Drug Res (Stuttg) 2020; 70(02/03): 101-106
DOI: 10.1055/a-1071-0849
Original Article
© Georg Thieme Verlag KG Stuttgart · New York

Validated HPLC Method for Quantification of a Novel Trk Inhibitor, Larotrectinib in Mice Plasma: Application to a Pharmacokinetic Study

Harsha K. Tripathy
1  Karnataka College of Pharmacy Bangalore, Medicinal Chemistry, Bangalore, India
,
S.V. Nair Manju
1  Karnataka College of Pharmacy Bangalore, Medicinal Chemistry, Bangalore, India
,
Ashok Zakkula
2  Jubilant Biosys, DMPK, Bangalore, India
,
Ram Murthi Bestha
2  Jubilant Biosys, DMPK, Bangalore, India
,
Sreekanth Dittakavi
2  Jubilant Biosys, DMPK, Bangalore, India
,
2  Jubilant Biosys, DMPK, Bangalore, India
› Author Affiliations
Further Information

Publication History

received 04 November 2019

accepted 26 November 2019

Publication Date:
13 January 2020 (online)

Abstract

Larotrectinib, is an orally active novel small molecule approved for the treatment of solid tumors in pediatrics and adult patients. It acts by inhibiting tropomyosin receptor kinase. In this paper, we report the development and validation of a high-performance liquid chromatography (HPLC) method for the quantitation of larotrectinib in mice plasma as per the FDA regulatory guideline. Plasma samples processing was accomplished through simple protein precipitation using acetonitrile enriched with internal standard (IS, enasidenib). The chromatographic analysis was performed using a gradient mobile phase comprising 10 mM ammonium acetate and acetonitrile at a flow-rate of 0.8 mL/min on an X-Terra Phenyl column. The UV detection wave length was set at λmax 262 nm. Larotrectinib and the IS eluted at 3.85 and 6.60 min, respectively with a total run time of 8.0 min. The calibration curve was linear over a concentration range of 0.20–5.00 μg/mL (r2=≥0.992). The intra- and inter-day precision and accuracy results were within the acceptable limits. Results of stability studies indicated that larotrectinib was stable on bench-top, in auto-sampler, up to three freeze/thaw cycles and long-term storage at −80°C. The validated HPLC method was successfully applied to a pharmacokinetic study in mice.