Identification of Iminium Intermediates Generation in the Metabolism of Tepotinib Using LC-MS/MS: In Silico and Practical Approaches to Bioactivation Pathway Elucidation
Abstract
:1. Introduction
2. Results and Discussion
2.1. Results of In Silico TEP Metabolites Prediction
2.2. Results of In Silico TEP Structural Alerts Sites and Toxicity Prediction
2.3. Identification of In Vitro Phase I TEP Metabolites
2.3.1. TEP Fragmentation Pattern
2.3.2. M1 Fragment Ions
2.3.3. M2 Fragment Ions
2.3.4. M3 Fragment Ions
2.3.5. M4 Fragment Ions
2.4. Reactive Metabolites
2.4.1. TEPCN518 Fragment Ions
2.4.2. TEPCN520 Fragment Ions
2.4.3. TEP534a and TEP534b Fragment Ions
2.5. Proposed TEP Bioactivation Mechanism
3. Chemicals and Methods
3.1. Chemicals
3.2. Chromatography Conditions
3.3. In Silico Prediction of TEP Metabolism Using WhichP450TM Module of StarDrop Software
3.4. In Silico Prediction of the Toxicity of TEP Metabolites and Reactivity Using DEREK Software and Xenosite Reactivity Model
3.5. HLM Incubation
3.6. Identification of TEP-Reactive Metabolites
4. Conclusions
Supplementary Materials
Supplementary File 1Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
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TEP and Its Phase I Metabolites | Phospholipidosis | Skin Sensitization | Nephrotoxicity | Chromosome Damage, Teratogenicity, Carcinogencity, Mutagenicity and Genotoxicity |
---|---|---|---|---|
Piperidine | Hydrazine | Halogenated nitrile | ||
TEP | Plausible | Equivocal | Equivocal | NA |
M1 | NA * | Plausible | Equivocal | NA |
M2 | NA | Equivocal | Equivocal | NA |
M3 | NA | Equivocal | Equivocal | NA |
M4 | NA | Equivocal | Equivocal | NA |
MS Scan | Fragment Ions | Elution Time (min) | Metabolic and Bioactivation Pathways | |
---|---|---|---|---|
TEP | 493 | 112 | 25.1 | Main drug |
Phase-I Metabolites | ||||
M1 | 493 | 382, 296, 185, 112, 72 | 26.6 | N-demethylation and α-oxidation at piperidine ring |
M2 | 479 | 382, 185, 98 | 24.6 | N-demethylation at piperidine ring |
M3 | 507 | 310, 126 | 28.3 | α-Oxidation at piperidine ring |
M4 | 509 | 128 | 28.4 | α-Hydroxylation at piperidine ring |
Reactive Metabolites | ||||
TEPCN518 | 518 | 491, 459, 137, 110, 104 | 39.2 | Cyano attack at bioactivated α-carbon of piperidine ring |
TEPCN520 | 520 | 493, 475, 278, 139 | 42.9 | N-demethylation, hydroxylation and cyano attack at bioactivated α-carbon of piperidine ring |
TEPCN534a | 534 | 516, 475, 296, 153 | 28.9 | Hydroxylation and cyano addition at α-carbon of piperidine ring |
TEPCN534b | 534 | 491, 382, 210, 153, 110 | Hydroxylation at methylene carbon attached to piperidine ring and cyano attack at bioactivated α-carbon of piperidine ring |
Sample Availability: Samples of the compounds are not available from the authors. |
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Abdelhameed, A.S.; Attwa, M.W.; Kadi, A.A. Identification of Iminium Intermediates Generation in the Metabolism of Tepotinib Using LC-MS/MS: In Silico and Practical Approaches to Bioactivation Pathway Elucidation. Molecules 2020, 25, 5004. https://doi.org/10.3390/molecules25215004
Abdelhameed AS, Attwa MW, Kadi AA. Identification of Iminium Intermediates Generation in the Metabolism of Tepotinib Using LC-MS/MS: In Silico and Practical Approaches to Bioactivation Pathway Elucidation. Molecules. 2020; 25(21):5004. https://doi.org/10.3390/molecules25215004
Chicago/Turabian StyleAbdelhameed, Ali S., Mohamed W. Attwa, and Adnan A. Kadi. 2020. "Identification of Iminium Intermediates Generation in the Metabolism of Tepotinib Using LC-MS/MS: In Silico and Practical Approaches to Bioactivation Pathway Elucidation" Molecules 25, no. 21: 5004. https://doi.org/10.3390/molecules25215004