# Design and Synthesis of Caffeine-Based Derivatives with Antioxidant and Neuroprotective Activity: In Vitro Evaluation and SwissADME Profiling

## TL;DR

Researchers designed new caffeine derivatives that show strong antioxidant and neuroprotective effects in lab tests, potentially useful for treating neurodegenerative diseases.

## Contribution

The study introduces novel caffeine-based compounds with enhanced antioxidant and neuroprotective activity compared to caffeine and standard antioxidants.

## Key findings

- Several caffeine derivatives showed higher antioxidant activity than caffeine and Trolox at low concentrations.
- Compound AL-7 provided up to 85% protection against glutamate-induced excitotoxicity in human neuroblastoma cells.
- SwissADME analysis predicted good oral bioavailability but limited blood–brain barrier permeability for the derivatives.

## Abstract

Oxidative stress and excitotoxicity are key contributors to neuronal damage in various neurodegenerative diseases. Caffeine, a widely used neuroactive compound with moderate antioxidant properties, may benefit from structural modifications to enhance its neuroprotective potential. In this study, a series of novel caffeine derivatives was synthesized and evaluated for antioxidant and potential neuroprotective relevance using in vitro models of oxidative stress and glutamate-induced excitotoxicity in SH-SY5Y human neuroblastoma cells. Antioxidant capacity was assessed using ABTS•+ radical cation decolorization and DPPH radical scavenging assays. Most derivatives exhibited strong free radical scavenging activity, surpassing both caffeine and the reference antioxidant Trolox at low concentrations (5 µM). Notably, compounds AL-7, AL-8, AL-9, and AL-10 demonstrated particularly high activity. Cytotoxicity evaluation using the MTT assay revealed low toxicity for all compounds, with calculated IC50 values above 500 µM. Intracellular reactive oxygen species (ROS) levels measured by the DCFH-DA assay showed that several derivatives, especially AL-4, significantly reduced H2O2-induced oxidative stress. In neuroprotection assays, compounds AL-0, AL-1, and AL-4 markedly protected against hydrogen peroxide-induced damage, restoring cell viability up to 73%, while AL-7 achieved up to 85% protection against L-glutamate-induced excitotoxicity, outperforming caffeine. In silico SwissADME analysis indicated favorable oral bioavailability, with predicted gastrointestinal absorption and limited blood–brain barrier permeability. Overall, these findings highlight structurally modified caffeine derivatives as promising antioxidant and neuroprotective agents warranting further mechanistic and therapeutic investigation.