Comparative inhibition of human alkaline phosphatase and diamine oxidase by bromo-levamisole, cimetidine and various derivatives

Analogues of bromo-levamisole and guanidine derivatives including cimetidine are examined in vitro in order to investigate their comparative inhibition, towards alkaline phosphatase (ALP) from human liver and diamine-oxidase (DAO) from human placenta. Bromo-levamisole, considered as a potent selective uncompetitive inhibitor of ALP (Ki, 2.8.10(-6) M at pH 10.5) is shown to be a noncompetitive inhibitor of DAO (Ki = 7.10(-4) M). According to the structure-inhibition relationship, the imidazole ring is important for ALP and DAO inhibition. The phenyl ring of bromo-levamisole is required for ALP inhibition but not for DAO inhibition, which is mediated mainly by aminoguanidine or guanidine groups. These results have allowed the selection of cimetidine, an H2-antagonist but also an immunomodulating compound, as inhibitor of these two enzymes. Cimetidine is an uncompetitive inhibitor of ALP (Ki = 3.2.10(-3) M at pH 10.5), and a good inhibitor of DAO (I50 = 3.8.10(-4) M). The Ki of ALP is commonly calculated at pH 10.5, but to study the role of the enzyme at the physiological pH, the inhibition has also been performed at pH 7.4. The Ki values are only slightly affected by this pH variation. So far several compounds, including levamisole, imidazole, theophylline and aminoguanidine are known to possess immunomodulating activities in vivo and/or in vitro and inhibit ALP and/or DAO. Therefore, it seems reasonable to assume that the inhibition of enzymes is involved in the immunomodulating effects of these drugs, when the ranges of active concentrations are similar for these properties.