JP3110132B2 - Novel β-alanine derivative, method for producing the same, and method for producing glutamate blocker - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a novel .beta.-alanine derivative useful as a raw material for introducing polyamine into amino acids, peptides and amine compounds, a method for producing the same, and a method for producing a glutamate blocking agent.

[0002]

2. Description of the Related Art In recent years, a compound in which an amino acid or peptide and a polyamine are bonded has been found as a component in a biotoxin. For example, glutamate receptor blockers, JSTXs, NSTXs, and nephylatoxins contained in slime spider venom are compounds containing cadaverine, a diamine, spermidine, a triamine, spermine, a tetraamine, and the like in a molecule `` Y. Aramaki et al., Pro
c. Acad. Jpn. Ser. B, 62, p359 (1986), T. Toki et.
al., Biomed. Res., 9, p421 (1988), T. Tokiet al.,
Jpn. J. Sanit. Zool., 41, p9 (1990) ". The present inventors have focused on the glutamate-blocking activity of spider venom components, and have described a derivative as a glutamate-blocking agent “
No. 294734, JP-A-2-256656 '', and amino acids,
Azide compounds as diamine introduction raw materials for peptides,
And a proposal for the introduction of cadaverine into the venom of the spider spider (see Japanese Patent Application No. 2-209043).

[0003]

However, some highly active spider venom components or their derivatives include compounds in which a diamine is further bound to a polyamine (JSTX-3, nefilatoxin-1, etc.).
-8 etc.) also exist. JSTX as an example of the synthesis of these compounds
-3 "Y. Hashimoto et al., Tetrahedron Lett., 28, p3
511 (1987) ". In this example, the diamine is reacted with acrylonitrile and then ethyl acrylate, the nitrile is reduced, the amino group is protected with a benzyloxycarbonyl group, the ester is converted into a carboxylic acid by saponification, and then reacted with the compound to be introduced. And many steps are required to obtain the target compound.

[0004]

Means for Solving the Problems The present inventors have conducted intensive studies on a more efficient polyamine introduction method in such a case, and as a result, have found a novel compound useful as a polyamine introduction raw material and completed the present invention. Reached. That is, the present invention provides a compound represented by the general formula (I):

[0005]

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Wherein R ₁ represents an alkyl group, an aryl group or an aralkyl group, Boc represents a t-butyloxycarbonyl group, and m represents an integer of 1 to 10. An alanine derivative; a method for producing the same; and a method for producing a glutamate blocker using the β-alanine derivative.

The β-alanine derivative of the present invention has the following general formula (VI) R ₁ O ₂ C (CH ₂ ) ₂ NH (CH ₂ ) _m NH (CH ₂ ) ₂ CO ₂ R ₁ (VI) Wherein R ₁ and m have the same meanings as described above, and a compound represented by the general formula (V) obtained by reacting a compound represented by the formula (V) with a t-butyloxycarbonylation reagent:

[0008]

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(Wherein R ₁ , Boc, m have the same meanings as described above), and further reacted with an alkali borohydride to obtain a compound represented by the general formula (III):

[0010]

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(Wherein R ₁ , Boc, and m have the same meaning as described above), which is further converted to a compound represented by the general formula (I
V) R ₂ -X (IV) (wherein, R ₂ represents an alkylsulfonyl group, an arylsulfonyl group or an aralkylsulfonyl group, and X represents a halogen atom). General formula (II)

[0012]

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(Wherein, R ₁ , R ₂ , Boc, and m have the same meanings as described above) and an alkali azide, whereby the compound can be produced in high yield.

The synthesis of the compound represented by the general formula (VI) is performed by cooling an alcohol solution of a diamine represented by the formula H ₂ N (CH ₂ ) _m NH ₂ (m has the same meaning as described above). Below, formula CH _{2
= CH-CO 2 R 1 (} R 1 is the same meaning as defined above) is carried out by dropwise an alcohol solution of an acrylic acid ester represented by. The reaction is usually completed in 30 minutes to several hours. General formula (VI) above
The introduction of a Boc group into the compound represented by the formula: 2-t-butyloxycarbonyl-oxyimino-2-phenylacetonitrile, St-butyloxycarbonyl-4,6-dimethyl-2-thiopyrimidine, di-t The reaction is performed using an existing t-butyloxycarbonylation reagent such as -butyl-dicarbonate. The reaction is usually carried out in the presence of a base such as triethylamine, sodium hydroxide or sodium carbonate, in an aqueous solution system or a mixed solvent system of water and an organic solvent such as dioxane. The t-butyloxycarbonylation reagent is used in an amount of 1 to 2 times the molar amount of the corresponding amino group, and the base is used in an amount of 1 to 1.5 times the amount of the t-butyloxycarbonylation reagent.
It is desirable to add a double molar amount. The reaction is completed at room temperature in 1 hour to 24 hours, and it is preferable that the reagents are mixed under ice cooling.

The reaction between the compound represented by formula (V) and the alkali borohydride is usually carried out in a mixed solvent system of water and an organic solvent such as tetrahydrofuran. The amount of the alkali borohydride to be added is desirably 5 to 6 times the mol of the compound represented by the general formula (V). Reaction 3
It is completed in 0 minutes to several hours, but it is desirable to add the alkali borohydride in 4 to 5 portions in order to suppress the by-product of diol.

The compound represented by the general formula (III) and the general formula
The reaction of the compound represented by (IV) is carried out in an organic solvent such as dichloromethane under basic conditions. As the base to be added, an organic base such as pyridine or triethylamine is desirable. The compound represented by the general formula (IV) is 1 to 3 times the molar amount of the corresponding hydroxyl group, and the amount of the base is the general formula
The amount of the compound represented by (IV) is desirably 1 to 2 times the molar amount. Although the addition of the reagent is desirably performed at 0 ° C. or lower, the reaction is completed in 30 minutes to several hours at room temperature. General formula (II)
The reaction of the compound represented by with an alkali azide is carried out by stirring in an organic solvent such as dimethylformamide at room temperature for 5 to 30 hours.

The product of each reaction can be purified by recrystallization or chromatography after solvent extraction, and the product can be identified by measuring elemental analysis, IR spectrum, NMR spectrum and Mass spectrum.

The method of introducing the compound represented by the general formula (I) into amino acids, peptides and other amine compounds is performed by converting the ester terminal of the compound represented by the general formula (I) into a carboxylic acid, Active ester method, amide bond formation method, dicyclohexylcarbodiimide (DCC)
It can be performed by the method or DCC-additive method. The conversion of the ester to the carboxylic acid is carried out by reacting the compound represented by the general formula (I) with an alkali hydroxide in a mixed solvent of alcohol and water. The amount of the alkali hydroxide to be added is desirably at least 1 times the molar amount of the compound represented by the general formula (I). Then, for example, the formula _{(VII) R 3 -Y- (Asn} ) n -NH (CH 2) p NH 2 ... (VII) ( wherein, R ₃ is A alkyl group, an aryl group or indolyl
Y represents a methyl group , Y represents a carbonyl group or a sulfonyl group, Asn represents an asparagine residue, n represents 0 or 1, and p represents an integer of 2 to 5. ) To react with a compound represented by the general formula (VIII)

[0019]

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(Where R ₃ , Y, Asn, Bo
c, m, n, and p have the above meanings. ), And subjecting this compound to a reduction reaction, a glutamate blocking agent represented by the general formula (IX) can be obtained. R ₃ -Y- (Asn) _n -NH (CH ₂ ) _p NHCO (CH ₂ ) ₂ NH (CH ₂ ) _m NH (CH ₂ ) ₃ NH ₂ … (IX) (where R ₃ , Y, Asn , m, n, and p have the above meanings.) The reduction of the azide group to the amino group can be carried out after the introduction by a catalytic reduction method using palladium carbon as a catalyst.
The removal of the Boc group is performed in an organic solvent such as dichloromethane.
It can be performed using a strong organic acid such as trifluoroacetic acid.

[0021]

EXAMPLES Hereinafter, the present invention will be described in detail based on examples, but the present invention is not limited to these examples.

Example 1 1,4-Diaminobutane (0.880 g) was dissolved in 10 ml of ethanol and cooled to -10 ° C. in an ice-salt bath. Methyl acrylate (1.808 g) was added dropwise to this solution over 30 minutes. The reaction mixture was stirred at −10 ° C. to 0 ° C. for another 2 hours,
The solvent was distilled off under reduced pressure, and the residue was dissolved in 10 ml of distilled water.
Sodium carbonate (3.180 g) and di-t-butyl-dicarbonate (6.547 g) were added to the solution, and the mixture was stirred at room temperature for 12 hours. The reaction mixture was extracted with ethyl acetate, washed twice with 10% citric acid and once with saturated brine, dried over anhydrous sodium sulfate, and the solvent was distilled off under reduced pressure. The residue was generated on a silica gel column (hexane / ethyl acetate = 4/1 to 2/1 by volume) to obtain 3.37 g of a colorless oily compound 1 having the following structural formula. FIG. 1 shows the ¹ H-NMR spectrum of the obtained compound 1 in deuterated chloroform, and FIG.
Shown in

[0023]

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Compound 1 (1.670 g) was dissolved in a mixed solution of 700 ml of tetrahydrofuran and 14 ml of distilled water, 80 mg of lithium borohydride was added to the solution, and 80 mg of hydrogen was added again to the reaction mixture stirred at room temperature for 1 hour. After adding lithium borohydride, the mixture was stirred at the same temperature for 1 hour. This operation was repeated four more times. The reaction mixture was filtered and the filtrate
After adding 10 ml of a 10% aqueous citric acid solution, the mixture was concentrated under reduced pressure. The residue was extracted with ethyl acetate, and the extract was washed twice with saturated saline, dried over anhydrous sodium sulfate, and the solvent was distilled off under reduced pressure. The residue is purified on a silica gel column (hexane /
(Ethyl acetate = 1/1 volume ratio) to give 0.889 g of the compound 2 having the following structural formula as a colorless oily substance.
FIG. 3 shows the ¹ H-NMR spectrum of the obtained compound 2 in deuterated chloroform, and FIG. 4 shows the IR spectrum.

[0025]

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Compound 2 (0.889 g) was dissolved in a mixed solution of 13.30 ml of pyridine and 4 ml of anhydrous dichloromethane, and cooled to 0 ° C. in an ice bath. To this solution was added 0.145 g of methanesulfonyl chloride, and the mixture was stirred at 0 ° C. for 1 hour and at room temperature for 12 hours. The reaction mixture was added to 60 ml of ice water, stirred for 30 minutes and extracted with ethyl acetate. The extract was washed five times with water and twice with a saturated saline solution, dried over anhydrous sodium sulfate, and the solvent was distilled off under reduced pressure. The residue was dissolved in 5 ml of dry dimethylformamide, 0.266 g of sodium azide was added to the solution, and the mixture was stirred at room temperature for 12 hours. 50 reaction mixture
After dilution with ml of ethyl acetate, the mixture was diluted twice with 50% saturated saline,
Washed once with water and once with saturated saline. After the organic layer was dried over anhydrous sodium sulfate, the solvent was distilled off under reduced pressure, and the residue was subjected to a silica gel column (hexane / ethyl acetate = 2 / 1-1 / 1 / hexane).
(1 volume ratio) to obtain 0.813 g of a colorless oily compound 3 having the following structural formula. FIG. 5 shows the ¹ H-NMR spectrum of the obtained compound 3 in deuterated chloroform, and FIG. 6 shows the IR spectrum.

[0027]

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Application Example 1 Synthetic compound 3 (100 mg) of a spider venom component, nefilatoxin-8, was dissolved in 5 ml of methanol, and 1 ml of 1N sodium hydroxide was added to the solution, followed by stirring at 55 ° C. for 2 hours. After adding 4 ml of 10% aqueous citric acid solution to the cooled reaction mixture, it was extracted with ethyl acetate. The extract was washed once with water and once with a saturated saline solution, dried over anhydrous sodium sulfate, and the solvent was distilled off under reduced pressure. The residue was dissolved in 4 ml of dichloroethane and the solvent was distilled off under reduced pressure. This operation was further repeated twice, and the remaining traces of methanol were completely distilled off. The residue was dissolved in 2 ml of dry ethyl acetate, and 46 mg of paranitrophenol and 91 mg of dicyclohexylcarbodiimide (DCC) were added to the solution, followed by stirring at room temperature for 12 hours. 3 ml of hexane was added to the reaction mixture, and the precipitated urea was filtered off. The filtrate was diluted with 5 ml of ethyl acetate and washed once with a 50% saturated saline solution and once with a saturated saline solution. The solvent was distilled off under reduced pressure to obtain 130 mg of crystals.

62 mg of 1-azido-5- (indole-3-acetylasparaginyl) aminopentane was dissolved in 2 ml of ethanol, and 10 mg of 10% by weight palladium carbon was added thereto.
(Pd / C) was added, and the mixture was stirred under a hydrogen atmosphere for 3 hours. Pd / C was filtered off from the reaction mixture, the filtrate was concentrated under reduced pressure, and the residue was dissolved in 2 ml of dry dimethylformamide.
To this solution, a solution of 130 mg of the above crystals dissolved in 1 ml of dimethylformamide and 45 mg of triethylamine were added, and the mixture was stirred at room temperature for 12 hours. The reaction mixture was diluted with 30 ml of ethyl acetate and washed successively twice with 50% saturated saline, once with water and once with saturated saline. The solvent was distilled off under reduced pressure, and the residue was purified by a silica gel column (dichloromethane / acetone / ethanol = 3/1 / 0.3 volume ratio) to give 69 mg of white crystals.
I got 20 mg of this white crystal was dissolved in 1 ml of acetic acid, 0.4 ml of trifluoroacetic acid and 7 mg of 10% Pd / C were added, and the mixture was stirred at room temperature for 4 hours under a hydrogen gas atmosphere. After the catalyst was filtered off, the solvent was distilled off under reduced pressure. The residue was dissolved in 1 ml of water, and reversed-phase high-performance liquid chromatography (manufactured by JASCO, Megapak SIL C-18).
Using a φ10.0 × 250 mm), a 15% aqueous acetonitrile solution containing 0.1% TFA was fractionated as a mobile phase to obtain 8 mg of nefilatoxin-8 represented by the following formula. FIG. 7 shows the ¹ H-NMR spectrum of the obtained nefilatoxin-8 in heavy water.

[0030]

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[0031]

The β-alanine derivative of the present invention is useful as a raw material for introducing polyamines into amino acids, peptides and other amine compounds. According to the present invention, it is possible to supply a large amount of a physiologically active substance containing a polyamine in a molecule. It was opened.

[Brief description of the drawings]

[1] ¹ in deuterochloroform of Compound 1 of Example 1
It is an H-NMR spectrum figure.

FIG. 2 is an IR spectrum of Compound 1 of Example 1.

[3] ¹ in deuterochloroform of Compound 2 of Example 1
It is an H-NMR spectrum figure.

FIG. 4 is an IR spectrum of Compound 2 of Example 1.

[5] ¹ in deuterochloroform of compound 3 of Example 1
It is an H-NMR spectrum figure.

FIG. 6 is an IR spectrum of Compound 3 of Example 1.

FIG. 7: Final product of application example 1, nefilatoxin-8
FIG. ¹ is a ¹ H-NMR spectrum diagram in heavy water.

──────────────────────────────────────────────────の Continued on the front page (51) Int.Cl. ⁷ Identification code FI C07C 303/28 C07C 303/28 309/65 309/65 309/67 309/67 309/72 309/72 C07D 209/18 C07D 209 / 18 (56) Reference Chem. Lett. , (6) 1993, p. 929-p. 932 (58) Fields surveyed (Int. Cl. ⁷ , DB name) CA (STN) CAOLD (STN) REGISTRY (STN)

Claims (3)

(57) [Claims] 1. A compound of the general formula (I) (Wherein, R ₁ represents an alkyl group, an aryl group or an aralkyl group, Boc represents a t-butyloxycarbonyl group, and m represents an integer of 1 to 10.) A novel β-alanine derivative represented by the following formula: . 2. A compound of the general formula (II) (Wherein, R ₁ represents an alkyl group, an aryl group or an aralkyl group, R ₂ represents an alkylsulfonyl group, an arylsulfonyl group or an aralkylsulfonyl group, and Boc represents t
Represents a -butyloxycarbonyl group, and m represents an integer of 1 to 10. The compound represented by the formula (I) according to claim 1, wherein the compound represented by the formula) is reacted with an alkali azide.
A method for producing a novel β-alanine derivative represented by the formula: 3. The β-alanine derivative represented by the general formula (I) according to claim 1, wherein the ester terminal is a carboxylic acid, and then a compound represented by the formula (VII) R ₃ -Y- (Asn) _n -NH (CH _{2) p NH 2 ... (VII} ) ( wherein, R ₃ is A alkyl group, an aryl group or indolyl
Y represents a methyl group , Y represents a carbonyl group or a sulfonyl group, Asn represents an asparagine residue, n represents 0 or 1, and p represents an integer of 2 to 5. ) To react with a compound represented by the general formula (VIII): (Wherein, R ₃ , Y, Asn, n, and p have the above meanings, and m is 1
To 10 and Boc represents a t-butyloxycarbonyl group. A method for producing a glutamate blocker represented by the general formula (IX), comprising obtaining a compound represented by the formula (1) and subjecting the compound to a reduction reaction. R ₃ -Y- (Asn) _n -NH (CH ₂ ) _p NHCO (CH ₂ ) ₂ NH (CH ₂ ) _m NH (CH ₂ ) ₃ NH ₂ … (IX) (where R ₃ , Y, Asn , m, n, and p have the above meanings.)