Metabolic Stability Assessment of Larotrectinib Using Liquid Chromatography Tandem Mass Spectrometry
Mohamed W Attwa1,2, Adnan A Kadi1, Hany W Darwish1,3
1Department of Pharmaceutical Chemistry, College of Pharmacy, King Saud University, Riyadh 11451, Saudi Arabia.
Introduction:
Larotrectinib (VITRAKVI) is an orally potent tropomyosin receptor kinase (Trk) inhibitor that acts by competitive inhibition of all corresponding receptor kinases. It demonstrated a marked response rate (75%) and robust anticancer activity in Trk fusion-positive patients. This response is independent of cancer type, age and gender.
Methods:
In this study, an efficient and accurate LC-MS/MS analytical method was developed for Larotrectinib (LRB) quantification in addition to evaluation of its metabolic stability. LRB and lapatinib (LTP) (which is chosen as an internal standard; IS) were eluted utilizing an isocratic mobile phase with a reversed phase elution system (C18 column).
Results And Discussion:
The linearity range of the established method was 5-500 ng/mL (r 2 ≥ 0.9999) in the human liver microsomes (HLMs) matrix. Various parameters were calculated to validate the method sensitivity (limit of quantification was 5 ng/mL) and reproducibility (inter and intra-day accuracy and precision were below 3% in all samples) of our methodology. For evaluation of LRB metabolic stability in HLMs matrix, in vitro half-life (48.8 min) and intrinsic clearance (14.19 µL/min/mg) were computed.
Conclusion:
Accordingly, we can conclude that LRB is a moderate extraction ratio drug when compared with other tyrosine kinase inhibitors (TKIs). According to our knowledge, the discussed procedure in this study is the first LC-MS/MS analytical method for evaluating LRB metabolic stability.
Insights
An LC-MS/MS method quantifies Larotrectinib (LRB), a Trk inhibitor, and assesses its metabolic stability. This method is crucial for understanding LRB
Area of Science:
- Pharmacology
- Analytical Chemistry
- Drug Metabolism
Background:
- Larotrectinib (VITRAKVI) is a potent tropomyosin receptor kinase (Trk) inhibitor with a 75% response rate in Trk fusion-positive cancers, irrespective of cancer type, age, or gender.
- Understanding the drug's metabolic profile is essential for optimizing its therapeutic use.
Purpose of the Study:
- To develop and validate an efficient and accurate LC-MS/MS analytical method for Larotrectinib (LRB) quantification.
- To evaluate the metabolic stability of Larotrectinib in human liver microsomes (HLMs).
Main Methods:
- Development of a reversed-phase LC-MS/MS method using a C18 column and lapatinib as an internal standard.
- Validation of the method for linearity, sensitivity (limit of quantification: 5 ng/mL), and reproducibility (accuracy and precision <3%).
- Assessment of LRB metabolic stability in HLMs to determine in vitro half-life and intrinsic clearance.
Main Results:
- The method demonstrated excellent linearity (5-500 ng/mL, r² ≥ 0.9999) and validated sensitivity and reproducibility.
- The in vitro half-life of LRB in HLMs was calculated as 48.8 minutes.
- The intrinsic clearance of LRB was determined to be 14.19 µL/min/mg.
Conclusions:
- The developed LC-MS/MS method is suitable for quantifying Larotrectinib and evaluating its metabolic stability.
- Larotrectinib exhibits moderate extraction ratio compared to other tyrosine kinase inhibitors (TKIs).
- This represents the first LC-MS/MS method for assessing Larotrectinib metabolic stability.
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