JPH06100559A - Trioxane and method for chemiluminescence of the same - Google Patents

Abstract

PURPOSE:To provide the subject new compound having luminescent colors except yellow color, thus useful as a detector for various kinds of diagnostics and as a light source for emergency use. CONSTITUTION:The objective compound of formula I (R1-R3 are each lower alkyl; X is H, halogen or lower alkyl), e.g. 5-(4-methyl-7-coumarylamino)-3- isopropyl-6,6-dimethyl-1,2,4-trioxane. The compound of the formula I can be obtained by reaction between a coumarylamine derivative of formula II, an aldehyde derivative 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, 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.

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

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, since the maximum emission wavelength of such a conventional trioxane compound is 560 nm, it is recognized as yellow by human vision. Therefore, in reality, the development of trioxane derivatives that emit light of other colors such as blue is strongly desired.

[0008]

The problem to be solved by the present invention is to provide a novel trioxane derivative having an emission color other than yellow.

Another object of the present invention is to provide a method for producing a novel trioxane derivative having an emission color other than yellow.

Further problems to be solved by the present invention are as follows.
It is intended to provide a chemiluminescence method characterized by chemiluminescence by mixing the novel trioxane derivative of the present invention with a base.

A further problem to be solved by the present invention is as follows.
The object is to provide a kit comprising a combination of the novel trioxane derivative of the present invention and a base, that is, a so-called “kit product”. According to the kit product of the present invention, chemiluminescence can be obtained by in situ mixing a novel trioxane derivative and a base.

Still another problem to be solved by the present invention is to establish a method for quantifying atoms or molecules involved in the reaction by utilizing the chemiluminescence reaction of the novel trioxane derivative of the present invention. To expand the use of derivatives.

[0013]

[Means for Solving the Problem] One means for solving the problem is represented by the general formula (1).

[0014]

[Chemical 7]

A trioxane derivative represented by the formula: wherein R ₁ , R ₂ and R ₃ are the same or different and each represents a lower alkyl group, and X represents a hydrogen atom, a halogen atom or a lower alkyl group. Is.

Another means for solving the problem is the general formula (2)

[0017]

[Chemical 8]

A coumarylamine derivative represented by the formula: wherein R ₃ represents a lower alkyl group, and X represents a hydrogen atom, a halogen atom or a lower alkyl group, and a general formula (3)

[0019]

[Chemical 9]

A general formula (1) comprising reacting an aldehyde derivative represented by the formula (wherein R ₁ and R ₂ are the same or different and each represents a lower alkyl group) with oxygen.

[0021]

[Chemical 10]

It is to provide a process for producing a trioxane derivative represented by the formula (wherein R ₁ , R ₂ , R ₃ and X have the same meaning as defined above).

A further means for solving the problem is the general formula (1)

[0024]

[Chemical 11]

(Wherein R ₁ , R ₂ and R ₃ are the same or different and each represents a lower alkyl group, and X represents a hydrogen atom, a halogen atom or a lower alkyl group), and a base. A chemiluminescent method is characterized in that chemiluminescence is achieved by mixing at a suitable ratio.

Another means for solving the problem is the general formula (1)

[0027]

[Chemical 12]

(Wherein R ₁ , R ₂ and R ₃ are the same or different and each represents a lower alkyl group, and X represents a hydrogen atom, a halogen atom or a lower alkyl group) and a base. A chemiluminescent kit comprising a combination of

The present invention will be described in more detail below.

Examples of the novel trioxane derivative represented by the general formula (1) of the present invention include 5- (4-methyl-).
7-coumarylamino) -3-isopropyl-6,6-dimethyl-1,2,4-trioxane, 5- (4,5-dimethyl-7-coumarylamino) -3-isopropyl-6,6-
Dimethyl-1,2,4-trioxane, 5- (8-fluoro-4-methyl-7-coumarylamino) -3- (1-
Propylbutyl) -6,6-dipropyl-1,2,4-trioxane and the like can be listed.

In the general formula (1), R ₁ , R ₂
And R ₃ are the same or different and each represents a methyl group, an ethyl group, a propyl group, a butyl group or a pentyl group.

In the general formula (1), X can be a hydrogen atom, a fluorine atom, a chlorine atom, a bromine atom, iodine, a methyl group, an ethyl group or a propyl group.

The most preferred coumarylamine derivative represented by the general formula (2) is 7-coumarylamine when X represents a hydrogen atom.

Examples of the aldehyde represented by the general formula (3) include isobutyraldehyde, dipropylacetaldehyde, dipentylacetaldehyde and the like.

The means and conditions for producing the novel trioxane derivative of the present invention according to the present invention can be widely varied. The amount of the starting materials of the present invention represented by the general formulas (2) and (3) is about 1: in a molar ratio of coumarylamine represented by the general formula (2) to the aldehyde derivative represented by the general formula (3). It can vary from 2 to about 1: 1000, preferably from about 1: 3 to about 1: 100. If the molar ratio is less than 2, the yield of the produced trioxane derivative tends to decrease, and if it exceeds 1000, unreacted aldehyde derivative remains after the reaction, which makes isolation of the target product difficult. Therefore, it is not preferable.

The reaction temperature used in the present invention is usually room temperature, but in some cases, the desired substance can be obtained in a good yield in a short time by reacting at room temperature after heating under reflux for several hours. it can.

The reaction of the present invention is carried out in an inert solvent. Various solvents are used provided they are inert under the reaction conditions and are easy to separate from the trioxane derivative formed. Particularly, ether, dimethoxyethane, tetrahydrofuran, hexane, benzene, toluene or a mixed solvent thereof is suitable. The amount of solvent used is not particularly limited, but is about 0.5 times to about 1 based on the total weight of the starting materials.
It can be varied in the range of 00 times.

The reaction of the coumarylamine derivative represented by the general formula (2) with the aldehyde derivative represented by the general formula (3) and oxygen according to the present invention is usually carried out under atmospheric pressure, but under other reaction conditions. Depending on the materials used, the desired reaction rate, etc., it can also be carried out under overpressure. Further, oxygen may be used in the state of a mixed gas of an inert gas such as nitrogen and argon, and air free of water is preferable.

The reaction of the present invention is carried out for a time sufficient to produce a crude product of the trioxane derivative. The exact reaction is determined according to the requirements such as reaction temperature, reaction pressure, and the ratio of starting materials, but it is usually about 1 hour to about 14 hours.
It is in the range of days, preferably about 10 hours to about 7 days.

The crude product of the produced trioxane derivative may be dissolved in an appropriate solvent such as acetone or dichloromethane or a mixed solvent thereof, and then precipitated as crystals by a conventional method or purified by a column chromatography method. it can.

In the chemiluminescence method of the present invention, the chemiluminescence phenomenon is induced not only in the organic solvent but also in the system in the presence of water. Examples of the base include potassium t-butoxy, sodium methoxy, potassium ethoxy, sodium hydroxide, potassium hydroxide, potassium carbonate, sodium carbonate,
Examples include sodium hydrogen carbonate and tetrabutylammonium hydroxide.

The organic solvent used in the chemiluminescence method of the present invention is preferably a polar solvent, such as dimethyl sulfoxide, dimethylformamide, HMPA, dioxane, acetonitrile,
Suitable are methanol, ethanol, t-butanol or nitromethane.

[0043]

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. In Example 1, as data for identifying the target trioxane derivative, IR-81 manufactured by JASCO Corporation
Infrared absorption spectrum data by 0 and D manufactured by JEOL Ltd.
The mass spectrometry data by X-303 is described in this order,
In Example 2 and Comparative Example 1, the chemiluminescence amount measurement count number using BLX-201 manufactured by Aloka Co. was described.

Example 1 5- (4-methyl-7-coumarylamino) -3-isopropyl-6,6-dimethyl-represented by the structural formula (4)
Synthesis of 1,2,4-trioxane

[0045]

[Chemical 13]

256 mg (1.46 mmol) of 4-methyl-7-aminocoumarin in an air atmosphere from which water was removed
Was added to a mixed solution of 1 ml (11.0 mmol) of isobutyraldehyde and 15 ml of dimethoxyethane-hexane (2: 1), and the mixture was stirred at room temperature for 1 week in the dark. The reaction solution was concentrated under reduced pressure, and the obtained residue was subjected to silica gel column chromatography. By developing with hexane-dimethoxyethane (100: 1) as an eluent,
(4-Methyl-7-coumarylamino) -3-isopropyl-6,6-dimethyl-1,2,4-trioxane 409
mg (1.23 mmol) was obtained.

IR (KBr) cm ^-1 3440 (H-
N), 1720 (C = O ), 1620 (C = C) MS m / z 333 (M +) Elemental analysis _{(C 18 H 23 NO 5)} C H N calc (%): 64.85 6.95 4.20 Measured value (%): 64.85 6.98 4.18

Example 2 Synthesis was carried out in Example 1 as a measurement compound.
(4-methyl-7-coumarylamino) -3-isopropyl-6,6-dimethyl-1,2,4-trioxane,
100 μl solution of dimethyl sulfoxide containing 10 ⁻⁷ M,
Injection was performed in a cell containing 900 μl of a solvent containing a base in a chemiluminescence measuring device, and immediately after that, the amount of light emission was measured for 10 seconds. At that time, a 10 mM dimethylsulfoxide solution of potassium t-butoxide was used as an organic solvent containing a base. The measurement results are shown in Table 1. The measurement count numbers in Table 1 are relative values of "no unit".

Example 3 In the same manner as in Example 1, 5- (4,5-dimethyl-7-coumarylamino) -3-isopropyl-6,6-dimethyl-
1,2,4-trioxane and 5- (8-fluoro-4)
-Methyl-7-coumarylamino) -3- (1-propylbutyl) -6,6-dipropyl-1,2,4-trioxane was prepared. The identification data are summarized below.

5- (4,5-Dimethyl-7-coumarylamino) -3-isopropyl-6,6-dimethyl-1,2,4
-Trioxane

IR (KBr) cm ^-1 3440 (H-
N), 1720 (C = O), 1620 (C = C) MS m / z 347 Elemental analysis (C ₁₉ H ₂₅ NO ₅ ) C H N Calculated value (%): 65.69 7.25 4.03 Measurement Value (%): 65.70 7.23 4.05

5- (8-fluoro-4-methyl-7-coumarylamino) -3- (1-propylbutyl) -6,6-
Dipropyl-1,2,4-trioxane

IR (KBr) cm ^-1 3460 (H-
N), 1725 (C = O ), 1630 (C = C) MS m / z 463 Elemental analysis _{(C 26 H 38 NFO 5)} C H N calc (%): 67.36 8.26 3.02 Measurement Value (%): 67.35 8.28 3.03

Comparative Example 1 5- (4-methyl-7-coumarylamino) -of Example 2
Instead of 3-isopropyl-6,6-dimethyl-1,2,4-trioxane, structural formula (5)

[0055]

[Chemical 14]

5- (9-anthrylamino) -3-isopropyl-6,6-dimethyl-1,2, represented by
The same measurement experiment as in Example 2 was conducted using 4-trioxane. The results are also shown in Table 1. Table 1
The measurement count number of is a relative value of "no unit".

[0057]

Table 1 Table 1 Compound emission amount (measurement count number) Maximum emission wavelength (nm) Example 2 3.87 × 10 ⁶ 470 Comparative Example 1 4.06 × 10 ⁶ 560

[0058]

Since the novel trioxane derivative of the present invention emits blue light having a maximum emission wavelength of 470 nm, it has an effect of expanding the use of the trioxane derivative as a chemiluminescent substance.

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Claims (4)

[Claims] 1. A compound represented by the general formula (1): (Wherein R ₁ , R ₂ and R ₃ are the same or different and each represents a lower alkyl group, and X represents a hydrogen atom, a halogen atom or a lower alkyl group). 2. A general formula (2): (Wherein R ₃ represents a lower alkyl group, and X represents a hydrogen atom, a halogen atom or a lower alkyl group), and a coumarylamine derivative represented by the general formula (3): (Wherein R ₁ and R ₂ are the same or different and each represents a lower alkyl group), and oxygen is reacted with a general formula (1) (Wherein R ₁ , R ₂ , R ₃ and X have the same meaning as defined above), and a method for producing the trioxane derivative. 3. A compound represented by the general formula (1): (Wherein R ₁ , R ₂ and R ₃ are the same or different and each represents a lower alkyl group, and X represents a hydrogen atom, a halogen atom or a lower alkyl group), and a base is mixed with the trioxane derivative. A chemiluminescence method characterized by causing chemiluminescence. 4. A compound represented by the general formula (1): (Wherein R ₁ , R ₂ and R ₃ are the same or different and each represents a lower alkyl group, and X represents a hydrogen atom, a halogen atom or a lower alkyl group), and a combination of a trioxane derivative and a base A chemiluminescent kit consisting of.