Nihal Mohamed Elmahdy ElsayyadOmar A. ElkadyMohamed M. SwidanHassan M. RashedTamer M. SakrAmr M. AbdelhamidMai A. ZaafanHanan M. El‑Laithy2025-07-102025-07-102025-07-05Elsayyad, N. M. E., Elkady, O. A., Swidan, M. M., Rashed, H. M., Sakr, T. M., Abdelhamid, A. M., Zaafan, M. A., & El-Laithy, H. M. (2025). Zonisamide nanodiamonds for brain targeting: A comprehensive study utilising in silico, in vitro, in vivo, and molecular investigation for successful nose-to-brain delivery for epilepsy management. Drug Delivery and Translational Research. https://doi.org/10.1007/s13346-025-01904-xhttps://doi.org/10.1007/s13346-025-01904-xhttps://repository.msa.edu.eg/handle/123456789/6468SJR 2024 1.175 Q1 H-Index 71The blood–brain barrier (BBB) is a stringent barrier that restricts the successful brain delivery of polar neurotherapeutics molecules. One such molecule is Zonisamide (ZNS), a hydrophilic centrally acting anti-epileptic drug. This study aims to overcome the poor ZNS BBB permeability using the nose-to-brain (NTB) carbon-based biocompatible nanodiamonds (ND) delivery system to deliver ZNS directly to the brain, bypassing the BBB, thereby enhancing its efcacy and reducing systemic side efects associated with oral ZNS currently available formulation in clinical practice. Intranasal (IN) ND-ZNS formulations were optimised using an Artifcial neuronal network (ANN) and assessed for particle size (PS), zeta potential, loading efciency (%LE), morphology, and in vitro release. The optimum radiolabelled ND-ZNS complex F1 biodistribution in diferent organs and its pharmacokinetics were compared to oral and IN-free ZNS in mice. Temporal lobe epilepsy (TLE) model in rats was used to compare the anti-epileptic activity of IN ND-ZNS F1 to IN free ZNS by assessing brain activity, epileptic biomarkers such as (brain neuronal specifc enolase (NSE), neuroflament light polypeptide (NEFL), and matrix metallopeptidase-9 (MMP-9)), hippocampal histopathology and the modulatory efect on epigenetic miR-199/SIRT-1 and PVT-1/BDNF pathways. Optimized ND-ZNS complex F1 consists of a ZNS:ND ratio of 1:2 and sonicated for 5 min exhibited the least PS (193.7±19.3 nm), adequate %LE (87.1±9.2%) similar to ANN predictions, with a biphasic in vitro release profle of ZNS, benefcial for both acute and chronic epilepsy treatment. The IN delivery of ND-ZNS complex F1 showed preferential higher in vivo brain uptake with minimal systemic exposure linked with higher brain/blood ratio and signifcant (p≤0.05) overall enhanced pharmacokinetics expressed by Cmax and AUC (0-120min) when compared to oral and IN free ZNS. Moreover, the TLE model confrmed the improved anti-epileptic activity of F1 compared to IN-free ZNS regarding brain activity and hippocampal histopathology, signifcant suppression of serum NSE, NEFL, MMP-9 levels, miR-199/ SIRT-1 pathway, and normalization of PVT-1/BDNF pathway. Therefore, ND used in this study could be a novel, promising carrier to target ZNS directly to the brain via the IN route for efective epilepsy management with less drug dosing and the least systemic side efects.en-USNeuronal networks · Molecular modelling · Zonisamide · Nanodiamonds · Nose-to-brain miR-199 · Temporal lobe epilepsy modelArticlehttps://doi.org/10.1007/s13346-025-01904-x