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Published on: March 28, 2017

A validated LC-MS/MS analytical method for the quantification of pemigatinib: metabolic stability evaluation in human

Mohamed W Attwa¹, Ali S Abdelhameed¹, Nawaf A Alsaif¹

¹Department of Pharmaceutical Chemistry, College of Pharmacy, King Saud University Riyadh Saudi Arabia halrabiah@ksu.edu.sa +966 1146 76220 +966 1146 70237.

|August 3, 2022

Abstract:

Pemigatinib (PMB) is a small molecule inhibitor of fibroblast growth factor receptor 1 (FGFR1), FGFR2 and FGFR3. On April 17, 2020, the US Food and Drug Administration granted accelerated approval for PMB for the treatment of adults with previously treated, unresectable metastatic or locally advanced cholangiocarcinoma with a fibroblast growth factor receptor 2 (FGFR2) fusion or other rearrangement. PMB is considered the first targeted treatment for cholangiocarcinoma approved in the US. In this study, in silico prediction of PMB metabolic stability was done using the WhichP450 module of the StarDrop software package. Further, an LC-MS/MS analytical method was developed for PMB quantification in human liver microsomes (HLM) to experimentally assess metabolic stability. PMB and flavopiridol (FVL), used as an internal standard IS, were resolved using an isocratic mobile phase and a C18 stationary phase. The LC-MS/MS method showed linearity in the range of 5 to 500 ng mL^-1 in an HLM matrix (R ² = 0.9995). The lower limit of quantification (LLOQ) was 5 ng mL^-1, indicating sensitivity. The inter- and intra-day accuracy and precision were within a variability of 10, confirming the reproducibility of the method. The measured in vitro half-life and intrinsic clearance of PMB were 27.29 min and 25.40 μL min^-1 mg^-1, respectively. PMB showed a moderate extraction ratio suggesting good bioavailability. The developed analytical method is the first LC-MS/MS method specific for PMB quantification with application to metabolic stability assessment.

Insights

Pemigatinib (PMB) is a targeted therapy for cholangiocarcinoma. This study developed a new LC-MS/MS method to assess its metabolic stability in human liver microsomes, revealing moderate bioavailability.

Area of Science:

Pharmacology
Analytical Chemistry
Drug Metabolism

Background:

Pemigatinib (PMB) is the first targeted therapy approved in the US for cholangiocarcinoma.
It functions as a small molecule inhibitor of fibroblast growth factor receptors (FGFR1, FGFR2, FGFR3).
Understanding PMB's metabolic stability is crucial for assessing its clinical efficacy and bioavailability.

Purpose of the Study:

To perform in silico prediction of Pemigatinib's metabolic stability.
To develop and validate a novel LC-MS/MS analytical method for quantifying PMB in human liver microsomes (HLM).
To experimentally assess PMB's metabolic stability and pharmacokinetic properties in vitro.

Main Methods:

In silico metabolic stability prediction using StarDrop software's WhichP450 module.
Development of a sensitive and reproducible LC-MS/MS method for PMB quantification in HLM.
Experimental assessment of PMB's in vitro half-life, intrinsic clearance, and extraction ratio using the validated method.

Main Results:

The developed LC-MS/MS method demonstrated excellent linearity (R² = 0.9995) and sensitivity (LLOQ = 5 ng mL⁻¹).
The method exhibited high accuracy and precision, confirming its reproducibility.
In vitro studies showed a half-life of 27.29 min and intrinsic clearance of 25.40 μL min⁻¹ mg⁻¹, with a moderate extraction ratio indicating good bioavailability.

Conclusions:

A novel, validated LC-MS/MS method for PMB quantification in HLM was successfully developed.
The study provides crucial in vitro data on Pemigatinib's metabolic stability and pharmacokinetic profile.
These findings support the use of Pemigatinib as a targeted therapy for cholangiocarcinoma.

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