title: Inhibition of alpha-synuclein aggregation by multifunctional dopamine agonists assessed by a novel in vitro assay and an in vivo Drosophila synucleinopathy model.
abstract: Aggregation of alpha synuclein (α-syn) leading to dopaminergic neuronal death has been recognized as one of the main pathogenic factors in the initiation and progression of Parkinson's disease (PD). Consequently, α-syn has been targeted for the development of therapeutics for PD. We have developed a novel assay to screen compounds with α-syn modulating properties by mimicking recent findings from in vivo animal studies involving intrastriatal administration of pre-formed fibrils in mice, resulting in increased α-syn pathology accompanying the formation of Lewy-body (LB) type inclusions. We found that in vitro generated α-syn pre-formed fibrils induce seeding of α-syn monomers to produce aggregates in a dose-and time-dependent manner under static conditions in vitro. These aggregates were toxic towards rat pheochromocytoma cells (PC12). Our novel multifunctional dopamine agonists D-519 and D-520 exhibited significant neuroprotection in this assay, while their parent molecules did not. The neuroprotective properties of our compounds were further evaluated in a Drosophila model of synucleinopathy. Both of our compounds showed protective properties in fly eyes against the toxicity caused by α-syn. Thus, our in vitro results on modulation of aggregation and toxicity of α-syn by our novel assay were further validated with the in vivo experiments.
authors.affiliation: Department of Pharmaceutical Sciences, Wayne State University, Detroit, MI 48202,USA.
authors.forename: Deepthi
authors.initials: D
authors.lastname: Yedlapudi
authors.affiliation: Department of Pharmacology, Wayne State University, Detroit, MI 48201, USA.
authors.forename: Gnanada S
authors.initials: GS
authors.lastname: Joshi
authors.affiliation: Department of Pharmaceutical Sciences, Wayne State University, Detroit, MI 48202,USA.
authors.forename: Dan
authors.initials: D
authors.lastname: Luo
authors.affiliation: Department of Pharmacology, Wayne State University, Detroit, MI 48201, USA.
authors.forename: Sokol V
authors.initials: SV
authors.lastname: Todi
authors.affiliation: Department of Pharmaceutical Sciences, Wayne State University, Detroit, MI 48202,USA.
authors.forename: Aloke K
authors.initials: AK
authors.lastname: Dutta
mesh_terms: D000818:Animals; D052160:Benzothiazoles / Q000737:chemistry / Q000494:pharmacology; D001681:Biological Assay / Q000379:methods*; D002942:Circular Dichroism; D004195:Disease Models, Animal; D018491:Dopamine Agonists / Q000737:chemistry / Q000494:pharmacology*; D004331:Drosophila melanogaster / Q000187:drug effects / Q000378:metabolism* / Q000648:ultrastructure; D005123:Eye / Q000378:metabolism; D016716:PC12 Cells; D000077487:Pramipexole; D066329:Protein Aggregates / Q000187:drug effects*; D017433:Protein Structure, Secondary; D051381:Rats; D012293:Rifampin / Q000737:chemistry / Q000494:pharmacology; D013764:Tetrahydronaphthalenes / Q000737:chemistry / Q000494:pharmacology; D051844:alpha-Synuclein / Q000737:chemistry* / Q000633:toxicity / Q000648:ultrastructure
chemical_list: D052160:Benzothiazoles; D018491:Dopamine Agonists; D066329:Protein Aggregates; D013764:Tetrahydronaphthalenes; D051844:alpha-Synuclein; D000077487:Pramipexole; C031349:7-hydroxy-2-N,N-dipropylaminotetralin; D012293:Rifampin