Circadian rhythms in CYP2A5 expression underlie the time-dependent effect of tegafur on breast cancer

The chemotherapeutic agent tegafur, a prodrug that prolongs the half-life of fluorouracil (5-FU), exerts antitumor effects against various cancers. Since tegafur is metabolized to 5-FU by CYP2A6 in the liver, the expression of CYP2A6 determines the effect of tegafur. Here, we report that the expression rhythm of Cyp2a5, a homolog of human CYP2A6, in female mice causes dosing time-dependent differences in tegafur metabolism. In the livers of female mice, CYP2A5 expression showed a circadian rhythm, peaking during the dark period. This rhythm is regulated by RORA, a core clock component, and abrogation of the CYP2A5 activity abolished the time-dependent difference in the rate of tegafur metabolism in female mice. Furthermore, administration of tegafur to mice transplanted with 4T1 breast cancer cells during the dark period suppressed increases in tumor size compared to female mice treated during the light period. Our findings reveal a novel relationship between 5-FU prodrugs and circadian clock machinery, potentially influencing antitumor effects, and contributing to the development of time-aware chemotherapy regimens for breast cancer. •Tegafur is a prodrug that is metabolized in the body to 5-fluorouracil by CYP2A6.•CYP2A5 function in the liver of BALB/c female mice exhibits circadian variation.•Antitumor effect of tegafur on female mice implanted with 4T1 cells is influenced by administration time.•Tegafur treatment protocols for breast cancer should consider the timing of administration to maximize effect.