A series of novel 1,4-dioxane analogues of the muscarinic acetylcholine receptor (mAChR) antagonist <b>2</b> was synthesized and studied for their affinity at M<sub>1</sub>-M<sub>5</sub> mAChRs. The 6-cyclohexyl-6-phenyl derivative <b>3b</b>, with a <i>cis</i> configuration between the CH<sub>2</sub>N<sup>+</sup>(CH<sub>3</sub>)<sub>3</sub> chain in the 2-position and the cyclohexyl moiety in the 6-position, showed p<i>K</i><sub>i</sub> values for mAChRs higher than those of <b>2</b> and a selectivity profile analogous to that of the clinically approved drug oxybutynin. The study of the enantiomers of <b>3b</b> and the corresponding tertiary amine <b>33b</b> revealed that the eutomers are (2<i>S</i>,6<i>S</i>)-(-)-<b>3b</b> and (2<i>S</i>,6<i>S</i>)-(-)-<b>33b</b>, respectively. Docking simulations on the M<sub>3</sub> mAChR-resolved structure rationalized the experimental observations. The quaternary ammonium function, which should prevent the crossing of the blood-brain barrier, and the high M<sub>3</sub>/M<sub>2</sub> selectivity, which might limit cardiovascular side effects, make <b>3b</b> a valuable starting point for the design of novel antagonists potentially useful in peripheral diseases in which M<sub>3</sub> receptors are involved.