TY - JOUR

T1 - Development and Validation of a Method of Liquid Chromatography Coupled with Tandem Mass Spectrometry for Quantification of ST-246 (Tecovirimat) in Human Plasma

AU - Oleinik, Galina A.

AU - Koval, Vladimir V.

AU - Usova, Svetlana V.

AU - Shishkina, Larisa N.

AU - Chernonosov, Alexander A.

N1 - Funding Information: This research was supported by the Russian Science Foundation (grant no. 22-24-00697).We thank the State Research Center of Virology and Biotechnology VECTOR for providing blood plasma samples from volunteers who participated in the clinical trial of NIOCH-14 for the practical application of the presented method. We are grateful to A.Ya. Tikhonov and B.A. Selivanov for kindly providing the reagents ST-246 and 2-hydroxy-N-{3,5-dioxo-4-azatetracyclo [5.3.2.02.6.08.10]dodec-11-en-4-yl}-5-methylbenzamide. Publisher Copyright: © 2022 by the authors. Licensee MDPI, Basel, Switzerland.

PY - 2022/6/1

Y1 - 2022/6/1

N2 - The aim of this work was to develop and validate a sensitive and robust method of liquid chromatography coupled with tandem mass spectrometry to quantitate ST-246 (tecovirimat) in plasma using an internal standard (2-hydroxy-N-{3,5-dioxo-4-azatetracyclo [5.3.2.02.6.08.10]dodec-11-en-4-yl}-5-methylbenzamide). The method was validated in negative multiple reaction monitoring mode following recommendations of the European Medicines Agency for the validation of bioanalytical methods. The calibration curve for the analyte was linear in the 10–2500 ng/mL range with determination coefficient R2 > 0.99. Intra-and inter-day accuracy and precision for three concentrations of quality control were <15%. Testing of long-term stability of ST-246 (tecovirimat) in plasma showed no degradation at −20◦ C for at least 3 months. The method was applied to a clinical assay of a new antipoxvirus compound, NIOCH-14. Thus, the proposed method is suitable for therapeutic drug monitoring of ST-246 (tecovirimat) itself and of NIOCH-14 as its metabolic precursor.

AB - The aim of this work was to develop and validate a sensitive and robust method of liquid chromatography coupled with tandem mass spectrometry to quantitate ST-246 (tecovirimat) in plasma using an internal standard (2-hydroxy-N-{3,5-dioxo-4-azatetracyclo [5.3.2.02.6.08.10]dodec-11-en-4-yl}-5-methylbenzamide). The method was validated in negative multiple reaction monitoring mode following recommendations of the European Medicines Agency for the validation of bioanalytical methods. The calibration curve for the analyte was linear in the 10–2500 ng/mL range with determination coefficient R2 > 0.99. Intra-and inter-day accuracy and precision for three concentrations of quality control were <15%. Testing of long-term stability of ST-246 (tecovirimat) in plasma showed no degradation at −20◦ C for at least 3 months. The method was applied to a clinical assay of a new antipoxvirus compound, NIOCH-14. Thus, the proposed method is suitable for therapeutic drug monitoring of ST-246 (tecovirimat) itself and of NIOCH-14 as its metabolic precursor.

KW - LC-MS/MS

KW - MRM

KW - NIOCH-14

KW - plasma

KW - ST-246

KW - tecovirimat

KW - Plasma

KW - Reproducibility of Results

KW - Humans

KW - Tandem Mass Spectrometry/methods

KW - Isoindoles

KW - Chromatography, High Pressure Liquid/methods

KW - Chromatography, Liquid

KW - Benzamides

UR - http://www.scopus.com/inward/record.url?scp=85131620866&partnerID=8YFLogxK

UR - https://www.mendeley.com/catalogue/7951768d-a199-37e9-8fa5-2b84c5af2c5f/

U2 - 10.3390/molecules27113577

DO - 10.3390/molecules27113577

M3 - Article

C2 - 35684513

AN - SCOPUS:85131620866

VL - 27

JO - Molecules

JF - Molecules

SN - 1420-3049

IS - 11

M1 - 3577

ER -