Design and Synthesis of Fluorescent Methylphenidate Analogues for a FRET-Based Assay of Synapsin III Binding.
ChemMedChem
; 15(14): 1330-1337, 2020 07 20.
Article
em En
| MEDLINE
| ID: mdl-32452650
ABSTRACT
We previously described synapsin III (Synâ
III) as a synaptic phosphoprotein that controls dopamine release in cooperation with α-synuclein (aSyn). Moreover, we found that in Parkinson's disease (PD), Synâ
III also participates in aSyn aggregation and toxicity. Our recent observations point to threo-methylphenidate (MPH), a monoamine re-uptake inhibitor that efficiently counteracts the freezing-gait characteristic of advanced PD, as a ligand for Synâ
III. We have designed and synthesised two different fluorescently labelled MPH derivatives, one with Rhodamine Red (RHOD) and one with 5-carboxytetramethylrhodamine (TAMRA), to be used for assessing MPH binding to Synâ
III by FRET. TAMRA-MPH exhibited the ideal characteristics to be used as a FRET acceptor, as it was able to enter into the SK-N-SH cells and could interact specifically with human green fluorescent protein (GFP)-tagged Synâ
III but not with GFP alone. Moreover, the uptake of TAMRA-MPH and co-localization with Synâ
III was also observed in primary mesencephalic neurons. These findings support that MPH is a Synâ
III ligand and that TAMRA-conjugated drug molecules might be valuable tools to study drug-ligand interactions by FRET or to detect Synâ
III in cytological and histological samples.
Palavras-chave
Texto completo:
1
Bases de dados:
MEDLINE
Assunto principal:
Desenho de Fármacos
/
Sinapsinas
/
Transferência Ressonante de Energia de Fluorescência
/
Corantes Fluorescentes
/
Metilfenidato
Tipo de estudo:
Prognostic_studies
Limite:
Animals
/
Humans
Idioma:
En
Revista:
ChemMedChem
Assunto da revista:
FARMACOLOGIA
/
QUIMICA
Ano de publicação:
2020
Tipo de documento:
Article
País de afiliação:
Itália