Neuroprotective propensity of N-(5-chloro-2-hydroxyphenyl)-2-(morpholin-4-yl-methyl) benzamide, an inventive calcineurin inhibitor, in biological models of Parkinson's disease

Abstract

The current work initially focused on the custom synthesis of ILB-162, an innovative calcineurin inhibitor, known chemically as N-(5-chloro-2-hydroxyphenyl)-2-(morpholin-4-yl-methyl) benzamide to get a yield of 25 mg with 90.73% purity and confirmed its significant calcineurin inhibitory potential in vitro having very low IC₅₀ value (0.057 µM). Further assessment in L929 cells revealed low cytotoxicity (LD₅₀ = 305.28 µg/mL) and the neuroprotection evaluated in SHSY 5Y cells induced with 6OHDA, identified 50 µg/mL as the optimal protective dose. The Parkinson's disease (PD) models in the current study were divided into three groups; Normal control (NC) without any treatment, Disease Control (DC) induced with chemical screens (6OHDA for SHSY 5Y cell line and Rotenone for C. elegans) and treated with ILB-162 group (induced cells treated with 50 µg/mL ILB-162), for accessing molecular as well as behavioral effects. In the Cell line models, ILB-162 treatment significantly reduced intracellular ROS as well as reversed α-synuclein overexpression, indicating its potential to reverse the molecular pathology underlying PD. Subsequent studies using C. elegans PD model assessed toxicity (LD₅₀ = 1436.39 µM) comparable to in vitro observations and demonstrated the outstanding capability of ILB-162 to restore the dopamine-dependent behaviors which were disrupted in DC. These findings suggest that ILB-162 can be a promising therapeutic candidate against PD, justifying the further development.

Keywords: Alpha synuclein expression; Basal slowing; Calcineurin phosphatase activity; Chemotaxis; Parkinson’s disease; Survivability analysis.