Alzheimer's disease (AD), the most common type of dementia, currently represents an extremely challenging and unmet medical need worldwide. Amyloid-β (Aβ) and Tau proteins are prototypical AD hallmarks, as well as validated drug targets. Accumulating evidence now suggests that they synergistically contribute to disease pathogenesis. This could not only help explain negative results from anti-Aβ clinical trials but also indicate that therapies solely directed at one of them may have to be reconsidered. Based on this, herein, we describe the development of a focused library of 2,4-thiazolidinedione (TZD)-based bivalent derivatives as dual Aβ and Tau aggregation inhibitors. The aggregating activity of the 24 synthesized derivatives was tested in intact Escherichia coli cells overexpressing Aβ42 and Tau proteins. We then evaluated their neuronal toxicity and ability to cross the blood-brain barrier (BBB), together with the in vitro interaction with the two isolated proteins. Finally, the most promising (most active, nontoxic, and BBB-permeable) compounds 22 and 23 were tested in vivo, in a Drosophila melanogaster model of AD. The carbazole derivative 22 (20 μM) showed extremely encouraging results, being able to improve both the lifespan and the climbing abilities of Aβ42 expressing flies and generating a better outcome than doxycycline (50 μM). Moreover, 22 proved to be able to decrease Aβ42 aggregates in the brains of the flies. We conclude that bivalent small molecules based on 22 deserve further attention as hits for dual Aβ/Tau aggregation inhibition in AD.

Biomedical Research Center University Hospital Hradec Kralove 500 00Hradec Kralove Czech Republic

Department of Life Health and Environmental Sciences University of L'Aquila Via Vetoio Coppito 2 67100L'Aquila Italy

Department of Neuroscience Laboratory of Prion Biology Scuola Internazionale Superiore di Studi Avanzati Via Bonomea 265 1 34136Trieste Italy

Department of Pharmacy and Biotechnology Alma Mater Studiorum University of Bologna Via Belmeloro 6 1 40126Bologna Italy

Department of Pharmacy and Pharmaceutical Technology and Physical Chemistry Faculty of Pharmacy and Food Science University of Barcelona Av Joan XXIII 27 31 E 08028Barcelona Spain

Laboratory of Medicinal Chemistry Av Joan XXIII 27 31 E 08028Barcelona Spain

Pharmaceutical Sciences Postgraduate Program Center of Health Sciences Universidade do Vale do Itajaí Rua Uruguai 458 88302 202Itajaí Santa Catarina Brazil

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