Cholinesterase inhibitors (ChEI) play a significant role in enhancing synaptic cholinergic activity avoiding cholinergic poisoning, and consequently have therapeutic relevance related to Alzheimer's disease (AD), myasthenia gravis, and glaucoma. In this sense, photochromic ChEI have become interesting due to the expansion of photopharmacological approach towards the remote control of acetylcholinesterase (AChE). Inspired in the concept of “azologization” as the rational introduction of the azobenzene group into the structure of a designed active compound, and based on our previous results, indicating aza-stilbene activity as dual ChEI, we synthesized several azoderivatives with potential application as reversible photochromic AChE inhibitors. In this work we present 1,2-bis(4-(4-(pyrrolidin-1-yl)butoxy)phenyl)diazene (di-PRLC₄OAzo), obtained by microwave assisted synthesis, which showed in vitro enzymatic response for its (E) isomer (IC₅₀: 1,08 µM) beside a stable photostationary state, indicating it might be an efficient photo-responsible probe to remote control the activity of the enzyme. Ellman´s assay was performed for measuring AChE activity in the presence of the reversible photoswitchable blocker, whose in vitro photoisomerization efficiency was evaluated by UV/Vis absorption spectroscopy.