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An<jats:italic>in vitro<\/jats:italic>CYP inhibition study in rat liver microsomes (RLM) was conducted using probe substrates for CYPs. Then, an<jats:italic>in vivo<\/jats:italic>pharmacokinetics of intravenous and oral buspirone (BUS), a probe substrate for CYP3A, was studied with the concurrent administration of oral GLY in rats. In the<jats:italic>in vitro<\/jats:italic>CYP inhibition study, CYP3A was involved in the metabolism of GLY. Moreover, GLY inhibited CYP3A activity with an IC<jats:sub>50<\/jats:sub>of 20.1\u2009\u00b1\u200910.7\u2009\u03bcM<jats:italic>via<\/jats:italic>a mixed inhibition mechanism. In the<jats:italic>in vivo<\/jats:italic>rat pharmacokinetic study, single oral GLY dose enhanced the area under plasma concentration\u2013time curve (AUC) of intravenous and oral BUS, but the extent of increase in AUC was only minimal (1.12\u20131.45 fold). These results indicate that GLY can inhibit the<jats:italic>in vitro<\/jats:italic>CYP3A\u2010mediated drug metabolism in RLM<jats:italic>via<\/jats:italic>a mixed inhibition mechanism. However, the impact of single oral GLY dose on the pharmacokinetics of BUS in rats was limited, showing that GLY could function as merely a weak inhibitor for CYP3A\u2010mediated drug metabolism<jats:italic>in vivo<\/jats:italic>. 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