Identification of human metabolites for Phase 1 radiolabeled drugs
Identification of human drug metabolites via combined ULPC-hrMS/MS technology. Early-stage metabolite identification provides valuable insights for further studies (DDI, impairment) and prevents late-stage surprises that require new preclinical safety assessments. As an additional benefit, animal metabolism studies can be waived.
Eliminate the risk of discovering unexpected human unique metabolites in late-stage studies
Metabolite identification reveals which metabolites are formed in the human body after drug administration. Traditionally, finding out how a new drug asset metabolizes inside the human body happens in late-stage clinical studies. The industry relies heavily on animal models, which frequently fail to predict human pathways. Late-stage findings of human unique metabolites require new preclinical safety testing and may cause significant delays in drug development.
Peregrion’s microtracer 14C-labeled metabolite identification services can be done from Phase 1. At this stage timely mitigation strategies can be executed. Furthermore, early-stage metabolite profiling via AMS eliminates the need to conduct radioactive animal metabolite profiling studies, saving animals, and time and money. Early information on human metabolites also guides the design of DDI studies specific to the pathways involved in metabolism and excretion.
Examples where early-stage metabolite identification accelerates your clinical development programs
Formation of active metabolites
Your compound is expected to metabolize into fractions with pharmacological activity. Risk mitigation strategies can be designed promptly
Drug interactions affect excretion
Drug-drug interactions are suspected play a role in the excretion of your compound’s metabolites. Early insights on the nature and quantity of metabolites support efficient design of DDI studies
Metabolite clearance is impaired
Anticipated metabolites may be difficult to clear in populations with renal and hepatic impairments. Later-stage impairment studies can effectively be designed using data on human metabolites
Practical study information for absolute bioavailability
| Parameter | Peregrion Approach | Client Benefit |
|---|---|---|
| Typical cohort size | 6-8 participants | limited number of participants |
| Sample matrices | whole blood, plasma, urine, feces, exhaled air, bile and vomit | full quantitative picture of metabolite abundances per matrix |
| Radiation dose | ≤ 1 µCi; 100-1,000 lower than conventional studies | studies are approved as early as Phase 1, as the radiological exposure falls in ICRP Class I due to the low radiological exposure |
| Sensitivity range | accurate quantification and identification of metabolites with abundances as low as 1% | complete picture of all human metabolites, including the minor with potential pharmacological effects |
| Match of spectra | a single injection for metabolite quantification and identification by coupled UPLC-hrMS/MS + AMS system | 100% match between AMS and hrMS data; no issues with shifts in retention time |
| Study design | recommendations on position of radiolabel(s) and microtracer dosing | scientist to scientist interactions resulting in optimal study design for evaluating drug metabolism and excretion |
| Interindividual differences and time dependencies | flexibility on additional analysis, recommendations on studies for selected individual or timepoint samples | selection of relevant metabolites for further analyses |
Spectra of metabolite quantification and identification are fully matched by a single injection
Integrated results are generated from a single injection by coupling UPLC-hrMS/MS+AMS systems
Benefits of combining studies
- Eliminate the need for animals, time and money on animal metabolism studies
- Full mass balance and metabolite data package from a single sample
- 100% match between metabolite identification and quantification
- Decreased interindividual variability
Why Peregrion for human metabolite identification services
Guided by Scientific Experts
Jelle Reinen
Jelle has a background in pharmaceutical sciences and holds a PhD in molecular toxicology. He has worked as a Study Director in the pharmaceutical industry since 2015. He has 3 years’ experience in the AMS field with his main focus on metabolite profiling and identification and absolute bioavailability studies.
Ioana Barbu
Ioana has a background in analytical chemistry and physics and holds a PhD in mass spectrometry. She has 5 years experience as a scientist in AMS with her focus on metabolite profiling/identification, mass balance and absolute bioavailability studies
Lotte van Andel
Lotte brings a bioanalysis background and a PhD (Netherlands Cancer Institute/NKI) in conventional high-dose human ADME studies. With six years of total industry experience, she has spent three years working as a scientist and three years as a project manager. She currently also serves as Team Lead of the Research Technician group.
Related Services
Absolute Bioavailability
Information on absolute bioavailability can help interpret mass balance data and understand the overall drug elimination pathways. For instance, if a large fraction of the drug is found unchanged in feces, bioavailability data supports understanding of the role of biliary and/or gut wall secretion to drug elimination.
Metabolite Profiling
Information on absolute bioavailability can help interpret mass balance data and understand the overall drug elimination pathways. For instance, if a large fraction of the drug is found unchanged in feces, bioavailability data supports understanding of the role of biliary and/or gut wall secretion to drug elimination.
Metabolite Profiling
Information on absolute bioavailability can help interpret mass balance data and understand the overall drug elimination pathways. For instance, if a large fraction of the drug is found unchanged in feces, bioavailability data supports understanding of the role of biliary and/or gut wall secretion to drug elimination.
Feces homogenization
Information on absolute bioavailability can help interpret mass balance data and understand the overall drug elimination pathways. For instance, if a large fraction of the drug is found unchanged in feces, bioavailability data supports understanding of the role of biliary and/or gut wall secretion to drug elimination.
Support
FAQs
How do I start a microtracer study with Peregrion?
That’s easy. Contact us via the Contact Page on the website or reach out to your existing contact person at Peregrion. After initial exploratory conversations, we can arrange a CDA to freely talk about the project and your needs, and suggested steps forward. Our scientists will be involved at an early stage to develop optimal study designs. After submission and approval of our proposal we will agree on a Study Agreement or Master Services Agreement. Your dedicated Project Manager will then guide you through the practicalities of getting started. When there is clarity on the dosing date(s) we will reserve time slots in our facility. You can contact us any time, we are happy to help.
What type of excreta are used in radiolabeled studies?
Total radioactivity measurements are done with human plasma, urine, whole blood, feces, exhaled air, bile and vomit. Very low sample volumes are required, i.e. 5 uL of plasma for a total radioactivity analysis and 50 uL of plasma for the generation of PK data or metabolite profling data. Samples are introduced via an automated sample combustion device that generates CO2, which is directed to the AMS via an interface. A single sample is completed in less than 10 minutes. The speed of the analysis supports discharge studies from a clinical site. You will receive the study results within 48 hours after sample receipt.
Which data can be generated with Peregrion’s AMS-enabled studies?
- Absolute bioavailability data
- PK profile (for parent drug and known metabolites)
- Metabolite profile including metabolite identification
- Mass balance data
- Routes of excretion
- Information on the first pass effect
- Fraction absorbed
- Volume of distribution
What is a microdosing study in the context of a microtracer study?
A microdosing study includes a very low dose of the drug product (e.g. 100 µg) with a 14C microtracer. Due to the low dose (considered as an impurity) there is no toxicological concern and only a limited pre-clinical package is required to conduct such a study. This comes with the advantage that fewer lab animals are needed. Multiple drugs can be administered in the same study in parallel groups to aid in PK based candidate selection. A typical microdosing study consists of approximately 6 volunteers per drug. The microdose can be administered via any route, e.g. oral or intravenous.
Is human ADME data that is generated with microtracer studies acceptable for regulatory submissions?
The answer is ‘Yes’. We deliver data to our clients to support their regulatory submissions with regulatory-required or requested data such as human metabolism, routes of excretion, absolute bioavailability, and fraction absorbed. The FDA guideline on Safety Testing of Drug Metabolites Guidance for Industry (CDER) March 2020 Pharmacology/Toxicology recommends performing human in vivo metabolic evaluation as early as possible. The level of radioactivity in a microtracer study is only 0.1-1µCi. Conventional studies generally apply 100µCi. Due to the 100-fold lower radioactivity levels, ethical committees approve the use of 14C microtracers in early stage clinical development.
Why is 14C labeling required for safety studies?
Incorporating radioactivity in the drug molecule is needed to ascertain that all metabolites will be found in the systemic circulation. Since virtually all drugs contain carbon, the highest scientific standard practice is to synthesize the drug with incorporation of radioactive carbon-14, and to dose this radioactive material to a small number of healthy participants or patients in a hADME study. Samples from excreta and blood or plasma can be analyzed to trace the drug and its metabolites. Analysis via AMS is up to 10,000-fold more sensitive than classical methods, for instance Liquid Scintillation Counting (LSC) and therefore very small amounts of radioactivity are sufficient.
What are the combined advantages of microtracer studies in clinical development?
Overall, the microtracer-based mass balance, metabolite profiling and absolute bioavailability studies in early clinical developments give an enormous enrichment of the data package available at early stage without the need for separate studies to determine human ADME data. The data richness allows better study designs for follow up clinical studies, it allows earlier assessment and risk mitigation strategies for unexpected human unique metabolites, and it eliminates the need to conduct radioactive animal mass balance/metabolite profiling studies, where many animals, and much time and money are being spent on non-relevant animal metabolites.
