Ameliorating Effect of 7,3´-Dihydroxyflavone in Paclitaxel Induced Neurotoxicity: An In-silico and In-vitro Study

Introduction: Paclitaxel is an antineoplastic agent belonging to taxane group. It stabilises microtubule structure and one of its major dose limiting side-effects is neurotoxicity. Flavones are promising compound to alleviate paclitaxel induced neurotoxicity as it has anti-nociceptive, anti-inflammatory effect, antioxidant and neuroprotective effect. Aim: To evaluate the neuroprotective property of 7,3´- dihydroxyflavone (7,3´-DHF) against Paclitaxel induced neurotoxicity in SH-SY5Y neuroblastoma cell line through a combined in-silico and in-vitro approach with a focus on its ability to modulate ion channels. Materials and Methods: The present in-vitro study employing human neuroblastoma cell line SH-SY5Y was conducted at Sri Ramachandra Medical College and Research Institute, Chennai, Tamil Nadu, India, from April to July 2024. Molecular docking was performed against Cav3.2, Nav1.7 and Kv7.2 protein targets using auto dock tools 1.5.7, followed by Molecular Dynamics (MD) analysis using PyMOL version 2.0. Flow cytometry was done to assess antioxidant effect of 7,3´-DHF by free radical scavenging activity. Reverse Transcription Polymerase Chain Reaction (RT-PCR) was done to study gene expression of CACNA1H gene associated with paclitaxel induced peripheral neuropathy. Results: Molecular docking scores of 7,3´-DHF with target was -6.66 kcal/mol for Nav1.7, -6.56 kcal/mol for Cav3.2 and -6.66 kcal/mol for Kv7.2. In MD, Root Mean Square Deviation (RMSD) stabilised around 5 Å for both Cav 3.2 and Kv 7.2 after 50 ns. Molecular Mechanics/Generalised Born Surface Area (MM/ GBSA) binding free energy (ΔG Bind) values were -62.2 kcal/ mol for Cav 3.2 and -66.1 kcal/mol for Kv 7.2. Flow cytometry showed reduced Reactive Oxygen Species (ROS) generation in cells treated with 7,3´-dihydroxyflavone in dose dependent manner. Gene expression showed that 7,3´-dihydroxyflavone down regulates the CACNA1H gene that encodes Cav 3.2 which is associated with paclitaxel induced peripheral neuropathy. Conclusion: This study adds evidence to the significant neuroprotective potential of 7,3´-dihydroxyflavone against paclitaxel-induced neurotoxicity, demonstrated through in-silico and in-vitro approaches and identifies it as one of the promising therapeutic candidates for mitigating neurotoxicity in paclitaxeltreatment. However further confirmation through in vivo study is needed.