Abstract: Background and Objective: Neurodegenerative diseases are complex and heterogeneous group of multifactorial and progressive disorders, which constitute a worldwide health problem. Plants are a potential source of multi-target neuroprotective agents for the treatment of multifaceted diseases such as neurological disorders. In this study, in vitro antioxidant and anticholinesterase screening of 81 ethanolic extracts of 43 Colombian species (Lauraceae, Piperaceae, Rutaceae and Myristicaceae) was performed in order to determine their neuroprotective potential. Materials and Methods: The preliminary activity was tested using TLC-bioautography assay with 2,2-Diphenyl-1-picrylhydrazyl (DPPH•), β-carotene and acetylcholinesterase (AChE). The inhibitory effect (IC50) of the most active extracts in each assay was quantified by a microplate colorimetric test. Results: From the TLC-bioautography assay, almost 36 extracts showed radical scavenging capacity and antioxidant activity by β-carotene bleaching mainly of Lauraceae, Myristicaceae and Piperaceae. In addition, 13 extracts exhibited anticholinesterase activity and the most potential extracts were characterized by the presence of active alkaloids. Extracts of Lauraceae and Rutaceae were more active against enzymatic inhibition than DPPH assay. Five extracts of Zanthoxylum (Rutaceae) and Ocotea (Lauraceae) presented strong inhibition of AChE with IC50 lower than 50 μg mL–1. However, most of these were inactive against butyrylcholinesterase. Conclusion: The most active extracts showed the presence of isoquinoline alkaloids, this characteristic may be related with the strong AChE inhibition.