JP2911287B2 - N-acyl amino acid compound and method for producing the same - 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 N-acyl amino acid compound and a method for producing the same. More specifically, antioxidant power, emulsifying power, antibacterial power, chelating power obtained by acylating histidine and a peptide containing the histidine with a long-chain fatty acid,
The present invention relates to a novel compound having various functions such as an ultraviolet absorbing power and a moisturizing power, and a method for producing the same.

[0002]

2. Description of the Related Art In recent years, it has been considered that active oxygen causes various oxidation reactions and is considered to be an important cause of various diseases such as aging, arteriosclerosis, and cancer. Has been actively researched. For example, in the food industry, BHT (3,5-t-butyl-4
-Hydroxytoluene) and synthetic antioxidants such as BHA (2, (3) -t-butyl-4-hydroxyanisole) have been used as inexpensive and efficient antioxidants.

[0003]

However, since these compounds, especially BHA, were reported to have a carcinogenic effect, concerns have been raised over their safety. Tocopherol, which is used as a naturally occurring antioxidant, has excellent antioxidant power, but is poorly soluble in water, and promotes oxidation at high concentrations. You will be limited if you think about it. Other polyphenols having antioxidant power are water-soluble and have weaknesses such as being difficult to dissolve in oil, so that their applications are limited.

Accordingly, it is an object of the present invention to provide a novel acyl compound and a method for producing the same.

Another object of the present invention is to provide an antioxidant power, an emulsifying power,
It is an object of the present invention to provide a novel compound having functions such as antibacterial activity, chelating activity, ultraviolet absorbing activity, and moisturizing activity, and a method for producing the same.

[0006]

These objects are achieved by the general formula (1) R-CO- (NH-X-CO) _n -OR ¹ (1) (wherein (NH-X-CO) Represents a carnosine residue, R-CO represents a saturated or unsaturated fatty acid residue having 6 to 24 carbon atoms, R ¹ represents a hydrogen atom, a sodium atom, a potassium atom or a methyl group, and n represents 1 or Which is an integer of 2 and contains at least one histidine as a constituent amino acid).

[0007] These objects are represented by the general formula (2)

[0008]

Embedded image

(Wherein, R-CO has 6 carbon atoms)
~ 24 saturated or unsaturated fatty acid residues. ) Is represented by the general formula (3) H- (NH-X-CO) _n -OR ¹ (3) (wherein (NH-X-CO) represents an amino acid residue) , (NH-X-CO) different .R-CO depending on the type of amino acid X is used in indicates a saturated or unsaturated fatty acid residue having 6 to 24 carbon atoms .R ¹ is hydrogen atom, sodium atom, potassium And n represents an atom or a methyl group, and n is an integer of 1 to 3, and includes at least one histidine as a constituent amino acid.) -CO- (NH-X-CO) _n -OR ¹ (1) (wherein, His, X, R, R ¹ and n are as described above). Yo It is also achieved.

R—CO— (NH—X—CO) _n —OR ¹ (1) (where, (NH—X—CO) represents an amino acid residue and X in (NH—X—CO) R-CO represents a saturated or unsaturated fatty acid residue having 6 to 24 carbon atoms, R ¹ represents a hydrogen atom, a sodium atom, a potassium atom or a methyl group, and n represents 1 Which is an integer of 1 to 3 and contains at least one histidine as a constituent amino acid).

R—CO— (NH—X—CO) _n —OR ¹ (1) (where, (NH—X—CO) represents a carnosine residue, and R—CO has 6 carbon atoms) .R ¹ illustrating a saturated or unsaturated fatty acid residue 24 include a hydrogen atom, a sodium atom, a potassium atom or a methyl group. also n is the constituent amino acids of at least one histidine integer of 1 or 2 This is also achieved by an emulsifier containing an N-acylamino acid compound represented by the formula (1) as an active ingredient.

[0012]

The N-acylamino acids, N-acylpeptide compounds, their salts and their methyl esters (hereinafter referred to as N-acylamino acid compounds) according to the present invention are compounds represented by the general formula (1) as described above. Wherein R-CO has 6 to 24 carbon atoms, preferably 8 to 2 carbon atoms.
0 saturated or unsaturated fatty acid residues, and the fatty acid residues include capryloyl (C ₈ ), caprinoyl (C ₁₀ ),
Lauroyl (C _12), myristoyl (C _14), palmitoyl (C _16), oleoyl (C _18: 1), linoleyl (C
_{18: 2} ), linolenyl (C _{18: 3} ), stearoyl (C ₁₈ ), arachidoyl (C ₂₀ ) and the like.

The (NH-X-CO) portion is an amino acid residue, wherein X differs depending on the amino acid used, and the amino acids include histidine (His) and glycine (Gl
y), tryptophan (Trp), isoleucine (Il)
e), phenylalanine (Phe), methionine (Me
t), cysteine (Cys), leucine (Leu), lysine (Lys), alanine, valine, proline, serine, threonine, tyrosine, asparagine, glutamine, arginine, aspartic acid, glutamic acid, β-
Alanine, dopa, creatine, ornithine, (D, L
Any of those having a body may be used). If the above conditions are satisfied, the peptide may be a branched peptide.
(In the following description, abbreviations are used in parentheses in the above amino acids.) N is an integer of 1 to 3, preferably 1 to 2 and constitutes at least one His. Contains as amino acids. Therefore, when n is 1, it is a His residue, when n is 2, 3 is a peptide residue, and when n is 2, -His-His, -Gl
y-His, -His-Gly, -His-Leu,-
Leu-His, -Trp-His, -His-Tr
p, -Ile-His, -His-Ile, -Phe-
His, -His-Phe, -Met-His, -Hi
s-Met, -Cys-His, -His-Cys, β
-Alanyl-3-methyl-L-His, γ-aminobutyryl-L-His, carnosine (β-alanyl-LH
is), and when n is 3, -His-His-H
is, -Gly-His-Lys, -Gly-His-
His, -Gly-Gly-His, -Ile-His
-His, -Ile-Ile-His, -Phe-Hi
s-His, -Phe-Phe-His, -Met-H
is-His, -Met-Met-His, -Cys-
His-His, -Cys-Cys-His, -Trp
-His-His, -Trp-Trp-His and the like.

That is, the N-acylamino acid compound of the present invention is a compound containing the above-mentioned fatty acid residue and amino acid residue as constituents, a salt thereof and a methyl ester thereof,
In particular, N-capryloyl-His, N-capryloyl-
His-His, N-Capryloyl-His-Leu,
N-Capryloyl-Leu-His, N-Capryloyl-β-alanyl-3-methyl-L-His, N-Capryloyl-γ-aminobutyryl-L-His, N-Capryloyl-carnosine, N-Capryloyl-His-Hi
s-His, N-capryloyl-Gly-His-Ly
s, N-Capryloyl-Gly-His-His, N-
Capryloyl-Gly-Gly-His, N-Caprinoyl-His, N-Caprinoyl-His-His, N
-Caprinoyl-His-Leu, N-Caprinoyl-
Leu-His, N-caprinoyl-β-alanyl-3
-Methyl-L-His, N-caprinoyl-γ-aminobutyryl-L-His, N-caprinoyl-carnosine, N-caprinoyl-His-His-His, N-
Caprinoyl-Gly-His-Lys, N-Caprinoyl-Gly-His-His, N-Caprinoyl-G
ly-Gly-His, N-lauroyl-His, N-
Lauroyl-His-His, N-Lauroyl-His
-Leu, N-lauroyl-Leu-His, N-lauroyl-β-alanyl-3-methyl-L-His, N-
Lauroyl-γ-aminobutyryl-L-His, N-lauroyl-carnosine, N-lauroyl-His-Hi
s-His, N-lauroyl-Gly-His-Ly
s, N-lauroyl-Gly-His-His, N-lauroyl-Gly-Gly-His, N-myristoyl-His, N-myristoyl-carnosine, N-palmitoyl-His, N-palmitoyl-His-His,
N-palmitoyl-His-Leu, N-palmitoyl-Leu-His, N-palmitoyl-β-alanyl-
3-methyl-L-His, N-palmitoyl-γ-aminobutyryl-L-His, N-palmitoyl-carnosine, N-palmitoyl-His-His-His, N-
Palmitoyl-Gly-His-Lys, N-palmitoyl-Gly-His-His, N-palmitoyl-G
ly-Gly-His, N-oleoyl-His, N-
Oleoyl-His-His, N-Oleoyl-His
-Leu, N-oleoyl-Leu-His, N-oleoyl-β-alanyl-3-methyl-L-His, N-
Oleoyl-γ-aminobutyryl-L-His, N-oleoyl-carnosine, N-oleoyl-His-Hi
s-His, N-oleoyl-Gly-His-Ly
s, N-oleoyl-Gly-His-His, N-oleoyl-Gly-Gly-His, N-linoleyl-
His, N-linoleyl-carnosine, N-linolenyl-His, N-linolenyl-carnosine, N-stearoyl-His, N-stearoyl-His-His, N
-Stearoyl-His-Leu, N-stearoyl-
Leu-His, N-stearoyl-β-alanyl-3
-Methyl-L-His, N-stearoyl-γ-aminobutyryl-L-His, N-stearoyl-carnosine, N-stearoyl-His-His-His, N-
Stearoyl-Gly-His-Lys, N-stearoyl-Gly-His-His, N-stearoyl-G
ly-Gly-His, N-arachidoyl-His, N
-Arachidoyl-carnosine and the like.

As described above, the N-acylamino acid compound according to the present invention is obtained by reacting an N-hydroxysuccinimide ester represented by the general formula (2) with an amino acid or peptide represented by the general formula (3). Can be

This N-hydroxysuccinimide ester can be prepared by reacting an organic solvent such as ethyl acetate with 6 to 6 carbon atoms.
It is obtained by reacting 24 saturated or unsaturated fatty acids with N-hydroxysuccinimide in the presence of dichlorohexylcarbodiimide.

By reacting the N-hydroxysuccinimide ester with an amino acid or peptide in an organic solvent such as tetrahydrofuran at room temperature under alkaline conditions, an N-acylamino acid compound can be easily obtained in high yield. At this time, the amount of the amino acid or peptide is 0.1 to 5 mol, preferably 0.5 to 2 mol, per 1 mol of N-hydroxysuccinimide ester.

The alkaline condition is pH 7 to 12, preferably 8 to 11, and is adjusted using sodium carbonate, potassium carbonate, sodium hydrogen carbonate, potassium hydrogen carbonate and the like. The thus obtained N-acyl-His and N-acyl-His-containing peptides, especially N-acyl-His-
Carnosine (carnosine: β-alanyl-L-His,
Present at high concentrations in human brain and muscle. )) Is an amphiphilic substance and has the following functionality. For example, the rate of peroxidation of a higher fatty acid methyl ester in a non-aqueous hexane-isopropyl alcohol solution can be suppressed. Also, in a water system, when a liposome is made with egg yolk lecithin using a biological membrane as a model and an oxidation reaction is promoted with ferrous-ascorbic acid, the oxidation rate can be suppressed by incorporating the N-acyl compound into the liposome. . From these results, N-acyl-His and N-acyl-His-containing peptides (particularly, N-acyl-His-
(Carnosine) was confirmed to have, for example, a chelating power of iron ions in addition to the antioxidant power of suppressing the production of peroxide. When the emulsifying power was measured in an oil-water system using the same compound, it was found that the emulsifying power was higher than that of commercially available emulsifiers such as casein, sugar ester, and Triton X100 under the same conditions. In addition, since the compound has a large number of double bonds on the amino acid residue side, it can be understood that the compound naturally has ultraviolet absorbing power.

[0019]

EXAMPLES The present invention will be described below with reference to examples, but it is needless to say that the scope of the present invention is not limited by these examples.

Reference Example 1 In 45 ml of ethyl acetate, 3.45 g (30 mmol) of N-hydroxysuccinimide was dissolved, and then 3.45 g (30 mmol) of oleic acid was dissolved. To this solution was added 10 ml of ethyl acetate and 6.1 ml of dicyclohexylcarbodiimide.
A solution of 8 g (30 mmol) was added, and the mixture was stirred at room temperature with a magnetic stirrer for 1 hour.
The reaction was performed for 5 hours. From the reaction solution thus obtained, about 7 g of highly pure N-hydroxysuccinimide ester of oleic acid was obtained by repeatedly subjecting the target substance, N-hydroxysuccinimide ester of oleic acid, to a crystallization method using ethanol. Obtained. Other fatty acids can be synthesized and purified in the same manner as described above.

Example 1 226 mg (1 mmol) of carnosine and 84 mg (1 mmol) of sodium hydrogen carbonate are dissolved in 10 ml of water. A solution prepared by dissolving 380 mg (1 mmol) of N-hydroxysuccinimide ester of high-purity oleic acid obtained in Reference Example in 10 ml of tetrahydroxyfuran was added to this solution, and the mixture was stirred at room temperature with a magnetic stirrer. After reacting for 15 hours, the reaction solution is acidified with 6N hydrochloric acid, extracted with ethyl acetate, and repeatedly crystallized with hexane / ethyl acetate (4: 1, v: v) to obtain high-purity N-oleoyl. -About 200 mg of carnosine was obtained. IR, ¹ H-NM of N-oleoyl-carnosine
The results of R and ¹³ C-NMR are shown. In the infrared spectrum, the absorption peak derived from oleic acid was 2920 cm ⁻¹ , 28
It can be confirmed near 50 cm ^-1 (CH stretching), and the absorption peak derived from imidazole of carnosine 1650 to 1400 cm
^-1 (C = C, C = N expansion and contraction) was also confirmed. Also, 1580 cm ^-1 (NH bending angle) seen in the carnosine chart
Was noticeably reduced. ^{In the 1} H-NMR spectrum, 5.30 ppm of a chemical shift value of C ₉ and C ₁₀ cis-form protons derived from oleic acid were confirmed. An amide bond between oleic acid and carnosine is considered from the integral ratio. From -COOH of oleic acid, the chemical shift value of the proton at the α-position carbon shifted from 2.30 ppm to 2.00 ppm, and the chemical shift value of the proton at the β-position carbon also shifted from 1.60 ppm to 1.45 ppm. It was recognized that. ^{In the 13} C-NMR spectrum, the chemical shift value of carbon of —COOH of oleic acid is 180.40.
A shift from 170 ppm to 170.59 ppm was observed. From these, it was confirmed that oleic acid was amide-bonded to the β-alanine residue of carnosine. FIG. 1 shows the infrared absorption spectrum.

Example 2 In the same manner as in Example 1, high-purity N-oleoyl-Hi
About 150 mg were obtained, and the infrared spectrum was examined. The absorption peak derived from oleic acid is 2920 cm ⁻¹ , 2850 c
It can be confirmed near m ^-1 (CH stretching) and absorption peak derived from His 1650-1400 cm ^-1 (C = C, C = N stretching)
Was also confirmed. In addition, the absorption peak of amide 3400-3
Since 100 cm ^-1 (NH stretching) was confirmed, N-
The structure of oleoyl-His was revealed. FIG. 2 shows the infrared absorption spectrum.

Examples 3 to 7 In the same manner as in Example 1, N-caprinoylcarnosine (Example 3), N-oleylcarnosine methyl ester (Example 4), N-caprinoylhistidine (Example 5), N-oleoyl histidyl leucine (Example 6)
And N-oleoylglycylglycylhistidine (Example 7). These infrared absorption spectra are shown in FIG. 3 (Example 3), FIG. 4 (Example 4), FIG. 5 (Example 5), FIG. 6 (Example 6), and FIG. 7 (Example 7).

For other peptides, N-
Acyl-peptides can be easily synthesized and their structures can be analyzed.

Example 8 The antioxidant power of N-oleoylcarnosine, N-caprinoylcarnosine and N-oleoylcarnosine methyl ester obtained in Examples 1, 3 and 4 was evaluated by radical chain autoxidation of methyl linoleate. The evaluation was performed by suppressing the above. 100 mM methyl linoleate, 10 mM fat-soluble radical generator (2,2'-azobis (2,4-dimethylvaleronitrile)), N-oleoylcarnosine, N-caprinoylcarnosine or N-oleoyl-carnosine at various concentrations After incubating hexane / isopropyl alcohol (1: 1, v / v) containing methyl ester at 37 ° C., sampling over time, and measuring the production amount of methyl linoleate hydroperoxide by high performance liquid chromatography. As shown in FIG. 8, it was found that the compound had antioxidant activity.

Example 9 The antioxidant power of N-oleoyl histidine and N-caprinoyl histidine obtained in Examples 2 and 5 was measured by the same method as in Example 8, and as shown in FIG. Has antioxidant activity.

Example 10 The antioxidant activity of N-oleoylhistidylleucine and N-oleoylglycylglycylhistidine obtained in Examples 6 and 7 was measured by the same method as in Example 8. As shown in FIG. 10, it was found to have antioxidant power.

Example 11 The antioxidant activity of N-oleoyl-carnosine and N-oleoyl-histidine obtained in Examples 1 and 2 was evaluated by inhibiting the radical chain autoxidation of multilayer liposomes. 10 mM Tris-HCl buffer (pH
7.4 mM egg yolk phosphatidylcholine, 0.4.
Forming a multilamellar liposome with 4 mM egg yolk phosphatidylcholine hydroperoxide, 10 μM N-oleoyl-carnosine or N-oleoyl-histidine, adding 0.1 mM ferrous sulfate and 1 mM ascorbic acid;
After incubation at 37 ° C, sample over time,
Malondialdehyde generated by decomposition of lipid peroxide by the TBA reaction was colored, and the amount of the produced malondialdehyde was measured by high performance liquid chromatography. As shown in FIG. 11, it was clear that it also had antioxidant and chelating ability. became.

Example 12 The emulsifying power of N-oleoyl-carnosine and N-oleoyl-histidine obtained in Examples 1 and 2 was examined. 50 m of N-acyl compound or commercially available emulsifier
Dissolve 0.2% in M phosphate buffer (pH 7), add 1/2 volume of soybean oil to the solution, sonicate at 30 ° C, leave at 30 ° C for a predetermined time, and separate the lower layer. Turbidity is O. When the emulsifying power was evaluated by measuring at D (500 nm), excellent emulsifying power of N-oleoyl-carnosine and N-oleoyl-histidine was recognized. Table 1 shows the results. The emulsifying power in Table 1 is represented by a relative emulsifying activity when the emulsifying power of N-oleoyl-carnosine is set to 100.

[0030]

[Table 1]

[0031]

Industrial Applicability The N-acylamino acid compound and the method for producing the same according to the present invention are not only novel as described above, but also a novel antioxidant having antioxidant activity in polar and non-polar solvents. Yes, these compounds have very strong emulsifying power, and also have antibacterial, chelating, ultraviolet absorbing, moisturizing, etc., making them very useful as raw materials for cosmetics, foods, pharmaceuticals, etc. It is.

[Brief description of the drawings]

FIG. 1 shows the N-acyl peptide N- according to the present invention.
Infrared absorption spectrum diagram showing oleoyl carnosi,

FIG. 2 shows N-acylamino acid N- according to the present invention.
Infrared absorption spectrum diagram showing oleoyl histidine,

FIG. 3 is an infrared absorption spectrum of N-caprinoylcarnosine.

FIG. 4 is an infrared absorption spectrum of N-oleoylcarnosine methyl ester.

FIG. 5 is an infrared absorption spectrum of N-caprinoyl histidine,

FIG. 6 is an infrared absorption spectrum of N-oleoyl histidyl leucine,

FIG. 7 is an infrared absorption spectrum of N-oleoylglycylglycylhistidine;

FIG. 8 is a graph showing the antioxidant power of the N-acylamino acid compound according to the present invention,

FIG. 9 is a graph showing the antioxidant power of the N-acylamino acid compound according to the present invention,

FIG. 10 is a graph showing the antioxidant activity of the N-acylamino acid compound according to the present invention;

FIG. 11 is a graph showing the antioxidant power of the N-acylamino acid compound according to the present invention.

Continuation of the front page (51) Int.Cl. ⁶ Identification symbol FI C09K 15/22 C09K 15/22 // C09K 3/00 108 3/00 108 (56) References International Publication 90/14429 (WO, A1) Biochemistry , 26 (12), 3267-76 (1987) (58) Fields investigated (Int. Cl. ⁶ , DB name) C07D 233/64 CA (STN) REGISTRY (STN)

Claims (4)

(57) [Claims] 1. A compound represented by the general formula (1) R-CO- (NH-X-CO) _n -OR ¹ (1) (wherein (NH-X-CO) represents a carnosine residue, and R-CO Represents a saturated or unsaturated fatty acid residue having 6 to 24 carbon atoms, R ¹ represents a hydrogen atom, a sodium atom, a potassium atom or a methyl group, and n is an integer of 1 or 2.) N-acyl amino acid compounds. 2. A compound of the general formula (2) (Wherein, R-CO is a saturated or unsaturated fatty acid residue having 6 to 24 carbon atoms) by converting the N-hydroxysuccinimide ester represented by the general formula (3) H- (NH-X-CO ) _N -OR ¹ (3) (where, (NH-X-CO) represents an amino acid residue, and X in (NH-X-CO) differs depending on the type of amino acid used. Represents a saturated or unsaturated fatty acid residue having 6 to 24 atoms, R ¹ represents a hydrogen atom, a sodium atom, a potassium atom or a methyl group, and n is an integer of 1 to 3 and constitutes at least one histidine. (1) R-CO- (NH-X-CO) _n -OR ¹ (1) (where His, X, R, R ¹ and Manufacturing method of the same meanings as above.) Represented by N- acyl amino acid compounds. 3. The general formula (1) R-CO- (NH-X-CO) _n -OR ¹ (1) (wherein (NH-X-CO) represents an amino acid residue and (NH-X-CO) X in (X—CO) varies depending on the type of amino acid used, R—CO represents a saturated or unsaturated fatty acid residue having 6 to 24 carbon atoms, and R ¹ represents a hydrogen atom, a sodium atom, a potassium atom or a methyl group. And n is an integer of 1 to 3 and contains at least one histidine as a constituent amino acid.) An antioxidant containing an N-acyl amino acid compound represented by the formula (1) as an active ingredient: 4. General formula (1) R-CO- (NH-X-CO) _n -OR ¹ (1) (wherein (NH-X-CO) represents a carnosine residue, and R-CO Represents a saturated or unsaturated fatty acid residue having 6 to 24 carbon atoms, R ¹ represents a hydrogen atom, a sodium atom, a potassium atom or a methyl group, n is an integer of 1 or 2 and at least one histidine An emulsifier comprising an N-acyl amino acid compound represented by the formula (1) as an active ingredient: