Metabolite Profiling and Characterization of LW6, a Novel HIF-1α Inhibitor, as an Antitumor Drug Candidate in Mice
Abstract
:1. Introduction
2. Results
2.1. Fragmentation of LW6
2.2. Metabolite Profiling of LW6 in Serial Mouse Plasma
2.3. Time Profiling and Relative Quantification of LW6 and its Metabolites
3. Discussion
4. Materials and Methods
4.1. Chemicals and Reagents
4.2. Animal Study
4.3. Sample Preparation
4.4. Chromatographic and Mass Spectrometric Methods
4.5. Predicted MRM-Enhanced Product Ion Scan (pMRM-EPI) Method
4.6. Data Processing Method and Set Criteria for Metabolite Hits
4.7. Semi-Quantitation of Targeted Metabolites
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
Sample Availability
References
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Metabolite | Retention Time (min) | [M-H]− | Major Fragments |
---|---|---|---|
LW6 (parent) | 17.46 | 434 | 227, 206,166 |
APA (M7) | 15.73 | 285 | 241, 227 |
M1 | 2.84 | 342 | 166, 151, 117 |
M2 | 10.62 | 301 | 257, 243 |
M3 | 10.96 | 626 | 450, 243, 206, 113, 85 |
M4 | 13.22 | 450 | 243, 206, 178, 166 |
M5 | 13.37 | 450 | 392, 243, 206, 178, 166 |
M6 | 14.59 | 610 | 434, 227, 206, 178, 166, 85 |
M8 | 15.76 | 461 | 285, 227 |
M9 | 16.09 | 420 | 227, 192 |
M10 | 16.81 | 450 | 227, 222, 180 |
M11 | 17.49 | 436 | 227, 206, 179, 166 |
M12 | 17.48 | 227 | 227, 169 |
Metabolite | i.v. | p.o. | ||||
---|---|---|---|---|---|---|
Average AUC0→t (Peak Intensity, Cps) | Relative % of Parent AUC0→t (i.v.) 1 | Relative % of Total Metabolite AUC0→t 2 | Average AUC0→t (Peak inTensity, cps) | Relative % of Parent AUC0→t (p.o.) 1 | Relative % of Total Metabolite AUC0→t 2 | |
LW6 (Parent) | 1,048,883 | 100.00 | - | 28,735 | 2.74 | - |
APA (M7) | 19,697,150 | 1877.92 | 92.09 | 17,688,080 | 1686.37 | 93.32 |
M1 | 42,982 | 4.10 | 0.20 | 66,545 | 6.34 | 0.35 |
M2 | 653,800 | 62.33 | 3.06 | 457,710 | 43.64 | 2.41 |
M3 | 23,904 | 2.28 | 0.11 | 10,404 | 0.99 | 0.05 |
M4 | 170,950 | 16.30 | 0.80 | 10,340 | 0.99 | 0.05 |
M5 | 116,583 | 11.11 | 0.55 | 11,845 | 1.13 | 0.06 |
M6 | 2445 | 0.23 | 0.01 | 1127 | 0.11 | 0.01 |
M8 | 622,040 | 59.30 | 2.91 | 708,375 | 67.54 | 3.74 |
M9 | 7672 | 0.73 | 0.04 | - | - | - |
M10 | 511 | 0.05 | 0.00 | - | - | - |
M11 | 17,015 | 1.62 | 0.08 | - | - | - |
M12 | 31,345 | 2.99 | 0.15 | - | - | - |
Type of Biotransformation | [M-H]− m/z | pMRM |
---|---|---|
Parent (LW6) | 434 | 434 > 227 |
Mono-oxidation | 450 | 450 > 227, 450 > 243 |
Di-oxidation | 466 | 466 > 227, 466 > 243 |
Glucuronidation | 610 | 610 > 227, 610 > 243 |
Mono-oxidation + glucuronidation | 626 | 626 > 450, 626 > 243 |
Ester hydrolysis | 420 | 420 > 227 |
Ester hydrolysis + mono-oxidation | 436 | 436 > 227, 436 > 243 |
Ester hydrolysis + mono-oxidation + glucuronidation | 612 | 612 > 227, 612 > 243 |
Ester hydrolysis + glucuronidation | 596 | 596 > 420, 596 > 227 |
Amide hydrolysis (amine part) | 166 | 166 > 166 |
Amide hydrolysis + ester hydrolysis | 152 | 152 > 152 |
Amide hydrolysis + glucuronidation | 342 | 342 > 166 |
Amide hydrolysis + ester hydrolysis + glucuronidation | 328 | 328 > 152 |
Amide hydrolysis (acid part) | 285 | 285 > 227 |
Amide hydrolysis + glucuronidation | 461 | 461 > 227 |
Amide hydrolysis + mono-oxidation | 301 | 301 > 243 |
Amide hydrolysis + mono-oxidation + glucuronidation | 477 | 477 > 301, 477 > 243 |
O-dealkylation (alcohol part) | 227 | 227 > 227 |
O-dealkylation + glucuronidation | 403 | 403 > 227 |
O-dealkylation + mono-oxidation | 243 | 243 > 243 |
O-dealkylation + mono-oxidation + glucuronidation | 419 | 419 > 243 |
O-dealkylation (aldehyde part) | 222 | 222 > 222 |
O-dealkylation + glucuronidation | 398 | 398 > 222 |
O-dealkylation + aldehyde oxidation | 238 | 238 > 238 |
O-dealkylation + aldehyde oxidation + glucuronidation | 414 | 414 > 238 |
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Lee, K.; Lee, J.-Y.; Lee, K.; Jung, C.-R.; Kim, M.J.; Kim, J.A.; Yoo, D.G.; Shin, E.J.; Oh, S.J. Metabolite Profiling and Characterization of LW6, a Novel HIF-1α Inhibitor, as an Antitumor Drug Candidate in Mice. Molecules 2021, 26, 1951. https://doi.org/10.3390/molecules26071951
Lee K, Lee J-Y, Lee K, Jung C-R, Kim MJ, Kim JA, Yoo DG, Shin EJ, Oh SJ. Metabolite Profiling and Characterization of LW6, a Novel HIF-1α Inhibitor, as an Antitumor Drug Candidate in Mice. Molecules. 2021; 26(7):1951. https://doi.org/10.3390/molecules26071951
Chicago/Turabian StyleLee, Kiho, Ji-Yoon Lee, Kyeong Lee, Cho-Rock Jung, Min Ju Kim, Jung Ah Kim, Dong Gu Yoo, Eun Jin Shin, and Soo Jin Oh. 2021. "Metabolite Profiling and Characterization of LW6, a Novel HIF-1α Inhibitor, as an Antitumor Drug Candidate in Mice" Molecules 26, no. 7: 1951. https://doi.org/10.3390/molecules26071951