Evaluation of Neurotoxicity of NBOH Derivatives
Rodrigo Foss da Silva, Lennon Machado Alves, Fernanda Conte, Bruno Pereira Dos Santos, Rodrigo Ligabue‐braun, Solange Cristina García, Tiago Franco de Oliveira, Hecson Jesser Segat, Silvana Peterini Boeira, Dieniffer Espinosa Janner, Ana Cristina Correa Carvalhal Ferreira, Ariane Correa Carvalhal Ferreira, Marcelo Dutra Arbo
Pharmaceuticals July 8, 2026 DOI: 10.3390/ph19071055 (opens in new tab)
Study at a glance
AI-extracted from the abstract| Characteristics | Experimental study Peer reviewed |
|---|---|
| Population | Differentiated SH-SY5Y cells and Drosophila melanogaster |
| Interventions | 25I-NBOH 25E-NBOH 25B-NBOH |
| Dose | 0.5, 1.0, 3.5, 5, and 10 nM for cells; 5, 10, and 50 nM for flies |
| Duration | 24-hour incubation for cells; 4-hour exposure for flies |
| Measures | MTT reduction assay, neutral red uptake assay, mitochondrial membrane potential, free radical production, negative geotaxis, catalase (CAT) activity, acetylcholinesterase (AChE) activity |
| Keywords | Neurotoxicity Viability assay Oxidative stress Aché Mitochondrion Oxidative phosphorylation Lactate dehydrogenase Acetylcholinesterase Pharmacology Membrane potential Biochemistry Cholinergic Cell Superoxide dismutase Mitochondrial toxicity Inner mitochondrial membrane Enzyme |
| Key findings | 25E-NBOH and 25B-NBOH decreased cell viability, induced mitochondrial hyperpolarization and oxidative stress in cells, and increased catalase and acetylcholinesterase activities in flies, indicating greater neurotoxicity than 25I-NBOH. |
Abstract
Background/Objectives: The study sought to investigate the neurotoxicity of NBOHs, which are a class of new psychoactive substances (NPSs) that act as agonists of serotonin 5-HT2A and 5-HT2C receptors, resulting in hallucinogenic effects similar to those of LSD, often found in paper blotters.
Methods: Differentiated SH-SY5Y cells were incubated for 24 h with 0.5, 1.0, 3.5, 5 and 10 nM of the derivatives 25I-NBOH, 25E-NBOH and 25B-NBOH. Cell viability was assessed by MTT reduction and neutral red uptake assays. Mitochondrial membrane potential and production of free radicals were also evaluated. In addition, Drosophila melanogaster flies were exposed to 5, 10 and 50 nM of each NBOH derivative for 4 h. Negative geotaxis, CAT, and AChE enzymatic activities were evaluated.
Results: Both 25E-NBOH and 25B-NBOH decreased cell viability. Furthermore, mitochondrial hyperpolarization of cells and an increase in oxidative species were observed in response to higher concentrations of 25E-NBOH and 25B-NBOH. In D. melanogaster, 25B-NBOH increased climbing time and both 25E-NBOH and 25B-NBOH increased CAT and AChE activities at 50 nM.
Conclusions: These findings indicate greater toxicity of the 25E- and 25B-NBOH compared to 25I-NBOH, suggesting metabolic activity or mitochondrial signaling, oxidative stress, and cholinergic dysregulation as a potential mechanism related to NBOH neurotoxicity.