Covalent Binding Mechanism of Furmonertinib and Osimertinib With Human Serum Albumin
Open accessAbstract
As third-generation tyrosine kinase inhibitors, furmonertinib and osimertinib exhibit better efficacy than first- and second-generation tyrosine kinase inhibitors in patients with advanced non-small cell lung cancer. However, radioactive pharmacokinetics studies showed that parent-related components remain in human plasma for at least 21 days after oral administration. Similar pharmacokinetic profiles were found in pyrotinib and neratinib, which have been identified to covalently bind with human serum albumin at Lys-190, leading to low extraction recovery in protein precipitation. However, the binding mechanism of furmonertinib and osimertinib in human plasma has not been confirmed. Comprehensive techniques were used to investigate the mechanism of this binding, including ultra high-performance liquid chromatography coupled with high-resolution mass spectrometry and online/offline radioactivity profiling. SDS-PAGE and further autoradiography were also used to detect drug-protein adducts. We found that most furmonertinib exists in the human plasma following ex vivo incubation in the form of protein-drug adducts. Only lysine-furmonertinb adducts were found in pronase digests. A standard reference of lysine-furmonertinib was synthesized and confirmed by NMR. Through peptide mapping analysis, we confirmed that furmonertinib almost exclusively binds with human serum albumin (HSA) in plasma following ex vivo incubation, via Michael addition at Lys-195 and Lys-199, instead of Lys-190. Two peptides found to bond with furmonertinib were ASSAKQR and LKCASLQK. Osimertinib was also found to bond with Lys-195 and Lys-199 of HSA via peptide mapping analysis. SIGNIFICANCE STATEMENT: Here we report that furmonertinib and osimertinib can covalently bind with human serum albumin at the site of Lys-195 and Lys-199 instead of Lys-190, potentially leading to the long duration of drug-protein adducts in the human body.
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This enhances alflutinib's activity and kinase selectivity while reducing off-target effects, leading to fewer side effects.
“This enhances alflutinib’s activity and kinase selectivity while reducing off-target effects, leading to fewer side effects.146–148”
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- Authors
- Wu, Y., Chen, L., Chen, J., Xue, H., He, Q., Zhong, D., & Diao, X.
- Affiliations
- Shanghai Medical College of Fudan University (CN)
- Venue
- Drug Metabolism and Disposition
- Publisher
- Elsevier BV
- Published
- 2023-01
- Volume
- 51
- Issue
- 1
- Pages
- 8-16
- ISSN
- 0090-9556 (ISSN-L)
- OpenAlex
- W4308183940
- Cited by
- 21
- References
- 22
- Citations by year
- 2026: 3 · 2025: 9 · 2024: 5 · 2023: 4
- FWCI
- 1.47
- Type
- journal article
- Language
- en
- Open access
- bronze
- doi.org
- Locations
- Drug Metabolism and Disposition (publishedVersion)
- PubMed (publishedVersion)
- Funders
- National Natural Science Foundation of China (81903701)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein Interaction Studies and Fluorescence Analysis
- Life Sciences › Pharmacology, Toxicology and Pharmaceutics › Pharmacology › Pharmacogenetics and Drug Metabolism
- Physical Sciences › Chemistry › Analytical Chemistry › Analytical Methods in Pharmaceuticals
- Keywords
- furmonertinib, osimertinib, human serum albumin, Michael addition, lysine adducts, non-small cell lung cancer, tyrosine kinase inhibitors, plasma protein binding, drug-protein adducts, pharmacokinetics
- MeSH terms
- Serum Albumin, Human, Acrylamides, Aniline Compounds, Carcinoma, Non-Small-Cell Lung, Humans, Indoles, Lung Neoplasms, Lysine, Pyridines, Pyrimidines, Serum Albumin
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