Male fertility depends on sperm motility, which enables sperm to traverse the female reproductive tract to the site of fertilization. Soluble adenylyl cyclase (sAC) is a key regulator of sperm motility defining it as a promising non-hormonal contraceptive target for women and men. To evaluate the efficacy of sAC inhibitors under conditions mimicking the female reproductive tract, we employed a capillary assay using cervical mucus isolated from humans or cows as well as methylcellulose, a viscoelastic liquid that simulates cervical mucus. Addition of a sAC inhibitor either directly to semen, modeling an on-demand male contraceptive, or to the cervical mucus, approximating a female contraceptive, effectively blocked sperm migration. These studies provide proof-of-concept for the efficacy of sAC inhibitors under physiologically relevant conditions.