A Simple LC-MS/MS Method for the Quantification of PDA-66 in Human Plasma.

Rico Schwarz,Dietmar Zechner,Anahit Pews-Davtyan,Christian Junghanß,Hugo Murua Escobar,Burkhard Hinz,Elisabeth R D Seiler,Matthias Beller,Sina Sender

doi:10.3390/molecules27030974

Rico Schwarz, Dietmar Zechner + Show 7 more

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https://doi.org/10.3390/molecules27030974

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Abstract

The treatment of cancer is one of the most important pharmacotherapeutic challenges. To this end, chemotherapy has for some time been complemented by targeted therapies against specific structures. PDA-66, a structural analogue of the inhibitor of serine–threonine kinase glycogen synthase kinase 3β SB216763, has shown preclinical antitumour effects in various cell lines, with the key pathways of its anticancer activity being cell cycle modulation, DNA replication and p53 signalling. For the monitoring of anticancer drug treatment in the context of therapeutic drug monitoring, the determination of plasma concentrations is essential, for which an LC-MS/MS method is particularly suitable. In the present study, a sensitive LC-MS/MS method for the quantification of the potential anticancer drug PDA-66 in human plasma with a lower limit of quantification of 2.5 nM is presented. The method was successfully validated and tested for the determination of PDA-66 in mouse plasma and sera.

Highlights

The treatment of different types of cancer has been the focus of scientific interest for decades
One example of potentially new targets for cancer therapy is the serine–threonine kinase glycogen synthase 3β (GSK3β), for which the maleimide derivative SB216763 has been described as an inhibitor [1]
Acridine formic acid from Honeywell Fluka (Seelze, Germany), S9 liver fraction (mouse and huorange and LiChrosolv® water were obtained from Merck Millipore (Darmstadt, German) and lyophilised mouse plasma from Merck Millipore (Darmstadt, Germany) and many), formic acid from Honeywell Fluka (Seelze, Germany), S9 liver fraction

Summary

Introduction

The treatment of different types of cancer has been the focus of scientific interest for decades. Based on the antitumour activity of SB216763, a structural analogue, PDA-66 (Figure 1), was developed, which, compared to SB216763, has an unprotected 2-methylindole moiety, a methylated maleimide group, and a 4-acetyl group instead of the 2,4-dichloro substituent [2]. Initial preclinical studies with PDA-66 showed antiproliferative and proapoptotic effects on human progenitor and cancer cells [3]. Later, these effects were confirmed in acute lymphoblastic leukaemia [4] and canine lymphoma cells [5], the latter being a model very similar to human high-grade non-Hodgkin’s lymphoma. Whole transcriptome sequencing analyses have clearly shown that the key pathways of PDA-66’s antitumour activity are cell cycle modulation, DNA replication and p53 signalling [5]. Kinetic studies require the establishment of a reliable method to quantify the substance from plasma or serum

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