Immunochemical Detection of Cytochrome P450 Enzymes in Small Intestine Microsomes of Male and Female Untreated Juvenile Cynomolgus Monkeys

Overview

Journal Xenobiotica

Publisher Informa Healthcare

Specialties Biochemistry
Toxicology

Date 2014 Mar 6

PMID 24593267

Citations 4

Authors

Shotaro Uehara

Norie Murayama

Yasuharu Nakanishi

Chika Nakamura

Takanori Hashizume

Darryl C Zeldin

Hiroshi Yamazaki

Yasuhiro Uno

Affiliations

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Abstract

The expression of small intestinal cytochromes P450 (P450s) has not been systematically measured in cynomolgus monkeys, which are widely used in preclinical drug studies to predict pharmacokinetics and toxicity in humans: therefore, P450 content of small intestine was quantified in 35 cynomolgus monkeys by immunoblotting using 11 selective antibodies. CYP2D, CYP2J2, CYP3A4 and CYP3A5 were detected in all 35 animals, while CYP1A and CYP2C9/19 were detected in 31 and 17 animals, respectively. CYP2C9 and CYP2C19 were detected with the same antibody. CYP1D, CYP2A, CYP2B6, CYP2C76 and CYP2E1 were not detected in any of the 35 animals examined. On analysis of pooled microsomes (35 animals), CYP3A (3A4+3A5) was most abundant (79% of total immunoquantified CYP1-3 proteins), followed by CYP2J2 (13%), CYP2C9/19 (4%), CYP1A (3%) and CYP2D (0.4%). On the analysis of individual microsome samples, each P450 content varied 2-to-6-fold between animals, and no sex differences were observed in any P450 content. These findings should help to increase the understanding of drug metabolism, especially the first-pass effect, in cynomolgus monkey small intestines.

Citing Articles

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Predicting Drug Extraction in the Human Gut Wall: Assessing Contributions from Drug Metabolizing Enzymes and Transporter Proteins using Preclinical Models.

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Immunochemical quantification of cynomolgus CYP2J2, CYP4A and CYP4F enzymes in liver and small intestine.

Uehara S, Murayama N, Nakanishi Y, Nakamura C, Hashizume T, Zeldin D Xenobiotica. 2014; 45(2):124-30.

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References

Nakanishi Y, Matsushita A, Matsuno K, Iwasaki K, Utoh M, Nakamura C . Regional distribution of drug-metabolizing enzyme activities in the liver and small intestine of cynomolgus monkeys. Drug Metab Pharmacokinet. 2011; 26(3):288-94. DOI: 10.2133/dmpk.DMPK-10-NT-101. View

Takahashi M, Washio T, Suzuki N, Igeta K, Fujii Y, Hayashi M . Characterization of gastrointestinal drug absorption in cynomolgus monkeys. Mol Pharm. 2008; 5(2):340-8. DOI: 10.1021/mp700095p. View

Ogasawara A, Utoh M, Nii K, Ueda A, Yoshikawa T, Kume T . Effect of oral ketoconazole on oral and intravenous pharmacokinetics of simvastatin and its acid in cynomolgus monkeys. Drug Metab Dispos. 2008; 37(1):122-8. DOI: 10.1124/dmd.108.022574. View

Shimada T, Yamazaki H, Mimura M, Inui Y, Guengerich F . Interindividual variations in human liver cytochrome P-450 enzymes involved in the oxidation of drugs, carcinogens and toxic chemicals: studies with liver microsomes of 30 Japanese and 30 Caucasians. J Pharmacol Exp Ther. 1994; 270(1):414-23. View

Nakanishi Y, Yamashita H, Yoshikawa T, Tominaga T, Nojiri K, Sunaga Y . Cytochrome P450 metabolic activities in the small intestine of cynomolgus macaques bred in Cambodia, China, and Indonesia. Drug Metab Pharmacokinet. 2013; 28(6):510-3. DOI: 10.2133/dmpk.dmpk-13-nt-031. View

King L, Ma J, Srettabunjong S, Graves J, Bradbury J, Li L . Cloning of CYP2J2 gene and identification of functional polymorphisms. Mol Pharmacol. 2002; 61(4):840-52. DOI: 10.1124/mol.61.4.840. View

Uno Y, Uehara S, Murayama N, Yamazaki H . CYP1D1, pseudogenized in human, is expressed and encodes a functional drug-metabolizing enzyme in cynomolgus monkey. Biochem Pharmacol. 2010; 81(3):442-50. DOI: 10.1016/j.bcp.2010.11.003. View

Uno Y, Hosaka S, Matsuno K, Nakamura C, Kito G, Kamataki T . Characterization of cynomolgus monkey cytochrome P450 (CYP) cDNAs: is CYP2C76 the only monkey-specific CYP gene responsible for species differences in drug metabolism?. Arch Biochem Biophys. 2007; 466(1):98-105. DOI: 10.1016/j.abb.2007.07.003. View

Nelson D, Zeldin D, Hoffman S, Maltais L, Wain H, Nebert D . Comparison of cytochrome P450 (CYP) genes from the mouse and human genomes, including nomenclature recommendations for genes, pseudogenes and alternative-splice variants. Pharmacogenetics. 2004; 14(1):1-18. DOI: 10.1097/00008571-200401000-00001. View

10.

Nakanishi Y, Matsushita A, Matsuno K, Iwasaki K, Utoh M, Nakamura C . Regional distribution of cytochrome p450 mRNA expression in the liver and small intestine of cynomolgus monkeys. Drug Metab Pharmacokinet. 2010; 25(3):290-7. DOI: 10.2133/dmpk.25.290. View

11.

Uno Y, Fujino H, Iwasaki K, Utoh M . Macaque CYP2C76 encodes cytochrome P450 enzyme not orthologous to any human isozymes. Curr Drug Metab. 2010; 11(2):142-52. DOI: 10.2174/138920010791110854. View

12.

Matsumoto S, Hirama T, Matsubara T, Nagata K, Yamazoe Y . Involvement of CYP2J2 on the intestinal first-pass metabolism of antihistamine drug, astemizole. Drug Metab Dispos. 2002; 30(11):1240-5. DOI: 10.1124/dmd.30.11.1240. View

13.

Wilkinson G . Drug metabolism and variability among patients in drug response. N Engl J Med. 2005; 352(21):2211-21. DOI: 10.1056/NEJMra032424. View

14.

Paine M, Hart H, Ludington S, Haining R, Rettie A, Zeldin D . The human intestinal cytochrome P450 "pie". Drug Metab Dispos. 2006; 34(5):880-6. PMC: 2222892. DOI: 10.1124/dmd.105.008672. View

15.

Uno Y, Iwasaki K, Yamazaki H, Nelson D . Macaque cytochromes P450: nomenclature, transcript, gene, genomic structure, and function. Drug Metab Rev. 2011; 43(3):346-61. DOI: 10.3109/03602532.2010.549492. View

16.

Lee C, Neul D, Clouser-Roche A, Dalvie D, Wester M, Jiang Y . Identification of novel substrates for human cytochrome P450 2J2. Drug Metab Dispos. 2009; 38(2):347-56. DOI: 10.1124/dmd.109.030270. View

17.

Omura T, Sato R . THE CARBON MONOXIDE-BINDING PIGMENT OF LIVER MICROSOMES. I. EVIDENCE FOR ITS HEMOPROTEIN NATURE. J Biol Chem. 1964; 239:2370-8. View

18.

Uno Y, Fujino H, Kito G, Kamataki T, Nagata R . CYP2C76, a novel cytochrome P450 in cynomolgus monkey, is a major CYP2C in liver, metabolizing tolbutamide and testosterone. Mol Pharmacol. 2006; 70(2):477-86. DOI: 10.1124/mol.106.022673. View

19.

Paine M, Schmiedlin-Ren P, Watkins P . Cytochrome P-450 1A1 expression in human small bowel: interindividual variation and inhibition by ketoconazole. Drug Metab Dispos. 1999; 27(3):360-4. View

20.

Nishimuta H, Sato K, Mizuki Y, Yabuki M, Komuro S . Species differences in intestinal metabolic activities of cytochrome P450 isoforms between cynomolgus monkeys and humans. Drug Metab Pharmacokinet. 2011; 26(3):300-6. DOI: 10.2133/dmpk.DMPK-10-SH-119. View