Synthesis, biological activity and HPLC validation of 1,2,3,4-tetrahydroacridine derivatives as acetylcholinesterase inhibitors

The synthesis and biochemical evaluation of new hybrids of tacrine (THA) and 4-fluorobenzoic acid (4-FBA) possessing activity towards acetylcholinesterase (AChE) and butyrylcholinesterase (BuChE) inhibition are presented. The compounds of interest were obtained from the reaction of activated 4-FBA and diamino derivatives of 1,2,3,4-tetrahydroacridine. The compounds C6-2KW/HCl, C6-4KW/HCl and C6-3KW/HCl have four-fold higher antiacetylcholinesterase activity than THA. All of the acquired compounds present higher selectivity towards AChE than THA and lower selectivity towards BuChE. In addition, a rapid, selective and stability-indicating HPLC method was developed and validated for the determination of C6-2KW/HCl, C6-3KW/HCl and C6-4KW/HCl. THA and 4-FBA were found to be the main impurities. Chromatographic separation was achieved isocratically on a Waters Symmetry C18 150 × 3.9 mm, 4 μm column with a mobile phase of acetonitrile/buffer (17 mM sodium dodecyl sulphate and 8.3 mM sodium dihydrogen phosphate, 50:50 v/v) (overall pH 4). A 1.5 ml/min flow rate and a 247 nm wavelength were chosen for this method. C6-2KW/HCl, C6-3KW/HCl and C6-4KW/HCl were subjected to acidic and basic hydrolysis, chemical oxidation, thermal exposition at 60 °C and intense UV light. The limits of detection (LOD) and quantification (LOQ) were less than 2 μg/ml and 6 μg/ml for C6-2KW/HCl, C6-3KW/HCl and C6-4KW/HCl, 0.04 μg/ml and 0.12 μg/ml for THA, 0.42 μg/ml and 1.41 μg/ml for 4-FBA, respectively.

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► Synthesis of new hybrids from 4-FBA and diamino derivatives of THA was provided.
► New compounds have four-fold higher antiacetylcholinesterase activity than THA.
► Higher selectivity towards AChE than BuChE was observed.
► Isocratic HPLC method for related compounds and content assay was proposed.
► New method was found to be selective, sensitive, precise and stability indicating.