In Vitro metabolism
In vitro metabolic screening offers a cost-effective and efficient way to evaluate compound metabolism during the hit-to-lead and lead optimisation stages of discovery. These tests help to narrow down the number of clinically relevant compounds selected for more extensive – and costly – in vivo PK profiling.
We can provide metabolic stability assessments in both microsomes and hepatocytes. They can be tailored to answer specific questions using relevant co-factors, as required.
We also offer a cross-species metabolic profiling comparison in human and preclinical hepatocytes. This has two important benefits. First, it increases confidence around the choice of toxicology species when moving to early development. And second, it can flag up in vitro human specific metabolites (HSMs) or any disproportionate human metabolites (DHMs) that might prove problematic as a molecule moves towards candidate nomination and development. Gaining maximum value from these data is important when de-risking a development candidate. Being aware of potential difficulties well ahead of time will greatly assist in mitigating risks, as dictated by industrial guidelines such as ICH M3(R2) and FDA Metabolites in Safety Testing (MIST).
If the metabolites that are formed in preclinical species are identified, they can give crucial insights to inform the later stages of drug discovery. They can provide critical information about how a molecule might be metabolised, and the routes by which it might be eliminated. These data can be particularly useful in elucidating PK/PD disconnects. They also provide valuable information to drive discussions around efficacy.
By combining relevant analytical approaches, including high-resolution mass spectrometry with highly skilled manual data interpretation and chemically intelligent software algorithms, we are able to enhance metabolic coverage and provide greater confidence in the characterisation of metabolites in plasma, urine, blood and bile samples.
Drug-induced toxicity is one of the main causes of failure in the clinic. The likelihood that a compound might cause these problems can be assessed via in vitro experiments using specific trapping reagents, and subsequent detection and characterisation of these reactive metabolites.
Tailored hepatocyte incubations can be used to stabilise potential reactive metabolites using glutathione, which may indicate potential liabilities around reactivity. These metabolites are detected and characterised using non-targeted high-resolution mass spectrometry, which give the ability to find metabolites with high selectivity based on chemical functionality and at trace levels, where often the initial detection of these trapped intermediates can be made more difficult by traditional targeted methods. This approach gives wider coverage and provides a more accurate assessment of overall liabilities around potential reactivity in a time-efficient manner. Subsequent structural characterisation of key metabolites can be performed, where relevant and where feasible, and forms part of the ongoing conversation to maximise impact of these data.
Where characterisation of a metabolite is required, either unequivocally or in more detail than high-resolution mass spectrometric analysis can provide, further analysis by nuclear magnetic resonance (NMR) spectroscopy can be instigated to give greater insights into the site of biotransformation, where feasible. These in-depth studies can provide early, unequivocal data around metabolite structure that can better inform PK/PD relationships, and inform early assessments around metabolite safety.
We have access to several cryoprobe enabled 500 – 800 MHz Bruker spectrometer, and in-house expertise to analyse the data it generates. Our scientific leadership in the field enables us to suggest confident, data-led outcomes in the context of the questions being asked.
We offer a tailored service to meet our customers’ needs. It is very much a conversation – we will listen to what they require and suggest how we can use all the techniques and tools at our disposal to achieve that. The process is always led by high-quality science, and it is important to focus on the question we are trying to answer. The starting point is a discussion about what is feasible and aligning this with realistic expectations.
Please contact us to discuss your needs, and how we can help you meet them.