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Synthesis, characterization and anticonvulsant activity of new azobenzene-containing VV-hemorphin-5 bio photoswitch

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Abstract

A novel analog of VV-hemorphin-5 containing azobenzene moiety has been synthesized and investigated for anticonvulsant activity in relation to its E → Z photophysical properties activated by long wavelength light at 365 nm. The synthesis was achieved by a modified SPPS by Fmoc-dimerization strategy. The electrochemical behavior before and after UV illumination was investigated using different voltammetric modes. The number of electrons transferred, heterogenic rate constant and diffusion coefficient for E- and Z-isomers were also evaluated. Revealing the governing principles involved in signaling and nerve pulse propagation requires the detailed characterization of the electrical properties of cell membranes. For probing the effect of synthesized azo-peptide on the membrane electrical properties, we measured the specific capacitance of lipid bilayers, representing a basic physical model of biomembranes with their simple reproducibility in laboratory conditions at controlled membrane composition and physicochemical parameters of the surrounding aqueous medium. Our results have shown reduced membrane capacitance in the presence of the azo-peptide, thus providing evidences for possible alterations in the dielectric permittivity of the bilayer. The (Val-Val-Tyr-Pro-Trp-Thr-Gln)2Azo peptide was explored also in vivo for preliminary anticonvulsant activity by using the 6-Hz seizure test and pentylenetetrazol (PTZ) seizure test in mice. The Z-isomer has exhibited higher potency compared to E-isomer most pronouncedly in the 6 Hz test for psychomotor seizures where the compound had activity at all three tested doses. It was found that the Z-isomer decrease the latency for onset of clonic seizures induced by PTZ. These results demonstrate that the Z-isomer deserves further evaluation in other screening tests for anticonvulsant activity.

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References

  • Aemissegger A, Krautler V, Gunsteren W, Hilvert D (2005) A photoinducible β-hairpin. J Am Chem Soc 127:2929–2936

    Article  CAS  PubMed  Google Scholar 

  • Angelova M, Dimitrov D (1986) Liposome electroformation. Faraday Discuss Chem Soc. 81:303–311

    Article  CAS  Google Scholar 

  • Antosiewicz JM, Shugar D (2016) UV–Vis spectroscopy of tyrosine side-groups in studies of protein structure. Part 2: selected applications. Biophys Rev 8:163–177

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Barton ME, Klein BD, Wolf HH, White HS (2001) Pharmacological characterization of the 6 Hz psychomotor seizure model of partial epilepsy. Epilepsy Res 47:217–227

    Article  CAS  PubMed  Google Scholar 

  • Bleger D, Hecht S (2015) Visible-light-activated molecular switches. Angew Chem Int Ed 54:11338

    Article  CAS  Google Scholar 

  • Blishchenko E, Sazonova O, Kalinina O, Yatskin O, Philippova M, Surovoy A, Karelin A, Ivanov V (2002) Family of hemorphins: co-relations between amino acid sequences and effects in cell cultures. Peptides 23:903–910

    Article  CAS  PubMed  Google Scholar 

  • Boulgue C, Lçweneck M, Renner Ch, Moroder L (2007) Redox potential of azobenzene as an amino acid residue in peptides. ChemBioChem 8:591–594

    Article  CAS  Google Scholar 

  • Çakir I, EBiçer E, Odabasoglu M, Albayrak Ç (2005) Electrochemical and spectroscopic study of 4-(Phenyldiazenyl)-2-{[tris-(hydroxymethyl)methyl]aminomethylene} cyclohexa-3,5-dien-1(2H)-one Mechanism of the azo and imine electroreduction. J Braz Chem Soc 16(4):711–717

    Article  Google Scholar 

  • Carroll EC, Berlin S, Levitz J, Kienzler MA, Yuan Z, Madsen D, Larsen DS, Isacoff EY (2015) Two-photon brightness of azobenzene photoswitches designed for glutamate receptor optogenetics. Proc Natl Acad Sci USA 112:E776–E785

    Article  CAS  PubMed  Google Scholar 

  • Cohen M, Fruitier-Arnaudin I, Piot JM (2004) Hemorphins: substrates and/or inhibitors of dipeptidyl peptidase IV Hemorphins N-terminus sequence influence on the interaction between hemorphins and DPPIV. Biochimie 86:31–37

    Article  CAS  PubMed  Google Scholar 

  • Corbett AD, Henderson G, McKnight AT, Paterson SJ (2006) 75 years of opioid research: the exciting but vain quest for the Holy Grai. Br J Pharmacol 147(1):153–162

    Google Scholar 

  • Dimova R (2012) Giant vesicles: a biomimetic tool for membrane characterization. In: Iglic A (ed) Advances in planar lipid bilayers and liposomes. Academic Press, Elsevier, Burlington

    Google Scholar 

  • Garreau I, Chansel D, Vandermeersch S, Fruitier I, Piot JM, Ardaillou R (1998) Hemorphins inhibit angiotensin IV binding and interact with aminopeptidase N. Peptides 19(8):1339–1348

    Article  CAS  PubMed  Google Scholar 

  • Genova J, Vitkova V, Bivas I (2013) Registration and analysis of the shape fluctuations of nearly spherical lipid vesicles. Phys Rev E 88:022707

    Article  CAS  Google Scholar 

  • Georgieva S, Nedeltcheva T, Nikolova L (2010) Study of labile complexes by differential pulse polarography. J Univ Chem Technol Metall 45:201

    CAS  Google Scholar 

  • Goodman M, Falxa ML (1967) Comformational aspects of polypeptide structure. XXIII. Photoisomerization of azoaromatic polypeptides. J Am Chem Soc 89:3863–3867

    Article  CAS  PubMed  Google Scholar 

  • Goodman M, Kossoy A (1966) Comformational aspects of polypeptide structure. XIX. Azoaromatic side-chain effects. J Am Chem Soc 88:5010–5015

    Article  CAS  Google Scholar 

  • Higuchi Y, Takano Y, Shimazaki H, Shimoigashi Y, Kodama H, Matsumoto H, Nonaka S, Saito R, Waki M, Kamiya H (1989) Dimeric substance P analogue shows a highly potent activity of the in vivo salivary secretion in the rat. Eur J Pharmacol 160:413–416

    Article  CAS  PubMed  Google Scholar 

  • Hoppmann C, Seedorff S, Richter A, Fabian H, Schmieder P, Ruck-Braun K, Beyermann M (2009) Light-directed protein binding of a biologically relevant beta-sheet. Angew Chem Int Ed 48:6636–6639

    Article  CAS  Google Scholar 

  • Ivanov VT, Karelin AA, Yatskin ON (2005) Generation of peptides by human erythrocytes: facts and artifacts. Biopolymers 80:332–346

    Article  CAS  PubMed  Google Scholar 

  • Jinsmaa Y, Yoshikawa M (2002) Release of hemorphin-5 from human hemoglobin by pancreatic elastase. Biosci Biotechnol Biochem 66(5):1130–1132

    Article  CAS  PubMed  Google Scholar 

  • Lakowicz JR (2006) Principles of fluorescence spectroscopy, 3rd edn. Springer, Boston, pp 443–530. ISBN 978-0387-31278-1

    Book  Google Scholar 

  • Malik WU, Goyal RN (1976) Polarographic reduction of some potential antidiabetic compounds with more than one reduction site. Talanta 23:705

    Article  CAS  PubMed  Google Scholar 

  • Mart RJ, Allemann RK (2016) Azobenzene photocontrol of peptides and proteins. Chem Commun 52:12262

    Article  CAS  Google Scholar 

  • Martinek K, Berzein IV (1979) Artificial light-sensitive enzymatic systems as chemical amplifiers of weak light signals. Photochem Photobiol 29:637–649

    Article  CAS  PubMed  Google Scholar 

  • Merino E, Ribagorda M (2012) Control over molecular motion using the cis–trans photoisomerization of the azo group. Beilstein J Org Chem 8:1071–1090

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Moisan S, Harvey N, Beaudry G, Forzani P, Burhop KE, Drapeau G, Rioux F (1998) Structural requirements and mechanism of the pressor activity of Leu-Val-Val- hemorphin-7, a fragment of hemoglo- bin beta-chain in rats. Peptides 19(1):119–131

    Article  CAS  PubMed  Google Scholar 

  • Mortensen UH, Raaschou-Nielsen M, Breddam K (1994) Recognition of C-terminal amide groups by (serine) carboxypeptidase Y investigated by site-directed mutagenesis. J Biol Chem 269:15528–15532

    CAS  PubMed  Google Scholar 

  • Needham D, Hochmuth R (1989) Electro-mechanical permeabilization of lipid vesicles. Role of membrane tension and compressibility, Biophys J 55:1001–1009

    CAS  PubMed  Google Scholar 

  • Petrov AG (1999) The lyotropic state of matter: molecular physics and living matter physics. Gordon & Breach Publ, Amsterdam

    Google Scholar 

  • Pieroni O, Fissi A (1992) Synthetic photochromic polypeptides: possible models for photoregulation in biology. J Photochem Photobiol B: Biol 12:125–140

    Article  CAS  Google Scholar 

  • Podewin T, Rampp MS, Turkanovic I, Karaghiosoff KL, Zinth W, Hoffmann-Roder A (2015) Photocontrolled chignolin-derived β-hairpin peptidomimetics. Chem Commun 51:4001–4004

    Article  CAS  Google Scholar 

  • Pogozheva ID, Przydzial MJ, Mosberg HI (2005) Homology modeling of opioid receptor-ligand complexes using experimental constraints. AAPS J 7:434–448

    Article  Google Scholar 

  • Rau H (2002) Photo reactive organic thin films. In: Zhao Y, Ikeda T, editors. Elsevier Sci Ltd, pp 3–48. ISBN: 9780080479972

  • Rau H (2003) Photochromism: molecules and systems. In: Duerr H, Bouas-Laurent H. Elsevier, Amsterdam, pp 165–192, ISBN: 9780080538839

  • Renner C, Moroder L (2006) Azobenzene as conformational switch in model peptides. ChemBioChem 7:869

    Article  CAS  Google Scholar 

  • Renner C, Behrendt R, Heim N, Moroder L (2002) Photomodulation of conformational states. III. Water-soluble bis-cysteinyl-peptides with (4-aminomethyl)phenylazobenzoic acid as backbone constituent. Biopolymers 63:382–393

    Article  CAS  PubMed  Google Scholar 

  • Rover P, Pegoraro S, Vigano S, Amato C, Vaccari L, Balestreri E, Felicioli R (1995) Solid- phase synthesis and dimerization of an azobenzene- containing peptide as photoisomerizable proteinase inhibitor. Lett Pept Sci 2:27–32

    Article  Google Scholar 

  • Salipante PF, Knorr R, Dimova R, Vlahovska P (2012) Electrodeformation method for measuring the capacitance of bilayer membranes. Soft Matter 8:3810–3816

    Article  CAS  Google Scholar 

  • Santurri P, Robbins F, Stubbings R (1960) 4,4′-Diaminoazobenzene. Org Synth Coll 40:18

    Article  CAS  Google Scholar 

  • Santurri P, Robbins F, Stubbings R (1973) 4,4′-Diaminoazobenzene. Org Synth Coll 5:341

    Google Scholar 

  • Schwan HP (1989) Dielectrophoresis and Rotation of Cells. In: Neumann E, Sowers AE, Jordan CA (eds) Electroporation and electrofusion in cell biology. Springer, Boston, pp 3–21

    Chapter  Google Scholar 

  • Scoto GM, Aricò G, Ronsisvalle S, Parenti C (2007) Blockade of the nociceptin/orphanin FQ/NOP receptor system in the rat ventrolateral periaqueductal gray potentiates DAMGO analgesia. Peptides 28(7):1441–1446

    Article  CAS  PubMed  Google Scholar 

  • Song CZ, Wang WQ, Song CC (2012) Hemorphin as a prognostic biomarker and potential drug for breast cancer? Int J Cancer 131:1011–1012

    Article  CAS  PubMed  Google Scholar 

  • Szymanski W, Beierle JM, Kistemaker HA, Velema WA, Feringa BL (2013) Reversible photocontrol of biological systems by the incorporation of molecular photoswitches. Chem Rev 113:6114

    Article  CAS  PubMed  Google Scholar 

  • Todorov P, Peneva P, Pechlivanova D, Georgieva S, Dzhambazova E (2018) Synthesis, characterization and nociceptive screening of new VV-hemorphin-5 analogues. Bioorg Med Chem Lett 28:3073–3079

    Article  CAS  PubMed  Google Scholar 

  • Uçar M, Polat K, Solak A, Toyc M, Aksu M (2010) The electrochemical behaviour of 2′-halogenated derivatives of 4-methoxyazobenzene at a mercury electrode. Dyes Pigments 87:55e61

    Article  CAS  Google Scholar 

  • Vitkova V, Antonova K, Popkirov G, Mitov M, Ermakov Y, Bivas I (2010) Electrical resistivity of the liquid phase of vesicular suspensions prepared by different methods. J Phys Conf Ser 253:012059

    Article  CAS  Google Scholar 

  • Vitkova V, Mitkova D, Antonova K, Popkirov G, Dimova R (2018) Sucrose alter the electric capacitance and dielectric permittivity of lipid bilayers. Coll Surf A Physicochem Eng Asp 557:51–57

    Article  CAS  Google Scholar 

  • Westmark P, Kelly JP, Smith BD (1993) Photoregulation of enzyme activity. Photochromic, transition-state-analogue inhibitors of cysteine and serine proteases. J Am Chem Soc 115:3416

    Article  CAS  Google Scholar 

  • Woolley GA (2005) Photocontrolling peptide α-helices. Acc Chem Res 38:486–493

    Article  CAS  PubMed  Google Scholar 

  • Yoshikawa M (2015) Bioactive peptides derived from natural proteins with respect to diversity of their receptors and physiological effects. Peptides 72:208–225

    Article  CAS  PubMed  Google Scholar 

  • Zhao Q, Plot JM (1997) Investigation of inhibition angiotensin-converting enzyme (ACE) activity and opioid activity of two hemorphins, LVV-hemorphin-5 and VV-hemorphin-5, isolated from a defined peptic hydrolysate of bovine haemoglobin. Neuropeptides 31:147–153

    Article  CAS  PubMed  Google Scholar 

  • Zhao Y, Lei B, Wang M, Wu S, Qi W, Su R, He Z (2018) A supramolecular approach to construct a hydrolase mimic with photo-switchable catalytic activity. J Mater Chem B 6:2444–2449

    Article  CAS  Google Scholar 

Download references

Acknowledgements

This work was financially supported by the Bulgarian National Scientific Fund project ДH 18/5 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 T. Todorov, Petia N. Peneva & Anton Georgiev

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

    Stela I. Georgieva

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

    Jana Tchekalarova

  4. Institute of Solid State Physics, Bulgarian Academy of Sciences, 1784, Sofia, Bulgaria

    Victoria Vitkova & Krassimira Antonova

  5. Department of Optical Metrology and Holography, Institute of Optical Materials and Technologies, Bulgarian Academy of Science, 1113, Sofia, Bulgaria

    Anton Georgiev

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  1. Petar T. Todorov
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  4. Jana Tchekalarova
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Correspondence to Petar T. 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 were no human participants.

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Todorov, P.T., Peneva, P.N., Georgieva, S.I. et al. Synthesis, characterization and anticonvulsant activity of new azobenzene-containing VV-hemorphin-5 bio photoswitch. Amino Acids 51, 549–563 (2019). https://doi.org/10.1007/s00726-018-02691-1

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