JPH06100554A - Trioxane derivative and method for chemiluminescence of the same - Google Patents

Abstract

PURPOSE:To provide the subject new compound useful as a detector for diagnostics, luminescent probes, or light sources for emergency use, and adaptable to the spectroscopic sensitivity of photomultiplier tubes. CONSTITUTION:The objective compound of formula I (R1 is halogen, 1-10C alkyl, COOH, OH, cyano, etc.,; R2 and R3 are each 1-3C alkyl; n is 0-11), e.g. 5-(3-perylenylamino)-6,6-dimethyl-3-isopropyl-1,2,4-trioxane. The compound of the formula I can be obtained by reaction between an aminoperylene derivative of formula II, an aldehyde of formula III and oxygen.

Description

Detailed Description of the Invention

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a novel trioxane derivative, its production method and chemiluminescence method. The derivative of the present invention is used as a detecting agent for various diagnostic agents, various luminescent probes, an emergency light source, and the like.

[0002]

2. Description of the Related Art Chemiluminescence refers to a phenomenon in which an atom or molecule is excited by energy generated by a chemical reaction to emit light. To date, compounds having various skeletons have been developed as chemiluminescent compounds, and acridinium ester, oxalate ester, etc. are known ("Bioluminescence and chemiluminescence" edited by Kazuhiro Imai, published by Hirokawa Shoten. ,
75-150, 1989). These compounds are used as a detecting agent for diagnostic agents or as a light source for emergencies.

[0003] For example, the reaction with luminol, lucigenin [bis (N-methylacridinium nitrate)], lophine (2,4,5-triphenylimidazole) or gallic acid, which are well known for the identification of blood stains, is well known. Has been.

Under alkaline conditions, they emit light in the presence of hydrogen peroxide and a metal such as Fe. By detecting the light generated here with a photomultiplier tube, the atoms or molecules involved in the reaction can be quantified. Many substances such as vitamin B ₁₂ and glucose have been measured.

Further, as another conventional technique, there is known a light emitting system in which an active intermediate produced by a reaction between an oxalate ester and hydrogen peroxide excites a fluorescent substance to emit light. A method for quantifying hydrogen peroxide generated from glucose by the action of glucose oxidase by this conventional technique and measuring the original glucose amount has been established.

On the other hand, 5- (9 having a trioxane skeleton
It is known that -anthrylamino) -3-isopropyl-6,6-dimethyl-1,2,4-trioxane also has chemiluminescent ability. (J.C.S. Chem. Co.
mm. , 180, 1976)

However, such a conventional trioxane compound emits light having a maximum at 550 nm, and depending on the type of the emission detector (photomultiplier tube), the sensitivity may not be satisfied at this wavelength. For this reason, there is a strong demand for chemiluminescent substances having various wavelengths suitable for luminescence detectors.

[0008]

The problem to be solved by the present invention is to provide a novel trioxane derivative having a spectral sensitivity of a photomultiplier tube. Another problem to be solved by the present invention is to provide a novel method for producing a trioxane derivative that is compatible with the spectral sensitivity of a photomultiplier tube. Further, the problem to be solved by the present invention is to provide a chemiluminescence method characterized in that the novel trioxane derivative of the present invention is mixed and reacted with a base to cause chemiluminescence. A further problem to be solved by the present invention is to provide a so-called “kit product”, which is composed of a combination of the novel trioxane derivative of the present invention and a base.

[0009]

According to the present invention, the general formula (1)

[0010]

[Chemical 7]

(In the formula, R ₁ is a halogen atom and has 1 to 1 carbon atoms.
10 represents an alkyl group or an alkoxy group, a carboxyl group, a hydroxyl group, a sulfonic acid group, a cyano group, an amino group or a nitro group, and R ₂ and R ₃ are the same or different,
Each represents an alkyl group having 1 to 3 carbon atoms. n is 0 to 1
A trioxane derivative (hereinafter referred to as trioxane 1) is provided.

According to the present invention, the general formula (2)

[0013]

[Chemical 8]

An aminoperylene derivative represented by the formula (wherein R ₁ has the same meaning as defined in the general formula 1) (hereinafter referred to as perylene 2), and a general formula (3)

[0015]

[Chemical 9]

A trioxane derivative (wherein R ₂ and R ₃ have the same meanings as defined in formula 1) and oxygen are reacted with each other (hereinafter referred to as aldehyde 3) A method for producing trioxane 1) is provided.

Further, according to the present invention, there is provided a chemiluminescent method characterized by reacting a trioxane derivative represented by the above general formula (1) with a base.

Furthermore, according to the present invention, there is provided a kit comprising a combination of the trioxane derivative represented by the general formula (1) and a base.

The present invention will be described in more detail below.

The trioxane derivative of the present invention is trioxane 1 represented by the above general formula (1). In the above trioxane 1, R ₁ is a halogen atom such as fluorine, chlorine, bromine or iodine, or an alkyl group having 1 to 10 carbon atoms such as methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, nonyl or decyl. ,
Alternatively, it represents an alkoxy group such as methoxy, ethoxy, or propoxy, a carboxyl group, a hydroxyl group, a sulfonic acid group, a cyano group, an amino group or a nitro group. R ₂ and R ₃ are the same or different and each represents an alkyl group having 1 to 3 carbon atoms such as methyl, ethyl and propyl. n shows the integer of 0-3.

Specific examples of the trioxane 1 include, for example, 5- (1-perylenylamino) -6,6-dimethyl-3-isopropyl-1,2,4-trioxane, 5-
(2-perylenylamino) -6,6-dimethyl-3-isopropyl-1,2,4-trioxane, 5- (3-perylenylamino) -6,6-dimethyl-3-isopropyl-1,2,4-trioxane, 3- (3-isopropyl-6,6-dimethyl-2,4,5-trioxacyclohexyl) amino-9-chloroperylene, 3- (3-isopropyl-6,6-dimethyl-2,4,5-trio (Xacyclohexyl) amino-9-methylperylene, 3-
(3-isopropyl-6,6-dimethyl-2,4,5-
Trioxacyclohexyl) amino-9-methoxyperylene, 3- (3-isopropyl-6,6-dimethyl-
2,4,5-Trioxacyclohexyl) amino-9-
Carboxyperylene, 3- (3-isopropyl-6,6
-Dimethyl-2,4,5-trioxacyclohexyl)
Amino-9-hydroxyperylene, 3- (3-isopropyl-6,6-dimethyl-2,4,5-trioxacyclohexyl) amino-9-sulfoperylene, 3- (3-
Isopropyl-6,6-dimethyl-2,4,5-trioxacyclohexyl) amino-9-cyanoperylene, 3
-(3-isopropyl-6,6-dimethyl-2,4,5
-Trioxacyclohexyl) amino-12-aminoperylene, 3- (3-isopropyl-6,6-dimethyl-
2,4,5-Trioxacyclohexyl) amino-9-
Nitroperylene and the like can be preferably mentioned.

In the method for producing the trioxane derivative of the present invention, the specific perylene 2 is reacted with the aldehyde 3 and oxygen to produce the trioxane 1 represented by the general formula (1).

The aminoperylene derivative used in the present invention is
It is perylene 2 represented by the general formula (2). Specific examples of the perylene 2 include 1-aminoperylene, 2-aminoperylene, 3-aminoperylene, 3-amino-9-chloroperylene, 3-amino-9-methylperylene and 3-amino-9-methoxyperylene. , 3-amino-9-carboxyperylene, 3-amino-9-hydroxyperylene, 3-amino-9-sulfoperylene, 3-
Preferable examples include amino-9-cyanoperylene, 3-amino-9-aminoperylene, 3-amino-9-nitroperylene and the like.

The aldehyde used in the present invention is aldehyde 3 represented by the above general formula (3). Specific examples of the aldehyde 3 include 2-methylpropanal and 2-methylpropanal.
Methylbutanal, 2-methylpentanal, 2-ethylpropanal, 2-ethylbutanal, 2-ethylpentanal, 2-propylpropanal, 2-propylbutanal, 2-propylpentanal and the like are preferably mentioned. You can

In the present invention, when the perylene 2 represented by the general formula (2) is reacted with the aldehyde 3 represented by the general formula (3), the charged molar ratio of the perylene 2 and the aldehyde 3 is 1: 2 to 1:10
00 is preferable, and particularly preferably 1: 3 to 1: 100. If the molar ratio is less than 2, the yield of the trioxane derivative formed tends to decrease,
When it exceeds, unreacted aldehyde remains after the reaction,
It is not preferable because isolation of the desired product becomes difficult. The reaction can be carried out at normal pressure, and the reaction temperature is generally -20 to 150 ° C, preferably -10.
It is preferably in the range of -100 ° C. If the reaction temperature is lower than -20 ° C, the reaction time tends to be long, while if it exceeds 150 ° C, the reaction operation is difficult, which is not preferable.

In the production method of the present invention, the desired trioxane 1 can be obtained by reacting the perylene 2 with the aldehyde 3 and oxygen under the various reaction conditions, but the reaction is smoother. It is desirable to use a solvent to do this. As the solvent, various solvents can be used as long as they are inert under the reaction conditions and can be easily separated from the produced trioxane derivative. Particularly, pentane, hexane, heptane, diethyl ether, dimethoxyethane, tetrahydrofuran, benzene, toluene, xylene, carbon tetrachloride or a mixed solvent thereof is suitable. The amount of the solvent used is not particularly limited, but is preferably 0.5 times to 100 times based on the total weight of the starting materials.

In the production method of the present invention, the target trioxane 1 can be obtained by reacting the perylene 2 with the aldehyde 3 and oxygen. The oxygen concentration is preferably 5 to 100%. . This reaction is carried out for a time sufficient to produce trioxane 1. The reaction time is determined depending on the conditions such as reaction temperature, reaction pressure, and the amount ratio of starting materials, and can be usually in the range of 1 hour to 14 days, but it is practically 10 hours to 7 days. It is desirable to set the conditions as follows.

The reaction product obtained by the production method of the present invention can be purified by a known method.

In the chemiluminescence method of the present invention, chemiluminescence can be obtained by mixing trioxane 1 and a base. Preferred examples of the base include sodium methoxide, potassium t-butoxide, sodium hydroxide, potassium hydroxide, potassium carbonate, sodium carbonate, sodium hydrogen carbonate, tetrabutylammonium hydroxide and the like.

In the chemiluminescence method of the present invention, trioxane 1 is used.
Although chemiluminescence can be obtained by mixing a base with a base, chemiluminescence can also be obtained by further mixing with another organic solvent. The organic solvent is preferably a polar solvent such as dimethyl sulfoxide, dimethylformamide, HMPA, dioxane, acetonitrile, methanol, ethanol, t-butanol or nitromethane.

[0031]

INDUSTRIAL APPLICABILITY The trioxane 1 of the present invention is a novel trioxane derivative suitable for the spectral sensitivity of a photomultiplier tube, and as a chemiluminescent substance having a high luminescence amount, it is a detection agent for various diagnostic agents and various luminescence. It can be used as a probe or an emergency light source. In addition, the production method of the present invention makes it possible to easily produce trioxane 1 in a short time with a high yield. Furthermore, the chemiluminescence method of the present invention can be applied to the above various uses.

[0032]

EXAMPLES The present invention will be described in more detail with reference to Examples and Comparative Examples, but the present invention is not limited thereto.

Example 1 0.13 g (0.60 mmol) of 6-aminoperylene was dissolved in 20 ml of benzene. Then isobutyraldehyde
Add 0.24 g (3.28 mmol) and room temperature at room temperature for 2
Reacted for days. After the reaction solvent was distilled off under reduced pressure, the residue was purified by silica gel chromatography. As a result, 5- (3-perylenylamino) -6,6-dimethyl-3-isopropyl-1,2,4-trioxane was obtained.

[0034]

[Chemical 10]

0.025 g (0.06 mmol) of was obtained. The spectrum data of this obtained compound are shown below.

¹ H NMR (CDCl ₃ ); δ = 1.01
(D, 6H, J = 4.6Hz), 1.30 (s, 3H), 1.67
(S, 3H), 1.92 (m, 1H), 4.52 (d, 1H, J
= 11.0Hz), 5.09 (d, 1H, J = 11.0Hz), 5.23
(D, 1H, J = 5.3Hz), 7.00-8.23 (m, 11
H) IR (KBr, cm ^-1 ); 3444, 2875, 150
4, 1390, 1062 MS; 425 (M ⁺ ) Elemental analysis C H N calculated value (%): 79.03 6.40 3.29 Measured value (%): 78.94 6.56 3.15

Example 2 5- (3-Perenylenylamino) -obtained in Example 1
10 to 50 μl of a solution of 6,6-dimethyl-3-isopropyl-1,2,4-trioxane in dimethylsulfoxide.
Chemoluminescence was obtained by adding 450 μl of a μmol / ml potassium t-butoxide-dimethylsulfoxide solution. The luminescence at this time was detected using a chemiluminescence detector CLD-100 (manufactured by Tohoku Denshi Sangyo Co., Ltd.). Table 1 shows the amount of emitted light and the maximum emission wavelength.

Comparative Example 1 5- (9-anthrylamino) -6,6-dimethyl-3
-Isopropyl-1,2,4-trioxane

[0039]

[Chemical 11]

10 μmo in the dimethylsulfoxide solution of
1 / ml of potassium t-butoxide-dimethylsulfoxide solution was added to cause chemiluminescence. The amount of emitted light and the maximum emission wavelength are also shown in Table 1.

[0041]

[Table 1] Table 1 Concentration Luminescence Quantity Luminescence maximum wavelength compound (mol / ml) (count number) (nm) Example 1 10 ^-13 1.0 × 10 ⁵ 640 Comparative Example 1 10 ^-13 6.0 × 10 ⁵ 550

Claims (4)

[Claims] 1. A compound represented by the general formula (1): (In the formula, R ₁ is a halogen atom, an alkyl group or an alkoxy group having 1 to 10 carbon atoms, a carboxyl group, a hydroxyl group,
It represents a sulfonic acid group, a cyano group, an amino group or a nitro group, and R ₂ and R ₃ are the same or different and each represents an alkyl group having 1 to 3 carbon atoms. and n represents an integer of 0 to 11). 2. A general formula (2): (In the formula, R ₁ is a halogen atom, an alkyl group or an alkoxy group having 1 to 10 carbon atoms, a carboxyl group, a hydroxyl group,
A sulfonic acid group, a cyano group, an amino group or a nitro group is shown. n represents an integer of 0 to 11) and an aminoperylene derivative represented by the general formula (3): (R ₂ and R ₃ are the same or different and each represent an alkyl group having 1 to 3 carbon atoms) and oxygen is reacted with oxygen. (Wherein R ₁ , R ₂ , R ₃ and n have the same meanings as defined above), and a method for producing the trioxane derivative. 3. A compound represented by the general formula (1): (In the formula, R ₁ is a halogen atom, an alkyl group or an alkoxy group having 1 to 10 carbon atoms, a carboxyl group, a hydroxyl group,
It represents a sulfonic acid group, a cyano group, an amino group or a nitro group, and R ₂ and R ₃ are the same or different and each represents an alkyl group having 1 to 3 carbon atoms. n represents an integer of 0 to 11), and a chemiluminescence method is characterized in that chemiluminescence is caused by mixing a trioxane derivative represented by the formula (3) with a base. 4. A compound represented by the general formula (1): (In the formula, R ₁ is a halogen atom, an alkyl group or an alkoxy group having 1 to 10 carbon atoms, a carboxyl group, a hydroxyl group,
It represents a sulfonic acid group, a cyano group, an amino group or a nitro group, and R ₂ and R ₃ are the same or different and each represents an alkyl group having 1 to 3 carbon atoms. A chemiluminescence kit comprising a combination of a trioxane derivative represented by (n represents an integer of 0 to 11) and a base.