A New Marmoset P450 4F12 Enzyme Expressed in Small Intestines and Livers Efficiently Metabolizes Antihistaminic Drug Ebastine

Abstract

Common marmosets (Callithrix jacchus) are attracting attention as animal models in preclinical studies for drug development. However, cytochrome P450s (P450s), major drug-metabolizing enzymes, have not been fully identified and characterized in marmosets. In this study, based on the four novel P450 4F genes found on the marmoset genome, we successfully isolated P450 4F2, 4F3B, 4F11, and 4F12 cDNAs in marmoset livers. Deduced amino acid sequences of the four marmoset P450 4F forms exhibited high sequence identities (87%-93%) to the human and cynomolgus monkey P450 4F homologs. Marmoset P450 4F3B and 4F11 mRNAs were predominantly expressed in livers, whereas marmoset P450 4F2 and 4F12 mRNAs were highly expressed in small intestines and livers. Four marmoset P450 4F proteins heterologously expressed in Escherichia coli catalyzed the v-hydroxylation of leukotriene B 4 . In addition, marmoset P450 4F12 effectively catalyzed the hydroxylation of antiallergy drug ebastine, a human P450 2J/4F probe substrate. Ebastine hydroxylation activities by small intestine and liver microsomes from marmosets and cynomolgus monkeys showed greatly higher values than those of humans. Ebastine hydroxylation activities by marmoset and cynomolgus monkey small intestine microsomes were inhibited (approximately 60%) by anti-P450 4F antibodies, unlike human small intestine microsomes, suggesting that contribution of P450 4F enzymes for ebastine hydroxylation in the small intestine might be different between marmosets/ cynomolgus monkeys and humans. These results indicated that marmoset P450 4F2, 4F3B, 4F11, and 4F12 were expressed in livers and/or small intestines and were functional in the metabolism of endogenous and exogenous compounds, similar to those of cynomolgus monkeys and humans.

Utility of non-human primates in drug development: Comparison of non-human primate and human drug-metabolizing cytochrome P450 enzymes

“…On the other hand, both marmoset P450 4F12 and cynomolgus monkey P450 4F12, expressed in small intestines and livers, more efficiently metabolized ebastine oxidation than marmoset and monkey P450 2J2 did [33]. These findings indicated that some marmoset or cynomolgus monkey P450 enzymes had apparently different functional characteristics to those of humans.…”

Expression of Transcripts in Marmoset Oocytes Retrieved during Follicle Isolation Without Gonadotropin Induction

“…Non-human primates can be classified into old-world and new-world monkeys [29]; the marmoset belongs to the class of new-world monkeys. Their features and application to various biomedical research studies have been mainly conducted in Japan, and recently, the wider usage of marmoset as a model is emerging [23,30,31,32,33,34]. From the perspective of reproductive physiology, non-human primates are closely related to human models; however, due to the high cost of the supply and maintenance of marmosets, a limited number of researchers are using this NHP model.…”

Utility of Common Marmoset (Callithrix jacchus) Embryonic Stem Cells in Liver Disease Modeling, Tissue Engineering and Drug Metabolism

“…To date, 36 C. jacchus CYP isoenzymes (cjCYP) have been identified and 24 of them, belonging to 1A, 2A, 2B, 2C, 2D, 2E, 3A, 4A, and 4F subfamilies, share a high degree of homology in the cDNA (>89%) and amino acid sequences (>85%) with corresponding human P450s [ 180 ]. Among these, CYP1A2, 2A6, 2B6, four 2C subfamily members (2C8, 2C18, 2C19 and 2C58), 2D6, 2D8, 2E1, 2J2, 3A4, 3A5, 4A11, 4F2, 4F12 and 7B1 are expressed in the marmoset liver [ 181 , 182 , 183 , 184 , 185 , 186 , 187 , 188 , 189 , 190 , 191 ]. Interestingly, CYP1D1 , encoded by a pseudogene in human liver is also pseudogenized in C. jacchus due to an incomplete open reading frame [ 180 ].…”