JPH10150923A - Feed additive composition for ruminant - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a composition which is stable in the paunches (lumens) of ruminants and is elutable in the abomasum and subsequent digestive organs by chemically modifying amino acid. SOLUTION: This composition contains at least one kind of the N-acyl- aspartyl dipeptide expressed by the formula (R is a lower fatty acid acyl group, such as formyl group, acetyl group or propionyl group, benzyloxycarbonyl group, ternary butyloxycarbonyl group, benzoyl group, paratoluenesulfonyl group; Asp is an aspartic acid residue; X is a methinion residue, glutamic residue, alanine residue, valine residue, isoleucine residue or phenyl alanine residue) and its salt. The compd. of the formula is obtd. by bringing N-acyl-aspartic anhydride and neutral amino acid into reaction in acetic acid.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

TECHNICAL FIELD The present invention relates to a ruminant amino acid composition. More particularly, it relates to an amino acid dipeptide derivative which is stable in the rumen of rumen, and an amino acid composition for ruminants containing the same.

[0002]

2. Description of the Related Art In ruminants such as cattle and sheep, when amino acids are directly administered orally, most of them are decomposed by microorganisms in the rumen and are not effectively used. Therefore, the amino acid is first
A ruminant bypass preparation for ruminants that protects against decomposition of microorganisms in the stomach and is digested and absorbed by the digestive organs of the abomasum and beyond is important in the fields of ruminant feed, nutrients, veterinary drugs, etc. .

[0003] As a method for imparting rumen resistance to amino acids, for example, taking a ruminant feed additive as an example,
A method of dispersing and granulating amino acids in a matrix composed of a hydrophobic substance such as an oil or fat or a protective substance such as a basic polymer compound (JP-A-60-168351, JP-B-59-107)
80) or a method of coating an amino acid or a nucleus containing an amino acid with a hydrophobic substance such as an oil or fat or an acid-sensitive substance such as a basic polymer compound (JP-A-63-317053,
JP-A-54-46823 has been proposed for some time.

However, in the method of dispersing an amino acid in a protective substance, since the amino acid is present near the particle surface, it is hard to say that the protective property is sufficient in view of the long-term residence in the lumen. In addition, the method of coating an amino acid or a nucleus containing an amino acid with an acid-sensitive polymer compound or a hydrophobic protective substance is versatile, for example, when a physical pressure is applied to the coating layer, a crack is easily generated and the protective property is reduced. Hard to say. Therefore, if the rumen resistance can be imparted by chemically modifying amino acids, the above-mentioned drawbacks can be solved and it can be said that this is a very versatile protection method, but at present, a good chemical modification method has not yet been found. Not. On the other hand, polypeptides are reported to be stable in the lumen (J. Nutr. 91: 314), but problems remain in terms of cost.

The compound N-formyl-L-aspartyl-L-phenylalanine is disclosed in Japanese Patent Publication No. 55-26133 as an intermediate for producing L-aspartyl-L-phenylalanine methyl ester as a sweetener. Here, there is no description regarding the physical properties and utilization of the present invention.

A higher fatty acid acylated product of a collagen hydrolyzate containing aspartic acid is disclosed in Japanese Patent Application No. 65-197197 as a part of a general formula as a detergent having a surfactant activity. This description does not disclose the compound of the invention of the present application alone, isolate or take up the compound alone, and further, does not disclose the specific amino acid composition of the peptide portion as well as the characteristics relating to the physical properties.

The compound of the present invention can be easily synthesized by treating aspartic acid with at least equimolar amount of formic acid and at least twice molar amount of acetic anhydride as described in JP-B-55-26133. A method for isolating N-formyl-aspartic anhydride from the dehydration reaction solution is also known.
That is, as described in JP-B-55-26133, the reaction solution is concentrated and excess formic acid is distilled off to obtain N-formyl-aspartic anhydride, or N-formyl-aspartic anhydride is obtained from the reaction solution as shown in JP-T-5-500364. There are methods for crystallizing and separating formyl-aspartic anhydride, but these methods have the drawback that they require concentration and separation steps.

[0008]

SUMMARY OF THE INVENTION An object of the present invention is to create a derivative of a stable amino acid in the rumen by chemically modifying the amino acid, and to provide a ruminant animal feed additive.

[0009]

Means for Solving the Problems The present inventors have conducted intensive studies on amino acid derivatives in order to achieve the above object, and as a result, the following general formula (1)

Embedded image (Where R is a lower fatty acid acyl group such as a formyl group, an acetyl group or a propionyl group, a benzyloxycarbonyl group, a tertiary butyloxycarbonyl group, a benzoyl group, a paratoluenesulfonyl group, Asp is an aspartic acid residue, and X is N-acyl-aspartyl dipeptide represented by methionine residue, glutamine residue, alanine residue, valine residue, isoleucine residue or phenylalanine residue) in rumen (ruminal) of ruminant animals. They found that they were stable, and completed the present invention.
That is, the present invention is a ruminant feed additive composition containing at least one compound represented by the general formula (1).

[0010]

BEST MODE FOR CARRYING OUT THE INVENTION The following general formula (3) which is a compound used in the present invention:

Embedded image (However, R is a lower fatty acid acyl group such as a formyl group, an acetyl group or a propionyl group, a benzyloxycarbonyl group, a tert-butyloxycarbonyl group, a benzoyl group, a paratoluenesulfonyl group, Asp is an aspartic acid residue, X ′ Is a neutral amino acid excluding phenylalanine.) The N-acyl-aspartyl dipeptide and salts thereof are novel compounds.

In the above general formula (3), the acyl group R is a formyl group, a lower fatty acid acyl group such as an acetyl group or a propionyl group, a benzyloxycarbonyl group, a tert-butyloxycarbonyl group, a benzoyl group, a paratoluenesulfonyl group. Among them, a formyl group and an acetyl group are particularly preferably used.

The compounds used in the present invention are N-lower fatty acyl-aspartyl dipeptides represented by the above general formulas (1) and (3) and salts thereof. On the other hand, N-higher fatty acid acyl-aspartyl dipeptides and salts thereof which are not used in the present invention other than lower fatty acid acyl groups are used because the higher fatty acid acyl groups in the compound are hydrophobic and the dipeptide portion is hydrophilic. The compound has a surfactant activity. Therefore, when a ruminant ingests the compound, it stays in the rumen and acts on the plutozoa inhabiting the rumen, adversely affecting rumen fermentation. Further, even after passing through the lumen, there is a concern that there is a concern that the ruminant animal itself may be absorbed into the body by the small intestine and exert an unfavorable effect on health.

The neutral amino acids used in the ruminant feed additive composition of the present invention include methionine, glycine, alanine, aminobutyric acid, leucine, isoleucine, and the like.
Serine, threonine, asparagine, glutamine, tryptophan, cystine, proline, tyrosine and phenylalanine residues and the like, for example, if used for feed, such as methionine, may be used amino acids according to the purpose, As described below, there is no particular limitation in terms of manufacturing. The aspartic acid and the neutral amino acid used in the present invention may be any of L-form, D-form and DL-form.

The compound used in the ruminant feed additive of the present invention is synthesized by reacting N-acyl-aspartic anhydride with a neutral amino acid in acetic acid.
The molar ratio of N-acyl-aspartic anhydride to neutral amino acid is not particularly limited, but from the viewpoint of cost, it is usually about 1 mol of neutral amino acid to 1 mol of N-acyl-aspartic anhydride. used. Acetic acid, which is a reaction solvent, may contain an organic solvent inert to the reaction raw materials. The temperature is not particularly limited, but if the temperature is too high, the problem of racemization appears. Therefore, the temperature may be in the range of 0 to 100 ° C, but usually in the range of 10 to 40 ° C.

The above-mentioned N-acyl-aspartic anhydride can be easily synthesized by treating N-acyl-aspartic acid with acetic anhydride in an organic solvent at 0 to 70 ° C. The amount of acetic anhydride may be at least equimolar to N-acyl-aspartic acid.

The compound represented by the general formula (2), which is one of the compounds used in the present invention:

Embedded image (However, F is a formyl group, Asp is an aspartic acid residue, X is a methionine residue, a glutamine residue, an alanine residue, a valine residue, an isoleucine residue or a phenylalanine residue.) The method for producing an anhydride of formyl-aspartyl dipeptide can be synthesized in a one-step reaction without using aspartic acid as a starting material and concentrating or isolating in the reaction step. The following method is effective as an industrial production method. That is, when aspartic acid is converted into an anhydride using 1.1-1.3 moles of formic acid and 2-2.1 times mole of acetic anhydride, N-formyl-aspartic anhydride can be isolated by concentration or separation. A neutral amino acid can be directly added to the obtained reaction solution to carry out a condensation reaction. All reactions are possible in one pot without the need for concentration or isolation. The temperature of the anhydration reaction is in the range of 0 to 70 ° C, but preferably 40 to 60 ° C and the reaction time of 3 to 7 hours in consideration of racemization.

As a result of an in vitro test using bovine rumen solution, the compound of the present invention was stable to rumen solution as shown in Examples 13 and 14.

Hereinafter, the present invention will be described in detail with reference to examples.

Example 1 6.0 g (0.13 mol) of formic acid and 22.5 g (0.22 mol) of acetic anhydride were mixed.
To this, 13.3 g (0.10 mol) of L-aspartic acid was added.
And 0.2 g (1 mmol) of calcium acetate monohydrate were added, and the mixture was heated and stirred at 55 ° C. for 5 hours. After returning the reaction solution to room temperature, 15 ml of toluene was added, and the mixture was stirred at room temperature for 2 hours. The precipitated crystals were collected by filtration, dried under reduced pressure, and dried with N-formyl-L-aspartic anhydride (12.2 g, 0.02 g).
85 mol). The obtained N-formyl-L-aspartic anhydride was dispersed in acetic acid (40 ml), L-methionine (12.7 g, 0.085 mol) was added, and the mixture was stirred at room temperature overnight. The reaction solution slurry was filtered, and the crystals were washed with 20 ml of ethyl acetate and dried under reduced pressure.
16.9 g of aspartyl-L-methionine was obtained. L-
The yield was 68.0% based on methionine.

Example 2 5.1 g (0.11 mol) of formic acid and 22.5 g (0.22 mol) of acetic anhydride were mixed.
To this, 13.3 g (0.10 mol) of L-aspartic acid was added.
And 0.2 g (1 mmol) of calcium acetate monohydrate were added, and the mixture was heated and stirred at 55 ° C. for 5 hours. Acetic acid 15m was added to the obtained N-formyl-L-aspartic anhydride slurry.
After adding l, 13.4 g of DL-methionine was added at room temperature.
(0.09 mol) and stirred overnight. The reaction solution was filtered under reduced pressure, and the crystals were washed with 30 ml of ethyl acetate. The obtained crystals were dried under reduced pressure to obtain 17.6 g of N-formyl-L-aspartyl-DL-methionine. The yield was 66.7% based on DL-methionine.

EXAMPLE 3 6.0 g (0.13 mol) of formic acid and 22.5 g (0.22 mol) of acetic anhydride were mixed.
To this, 13.3 g (0.10 mol) of L-aspartic acid was added.
And 0.2 g (1 mmol) of calcium acetate monohydrate were added, and the mixture was heated and stirred at 55 ° C. for 5 hours. After returning the reaction solution to room temperature, 15 ml of toluene was added, and the mixture was stirred at room temperature for 2 hours. The precipitated crystals were collected by filtration, dried under reduced pressure, and dried with N-formyl-L-aspartic anhydride (12.2 g, 0.02 g).
85 mol). 2.38 g (0.02 mol) of N-formyl-L-aspartic anhydride was dispersed in 10 ml of acetic acid, and 2.34 g (0.02 mol) of L-valine was added thereto.
Was added and stirred at room temperature overnight. The reaction solution was concentrated under reduced pressure, and the residue was crystallized by adding 50 ml of ethyl acetate. After filtering the crystals, the crystals were dried under reduced pressure to obtain 4.56 g of N-formyl-L-aspartyl-L-valine. The yield was 85.7% based on L-valine.

Example 4 2.86 g (0.02 mol) of N-formyl-L-aspartic anhydride was added to 10 m of acetic acid.
1 and 2.38 g of L-threonine (0.
02 mol) and stirred at room temperature overnight. The reaction solution was concentrated under reduced pressure, and the residue was crystallized by adding 50 ml of ethyl acetate. After filtering the crystals, the crystals were dried under reduced pressure to obtain 4.4 g of N-formyl-L-aspartyl-L-threonine. L
The yield was 83.9% based on -threonine.

Example 5 13.4 g (0.05 mol) of N-benzyloxycarbonyl-L-aspartic acid was dispersed in 30 ml of toluene, and 0.1 g (0.5 mmol) of calcium acetate monohydrate was added. added. Under stirring, 5.4 g (0.053 mol) of acetic anhydride was added, and the mixture was heated and stirred at 50 ° C. for 3 hours. After returning to room temperature and stirring for 2 hours, the obtained slurry was filtered. After drying under reduced pressure, N-benzyloxycarbonyl-L-aspartic anhydride 10.2 g
(0.04 mol). 2.0 g (0.008 mol) of N-benzyloxycarbonyl-L-aspartic anhydride was dispersed in 10 ml of acetic acid to give L-methionine 1.2 g.
g (0.008 mol) was added and the mixture was stirred at room temperature overnight. The reaction solution was concentrated under reduced pressure, and 50 ml of hexane was added to the residue for crystallization. After filtration, the filtrate was dried under reduced pressure and N-benzyloxycarbonyl-L-aspartyl-L-methionine 2.7
g was obtained. The yield was 84.7% based on L-methionine.

Example 6 4.7 g (0.02 mol) of N-benzoyl-L-aspartic acid was dispersed in 10 m of acetic acid, and 0.04 g (0.2 mmol) of calcium acetate monohydrate was added. Acetic anhydride 2.1 g (0.02
1 mol), and the mixture was heated and stirred at 50 ° C. for 4 hours. After returning to room temperature, 2.7 g (0.018 mol) of L-methionine was added, and the mixture was stirred at room temperature overnight. The reaction solution was concentrated under reduced pressure, and 50 ml of hexane was added to the residue for crystallization. After filtration, the filtrate was dried under reduced pressure to obtain 5.2 g of N-benzoyl-L-aspartyl-L-methionine. The yield was 78.4% based on L-methionine.

Example 7 5.1 g (0.11 mol) of formic acid and 22.5 g (0.22 mol) of acetic anhydride were mixed.
To this, 13.3 g (0.10 mol) of L-aspartic acid was added.
And 0.2 g (1 mmol) of calcium acetate monohydrate were added, and the mixture was heated and stirred at 55 ° C. for 5 hours. Acetic acid 15m was added to the obtained N-formyl-L-aspartic anhydride slurry.
After adding l, 11.8 g of L-isoleucine was added at room temperature.
(0.09 mol) and stirred overnight. The reaction solution was concentrated under reduced pressure, and the residue was crystallized by adding 50 ml of ethyl acetate. After filtration under reduced pressure, the resulting crystals were dried under reduced pressure, and N-formyl-L-aspartyl-L-isoleucine was used.
9 g were obtained. The yield was 81.6% based on L-isoleucine.

Example 8 5.1 g (0.11 mol) of formic acid and 22.5 g (0.22 mol) of acetic anhydride were mixed.
To this, 13.3 g (0.10 mol) of L-aspartic acid was added.
And 0.2 g (1 mmol) of calcium acetate monohydrate were added, and the mixture was heated and stirred at 55 ° C. for 5 hours. Acetic acid 15m was added to the obtained N-formyl-L-aspartic anhydride slurry.
After adding l, 13.2 g of L-glutamine was added at room temperature.
(0.09 mol) and stirred overnight. The reaction solution was filtered under reduced pressure, and the crystals were washed with 30 ml of ethyl acetate. The obtained crystals were dried under reduced pressure to obtain 17.1 g of N-formyl-L-aspartyl-L-glutamine. The yield was 69.8% based on L-glutamine.

Example 9 5.1 g (0.11 mol) of formic acid and 22.5 g (0.22 mol) of acetic anhydride were mixed.
To this, 13.3 g (0.10 mol) of L-aspartic acid was added.
And 0.2 g (1 mmol) of calcium acetate monohydrate were added, and the mixture was heated and stirred at 55 ° C. for 5 hours. After adding 15 ml of acetic acid to the obtained N-formyl-L-aspartic acid slurry, 8.1 g (0.09 mol) of L-alanine was added at room temperature and stirred overnight. The reaction solution was filtered under reduced pressure, and the crystals were washed with 30 ml of ethyl acetate. The obtained crystals were dried under reduced pressure to obtain 13.3 g of N-formyl-L-aspartyl-L-alanine. The yield based on L-alanine is 63.6.
%Met.

Example 10 3.5 g (0.02 mol) of N-acetyl-L-aspartic acid was dispersed in 10 ml of acetic acid, and 0.04 g (0.2 mmol) of calcium acetate monohydrate was added. Under stirring, 2.1 g of acetic anhydride (0.0 g
21 mol), and the mixture was heated and stirred at 50 ° C. for 4 hours. After returning to room temperature, 2.7 g of DL-methionine (0.018
Mol) and stirred at room temperature overnight. The reaction solution was filtered under reduced pressure, and the crystals were washed with 10 ml of ethyl acetate. The obtained crystals were dried under reduced pressure to obtain 4.6 g of N-acetyl-L-aspartyl-DL-methionine. 83.4% yield based on DL-methionine

(Example 11) 2.86 g (0.02 mol) of N-formyl-L-aspartic anhydride was added to acetic acid 10
and L-phenylalanine 3.30.
g (0.02 mol) was added and the mixture was stirred at room temperature overnight. The slurry reaction solution was filtered under reduced pressure, and the crystals were collected in 10 ml of ethyl acetate.
After washing with, the obtained crystals were dried under reduced pressure, and N-formyl-L-aspartyl-L-phenylalanine 4.3 was obtained.
5 g were obtained. 70. Yield based on L-phenylalanine.
5%.

Example 12 3.8 g (0.02 mol) of N-propionyl-L-aspartic acid was added to 10 ml of acetic acid.
And 0.04 g of calcium acetate monohydrate (0.
2 mmol) was added. Acetic anhydride 2.1 g with stirring
(0.021 mol), and the mixture was heated and stirred at 50 ° C. for 4 hours. After returning to room temperature, 2.7 g of DL-methionine
(0.018 mol), and the mixture was stirred at room temperature overnight. The reaction solution was filtered under reduced pressure, and the crystals were washed with 10 ml of ethyl acetate. The obtained crystals were dried under reduced pressure to obtain 4.7 g of N-propionyl-L-aspartyl-DL-methionine. The yield was 81.5% based on DL-methionine.

(Example 13) 50 ml of bovine rumen solution was placed in a 100 ml Erlenmeyer flask, and 50 mg of N-formyl-L-aspartyl-DL-methionine was added thereto.
Shake at 39 ° C for 24 hours. HPLC of N-formyl-L-aspartyl-DL-methionine in the shaking solution
As a result, the residual ratio after 24 hours was 92.3%.

Example 14 In the same manner as in Example 12, the remaining ratio of the other N-acyl-aspartyl dipeptide in the rumen solution after 24 hours was measured to examine the stability. The results are shown in Table 1.

[0033]

[Table 1]

[0034]

EFFECT OF THE INVENTION By chemically modifying an amino acid, a derivative of an amino acid which is an N-acyl-asparatyl dipeptide which is stable in the rumen and can be eluted by the lower abdominal lower digestive organ is created, and is useful for ruminants. Feed additives could be provided.

Claims (6)

[Claims] 1. The following general formula (1), which is stable in the rumen of ruminants and can be eluted in the lower digestive organs of the rumen and lower stomach. (Where R is a lower fatty acid acyl group such as a formyl group, an acetyl group or a propionyl group, a benzyloxycarbonyl group, a tertiary butyloxycarbonyl group, a benzoyl group, a paratoluenesulfonyl group, Asp is an aspartic acid residue, and X is A ruminant feed containing at least one N-acyl-aspartyl dipeptide represented by methionine residue, glutamine residue, alanine residue, valine residue, isoleucine residue or phenylalanine residue and a salt thereof. Additive composition. 2. N-formyl aspartic anhydride obtained by treating aspartic acid with 1.1 to 1.3 times mol of formic acid and 2.0 to 2.1 times mol of acetic anhydride with respect to aspartic acid. Wherein a neutral amino acid is added to a reaction solution containing (However, F is a formyl group, Asp is an aspartic acid residue, X is a methionine residue, a glutamine residue, an alanine residue, a valine residue, an isoleucine residue or a phenylalanine residue.) A method for producing formyl-aspartyl dipeptide. 3. The following general formula (3): (However, R is a lower fatty acid acyl group such as a formyl group, an acetyl group or a propionyl group, a benzyloxycarbonyl group, a tert-butyloxycarbonyl group, a benzoyl group, a paratoluenesulfonyl group, Asp is an aspartic acid residue, X ′ Is a neutral amino acid excluding phenylalanine.) N-acyl-aspartyl dipeptide and salts thereof. 4. The N-acyl-aspartyl dipeptide and a salt thereof according to claim 3, wherein R is a formyl group and X is a methionine residue. 5. The N-acyl-aspartyl dipeptide according to claim 3, wherein R is an acetyl group and X is a methionine residue. 6. The N-acyl-acetylase according to claim 3, wherein X is each residue of methionine, glycine, alanine, aminobutyric acid, leucine, isoleucine, serine, threonine, asparagine, glutamine, tryptophan, cystine, proline and tyrosine. Aspartyl dipeptide and salts thereof.