Research Article
Development and validation of LC–MS/MS with in-source collision-induced dissociation for the quantification of pegcantratinib in human skin tumors
Aim: Pegcantratinib is a mini-PEGylated K252a derivative, under clinical evaluation as an anticancer agent acting through inhibition of the tropomyosin receptor kinase. A method for quantifying pegcantratinib in skin tumor biopsies of patients was required to determine tumor drug penetration. Methods & results: A sensitive and PEGylated molecule specific HPLC–MS/MS method coupled with in-source collision-induced dissociation was developed. The method exhibited excellent precision (coefficient of variation ≤8.5%), accuracy in the range 95–102%, high and consistent recovery and no matrix effect. The assay was linear across a range of 1–500 ng/ml, with a limit of quantitation of 2.5 ng/ml. Conclusion: We have developed and validated a method for analyzing pegcantratinib in human tumor biopsies, with the approach successfully applied to clinical trial samples.
First draft submitted: 22 July 2016; Accepted for publication: 1 December 2016; Published online: 23 January 2017
Keywords: LC–MS/MS • pegcantratinib • PEGylation • tropomyosin receptor kinase A • tumor
Pegcantratinib is the first topically applied tropomyosin receptor kinase (TRK) inhibi- tor that contains mini-PEGylated K252a, which exerts its effects at nanomolar concen- trations by inhibiting the intracellular kinase domain of TRK [1]. Unlike other TRK inhib- itors, which are available only in oral for- mulations, the conjugation process of small molecules to PEG gives rise to physicochemi- cal and pharmacological characteristics that optimize these molecules for topical routes of administration. This facilitates the delivery of high levels of the active ingredient to skin tumor cells, while avoiding the systemic side effects of oral applications [2]. Pegcantratinib has recently been evalu-
ated in a Phase llb clinical study setting as a potential treatment in common inflamma- tory skin conditions, including psoriasis and atopic dermatitis, with a good safety profile observed [3]. The drug is now entering clini- cal development in patients with germline
10.4155/bio-2016-0199 © Gareth Veal
mutations in the tumor suppressor gene CYLD, which include multiple clinical pre- sentations of rare, skin appendage tumors: Brooke–Spiegler syndrome, familial cyl- indromatosis and multiple familial tricho- epitheliomas [4–6]. Patients carrying CYLD mutations face repeated disfiguring surgery to control tumor burden, with approximately one in four affected patients undergoing total scalp removal, due to the lack of an available pharmacological treatment [7]. Studies per- formed in 3D primary cell cultures of cylin- droma, treated with salicylic acid and TRK inhibitors, showed them to be particularly sensitive to K252a, with a surviving fraction of 50% (SF50
) of 366 nM [8]. As part of an ongoing early phase clini-
cal trial, investigations of pegcantratinib tumor penetration and potential correlations between tumor exposure and response are warranted. Therefore, an analytical method was required to quantify pegcantratinib con-
Bioanalysis (2017) 9(3), 279–288 ISSN 1757-6180 Monique Zangarini1
& Gareth J Veal*,1 1
, Neil
Rajan2, Marina Danilenko2 Philip Berry1, Silvio Traversa3
,
Newcastle Cancer Centre Pharmacology Group, Newcastle University, Newcastle
upon Tyne, NE2 4HH, UK 2
NE2 4HH, UK 3
Institute of Genetic Medicine, Newcastle University, Newcastle upon Tyne,
Creabilis Therapeutics, Colleretto
Giacosa, Italy *Author for correspondence: Tel.: +44 0191 208 4332 Fax: +44 0191 208 3452
G.J.Veal@
ncl.ac.uk
part of
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