- (dimethylamino) naphthalene-1-sulfonyl chloride (80 mg, 0.3 mmol) and triethylamine (80 μL, 0.6 mmol) were added to anhydrous dimethylformamide (3 mL) After stirring for about 4 hours, the reaction solution was filtered, and the filtered resin was washed several times with dimethylformamide and methanol several times to prepare the desired compound (13).

Step 5: (S) -5-Amino-4 - ((S) -4- Carboxy -2 - ((S) -2- (5- ( Dimethylamino ) Naphthalene-1-sulfonamido) -3-hydroxypropanamido) butanamido) -5-oxopentanoic acid Acid  Produce

The compound of Formula 13 obtained in Step 4 was reacted with a mixed solution of trifluoroacetic acid: tripropylsilane: distilled water (95: 2.5: 2.5, v / v / v) at room temperature for 4 hours. After the reaction, the reaction solution was filtered to remove the t-butyl protecting group, and the desired compound was isolated from the resin. The filtered separation solution was passed through nitrogen gas to remove the trifluoroacetic acid and purified by reverse phase high performance liquid chromatography (HPLC, C-18 column delta pak C18-300A, 1.9 X 30 cm) (eluent: 0.1% trifluoroacetic acid (48 mg, yield 80%) was obtained as a white solid in a yield of 80% after purification by column chromatography using distilled water / acetonitrile (1: 1, v / v) .

The molecular weight of the purified tripeptide derivative was confirmed using an ESI mass spectrometer, and the results are as follows.

^{1 H NMR (400 MHz, 50} % CD 3 CN / D 2 O): δ 9.32 (m, 1H), 9.01 (m, 1H), 8.94 (m, 1H), 8.51 (m, 1H), 8.42 (m 2H), 2.40 (s, 3H), 2.45 (m, 2H) 2H), 2.22 (m, 2H).

^{13 C NMR (100 MHz, 50} % CD 3 CN / D 2 O): δ 176.7, 176.0, 172.8, 171.0, 163.1, 138.8, 135.2, 131.4, 128.5, 126.3, 119.9, 118.0, 115.4, 112.1, 62.0, 58.0 , 52.8, 47.2, 29.9, 26.1

ESI-MAss (m / z) : [M + H] + C 25 H 33 N 5 O 10 S: 595.19 ( calcd), 596.20 (measured value).

< Example  2 - ((S) -2- (5- (dimethylamino) naphthalene-1-sulfonamido) -3-hydroxypropanamido) Butanamido) Preparation of pentanedioic acid

(S) -2- ((S) -2- (5- (Dimethylamino) naphthalene-1-sulfonamido) -3-hvdroxy- Roxy-propanamido) butanamido) pentanedioic acid was prepared.

[Reaction Scheme 4]

Step 1: Preparation of compound of formula 10a

First, 2-chlorotrityl chloride resin (61 mg, 0.1 mmol) of formula (9a) was introduced into a libra tube together with anhydrous dichloromethane (3 ml) Swelled. Next, to the swollen resin was added Fmoc-L-glutamic acid (Glu) (OtBu) -OH (128 mg, 0.3 mmol) of formula 3a and 6 equivalents of diisopropylethylamine (DIPEA, 105 μL, 0.6 mmol ) Was added thereto, and the mixture was reacted for about 5 hours.

After the reaction, the reaction solution was filtered, and the filtered resin was washed several times with dichloromethane, dimethylformamide and methanol several times to prepare the desired compound (10a).

Step 2: Preparation of compound of formula 11a

To the compound of formula (10a) obtained in the above step 1, 25% piperidine / dimethylformamide mixed solution (3 mL) was added and activated for 15 minutes to remove the Fmoc protecting group at the terminal of the amino group. Then, Fmoc-L-glutamic acid (Glu) (OtBu) -OH (128 mg, 0.3 mmol), diisopropylcarbodiimide (DIC, 47 μL, 0.3 mmol) in the anhydrous dimethylformamide mmol) and hydroxybenzotriazole (HOBt, 40 mg, 0.3 mmol) were added and reacted with the solution in which the amino terminal Fmoc protecting group, which had been activated for 15 minutes in advance, was removed for about 4 hours.

After the reaction, the reaction solution was filtered, and the filtered resin was washed several times with dimethylformamide and methanol, and then a kaiser test was conducted. When the Kaiser test was positive, the above-mentioned activated solution was added again, and when it was negative, the reaction was stopped to prepare the compound of formula (11a) as a target compound.

Step 3: Preparation of the compound of formula (12a)

To the compound of Formula 11a obtained in Step 2, 25% piperidine / dimethylformamide mixed solution (3 mL) was added and activated for 15 minutes to remove the Fmoc protecting group at the terminal of the amino group. Then, Fmoc-L-serine (Ser) (tBu) -OH (128 mg, 0.3 mmol), diisopropylcarbodiimide (DIC, 47 μL, 0.3 mmol) of the formula 6a in anhydrous dimethylformamide (3 mL) mmol) and hydroxybenzotriazole (HOBt, 40 mg, 0.3 mmol) were added and reacted with the solution in which the amino terminal Fmoc protecting group, which had been activated for 15 minutes in advance, was removed for about 4 hours.

After the reaction, the reaction solution was filtered, and the filtered resin was washed several times with dimethylformamide and methanol, and then a kaiser test was conducted. When the Kaiser test was positive, the above-mentioned activated solution was added again, and when it was negative, the reaction was stopped to prepare the compound of formula (12a) as the target compound.

Step 4: Preparation of the compound of formula (13a)

To the compound of Formula 12a obtained in Step 3, 25% piperidine / dimethylformamide mixed solution (3 mL) was added and the mixture was activated for 15 minutes to remove the Fmoc protecting group at the terminal of the amino group. Then, 5- (dimethylamino) naphthalene-1-sulfonyl chloride (80 mg, 0.3 mmol) and triethylamine (80 μL, 0.6 mmol) were added to anhydrous dimethylformamide (3 mL) After stirring for about 4 hours, the reaction solution was filtered, and the filtered resin was washed several times with dimethylformamide and methanol several times to prepare the desired compound (13a).

Step 5: (S) -2 - ((S) -4-Carboxy-2 - ((S) -2- (5- (dimethylamino) naphthalene- 1 -sulfonamido) -3- ) Butanamido) &lt; / RTI &gt; pentanedioic acid &lt; RTI ID =

The compound of Formula 13a obtained in Step 4 was reacted with a mixed solution of trifluoroacetic acid: tripropylsilane: distilled water (95: 2.5: 2.5, v / v / v) at room temperature for 4 hours. After the reaction, the reaction solution was filtered to remove the t-butyl protecting group, and the desired compound was isolated from the resin. The filtered separation solution was passed through nitrogen gas to remove the trifluoroacetic acid and purified by reverse phase high performance liquid chromatography (HPLC, C-18 column delta pak C18-300A, 1.9 X 30 cm) (eluent: 0.1% trifluoroacetic acid (46 mg, yield 77%) was obtained as a white solid in the form of a white solid by distillation under reduced pressure using distilled water / acetonitrile (1: 1, v / v) .

The molecular weight of the purified tripeptide derivative was confirmed using an ESI mass spectrometer and the results are as follows.

^{1 H NMR (400 MHz, 50} % CD 3 CN / D 2 O): δ 9.32 (m, 1H), 9.01 (m, 1H), 8.94 (m, 1H), 8.51 (m, 1H), 8.42 (m 2H), 2.89 (m, 2H), 2.71 (m, 1H), 4.75 (m, 2H), 2.40 (m, 2H), 2.22 (m, 2H).

¹³ C NMR (100 MHz, 50% CD ₃ CN / D ₂ O): δ 177.5, 176.8, 172.8, 171.0, 163.1, 138.8, 135.2, 131.4, 128.5, 126.3, 119.9, 118.0, 115.4, 112.1, 62.0, 58.0 , 52.8, 47.2, 29.9, 26.1

ESI-MAss (m / z) : [M + H +] + C 25 H 32 N 4 O 11 S: 597.18 ( calculated), 597.0 (measured value).

< Experimental Example  1> Fluorescence Sensitive Chemical Sensor - Aluminum Ion ( Al ^{3 +} ) Preparation of complex

The following experiments were carried out to analyze whether the aluminum and trivalent metal ions (M ³⁺ ) represented by the formula (1) according to the present invention and the aluminum ion (Al ^{3 +} ) bond and the compound state were analyzed.

The compound prepared in Example 1 according to the present invention was dissolved in distilled water to prepare a 1.0 mM fluorescence sensitized chemical sensor reagent. Aluminum ion (Al ^{3 +} ) reference solution was prepared by dissolving perchlorate salt in distilled water to 5 mM. In addition, a test solution was prepared by diluting the test tube with distilled water and acetonitrile (2: 8, v / v) so that the concentration of the fluorescent sensitized chemical sensor prepared above was 100 μM and that of aluminum ion was 500 μM. The test solution prepared above was measured by ESI-MS to confirm whether a compound represented by formula (1) and aluminum ion (Al ^{3 +} ) were formed or not. The results are shown in FIG.

As shown in Fig. 2, 596.4 m / e and 620.3 m / e were observed in the ESI-MS spectrum. At this time, 596.4 m / e is a form in which the compound prepared in Example 1 is bound to a proton, and the compound of Example 1 does not form a complex with aluminum ion (Al ^{3 +} ), and 620.3 m / e Example 1 is a value obtained by removing two protons from a compound and forming a complex with an aluminum ion (Al ³⁺ ). From this, it can be seen that the compound represented by the formula (1) according to the present invention is excellent in the binding property with the aluminum ion (Al ^{3 +} ) and can form a stable complex.

Therefore, since the aluminum (Al ^{3 +)} fluorescence-sensitive chemical sensors represented by the general formula (1) according to the invention the binding of the aluminum ions (Al ^{3 +)} excellent in an aqueous solution, to the ground water, interfering substances in aqueous environments such as rivers in the presence of other metal ions which may act also aluminum ions (Al ^{3 +),} gallium ion (Ga ^{3 +),} indium ion (in ^{3 +)} and iron ion (Fe ^{3 +)} Alternatively, quantitatively and simultaneously or the Can be detected at this time. In addition, the fluorescence by coupling of the living cells to facilitate penetration into the aluminum ions in living cells (Al ^{3 +),} gallium ion (Ga ^{3 +),} indium ion (In ^{3 +)} and iron ion (Fe ^{3 +)} It can be useful as a fluorescence-sensitive chemical sensor capable of detection through intensity change.

< Experimental Example  2> Fluorescence Sensitive Chemical Sensor Metal ion selectivity  evaluation

The following experiments were conducted to evaluate the selectivity of aluminum and trivalent metal ions (M ³⁺ ) according to the present invention to aluminum and trivalent metal ions (M ³⁺ ).

In order to evaluate the selectivity of the compound prepared in Example 1 according to the present invention on aluminum and trivalent metal ion (M ³⁺ ), various metal ions were tested. The metal ions used in the experiments is a nickel ion (Ni ^{2 +),} Pb (Pb ^{2 +),} cadmium ion (Cd ^{2 +),} silver ions (Ag ^+), calcium ions (Ca ^{2 +),} iron ion (Fe ^{2 +)} and chromium ions (Cr ^{+ 3),} aluminum ion (Al ^3+), cobalt ion (Co ^2+), a copper ion (Cu ^2+), manganese ion (Mn ^2+), magnesium ion (Mg ²⁺ ), zinc ion (Zn ^{2 +),} iron ions (Fe ^{3 +)} and mercury ions (Hg ^{2 +)} a perchlorate salt and a potassium ion (K ⁺⁾ and chloride salts and gallium ions of the sodium ion (Na ⁺⁾ (Ga ^{3 +} ), and indium ion (In ^{3 +} ), the metal ions were dissolved in distilled water to prepare a 10 mM metal ion reference solution. In addition, aluminum ions (Al ^{+ 3)} and other metal ions of 10 mM of aluminum ions, including along the (Al ³⁺⁾ - were also prepared metal ion based solution.

10 mM hexamine buffer solution (pH 5.5, 1 mL) was injected into the test tube, and the compound prepared in the above example was dissolved in distilled water to prepare a 10 μM fluorescent sensitized chemical sensor reagent. After adding 10 μL each of the standard solution of metal standard solution or aluminum ion (Al ^{3 +} ) - metal ion standard, distilled water was injected so that the total solution amount was 2 mL. The prepared test solution was adjusted to have an excitation wavelength of 380 nm, a width of the excitation slit and an emission slit of 12 nm and 8 nm, respectively, and the fluorescence spectrum was measured. The results are shown in FIG. 3 to FIG.

As shown in FIGS. 3 to 5, the compound represented by Formula 1 according to the present invention selectively binds to aluminum and a trivalent metal ion (M ³⁺ ) even in the presence of other metal ions.

More specifically, the fluorescence spectrum of the test solution in which the compound prepared in Example 1 according to the present invention was mixed with various metal ions was measured, and it was found that fluorescence at 516 nm was greatly increased with respect to aluminum ion (Al ³⁺ ) (See FIG. 3). As a result of measurement of the fluorescence spectrum of the test solution in which the compound prepared in Example 1 and the trivalent metal ion were mixed, fluorescence of 516 nm was greatly increased with respect to gallium ion (Ga ³⁺ ) and indium ion (In ^{3 +} ) , The fluorescence of 443 nm was increased and the fluorescence of 550 nm was decreased with respect to the iron ion (Fe ^{3 +} ) (see FIG. 4).

Further, in the case of the test solution in which the compound prepared in Example 1 and a solution of an aluminum ion (Al ^{3 +} ) - metal ion reference solution containing aluminum ion (Al ^{3 +} ) and various metal ions are mixed, ions (copper ions (Cu ^2+), ionic mercury (Hg ^{+ 2)} and chromium ion (except for Cr ^{+ 3))} without being affected by the optionally made of aluminum ions (Al ^{+ 3)} and the conjugate is causing the change in fluorescence (See FIG. 5).

From this, it can be seen that the compound represented by formula (I) according to the present invention causes fluorescence intensity change by selectively binding with aluminum ion (Al ^{3 +} ) to form a complex even in the presence of other metal ions.

Therefore, the aluminum and trivalent metal ion (M ³⁺ ) fluorescence-sensitized chemical sensor represented by the formula (1) according to the present invention can also selectively react with aluminum and a trivalent metal ion (M ³⁺ ) in the presence of other transition metal ions (M ³⁺ ), it is possible to detect aluminum ions (M ³⁺ ) even in the presence of other metal ions which can act as an interfering substance in aquatic environment such as groundwater and river Al ^{+ 3),} gallium ion (Ga ^{+ 3),} indium ion (in ^{+ 3)} and iron ion (Fe ^{+ 3)} Alternatively, it can be detected in a quantitative and simultaneous or Ishikawa. In addition, the fluorescence by coupling of the living cells to facilitate penetration into the aluminum ions in living cells (Al ^{3 +),} gallium ion (Ga ^{3 +),} indium ion (In ^{3 +)} and iron ion (Fe ^{3 +)} It can be useful as a fluorescence-sensitive chemical sensor capable of detection through intensity change.

< Experimental Example  3> Aluminum and trivalent metal ions of fluorescent sensitized chemical sensor (M ³⁺ ) Detection ability according to concentration

The following experiment was conducted to evaluate the detection ability of the fluorescent sensitized chemical sensor according to the present invention on the concentration of aluminum and trivalent metal ions (M ³⁺ ).

First, perchlorate salts of aluminum (Al ^{3 +} ) and iron (Fe ^{3 +} ) ions and nitrate salts of gallium (Ga ^{3 +} ) and indium (In ^{3 +} ) ions were dissolved in distilled water to prepare 0.5 mM trivalent metal ions (M &lt; ^{3 +} &gt;) reference solution. 10 mM hexamine buffer solution (pH 5.5, 1 mL) was injected into the test tube, and the compound prepared in Example 1 was dissolved in distilled water and 1.0 mM of fluorescent sensitized chemical sensor reagent was added thereto so as to have a concentration of 10 μM. Thereafter, a trivalent metal ion (M ³⁺ ) standard solution was added to each reaction solution so as to be gradually increased from 0 equivalent to 0.5 equivalent of the compound of Example 1 which is a fluorescent sensitized chemical sensor present in the mixed solution, Solution. The test solution was subjected to measurement of the absorption and fluorescence spectra under the same conditions as in Experimental Example 2. The results are shown in Figs. 6 to 11.

As shown in FIG. 6 to FIG. 10, the compound represented by the formula (1) according to the present invention shows a change in intensity of absorption and fluorescence depending on the concentration of aluminum and trivalent metal ion (M ³⁺ ).

Specifically, the fluorescence spectrum of the compound represented by the formula (1) according to the present invention was measured according to the aluminum ion (Al ³⁺ ) concentration. As a result, the equivalent of aluminum ion (Al ³⁺ ) It was confirmed that the intensity and the wavelength of the fluorescence and the light absorption gradually change (see FIGS. 6 and 10). In the case of gallium ion (Ga ^{3 +} ) and indium ion (In ^{3 +} ), fluorescence intensities and wavelengths were gradually changed as the equivalence was increased (see FIGS. 7 and 8). It was confirmed that the fluorescence of 443 nm and the fluorescence of 550 nm decrease with respect to the iron ion (Fe ^{3 +} ) (see FIG. 9).

Further, embodiments of aluminum ions of a compound (Al ^{3 +)} Aluminum ion through a fluorescence intensity change of 516nm in accordance with the concentration of (Al ^{3 +)} was calculated, the detection limit, exemplary aluminum in Example 1, the compounds according to the invention ions (Al ^{+ 3),} the detection limit was found to be 0.236μM (see Fig. 7). Therefore, the compound of formula (I) according to the present invention has a greater degree of change in fluorescence intensity depending on the concentration of aluminum and trivalent metal ion (M ³⁺ ). Therefore, the compound of formula (M &lt; ^{3 +} &gt;).

Accordingly, the aluminum and trivalent metal ion (M ³⁺ ) fluorescent-sensitized chemical sensor represented by Formula 1 according to the present invention is superior in sensitivity to aluminum and trivalent metal ions (M ³⁺ ) under-bonding is remarkably excellent, and the presence of other metal ions, and optionally the metal ion to the aluminum ion and trivalent metal (M ³⁺⁾ and form complexes with aluminum, and 3 via the fluorescence intensity changes in the chemical sensors ⁽³ M ⁺⁾ detection is possible because, ground water, in the presence of other metal ions which may act as interfering substances in an aqueous environment, such as rivers also aluminum ions (Al ^{3 +),} gallium ion (Ga ^{3 +),} indium ion (in ^{3 +} ) And iron ions (Fe ³⁺ ) can be detected selectively and quantitatively. In addition, the fluorescence by coupling of the living cells are easy to infiltrate into the aluminum ions in living cells (Al ^{+ 3),} gallium ion (Ga ^{+ 3),} indium ion (In ³⁺⁾ and iron ion (Fe ^{+ 3)} It can be useful as a fluorescence-sensitive chemical sensor capable of detection through intensity change.

< Experimental Example  4> Fluorescent chemical sensor and aluminum ion ( Al ^{3 +} )of Complex  formation mode  Confirm

Job's plot method was used to confirm the complex formation ratio of the compound of Example 1 to aluminum ion (Al ^{3 +} ). The plot of the plot of the plot of the work shows that the maximum point is when the mole fraction is 0.5, which is the typical ligand mole fraction when the ratio of ligand to metal complex is 1: 1 (see FIG. 8).

Example 1 Example 1 compound in the compound and _{50% CD 3 CN / D 2} O with aluminum ions (Al ^{3+) 1} H NMR spectroscopy also complexes of the Example 1 compound for aluminum ions (Al ^{+ 3)} complexes of Formation mode, and the difference in spectra is shown in FIG.

Signals of H ₁ , H ₆ , and H ₈ were down-shifted by about 0.02 ppm while signals of other protons showed little change. These results show that the aluminum ion (Al ^{3 +} ) together with the compound of Example 1 forms a 1: 1 complex with the hydroxy and carbonyl groups and is bonded.

From all these spectral data, it can be concluded that the metal binding to the hydroxy and carbonyl groups of the compound of Example 1 causes the absorption band of the compound of Example 1 to be redshifted by ICT (intramolecular charge transfer). It is believed that the enhancement of the fluorescence intensity of the compound of Example 1 is due to the bonding of the metal with the hydroxy and carbonyl groups accompanied by CHEF, i. E., PET inhibition. The predictable complex formation mode is shown in FIG.

< Experimental Example  5> Fluorescence Sensitive Chemical Sensor Within the sample  pH-dependent Detectability  evaluation

The invention the following experiment with aluminum ions (Al ^{+ 3)} to to evaluate the aluminum ions (Al ^{+ 3)} detection function according to the sample within the pH-sensitive fluorescent chemical sensor was carried out according to. aluminum ions according to the pH (Al ^{3 +)} 20 of pH 3.3 in 20mM MES buffer solution, 20 mM HEPES pH hexahydro 20mM of buffer min pH6.5-7.4 buffer solution of pH 8.5-11.5 1.0-6.0 In order to evaluate the detection performance mM CHES buffer solution was used. To the test tube, 20 mM buffer solution (1 mL) corresponding to each pH was added, and 20 μL of a 1.0 mM solution of the fluorescent sensitization chemical sensor reagent prepared in Experimental Example 1 was added. Then, 10 mM aluminum ion (Al ^{3 +} ) solution (10 μM) was added, and each test solution was diluted with distilled water to make the total solution amount to 2 mL. At this time, the concentration of the test solution was adjusted so that the concentration of the sensor was 10 μM, the concentration of the aluminum ion was 50 μM, and the concentration of the buffer solution was 10 mM. Then, the fluorescence spectrum of the test solution was measured, and the results are shown in FIG.

As shown in FIG. 10, aluminum ion (Al ^{3 +} ) was measured at various pHs in the sample for the compound of Example 1 according to the present invention. As a result, it was found that the compound of Example 1 contained aluminum ions Al ^&lt; ^{3 + &} gt ^; ). From this, it can be seen that the compound represented by the formula (1) according to the present invention is easy to detect aluminum ion (Al ^{3 +} ) in the range of pH 4.0 to pH 7.4.

Accordingly, the aluminum and trivalent metal ion (M ³⁺ ) fluorescent-sensitized chemical sensor represented by Formula 1 according to the present invention has remarkably excellent bonding property with aluminum and trivalent metal ion (M ³⁺ ) in an aqueous solution, Aluminum and trivalent metal ions (M ³⁺ ) can be detected in the pH range of pH 4.0 to 7.4 where the solubility of the trivalent metal ion (M ³⁺ ) is high and can act as an interfering substance in aquatic environment such as ground water and river in the presence of other metal ions aluminum ions (Al ^{+ 3),} gallium ion (Ga ^{3 +),} indium ion (in ^{+ 3)} and iron ion (Fe ³⁺⁾ with can be selectively detected or quantitatively that . In addition, the fluorescence by coupling of the living cells are easy to infiltrate into the aluminum ions in living cells (Al ^{+ 3),} gallium ion (Ga ^{+ 3),} indium ion (In ³⁺⁾ and iron ion (Fe ^{+ 3)} It can be useful as a fluorescence-sensitive chemical sensor capable of detection through intensity change.

Claims (10)

A tripeptide derivative represented by the following formula (1), or an optical isomer thereof:
[Chemical Formula 1]

In Formula 1,
R is -NH ₂ or -OH; And
n is 1 to 4;
The method according to claim 1,
The tripeptide derivative represented by the above formula (1)
(S) -2- (5- (dimethylamino) naphthalene-1-sulfonamido) -3-hydroxypropanamide Butanamido) -5-oxopentanoic &lt; / RTI &gt;acid;
(S) -2- ((S) -2- (5- (dimethylamino) naphthalene-1-sulfonamido) -3-hydroxypropanamido) butane Amide Pentanedioic acid;
(S) -2- (5- (dimethylamino) naphthalene-1-sulfamoido) -3-hydroxypropanamide ) Propanamido) -4-oxobutanoic &lt; / RTI &gt;acid; And
(S) -2 - ((S) -3-carboxy-2 - ((S) -2- (5- (dimethylamino) naphthalene- 1 -sulfonamido) -3- hydroxypropanamido) Wherein the tripeptide derivative is one selected from the group consisting of:
Lt; RTI ID = 0.0 &gt; 1, &lt;
Reacting a compound supported on a solid phase represented by formula (2) with a compound represented by formula (3) to prepare a compound represented by formula (4) (step 1);
Reacting the compound of Formula 4 prepared in Step 1 with a compound of Formula 3 to prepare a compound of Formula 5 (Step 2);
Reacting the compound of Formula 5 prepared in Step 2 with a compound of Formula 6 to prepare a compound of Formula 7 (Step 3);
Reacting the compound of Formula 7 prepared in Step 3 with 5- (dimethylamino) naphthalene-1-sulfonyl chloride to prepare a compound of Formula 8 (Step 4); And
(Step 5) of removing the t-butyl protecting group from the compound of formula (8) prepared in step 4 and separating from the solid phase to prepare the tripeptide derivative represented by formula (1a). Method for preparing tripeptide derivatives:
[Reaction Scheme 1]

In the above Reaction Scheme 1,
n is from 1 to 4;

Is selected from the group consisting of methylbenzhydrylamine (MBHA) resin, Wang resin, polyethylene glycol-polystyrene (PEG-PS) resin, silica nanoparticles, titanium oxide nanoparticles and chitosan in the solid phase 1 &lt; / RTI &gt; And
Fmoc is a protecting group, fluorenylmethyloxycarbonyl chloride; And
(1a) is a derivative of the formula (1).
Lt; RTI ID = 0.0 &gt; 2,
Reacting a compound supported on a solid phase represented by formula (2a) with a compound represented by formula (3) to prepare a compound represented by formula (4a) (step 1);
Reacting the compound of formula (4a) prepared in step 1 with a compound of formula (3) to prepare a compound of formula (5a) (step 2);
Reacting the compound of formula 5a prepared in step 2 with a compound of formula 6 to prepare a compound of formula 7a (step 3);
Reacting the compound of formula (7a) prepared in step 3 with 5- (dimethylamino) naphthalene-1-sulfonyl chloride to prepare a compound of formula (8a) (step 4); And
Removing the t-butyl protecting group from the compound of formula (8a) prepared in step 4 and separating from the solid phase to prepare a tripeptide derivative represented by formula (1b) (step 5). Method for preparing tripeptide derivatives:
[Reaction Scheme 2]

In the above Reaction Scheme 2,
X is halogen;
n is from 1 to 4;

Is selected from the group consisting of 2-chlorotrityl chloride resin, Wang resin, polyethylene glycol-polystyrene (PEG-PS) resin, silica nanoparticles, titanium oxide nanoparticles and chitosan as solid phase. And is characterized in that:
Fmoc is a protecting group, fluorenylmethyloxycarbonyl chloride; And
Formula (1b) is a derivative of Formula (1).
A tripeptide derivative according to claim 1,
Aluminum ions (Al ^{3 +),} gallium ion (Ga ^{3 +),} indium ion (In ^{3 +)} and iron ion (Fe ^{3 +)} with fluorescence-sensitive chemical that is capable of detecting the one or more detection subject is selected from the group consisting of sensor.
Tripeptide derivative of claim 1 for the aluminum ion of the formula 1 (Al ^3+), gallium ion (Ga ^3+), indium ion (In ³⁺⁾ and iron ions 1 selected from the group consisting of (Fe ³⁺⁾ (Step 1); And
Measuring a fluorescence signal that is the reaction product of a tripeptide derivative of the first detection target in the general formula (1) or more species present in the sample of step 1 occurs (step 2); aluminum ion (Al ³⁺⁾ containing , Gallium ion (Ga ³⁺ ), indium ion (In ³⁺ ) and iron ion (Fe ³⁺ ).
The method according to claim 6,
The detection method is characterized in that at least one detection target selected from the group consisting of aluminum ions (Al ³⁺ ), gallium ions (Ga ³⁺ ), indium ions (In ³⁺ ) and iron ions (Fe ³⁺ ) (Al ³⁺ ), gallium ion (Ga ³⁺ ), indium ion (In ³⁺ ), or the like, wherein the tripeptide derivative represented by the formula (1) is selectively sensitized and detected simultaneously by the difference of the respective fluorescence signals. And iron ions (Fe &lt; ^{3 +} &gt;).
The method according to claim 6,
Wherein the sample is at least one selected from the group consisting of aluminum ions (Al ³⁺ ), gallium ions (Ga ³⁺ ), indium ions (In ³⁺ ) and iron ions (Fe ³⁺ ) A method for detecting an object.
9. The method of claim 8,
Wherein the aqueous solution is an aqueous solution phase containing at least one organic solution selected from the group consisting of methanol, ethanol, dimethylformamide (DMF) and acetonitrile, and the aluminum ion (Al ³⁺ ), gallium ion (Ga ³⁺ ) , Indium ion (In ³⁺ ), and iron ion (Fe ³⁺ ).
The method according to claim 6,
(Al ³⁺ ), gallium ions (Ga ³⁺ ), indium ions (In ³⁺ ), and iron ions (Fe ³⁺ ), which are characterized by being water, sewage, wastewater, ). &Lt; / RTI &gt;

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