A new MAP-Rasagiline conjugate reduces α-synuclein inclusion formation in a cell model

  • Nuno ValeEmail author
  • Cláudia Alves
  • Vaishali Sharma
  • Diana F. Lázaro
  • Sara Silva
  • Paula Gomes
  • Tiago Fleming Outeiro
Short Communication



Parkinson’s disease (PD) is the second most common neurodegenerative disease of the elderly. Current therapies are only symptomatic, and have no disease-modifying effect. Therefore, disease progresses continuously over time, presenting with both motor and non-motor features. The precise molecular basis for PD is still elusive, but the aggregation of the protein alpha-synuclein (α-syn) is a key pathological hallmark of the disease and is, therefore, a major focus of current research. Considering the intrinsic properties of cell-penetrating peptides (CPPs) for mediating drug delivery of neurotherapeutics across the blood brain barrier (BBB), these might open novel opportunities for the development of new solutions for the treatment of brain-related aspects of PD and other neurodegenerative disorders.


Here, we synthesized solid-phase CPPs using an amphipathic model peptide (MAP) conjugated with the drug Rasagiline (RAS), which we named RAS-MAP, and evaluated its effect on α-syn inclusion formation in a human cell-based model of synucleinopathy.


We found that treatment with RAS-MAP at low concentrations (1–3 µM) reduced α-syn aggregation in cells.


For the first time, we report that conjugation of a current drug used in the therapy of PD with CPP reduces α-syn aggregation, which might prove beneficial in PD and other synucleinopathies.

Graphic abstract


Alpha-synuclein Cell-penetrating peptides Lewy bodies Parkinson’s disease Rasagiline 





Blood brain barrier


Central nervous system


Cell-penetrating peptide






High-pressure liquid chromatography


Monoamide oxidase B


Amphipathic model peptide


Non-motor symptoms


Parkinson’s disease




Reverse-phase medium-pressure liquid chromatography


Standard deviation


TVP-1012 in early monotherapy for PD outpatients


Attenuation of disease progression with Azilect given once—daily


Parkinson’s Rasagiline: efficacy and safety in the treatment of “Off”


Lasting effect in adjunct therapy with Rasagiline given once—daily



This work was financed by FEDER—Fundo Europeu de Desenvolvimento Regional funds through the COMPETE 2020—Operacional Programme for Competitiveness and Internationalisation (POCI), Portugal 2020, and by Portuguese funds through FCT, in the framework of the project “Institute for Research and Innovation in Health Sciences” (POCI-01-0145-FEDER-007274), and through project IF/00092/2014/CP1255/CT0004. SS thanks her PhD Grant (PD/BD/135456/2017) from FCT. NV thanks FCT by IF position, Fundação Manuel António da Mota (FMAM, Portugal) and Pfizer Portugal by support Nuno Vale Lab. NV and PG thank Dr. Marco Colombo, from Dipharma, for the supply of Rasagiline used in the first phase of the work. The contents of this report are solely the responsibility of the authors and do not necessarily represent the official views of the FCT, FMAM or Pfizer Portugal. TFO was supported by the DFG Center for Nanoscale Microscopy and Molecular Physiology of the Brain (CNMPB).


This work has been financed by the Portuguese founds from Fundação para a Ciência e a Tecnologia (FCT), through of Grant Number IF/00092/2014/CP1255/CT0004.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interests.

Supplementary material

43440_2019_32_MOESM1_ESM.pdf (1.2 mb)
Supplementary material 1 (PDF 1187 kb)


  1. 1.
    Gasser T. Molecular pathogenesis of Parkinson disease: insights from genetic studies. Expert Rev Mol Med. 2009. Scholar
  2. 2.
    Ozansoy M, Başak AN. The central theme of Parkinson’s disease: α-synuclein. Mol Neurobiol. 2012;42:460–5. Scholar
  3. 3.
    Dickson DW, Fujishiro H, Orr C, DelleDonne A, Josephs KA, Frigerio R, et al. Neuropathology of non-motor features of Parkinson disease. Park Relat Disord. 2009. Scholar
  4. 4.
    Wolters EC. Non-motor extranigral signs and symptoms in Parkinson’s disease. Park Relat Disord. 2009;15:S6–12. Scholar
  5. 5.
    Baba M, Nakajo S, Tu PH, Tomita T, Nakaya K, Lee VM, et al. Aggregation of alpha-synuclein in Lewy bodies of sporadic Parkinson’s disease and dementia with Lewy bodies. Am J Pathol. 1998;152:879–84. Scholar
  6. 6.
    Spillantini MG, Schmidt ML, Lee VMY, Trojanowski JQ, Jakes R, Goedert M. Alpha-synuclein in Lewy bodies. Nature. 1997;388:839–40. Scholar
  7. 7.
    Mor DE, Tsika E, Mazzulli JR, Gould NS, Kim H, Daniels MJ, et al. Dopamine induces soluble α-synuclein oligomers and nigrostriatal degeneration. Nat Neurosci. 2017;20:1560–8. Scholar
  8. 8.
    Kahle PJ. Alpha-synucleinopathy models and human neuropathology: similarities and differences. Acta Neuropathol. 2008;115:87–95. Scholar
  9. 9.
    Pinho R, Paiva I, Jercic KG, Fonseca-Ornelas L, Gerhardt E, Fahlbusch C, Garcia-Esparcia P, et al. Nuclear localization and phosphorylation modulate pathological effects of alpha-synuclein. Hum Mol Genet. 2019;28(1):31–50. Scholar
  10. 10.
    Liu H, Wang X. Alpha-synuclein and Parkinson disease. Neural Regen Res. 2007;2:239–43. Scholar
  11. 11.
    Stefanis L. Alpha-synuclein in Parkinson’s disease. Cold Spring Harb Perspect Med. 2012;2:1–23. Scholar
  12. 12.
    Chung CY, Koprich JB, Siddiqi H, Isacson O. Dynamic changes in presynaptic and axonal transport proteins combined with striatal neuroinflammation precede dopaminergic neuronal loss in a rat model of AAV-synucleinopathy. J Neurosci. 2009;29:3365–73. Scholar
  13. 13.
    Alim MA, Ma Q-L, Takeda K, Aizawa T, Matsubara M, Nakamura V, et al. Demonstration of a role for α-synuclein as a functional microtubule-associated protein. J Alzheimer’s Dis. 2004;6:435–42. Scholar
  14. 14.
    Giasson BI, Forman MS, Higuchi M, Golbe LI, Graves CL, Kotzbauer PT, et al. Initiation and synergistic fibrillization of tau and alpha-synuclein. Science. 2003;300:636–40. Scholar
  15. 15.
    Kamp F, Exner N, Lutz AK, Wender N, Hegermann J, Brunner B, et al. Inhibition of mitochondrial fusion by α-synuclein is rescued by PINK1, Parkin and DJ-1. EMBO J. 2010;29:3571–89. Scholar
  16. 16.
    Kang SS, Zhang Z, Liu X, Manfredsson FP, Benskey MJ, Cao X, et al. TrkB neurotrophic activities are blocked by α-synuclein, triggering dopaminergic cell death in Parkinson’s disease. Proc Natl Acad Sci. 2017;114:201713969. Scholar
  17. 17.
    Lázaro DF, Rodrigues EF, Langohr R, Shahpasandzadeh H, Ribeiro T, Guerreiro P, et al. Systematic comparison of the effects of alpha-synuclein mutations on its oligomerization and aggregation. PLoS Genet. 2014. Scholar
  18. 18.
    Youdim MBH. Rasagiline: an anti-Parkinson drug with neuroprotective activity. Expert Rev Neurother. 2003;3:737–49. Scholar
  19. 19.
    Finberg JP, Takeshima T, Johnston JM, Commissiong JW. Increased survival of dopaminergic neurons by rasagiline, a monoamine oxidase B inhibitor. NeuroReport. 1998;9:703–7.CrossRefGoogle Scholar
  20. 20.
    Finberg JPM. Pharmacology of rasagiline, a new MAO-B inhibitor drug for the treatment of Parkinson’s disease with neuroprotective potential. Rambam Maimonides Med J. 2010;1:1–10. Scholar
  21. 21.
    Rabey JM, Sagi I, Huberman M, Melamed E, Korczyn A, Giladi N, et al. Rasagiline mesylate, a new MAO-B inhibitor for the treatment of Parkinson’s disease: a double-blind study as adjunctive therapy to levodopa. Clin Neuropharmacol. 2000;23:324–30. Scholar
  22. 22.
    Stocchi F, Fossati C, Torti M. Rasagiline for the treatment of Parkinson’s disease: an update. Expert Opin Pharmacother. 2015;16:2231–41. Scholar
  23. 23.
    Weinreb O, Amit T, Bar-Am O, Youdim MBH. Rasagiline: a novel anti-Parkinsonian monoamine oxidase-B inhibitor with neuroprotective activity. Prog Neurobiol. 2010;92:330–44. Scholar
  24. 24.
    Kakish J, Tavassoly O, Lee JS. Rasagiline, a suicide inhibitor of monoamine oxidases, binds reversibly to α-synuclein. ACS Chem Neurosci. 2015;6:347–55. Scholar
  25. 25.
    Kang SS, Ahn EH, Zhang Z, Liu X, Manfredsson FP, Sandoval IM, et al. α-Synuclein stimulation of monoamine oxidase-B and legumain protease mediates the pathology of Parkinson’s disease. EMBO J. 2018;37:e98878. Scholar
  26. 26.
    Guidotti G, Brambilla L, Rossi D. Cell-penetrating peptides: from basic research to clinics. Trends Pharmacol Sci. 2017;38:406–24. Scholar
  27. 27.
    Lönn P, Kacsinta AD, Cui XS, Hamil AS, Kaulich M, Gogoi K, Dowdy SF. Enhancing endosomal escape for intracellular delivery of macromolecular biologic therapeutics. Sci Rep. 2016;6:32301. Scholar
  28. 28.
    Lee HJ, Huang YW, Chiou SH, Aronstam RS. Polyhistidine facilitates direct membrane translocation of cell-penetrating peptides into cells. Sci Rep. 2019;9:9398. Scholar
  29. 29.
    Zong T, Mei L, Gao H, Shi K, Chen J, Wang Y, Zhang Q, Yang Y, He Q. Enhanced glioma targeting and penetration by dual-targeting liposome co-modified with T7 and TAT. J Pharm Sci. 2014;103:3891–901. 2014 Oct 22).CrossRefPubMedGoogle Scholar
  30. 30.
    Srimanee A, Regberg J, Hallbrink M, Vajragupta O, Langel U. Role of scavenger receptors in peptide-based delivery of plasmid DNA across a blood-brain barrier model. Int J Pharm. 2016;500:128–35. Scholar
  31. 31.
    Vale N, Ferreira A, Fernandes I, Alves C, Araújo MJ, Mateus N, Gomes P. Gemcitabine anti-proliferative activity significantly enhanced upon conjugation with cell-penetrating peptides. Bioorg Med Chem Lett. 2017;27:2898–901. Scholar
  32. 32.
    Kamei N, Yamaoka A, Fukuyama Y, Itokazu R, Takeda-Morishita M. Noncovalent strategy with cell-penetrating peptides to facilitate the brain delivery of insulin through the blood-brain barrier. Biol Pharm Bull. 2018;41:546–54. Scholar
  33. 33.
    Hällbrink M, Florén A, Elmquist A, Pooga M, Bartfai T, Langel Ü. Cargo delivery kinetics of cell-penetrating peptides. Biochim Biophys Acta Biomembr. 2001;1515:101–9. Scholar
  34. 34.
    Oehlke J, Scheller A, Wiesner B, Krause E, Beyermann M, Klauschenz E, et al. Cellular uptake of an α-helical amphipathic model peptide with the potential to deliver polar compounds into the cell interior non-endocytically. Biochim Biophys Acta Biomembr. 1998;1414:127–39. Scholar
  35. 35.
    Oehlke J, Wallukat G, Wolf Y, Ehrlich A, Wiesner B, Berger H, et al. Enhancement of intracellular concentration and biological activity of PNA after conjugation with a cell-penetrating synthetic model peptide. Eur J Biochem. 2004;271:3043–9. Scholar
  36. 36.
    Heitz F, Morris MC, Divita G. Twenty years of cell-penetrating peptides: from molecular mechanisms to therapeutics. Br J Pharmacol. 2009;157:195–206. Scholar
  37. 37.
    Zigoneanu IG, Pielak GJ. Interaction of α-synuclein and a cell penetrating fusion peptide with higher eukaryotic cell membranes assessed by 19F NMR. Mol Pharm. 2012;9:1024–9. Scholar
  38. 38.
    Nagel F, Falkenburger BH, Tönges L, Kowsky S, Pöppelmeyer C, Schulz JB, et al. Tat-Hsp70 protects dopaminergic neurons in midbrain cultures and in the substantia nigra in models of Parkinson’s disease. J Neurochem. 2008;105:853–64. Scholar
  39. 39.
    Borsello T, Bonny C. Use of cell-permeable peptides to prevent neuronal degeneration. Trends Mol Med. 2004;10:239–44. Scholar
  40. 40.
    Lázaro DF, Dias MC, Carija A, Navarro S, Madaleno CS, Tenreiro S, et al. The effects of the novel A53E alpha-synuclein mutation on its oligomerization and aggregation. Acta Neuropathol Commun. 2016;4(1):128.CrossRefGoogle Scholar
  41. 41.
    Lázaro DF, Rodrigues EF, Langohr R, Shahpasandzadeh H, Ribeiro T, Guerreiro P, et al. Systematic comparison of the effects of alpha-synuclein mutations on its oligomerization and aggregation. PLoS Genet. 2014;10(11):e1004741. Scholar
  42. 42.
    Kenien R, Zaro JL, Shen W-C. MAP-mediated nuclear delivery of a cargo protein. J Drug Target. 2012;20(4):329–37. Scholar
  43. 43.
    Grasso G, Muscat S, Rebella M, Morbiducci U, Audenino A, Danani A, Deriu MA. Cell penetrating peptide modulation of membrane biomechanics by molecular dynamics. J Biomech. 2018;73:137–44. Scholar
  44. 44.
    Pujals S, Fernández-Carneado J, López-Iglesias C, Kogan MJ, Giralt E. Mechanistic aspects of CPP-mediated intracellular drug delivery: relevance of CPP self-assembly. Biochim Biophys Acta. 2006;1758(3):264–79. 16545772).CrossRefPubMedGoogle Scholar

Copyright information

© Maj Institute of Pharmacology Polish Academy of Sciences 2020

Authors and Affiliations

  • Nuno Vale
    • 1
    • 2
    • 3
    • 4
    Email author
  • Cláudia Alves
    • 5
  • Vaishali Sharma
    • 6
  • Diana F. Lázaro
    • 6
  • Sara Silva
    • 1
    • 2
    • 3
  • Paula Gomes
    • 5
  • Tiago Fleming Outeiro
    • 6
    • 7
    • 8
  1. 1.Laboratory of Pharmacology, Department of Drug Sciences, Faculty of PharmacyUniversity of PortoPortoPortugal
  2. 2.Institute of Molecular Pathology and Immunology of the University of Porto (IPATIMUP)PortoPortugal
  3. 3.Instituto de Investigação e Inovação em Saúde (i3S)University of PortoPortoPortugal
  4. 4.Department of Molecular Pathology and Immunology, Institute of Biomedical Sciences Abel Salazar (ICBAS)University of PortoPortoPortugal
  5. 5.Department of Chemistry and Biochemistry, Faculty of Sciences, LAQV/REQUIMTEUniversity of PortoPortoPortugal
  6. 6.Department of Experimental Neurodegeneration, Center for Biostructural Imaging of Neurodegeneration, Center for Nanoscale Microscopy and Molecular Physiology of the BrainUniversity Medical Center GöttingenGöttingenGermany
  7. 7.Max Planck Institute for Experimental MedicineGöttingenGermany
  8. 8.Institute of Neuroscience, The Medical SchoolNewcastle UniversityNewcastle upon TyneUK

Personalised recommendations