P450 Enzymes: Answering Key Questions with INDIGO Biosciences’ qPCR-based assay
By Robert Deyes
Customer Solutions Manager, MyBio Ltd
Drug developers are very interested in a class of enzymes called the P450 (CYP) enzymes because of the critical role that they play, primarily in the liver, in metabolising drugs/xenobiotics (1,2). From a drug development standpoint, it is fundamentally important to understand not only what impact specific drugs can have on P450 enzyme activity, but how effectively the P450 isozymes clear drugs from the human body (Of the 57 known P450 isozymes, 6 of them are responsible for 90% of drug metabolism - CYP1A2, CYP2C9, CYP2C19, CYP2D6, CYP2E1, and CYP3A4 (1)).
The efficacy of drug treatment can be severely impacted if a specific drug is metabolized by P450 isozymes before it reaches a therapeutically beneficial level (1). In contrast, a drug that causes P450 isozyme levels to drop might lead to the unhealthy accumulation of other drugs, eventually to levels that are cytotoxic and detrimental to health (1).
P450 enzymes have been described as ‘the backbone of clinical research’ (1). Many of them are intimately associated with human disease (for example, Cyp 1B1 is linked to the onset of primary congenital glaucoma; Cyp 4A11 associated with hypertension and coronary artery disease (2)). In the words of one review, “one or several genes in the CYP1, CYP2, CYP3 or CYP4 families are shown to be significantly associated with increased risk of a particular human complex disease” (2). Assays directed at studying P450s should therefore be a key part of the toolbox for any laboratory that is focused on drug metabolism in the context of P450 regulation of expression and metabolism.
CYP enzymes are found in two cellular organelles: the Endoplasmic Reticulum (ER Membrane) and the Mitochondrion (1). The mitochondrial enzymes are responsible for metabolism of internal targets such as steroid hormones, whereas the ER Membrane- associated isozymes are responsible for drug metabolism (1). Activation and down-regulation occurs at the mRNA level (1), making qPCR methods of analysis valuable assets for monitoring CYP expression levels.
INDIGO Biosciences’ kit for Expression Profiling of Clinically Relevant CYPs contains optimized reagents for the culturing and treatment of upcyte® hepatocytes, and validated qPCR primers for quantifying drug-induced changes in the expression of CYP3A4, CYP1A1, CYP2B6, CYP2C8, CYP2C9, CYP2C19, and CYP2E1. For more information, visit: Expression Profiling of Clinically Relevant CYPs Assay Kit - Indigo Biosciences
References
Brian Gilani, Manouchkathe Cassagnol (2023), Biochemistry, Cytochrome P450 - StatPearls - NCBI Bookshelf
Daniel W. Nebert, Kjell Wikvall, and Walter L. Miller (2018) Human cytochromes P450 in health and disease, Philos Trans R Soc Lond B Biol Sci, 2013 Feb 19;368(1612):20120431. doi: 10.1098/rstb.2012.0431
