Synthesis of the tripeptide H–ProAla–Glu–OH and an investigation of its toxicological properties

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DOI: https://doi.org/10.29296/25419218-2020-03-02
Issue: 
3
Year: 
2020

N.D. Bunyatyan(1, 4), G.M. Bobizoda(2), O.A. Popova(1), M.M. Sapovsky4, M. Samiev(2), A.B. Prokofyev(1, 4), I.P. Remezova(3), V.A. Evteev(1) 1-Research Center for Examination of Medical Products, Ministry of Health of the Russian Federation, 8, Petrovsky Boulevard, Moscow 127051, Russian Federation; 2-S. Aini Tajik State Pedagogical University, 121, Rudaki St., Dushanbe 734003, Republic of Tajikistan; 3-Pyatigorsk Medical and Pharmaceutical Institute, Branch, Volgograd State Medical University, 11, Kalinin Prospect, Pyatigorsk 357532, Russian Federation; 4-I.M. Sechenov First Moscow State Medical University (Sechenov University), 8, build. 2, Trubetskaya St., Moscow 119991, Russian Federation

Introduction. The biological activity of peptides depends on the amino acid composition; therefore, low-molecular-weight peptides have recently occupied the most prominent place among a large number of drugs. This is attributable to the fact that amino acids are not foreign to a living organism and that they do not cause adverse reactions. The important conditions for the synthesis of peptides are the choice of side blocking groups that are easily removable, as well as that of the amino acid composition. It has been suggested that an increased number of functional groups in the structure of a dipeptide will lead to the enhanced immunobiological activity of a tripeptide. The amino acid alanine has been chosen as a bridge to extend the chain between proline and glutamic acid. Objective: to develop the most optimal scheme for the synthesis of a tripeptide with the amino acid sequence H–Pro–Ala–Glu–OH and to investigate the properties of the synthesized compound. Material and methods. A method of mixed anhydrides and activated esters was used to prepare a tripeptide. The synthesis involved three steps: the formation of the protected dipeptide Boc–Ala–Glu-(γ-Bzl)–OH, its preparation for the synthesis of a tripeptide, and per se the synthesis of the tripeptide. UV and IR spectroscopies and HPLC were employed to identify the synthesized tripeptide, the acute and chronic toxicities of which were determined. Results. The tripeptide was synthesized through peptide elongation from the C-terminus. The amino groups of the amino acids proline and alanine were protected by a tert-butyloxycarbonyl group that is better suited for the above amino acids; the γ-carbonyl group of glutamic acid was protected by a benzyl group that is easily removed by hydrogenation. The synthesized tripeptide with the amino acid sequence H–Pro–Ala–Glu–OH at a concentration of 1∙10-3 mol/L was found to be a low toxic compound. Conclusion. A new low toxic tripeptide with the amino acid sequence H–Pro–Ala–Glu–OH was synthesized.
Keywords: 
immunoactivity
peptide synthesis
amino acids
peptide bioregulators
proline
alanine
acute and chronic toxicity
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