A novel class of fluoro-substituted tris-chalcones derivatives (5a-5i) was synthesized from phloroglucinol and corresponding benzaldehydes. A three step synthesis method was followed for the production of these trischalcone compounds. The structures of the newly synthesized compounds (5a-5i) were confirmed on the basis of IR, H-1 NMR, C-13 NMR, and elemental analysis. The compounds' inhibitory activities were tested against human carbonic anhydrase I and II isoenzymes (hCA I and hCA II), acetylcholinesterase (AChE), butyrylcholinesterase (BChE), and alpha-glycosidase (alpha-Gly). These chalcone derivatives had K, values in the range of 19.58-78.73 nM for hCA I, 12.23-41.70 nM for hCA II, 1.09-6.84 nM for AChE, 8.30-32.30 nM for BChE and 0.93 +/- 0.20-18.53 +/- 5.06 nM against alpha-glycosidase. These results strongly support the promising nature of the tris-chalcone scaffold as selective carbonic anhydrase, acetylcholinesterase, butyrylcholinesterase, and aglycosidase inhibitor. Overall, due to these derivatives' inhibitory potential on the tested enzymes, they are promising drug candidates for the treatment of diseases like glaucoma, leukemia, epilepsy; Alzheimer's disease; type-2 diabetes mellitus that are associated with high enzymatic activity of carbonic anhydrase, acetylcholine esterase, butyrylcholinesterase, and alpha-glycosidase.