{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T01:10:31Z","timestamp":1760145031533,"version":"build-2065373602"},"reference-count":43,"publisher":"MDPI AG","issue":"6","license":[{"start":{"date-parts":[[2024,6,12]],"date-time":"2024-06-12T00:00:00Z","timestamp":1718150400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Bulgarian National Scientific Fund project","award":["\u041a\u041f-06-\u0414\u041e-\u041e2\/3"],"award-info":[{"award-number":["\u041a\u041f-06-\u0414\u041e-\u041e2\/3"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Pharmaceuticals"],"abstract":"<jats:p>The discovery of new peptides and their derivatives is an outcome of ongoing efforts to identify a peptide with significant biological activity for effective usage as a possible therapeutic agent. Spinorphin peptides have been documented to exhibit numerous applications and features. In this study, biologically active peptide derivatives based on novel peptide analogues of spinorphin conjugated with 5,5\u2032-dimethyl (Dm) and 5,5\u2032-diphenyl (Ph) hydantoin derivatives have been successfully synthesized and characterized. Scanning electron microscopy (SEM) and spectral methods such as UV-Vis, FT-IR (Fourier Transform Infrared Spectroscopy), CD (Circular Dichroism), and fluorimetry were used to characterize the microstructure of the resulting compounds. The results revealed changes in peptide morphology as a result of the restructuring of the aminoacidic sequences and aromatic bonds, which is related to the formation of intermolecular hydrogen bonds between tyrosyl groups and the hydantoin moiety. Electrochemical and fluorescence approaches were used to determine some physicochemical parameters related to the biological behavior of the compounds. The biological properties of the spinorphin derivatives were evaluated in vivo for anticonvulsant activity against the psychomotor seizures at different doses of the studied peptides. Both spinorphin analog peptides with Ph and Dm groups showed activity against all three phases of the seizure in the intravenous Pentylenetetrazole Seizure (ivPTZ) test. This suggests that hydantoin residues do not play a crucial role in the structure of spinorphin compounds and in determining the potency to raise the seizure threshold. On the other hand, analogs with a phenytoin residue are active against the drug-resistant epilepsy test (6-Hz test). In addition, bioactivity analyses revealed that the new peptide analogues have the potential to be used as antimicrobial and antioxidant compounds. These findings suggest promising avenues for further research that may lead to the development of alternative medicines or applications in various fields beyond epilepsy treatment.<\/jats:p>","DOI":"10.3390\/ph17060770","type":"journal-article","created":{"date-parts":[[2024,6,12]],"date-time":"2024-06-12T06:47:30Z","timestamp":1718174850000},"page":"770","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Chemical Behavior and Bioactive Properties of Spinorphin Conjugated to 5,5\u2032-Dimethyl- and 5,5\u2032-Diphenylhydantoin Analogs"],"prefix":"10.3390","volume":"17","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-8519-6214","authenticated-orcid":false,"given":"Stela","family":"Georgieva","sequence":"first","affiliation":[{"name":"Department of Analytical Chemistry, University of Chemical Technology and Metallurgy, 1756 Sofia, Bulgaria"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8050-0149","authenticated-orcid":false,"given":"Petar","family":"Todorov","sequence":"additional","affiliation":[{"name":"Department of Organic Chemistry, University of Chemical Technology and Metallurgy, 1756 Sofia, Bulgaria"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1640-5099","authenticated-orcid":false,"given":"Jana","family":"Tchekalarova","sequence":"additional","affiliation":[{"name":"Institute of Neurobiology, Bulgarian Academy of Sciences, 1113 Sofia, Bulgaria"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0307-5396","authenticated-orcid":false,"given":"Subaer","family":"Subaer","sequence":"additional","affiliation":[{"name":"Material Physics Laboratory, Physics Department, Universitas Negeri Makassar (UNM), Makassar 90223, Indonesia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Petia","family":"Peneva","sequence":"additional","affiliation":[{"name":"Department of Organic Chemistry, University of Chemical Technology and Metallurgy, 1756 Sofia, Bulgaria"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Kalin","family":"Chakarov","sequence":"additional","affiliation":[{"name":"Department of Analytical Chemistry, University of Chemical Technology and Metallurgy, 1756 Sofia, Bulgaria"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Hartati","family":"Hartati","sequence":"additional","affiliation":[{"name":"Material Physics Laboratory, Physics Department, Universitas Negeri Makassar (UNM), Makassar 90223, Indonesia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Sitti","family":"Faika","sequence":"additional","affiliation":[{"name":"Material Physics Laboratory, Physics Department, Universitas Negeri Makassar (UNM), Makassar 90223, Indonesia"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2024,6,12]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"14015","DOI":"10.1021\/acs.chemrev.7b00522","article-title":"Small bioactive peptides for biomaterials design and therapeutics","volume":"117","author":"Hamley","year":"2017","journal-title":"Chem. Rev."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"535","DOI":"10.1007\/s13197-018-3549-4","article-title":"Bioactive food derived peptides: A review on correlation between structure of bioactive peptides and their functional properties","volume":"56","author":"Karami","year":"2019","journal-title":"J. Food Sci. Technol."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"139","DOI":"10.1007\/s10989-019-09823-5","article-title":"A review on bioactive peptides: Physiological functions, bioavailability and safety","volume":"26","author":"Bhandari","year":"2020","journal-title":"Int. J. Pept. Res. Ther."},{"key":"ref_4","doi-asserted-by":"crossref","unstructured":"Daliri, E.B.-M., Oh, D.H., and Lee, B.H. (2017). Bioactive peptides. Foods, 6.","DOI":"10.3390\/foods6050032"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"19","DOI":"10.2174\/1568026043451663","article-title":"New Trends in the Development of Opioid Peptide Analogues as Advanced Remedies for Pain Relief","volume":"4","author":"Gentilucci","year":"2004","journal-title":"Curr. Top. Med. Chem."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"5198","DOI":"10.1021\/jm0498811","article-title":"Synthesis and evaluation of the affinity toward \u03bc-opioid receptors of atypical, lipophilic ligands based on the sequence c [-Tyr-Pro-Trp-Phe-Gly-]","volume":"47","author":"Cardillo","year":"2004","journal-title":"J. Med. Chem."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"e23","DOI":"10.1016\/j.ddtec.2011.07.007","article-title":"Opioid peptides: Potential for drug development","volume":"9","author":"Aldrich","year":"2012","journal-title":"Drug Discov. Today Technol."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"1698","DOI":"10.1002\/pro.3907","article-title":"Harnessing synthetic biology to enhance heterologous protein expression","volume":"29","author":"Zarzhitsky","year":"2020","journal-title":"Protein Sci."},{"key":"ref_9","doi-asserted-by":"crossref","unstructured":"Todorov, P., Georgieva, S., Tchekalarova, J., Subaer, S., Peneva, P., and Hartati, H. (2022). Synthesis, Characterization and Biological Investigation of New N-Modified Spinorphin Analogs. Pharmaceuticals, 15.","DOI":"10.3390\/ph15101251"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"6609","DOI":"10.4049\/jimmunol.167.11.6609","article-title":"The endogenous opioid spinorphin blocks fMet-Leu-Phe-induced neutrophil chemotaxis by acting as a specific antagonist at the N-formylpeptide receptor subtype FPR","volume":"167","author":"Liang","year":"2001","journal-title":"J. Immunol."},{"key":"ref_11","first-page":"1497","article-title":"Spinorphin, a new inhibitor of enkephalin-degrading enzymes derived from the bovine spinal cord. Masui","volume":"42","author":"Nishimura","year":"1993","journal-title":"Jpn. J. Anesthesiol."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"713","DOI":"10.1006\/bbrc.1993.1880","article-title":"Isolation and identification of an endogenous inhibitor of enkephalin-degrading enzymes from bovine spinal cord","volume":"194","author":"Nishimura","year":"1993","journal-title":"Biochem. Biophys. Res. Commun."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"695","DOI":"10.1016\/S0006-2952(97)00221-9","article-title":"Inhibitory effects of spinorphin, a novel endogenous regulator, on chemotaxis, O\u2212 2 generation, and exocytosis by N-formylmethionyl-leucyl-phenylalanine (FMLP)-stimulated neutrophils","volume":"54","author":"Yamamoto","year":"1997","journal-title":"Biochem. Pharmacol."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"1215","DOI":"10.1016\/S0196-9781(00)00262-X","article-title":"Complete inhibition of purinoceptor agonist-induced nociception by spinorphin, but not by morphine","volume":"21","author":"Ueda","year":"2000","journal-title":"Peptides"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"263","DOI":"10.1254\/jjp.79.263","article-title":"In vivo molecular signal transduction of peripheral mechanisms of pain","volume":"79","author":"Ueda","year":"1999","journal-title":"Jpn. J. Pharmacol."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"587","DOI":"10.2174\/1389203023380404","article-title":"Spinorphin as an endogenous inhibitor of enkephalin-degrading enzymes: Roles in pain and inflammation","volume":"3","author":"Yamamoto","year":"2002","journal-title":"Curr. Protein Pept. Sci."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"261","DOI":"10.1254\/jjp.87.261","article-title":"Spinorphin, an endogenous inhibitor of enkephalin-degrading enzymes, potentiates leu-enkephalin-induced anti-allodynic and antinociceptive effects in mice","volume":"87","author":"Honda","year":"2001","journal-title":"Jpn. J. Pharmacol."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"2198","DOI":"10.1039\/D1NJ05235G","article-title":"Synthesis and characterization of new 5, 5\u2032-dimethyl-and 5,5\u2032-diphenylhydantoin-conjugated hemorphin derivatives designed as potential anticonvulsant agents","volume":"46","author":"Todorov","year":"2022","journal-title":"New J. Chem."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"3485","DOI":"10.2147\/DDDT.S169621","article-title":"The retinoprotective role of phenytoin","volume":"12","author":"Bartollino","year":"2018","journal-title":"Drug Des. Dev. Ther."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"127630","DOI":"10.1016\/j.molstruc.2019.127630","article-title":"Synthesis of a novel phenytoin derivative: Crystal structure, Hirshfeld surface analysis and DFT calculations","volume":"1205","author":"Guerrab","year":"2020","journal-title":"J. Mol. Struct."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"2740","DOI":"10.1039\/C8NJ05748F","article-title":"Photochromic and molecular switching behaviour of new Schiff bases containing hydantoin rings: Synthesis, characterization and crystal structures","volume":"43","author":"Todorov","year":"2019","journal-title":"New J. Chem."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"567","DOI":"10.1007\/s00726-020-02836-1","article-title":"Potential anticonvulsant activity of novel VV-hemorphin-7 analogues containing unnatural amino acids: Synthesis and characterization","volume":"52","author":"Todorov","year":"2020","journal-title":"Amino Acids"},{"key":"ref_23","first-page":"11","article-title":"Contrasting solution conformations of peptides containing \u03b1, \u03b1-dialkylated residues with linear and cyclic side chains","volume":"35","author":"Prasad","year":"1995","journal-title":"Biopolym. Orig. Res. Biomol."},{"key":"ref_24","first-page":"783","article-title":"Rapid characterization of peptide secondary structure by FT-IR spectroscopy","volume":"56","author":"Adochitei","year":"2011","journal-title":"Rev. Roum. Chim."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"1871","DOI":"10.1007\/s10482-018-1080-2","article-title":"In silico design of polycationic antimicrobial peptides active against Pseudomonas aeruginosa and Staphylococcus aureus","volume":"111","author":"Giraldo","year":"2018","journal-title":"Antonie Van Leeuwenhoek"},{"key":"ref_26","doi-asserted-by":"crossref","unstructured":"Saraiva, M.A. (2020). Interpretation of \u03b1-synuclein UV absorption spectra in the peptide bond and the aromatic regions. J. Photochem. Photobiol. B Biol., 212.","DOI":"10.1016\/j.jphotobiol.2020.112022"},{"key":"ref_27","first-page":"171","article-title":"Lipinski\u2019s rule of five, famous extensions and famous exceptions","volume":"3","year":"2020","journal-title":"Pop. Sci. Artic."},{"key":"ref_28","doi-asserted-by":"crossref","unstructured":"Enache, T., and Oliveira-Brett, A. (2013). Peptide methionine sulfoxide reductase A(MsrA): Direct electrochemical oxidation on carbon electrodes. Bioelectrochemistry, 89.","DOI":"10.1016\/j.bioelechem.2012.08.004"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"23e29","DOI":"10.1016\/j.aca.2014.06.025","article-title":"Electrochemical evaluation of Abelson tyrosineprotein kinase 1 activity and inhibition by imatinib mesylate and danusertib","volume":"845","author":"Diculescu","year":"2014","journal-title":"Anal. Chim. Acta"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"9e16","DOI":"10.1016\/j.jelechem.2011.02.022","article-title":"Phenol and para-substituted phenols electrochemical oxidation pathways","volume":"655","author":"Enache","year":"2011","journal-title":"J. Electroanal. Chem."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"1337e1344","DOI":"10.1002\/elan.201000671","article-title":"Pathways of electrochemical oxidation of indolic compounds","volume":"23","author":"Enache","year":"2011","journal-title":"Electroanalysis"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"36","DOI":"10.1016\/j.aca.2016.04.015","article-title":"Amyloid\u2013\u03b2 peptides time-dependent structural modifications: AFM and voltammetric characterization","volume":"926","author":"Enache","year":"2016","journal-title":"Anal. Chim. Acta"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"11892e11893","DOI":"10.1021\/ja052522q","article-title":"A rapid label-free electrochemical detection and kinetic study of Alzheimer\u2019s amyloid beta aggregation","volume":"127","author":"Vestergaard","year":"2005","journal-title":"J. Am. Chem. Soc."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"409","DOI":"10.1111\/j.1528-1157.1978.tb04507.x","article-title":"Antiepileptic drug development: II. Anticonvulsant drug screening","volume":"19","author":"Krall","year":"1978","journal-title":"Epilepsia"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"308","DOI":"10.1016\/j.tifs.2020.09.019","article-title":"Plant protein-derived antioxidant peptides: Isolation, identification, mechanism of action and application in food systems: A review","volume":"105","author":"Wen","year":"2020","journal-title":"Trends Food Sci. Technol."},{"key":"ref_36","doi-asserted-by":"crossref","unstructured":"Akbarian, M., Khani, A., Eghbalpour, S., and Uversky, V.N. (2022). Bioactive peptides: Synthesis, sources, applications, and proposed mechanisms of action. Int. J. Mol. Sci., 23.","DOI":"10.3390\/ijms23031445"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"e12667","DOI":"10.7717\/peerj.12667","article-title":"Antimicrobial peptides properties beyond growth inhibition and bacterial killing","volume":"10","year":"2022","journal-title":"PeerJ"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"535","DOI":"10.1007\/s00217-004-1074-3","article-title":"Antioxidative activity of a low molecular weight peptide derived from the sauce of fermented blue mussel, Mytilus edulis","volume":"220","author":"Jung","year":"2005","journal-title":"Eur. Food Res. Technol."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"100183","DOI":"10.1016\/j.fochx.2021.100183","article-title":"Conventional and in silico approaches to select promising food-derived bioactive peptides: A review","volume":"13","year":"2022","journal-title":"Food Chem. X"},{"key":"ref_40","doi-asserted-by":"crossref","unstructured":"Todorov, P., Georgieva, S., and Tchekalarova, J. (2022). Recent Synthesis, Characterization, and Pharmacological Evaluation of Multifunctional Hemorphins Containing Non-Natural Amino Acids with Potential Biological Importance. Pharmaceuticals, 15.","DOI":"10.3390\/ph15111425"},{"key":"ref_41","unstructured":"Murray, P.R., Barron, E.J., Jorgensen, J.H., Landry, M.L., and Pfaller, M.A. (2007). Antibacterial agents and susceptibility test methods. Manual of Clinical Microbiology, ASM Press. [9th ed.]."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"556","DOI":"10.1016\/j.jep.2009.05.003","article-title":"Antimicrobial activity of the crude extract, fractions and compounds from stem bark of Ficus ovata (Moraceae)","volume":"124","author":"Kuete","year":"2009","journal-title":"J. Ethnopharmacol."},{"key":"ref_43","first-page":"14","article-title":"Antioxidant activity and phenolic content of Orthosiphon stamineus Benth from different geographic","volume":"1","author":"Khamsah","year":"2006","journal-title":"J. Sustain. Sci. Manag."}],"container-title":["Pharmaceuticals"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1424-8247\/17\/6\/770\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T14:57:20Z","timestamp":1760108240000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1424-8247\/17\/6\/770"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2024,6,12]]},"references-count":43,"journal-issue":{"issue":"6","published-online":{"date-parts":[[2024,6]]}},"alternative-id":["ph17060770"],"URL":"https:\/\/doi.org\/10.3390\/ph17060770","relation":{},"ISSN":["1424-8247"],"issn-type":[{"type":"electronic","value":"1424-8247"}],"subject":[],"published":{"date-parts":[[2024,6,12]]}}}