Investigating amyloid aggregation and its inhibition: implication of reversible and non-reversible adducts formation

Vincenza Andrisano, For Life Quality Studies, University of Bologna Italy, Rimini, Italy (vincenza.andrisano@unibo.it)
Angela De Simone, Department for Life Quality Studies, Rimini Campus, Alma Mater Studiorum Università di Bologna, Italy
Marina Naldi, Department of Pharmacy and Biotechnology, Alma Mater Studiorum Università di Bologna, Italy
Center for Applied Biomedical Research (C.R.B.A.), S. Orsola-Malpighi Hospital, Bologna, Italy
Daniele Tedesco, Department of Pharmacy and Biotechnology, Alma Mater Studiorum Università di Bologna, Italy
Manuela Bartolini, Department of Pharmacy and Biotechnology, Alma Mater Studiorum Università di Bologna, Italy
Vincenza Andrisano, Department for Life Quality Studies, Rimini Campus, Alma Mater Studiorum Università di Bologna, Italy

Inhibition of aggregation of protein and peptides is a focus of therapeutic development for several diseases [1]. In particular, amyloid peptide 1-42 (Ab_1-42) self-assembly is a central event in the pathogenesis of Alzheimer’s disease, leading to the formation of amyloid plaques, involving several toxic intermediates.  In this talk, a multi-methodological approach will be introduced, which allowed the elucidation of the diverse assembly species formed during the Ab_1-42 aggregation process [2,3]. The combination of mass spectrometry (MS), circular dichroism (CD) and fluorescence spectroscopy and atomic force microscopy (AFM), as a fundamental part of this multi-methodological approach, enabled the setup of a reliable experimental protocol, whichwas applied to the chemical and morphological characterization of the self-assembly species during the Ab aggregation process and inhibition [4-6].

This approach resulted to be a successful tool to elucidate the mechanism of action of known and new inhibitors, because could highlight the formation of reversible and irreversible adducts at the basis of the inhibition process. A discussion of several examples of inhibitors will follow [7].

[1] Bartolini M, Andrisano V., Chembiochem. 2010;11(8):1018-35.

[2] Bartolini M., Bertucci C., Bolognesi M.L., Cavalli A., Melchiorre C., Andrisano V., Chembiochem, 2007; 8: 2152.

[3] Bartolini M., Naldi M., Fiori J., Valle F., Biscarini F., Nicolau D.V., Andrisano V., Anal. Biochem., 2011; 414: 215 .

[4] Fiori J, Naldi M, Bartolini M, Andrisano V,. Electrophoresis. 2012, 33(22):3380-6

[5] Fiori J, Naldi N, Andrisano V., Eur J Mass Spectrom (Chichester). 2013;19(6):483-90.

[6] Naldi M, Fiori J, Pistolozzi M, Drake AF, Bertucci C, Wu R, Mlynarczyk K, Filipek S, De Simone A, Andrisano V., ACS Chem. Neurosci. 2012;3(11):952-62.

[7] Pérez-Areales FJ, Betari N, Viayna A, Pont C, Espargaró A, Bartolini M, De Simone A, Rinaldi Alvarenga JF, Pérez B, Sabate R, Lamuela-Raventós RM, Andrisano V, Luque FJ, Muñoz-Torrero D., Future Med Chem. 2017;9(10):965-981.

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Abstract Reference & Short Personal Biography of Presenting Author

Vincenza Andrisano received her 5 years Degree in Chemistry and Pharmaceutical Technology from the University of Bologna (Italy) and a 2 years post-lauream Diploma from Scuola di specializzazione in Scienze e Tecnologie Cosmetiche Università di Ferrara (Italy). As part of her postdoctoral training, she spent two years as a research assistant at the Sydney University Australia (Department of Biochemistry), researching on the enzymatic properties of new mechanism based substrates and inhibitors for dihydrofolate reductase in the search of anticancer drugs (1987-88). She has been visiting professor at McGill University, Division de Pharmacocinétique, Department D'Oncologie, Montréal (Canada) (1995) (stereoselective interaction between drug-target protein by biochromatography) and at the Department of Pharmacology Georgetown University Medical Center Washington DC (USA) (2000)(enzyme immobilization for the development of glyceraldheyde 3-phosphate dehydrogenase inhibitors in the search of anti-tripanosomial drugs). 

Since 2012 she is full professor in medicinal chemistry at the Department for the Life Quality Studies, Rimini Campus, University of Bologna.

VA's present research covers four main lines: (i) structural characterization of protein targets (i.e. amyloid peptides) and candidate leads by circular dicroism in solution and HPLC-MS, for the development of new drugs for the treatment of degenerative diseases (Alzheimer's disease and cancer) : enzyme kinetics: determination of mechanism of action of new potential acetylcholinesterase, beta-secretase, Glicogeno Synthase Kinase 3 beta inhibitors with dual function, inhibition studies of b-amyloid fibril formation by circular dichroism, fluorescence spectroscopy, mass spectrometry (ii) development of analytical methodologies in the design, synthesis and biological evaluation of new bioactive compounds (iii) characterization of the ligand/target peptide/protein/enzyme interactions (classical and multiwells spectroscopic kinetic methods, biochromatography, analysis through optical biosensor, HPLC-MS) (iv) Immobilisation of target enzymes on solid matrices which are then inserted in fluidic and chromatographic systems for binding studies.