A carregar...
Publicação
Pharmacological modulators of Parkinson’s disease-associated proteins
| Nome: | Descrição: | Tamanho: | Formato: | |
|---|---|---|---|---|
| 21.61 MB | Adobe PDF |
Autores
Resumo(s)
Parkinson’s disease (PD) is the second most prevalent neurodegenerative disease worldwide, affecting approximately 1.5% of the population above 60 years of age.
Although PD is primarily a sporadic disorder of unclear etiology, it is now clear that genetic factors contribute to the pathogenesis of the disease. Mutations in the leucine-rich repeat kinase 2 (LRRK2) gene, which encodes a brain protein with the same name, are strongly linked to autosomal dominant forms of PD, and have been also considered as a risk factor for sporadic cases. On the other hand, mutations in the genes encoding PTEN-induced kinase 1 (PINK1) and E3 ubiquitin ligase Parkin proteins are associated with the most common forms of early-onset familial autosomal-recessive PD. Although the physiologic function of these proteins has not yet been fully established, it is now evident that they can provide alternative targets and approaches for the treatment of PD.
With the aim of discovering new and innovative small molecules that can modulate LRRK2 and be further used in the treatment of PD, a computer-aided drug design strategy combining structure-based drug design and molecular docking was developed. The lack of the LRRK2 crystallographic structure presented a major challenge in the implementation of this protocol, having defined the course of the working plan. Homology modelling was applied to predict the tridimensional structure of the LRRK2 kinase domain and molecular docking studies were developed. The previously constructed homology model was used to guide the rational design of novel LRRK2 inhibitors, exploring selectivity and additional binding interactions of small molecules to the LRRK2 ATP-binding site. Additionally, in order to further develop a high-throughput screening assay to evaluate the pharmacological inhibition of LRRK2, a yeast-based phenotypic assay was implemented and the biological activity of the newly synthesized compounds evaluated. In order to discover innovative approaches to modulate LRRK2 levels in PD patients, promoting LRRK2 selective and controlled degradation, a series of PROeolysis TArgeting Chimeras (PROTACs) targeting LRRK2 was developed. The previously constructed homology model was used to guide the rational design of PROTACs. In order to evaluate the degradation activity of LRRK2 targeting PROTACs, the newly synthesized compounds were screened in three different cell lines – WT-LRRK2 and pathogenic PD-associated mutants G2019S and R1441CLRRK2.
With the purpose of discovering novel biomarkers for Parkin activation, providing the bases for highthroughput screening campaigns, novel activity-based probes (ABPs) for profiling Parkin activity were designed and are currently under development.
Descrição
Palavras-chave
Doença de Parkinson LRRK2 Parkin Modelo de homologia Inibidores de cinase PROTACs Sondas Baseadas na Atividade Parkinson’s Disease Homology Model Kinase inhibitors Activity-Based Probes