Creative Biolabs has worked extensively on a wide variety of metabolism studies. As part of our in vitro metabolism service, we provide reactive metabolite analysis service for screening and assessing reactive metabolite of drugs.
Reactive metabolites are those metabolites which may be biologically active and confer to idiosyncratic adverse drug reactions (often associated with drug-induced skin, liver and hematopoietic toxicities) and/or pharmacology effects. Reactive metabolites may have positive effects. For instance, in pro-drugs, the administered parent compound is inactive. After a pro-drug entering to the body, the inactive drug is metabolized to become the pharmacologically active molecule. This process is called metabolic activation. However, in many cases, reactive metabolites are considered to be a key component of many idiosyncratic adverse drug reactions. As reactive metabolites are electrophilic species, it can bind covalently to macromolecules such as proteins and DNA, affecting their function and leading to toxicity. In addition, the reactive metabolites of a drug might stimulate innate immunity, or might bind to protein to generate neoantigens for cellular and humoral responses.
Creative Biolabs provides reactive metabolites characterization service to avoid unexpected idiosyncratic drug-induced toxicity. Due to the instincts of high reactivity and short half-life, reactive metabolites cannot be directly detected by traditional analytical methods. We provide detecting method by nucleophilic chemical trapping such as glutathione tri-peptide (GSH; most often used), potassium cyanide (KCN; used for hard electrophiles) or semicarbazide, which can form stable adducts with many reactive species and then detected by UPLC-TOFMS methods. Multivariate data analysis (MDA) was then conducted to screen trapped reactive metabolites.
Time-of-Flight Mass Spectrometry (TOFMS)
TOFMS is a method of mass spectrometry in which an ion's mass-to-charge ratio is determined via a time measurement. This method enables simultaneously acquiring in-source fragment ion data, providing structural determination of the detected trapped reactive metabolites. The detection sensitivity of scanning mode data is better than those of triple quadrupole MS.
Immune Activation by Drugs/Reactive Metabolites
Drugs or their reactive metabolites might stimulate innate immunity. Traditionally, drugs are thought to activate T cells via 2 pathways. One is covalently formed drug-protein adducts, followed by protein processing and release of drug-modified peptides that directly bind to MHC molecules prior to triggering T cells. The other way is direct binding to the peptide-loaded MHC molecules to form drug-MHC binding interaction and trigger T cell responses. Creative Biolabs provides service to evaluate idiosyncratic drug hypersensitivity reactions.
Figure 1. Integrated Reactive Metabolite Risk Assessments (Thompson et al. 2016)
Toxicity Testing
Creative Biolabs are pleased to introduce our impeccable service portfolio of drug in vitro toxicity and in vivo toxicity test to evaluate the toxicology potential of drug active metabolites for early lead optimization. For in vitro toxicity assays, we provide HPH and HepG2 cell line as hepatotoxicity evaluating models to minimize liability of drug-induced liver injuries. We also provide gene toxicity and cardiotoxicity test for drugs/active metabolites. For in vivo toxicity assays, we provide species of mice, rats, rabbits, guinea pigs, dogs, mini pigs and nonhuman primates for toxicity testing.
For more detailed information, please feel free to contact us or directly sent us an inquiry.
Reference
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