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Structure–activity relationship study on new hemorphin-4 analogues containing steric restricted amino acids moiety for evaluation of their anticonvulsant activity

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Abstract

In the present study, several new analogues of hemorphin-4, modified with unnatural conformationally restricted amino acids followed the structure Aaa–Tyr–Xxx–Trp–Thr–NH2, where Aaa is the low-molecular-weight lipophilic adamantyl building block, and Xxx is Ac5c (1-aminocyclopentanecarboxylic acid) or Ac6c (1-aminocyclohexane carboxylic acid) was synthesized, characterized and investigated for anticonvulsant activity in three seizure tests, the maximal electroshock test (MES), 6-Hz psychomotor seizure test and timed intravenous pentylenetetrazole infusion (ivPTZ) test. The acute neurological toxicity was determined using the rota-rod test. The new synthetic neuropeptide analogues were prepared by solid-phase peptide synthesis—Fmoc chemistry and were evaluated in three doses of 1, 3 and 5 µg, respectively, administered intracerebroventricularly in male ICR mice. The physicochemical properties of these peptide analogues were evaluated as pKa and pI values were calculated using potentiometry. The IR spectrum of the compounds was recorded and the characteristic lines of both adamantane moiety and the peptide backbone were registered in the wavelength range from 4000 to 400 cm−1. The hexapeptide Ang IV was used as a positive control. From the six synthesized peptide analogues, the P4-5 was the most active at doses of 1 and 3 µg in the three seizure tests. The order of potency of other peptides was as follows: P4 > P4-3 = P4-4 > P4-2 > Ang IV in MES, P4-4 ≥ P4-1 > P4-3 > P4-2 > P4 > Ang IV in 6-Hz test and P4-4 = P4-3 > P4-2 = P4 > Ang IV in ivPTZ test. None of the peptides displayed neurotoxicity in the rota-rod test. Docking study results suggest that direct H-bonding and ionic interactions between our synthetic ligands and residues, responsible for coordination of Zn2+ along with hydrophobic interactions between our ligands and IRAP active site are the most important for the ligand binding. The results propose that incorporation of adamantane and cycloalkane building blocks in the peptide chain of the hemorphin-4 scaffold is important for the potential high biological activity.

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Acknowledgements

This work was financially supported by the Bulgarian National Scientific Fund project КП-06-OПP 03/3 of the Ministry of Education and Science, Bulgaria.

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Authors and Affiliations

  1. Department of Organic Chemistry, University of Chemical Technology and Metallurgy, 1756, Sofia, Bulgaria

    Petar Todorov & Petia Peneva

  2. Institute of Neurobiology, Bulgarian Academy of Sciences, 1113, Sofia, Bulgaria

    Jana Tchekalarova

  3. Department of Analytical Chemistry, University of Chemical Technology and Metallurgy, 1756, Sofia, Bulgaria

    Stela Georgieva

  4. Institute of Organic Chemistry with Centre of Phytochemistry, Bulgarian Academy of Sciences, 1113, Sofia, Bulgaria

    Miroslav Rangelov

  5. Institute of Biodiversity and Ecosystem Research, Bulgarian Academy of Sciences, 1113, Sofia, Bulgaria

    Nadezhda Todorova

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  1. Petar Todorov
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All procedures were performed in agreement with the European Communities Council Directive 2010/63/EU. The experimental design was approved by the Institutional Ethics Committee. There are no human participants.

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Todorov, P., Peneva, P., Tchekalarova, J. et al. Structure–activity relationship study on new hemorphin-4 analogues containing steric restricted amino acids moiety for evaluation of their anticonvulsant activity. Amino Acids 52, 1375–1390 (2020). https://doi.org/10.1007/s00726-020-02898-1

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