JPS6035358B2 - Method for producing pyroglutamylglycylamino acid - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to a method for producing tribeptide, and more particularly pyroglutamyl glycylamino acid.

The present inventors have separately discovered that by simply heating glutamic acid (or pyroglutamic acid) and another amino acid different from glutamic acid, only a single type of pyroglutamyl peptide can be formed. In addition to the new findings, simply heating glutamic acid (or pyroglutamic acid), glycine, and other amino acids different from these at a specific molar ratio of the three results in a one-step reaction to form pyroglutamylglycylamino acid (tribeptide). We have found that this can be achieved efficiently. The present invention was completed based on this new knowledge. That is, the present invention provides a pyroglutamyl glycylamino acid, which is characterized in that glutamic acid (or pyroglutamic acid), glucine, and other amino acids different from these are heated in a molar ratio of 1 to 2:1:3 to 5. The present invention provides a method for manufacturing.

In the thermal condensation reaction of glutamic acid (or pyroglutamic acid) and glycine, pyroglutamylglycine (dibeptide), pyroglutamylglycylglycine (tribeptide), and pyroglutamylglycylglycylglycine (tetrabeptide) are produced, but glutamic acid ( In the thermal condensation reaction of pyroglutamic acid (or pyroglutamic acid) and other amino acids other than glycine, only pyroglutamyl amino acid (dipeptide) is produced, and in this case, tribeptide and tetrapeptide are not obtained.

By utilizing the different phenomena in these two cases, the present inventors used glutamic acid (or pyroglutamic acid), glycine, and other different amino acids in combination under specific molar ratio conditions, and by mere heating, They succeeded in obtaining pyroglutamylglycylamino acid (tribeptide) in a one-step reaction. The method of the present invention is characterized in that there is no need to use any additional components (substances) such as catalysts in addition to the raw materials glutamic acid (or pyroglutamic acid), glycine, and amino acids different from these.

However, when the reaction temperature is low, the addition of a small amount of water is effective as a reaction initiator. Although it is possible to use an inert solvent as the solvent, it is preferable to use no solvent. The above reaction is preferably carried out in an atmosphere of an inert gas such as nitrogen or carbon dioxide. The reaction temperature is usually 80 to 200 degrees Celsius, preferably 130 to 180 degrees Celsius. The water vapor generated during the reaction can be continuously removed, but this is not an essential condition in practice, but since the glutamic acid used as a raw material can easily be converted to pyroglutamic acid by heating, it is possible to remove either of the two. yields the same result. Examples of the above-mentioned "other amino acids" used in the method of the present invention include alanine and valine.

Isine, isoleucine, phenylalanine, tyrosine,
All common amino acids such as serine, threonine, lysine, proline, hydroxyproline, methionine, cysteine, 8-alanine, Q-aminobutyric acid, Q-aminoisobutyric acid, y-aminobutyric acid, norvaline, norleucine, and sarcosine can be mentioned. As in the method of the present invention, 3
Conventionally, there has been no known method for obtaining tripeptides in a one-step reaction by bonding seed amino acids with peptides in a predetermined order by simply heating. Moreover, such a method was completely unexpected. Moreover, not only is the operation simple, but the target tripeptide can be easily separated and purified by passing the product as an aqueous solution through an ion exchange resin, so it is an extremely excellent method from a practical standpoint.

The pyroglutamyl glycylamino acids that can be produced by the method of the present invention include all substances with various usefulness and intermediate raw materials for the synthesis of peptides used for various purposes. Its practical value is extremely high. The present invention will be explained below using Examples and Comparative Examples, but the present invention is not limited by the Examples.

Example 1 3.23 moles (0.025 moles) of pyroglutamic acid, 0.95 moles (0.013 moles) of glycine, and 3.35 moles (0.038 moles) of alanine were thoroughly ground in a mortar and mixed.
While flowing N2 gas in a glass reaction tube, the temperature was 3 at 175℃.
After reacting for 5 minutes, 0.6 moles (0.002 mol) of pyroglutamylglycylalanine was obtained.

In addition, pyroglutamylalanine 1.2 evenings (0.006
mole) and pyroglutamylglycine 1.0 mole (0.005 mole)
mol) was obtained and 1.6 pyroglutamic acid was recovered.
The conversion rate of pyroglutamic acid was 52%, the yield of pyroglutamylglycylalanine was 8%, and the selectivity was 15.4%.

Separation and fractionation of the product can be carried out in aqueous solution using Shode
The analysis was performed by high performance liquid chromatography using a x Ion-Exchanger HC-8-10 column. Comparative example
1 Pyroglutamic acid, glycine and alanine 1:1:1
When they were mixed in the same molar ratio and subjected to the same reaction as above, the main products were pyroglutamylglycine and pyroglutamylglycylglycine, and in addition, pyroglutamylglycylglycylglycine and pyroglutamylalanyl. Glycine was also created, and the yield and selectivity of the target pyroglutamylglycylalanine were extremely low.

Example 2 Crystals of 1.03 moles (0.008 moles) of pyroglutamic acid, 0.63 moles (0.008 moles) of glycine, and 2.23 moles (0.025 moles) of 8-alanine were collected by precise weighing and placed in a mortar. The mixture was thoroughly ground and mixed, sealed in a glass tube sealed with carbon dioxide gas, and reacted between the four powders at 16,000 yen.

After the reaction was completed, the product was dissolved in 0.1% H3P04 aqueous solution and separated by high performance liquid chromatography using a ShodexIonpac C-811 column to obtain pyroglutamylglycyl-8-alanine 0.26 (0.001
mole) was obtained. In addition, pyroglutamyl-8-alanine 0
．． 4 (0.002 mol), pyroglutamylglycine 0
．． 0.37 (0.002 mol) of pyroglutamic acid was obtained.
．． 38 evenings were collected.

The conversion rate of pyroglutamic acid was 63%, the yield of pyroglutamylglycyl-8-alanine was 13%, and the selectivity was 21%. Comparative Example 2 Pyroglutamic acid, alanine, and 8-alanine at 1:1:
When they were mixed at a molar ratio of 1 and reacted under exactly the same conditions as above, only pyroglutamylalanine and pyroglutamyl-8-alanine were obtained, and no formation of tribeptide was observed.

Example 3 Pyroglutamic acid 2.06 moles (0.016 moles), glycine 0.63 moles (0.008 moles), tyrosine 4.53 moles
The crystals (0.025 mol) were collected on a weighing scale, mixed, sealed in a glass tube purged with nitrogen, and reacted at 18,000 ml for 20 minutes.

As a result of analysis in the same manner as in Example 2, 0.79 (0.002 mol) of pyroglutamylglycyltyrosine was obtained. Other products include pyroglutamyl tyrosine 0.58
(0.003 mol), pyroglutamylglycine 0.3
(0.002 mol). Conversion rate of pyroglutamic acid: 43.8% Yield of pyroglutamylglycyltyrosine: 25% selectivity
It was 28.6%.

Claims (1)

[Claims] 1. A method for producing pyroglutamyl glycylamino acid, which comprises heating glutamic acid or pyroglutamic acid, glycine, and another amino acid different therefrom at a molar ratio of 1 to 2:1:3 to 5.