Genetic variants of CYP3A4 and CYP3A5 in cynomolgus and rhesus macaques.
Cynomolgus and rhesus macaques are frequently used in preclinical trials due to their close evolutionary relationships to humans. We conducted an initial screening for genetic variants in cynomolgus and rhesus macaque genes orthologous to human CYP3A4 and CYP3A5. Genetic screening of 78 Indochinese and Indonesian cynomolgus macaques and 34 Chinese rhesus macaques revealed a combined total of 42 CYP3A4 genetic variants, including 12 nonsynonymous variants, and 34 CYP3A5 genetic variants, including nine nonsynonymous variants. Four of these nonsynonymous variants were located at substrate recognition sites or the heme-binding region, domains essential for protein function, including c.886G>A (V296M) and c.1310G>A (S437N) in CYP3A4 and c.1437C>G (N479K) and c.1310G>C (T437S) in CYP3A5. The mutant proteins of these genetic variants were expressed in Escherichia coli and purified. Metabolic activity of these proteins measured using midazolam and nifedipine as substrates showed that none of these protein variants substantially influences the drug-metabolizing capacity of CYP3A4 or CYP3A5 protein. In Indonesian cynomolgus macaques, we also found IVS3+1delG in CYP3A4 and c.625A>T in CYP3A5, with which an intact protein cannot be produced due to a frameshift generated. Screening additional genomes revealed that two of 239 animals and three of 258 animals were heterozygous for IVS3+1delG of CYP3A4 and c.625A>T of CYP3A5, respectively. Some genetic variants were unevenly distributed between Indochinese and Indonesian cynomolgus macaques and between cynomolgus and rhesus macaques. Information on genetic diversity of macaque CYP3A4 and CYP3A5 presented here could be useful for successful drug metabolism studies conducted in macaques.
Uno
Yasuhiro
Y
Genome Research Group, Pharmacokinetics and Bioanalysis Center, Shin Nippon Biomedical Laboratories, Ltd., 16-1 Minami Akasaka, Kainan, Wakayama 642-0017, Japan. uno-yasuhiro@snbl.co.jp
Matsushita
Akinori
A
Osada
Naoki
N
Uehara
Shotaro
S
Kohara
Sakae
S
Nagata
Ryoichi
R
Fukuzaki
Koichiro
K
Utoh
Masahiro
M
Murayama
Norie
N
Yamazaki
Hiroshi
H
D000818:Animals; D001210:Asia, Southeastern; D002681:China; D051544:Cytochrome P-450 CYP3A / Q000235:genetics* / Q000378:metabolism; D004247:DNA / Q000235:genetics / Q000302:isolation & purification; D014644:Genetic Variation; D006801:Humans; D007438:Introns / Q000235:genetics; D007527:Isoenzymes / Q000235:genetics / Q000378:metabolism; D008252:Macaca fascicularis / Q000235:genetics* / Q000378:metabolism; D008253:Macaca mulatta / Q000235:genetics* / Q000378:metabolism; D008874:Midazolam / Q000378:metabolism; D009543:Nifedipine / Q000378:metabolism; D011994:Recombinant Proteins / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017422:Sequence Analysis, DNA
D007527:Isoenzymes; D011994:Recombinant Proteins; D004247:DNA; D051544:Cytochrome P-450 CYP3A; D009543:Nifedipine; D008874:Midazolam