Reaction of several aromatic/heterocyclic sulfonamides containing a free amino, imino, hydrazino, or hydroxyl group, with 2, 3-pyridinedicarboxylic anhydride or 2,6-pyridinedicarboxylic acid in the presence of carbodiimide derivatives, afforded two series of water-soluble (as hydrochloride, triflate, or carboxylate salts) compounds. The new derivatives were assayed as inhibitors of the zinc enzyme carbonic anhydrase (CA) and more precisely of three of its isozymes, CA I, II (cytosolic forms), and IV (membrane-bound form), involved in important physiological processes. Efficient inhibition was observed against all three isozymes, but especially against CA II and IV (in nanomolar range), the two isozymes known to play a critical role in aqueous humor secretion within the ciliary processes of the eye. Some of the best inhibitors synthesized were applied as 2% water solutions directly into the eye of normotensive and glaucomatous albino rabbits. Very strong and long-lasting intraocular pressure (IOP) lowering was observed with many of them. This result prompted us to reanalyze the synthetic work done by other groups for the design of water-soluble, topically effective antiglaucoma sulfonamides. According to these researchers, the IOP-lowering effect is due to the intrinsic nature of the specific heterocyclic sulfonamide considered, among which the thienothiopyran-2-sulfonamide derivatives represent the best-studied case. Indeed, the first agents developed for topical application, such as dorzolamide, are derivatives of this ring system. To prove that the tail (in this case the pyridinecarboxylic moieties) conferring water solubility to a sulfonamide CA inhibitor is more important than the ring to which the sulfonamido group is grafted, we also prepared dorzolamide derivatives incorporating such moieties. These new compounds possess good water solubility as hydrochloride or carboxylate salts, balanced by a relatively modest lipid solubility. They are strong CA II inhibitors and are able to lower IOP in experimental animals more than the parent derivatives. Our conclusion is that the tail conferring water solubility to such an enzyme inhibitor is more important for topical activity as an antiglaucoma drug, than the heterocyclic/aromatic ring to which the sulfonamido moiety is grafted.