Objective: To evaluate the effectiveness of a candidate compound identified in a high-throughput screening (HTS) assay carried out in a Drosophila model of Parkinson’s disease (PD) and validated in neuron-like human cells as PD cell model.

Background: There is no cure for PD, and current therapies are mainly addressed to relieve motor symptoms. However, they are not able to stop, delay or counteract the PD progression. To identify new therapeutic drugs, we carried out a HTS assay testing compounds from the Prestwick Chemical Library (1120 compounds) in a fly PD model based on DJ-1 deficiency, a causative gene of familial PD highly related to sporadic cases of the disease. Among the compounds able to suppress locomotor defects in PD model flies that were also shown to reduce oxidative stress (OS)-induced lethality in PD model cells, we focused on compound 118 (C118) since it was the most effective in DJ-1-deficient cells. Interestingly, one of its targets has never been proposed for therapeutic intervention in PD.

Method: We analyzed protein carbonylation levels using 2,4-dinitrophenylhydrazine and H₂O₂ levels using the commercial kit Amplex Red in extracts of 5-day-old treated PD model flies. Subsequently, we established the optimal dose of C118 by carrying out an MTT viability assay in DJ-1 mutant cells. Next, western-blots were performed to study Akt and JNK phosphorylation (anti-apoptotic and pro-apoptotic factors respectively) in C118 treated PD cells. Mitochondrial viability of cells was studied using the fluorescence dye MitoTracker. The glycolytic rate in treated cells was tested by quantifying the activity of key enzymes of this pathway.

Results: We found that C118 was able to reduce H₂O₂ and protein carbonylation levels in 5-day-old DJ-1β mutant flies. C118 exerted a neuroprotective effect in PD model cells by activating Akt and inhibiting JNK signaling. In addition, C118 was also able to increase mitochondrial viability and to enhance the glycolytic pathway in DJ-1-silenced cells.

Conclusion: In this work, we have demonstrated the effectiveness of C118 in suppressing PD-related phenotypes in preclinical models of the disease. In addition, C118 is able to increase the glycolytic rate, a strategy that has been recently proposed as a promising therapeutic target in PD. Interestingly, this compound presents two different targets with interest in PD treatment, hence widening the therapeutic landscape.

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MDS Abstracts - https://www.mdsabstracts.org/abstract/evaluation-of-a-double-target-therapeutic-in-preclinical-models-of-parkinsons-disease/