Abstract C179: Preclinical characterization of GPA512: A prodrug of a direct STAT3 inhibitor for the treatment of prostate cancer
Authors: Hellsten Rebecka , Escobar Zilma , Bjartell Anders , Canesin Giacomo , Evans Axelsson Susan , Sterner Olov , Johansson Martin 
Published: May 2016

Abstract
Background: The transcription factor STAT3 is a promising target for the treatment of castration-resistant prostate cancer (CRPC) as STAT3 is implicated in drug resistance and castration resistance as well as metastatic spread, tumor growth and immune escape. Galiellalactone (GL) is a direct inhibitor of STAT3 that prevents DNA binding, inhibits proliferation of prostate cancer cells expressing active STAT3 and induces apoptosis by down-regulation of STAT3 activated genes. In this study we aimed to characterize a prodrug of GL, GPA512, with improved drug-like properties and to demonstrate its effect on tumor growth in a xenograft model of prostate cancer following oral administration.
Methods: Stability studies of prodrugs based on GL were performed in 0.1 M PBS buffer (pH 7.4) and in plasma at 37°C. In vitro efficacy of prodrugs was studied by WST-1 proliferation assay in DU145 prostate cancer cells expressing active STAT3. The systemic exposure of GL in mice was studied following a single oral dose of GPA512 or GL (both 10 mg/kg). The plasma concentrations of GL were determined by LC-MS/MS. For the xenograft study NMRI-nude male mice were inoculated subcutaneously with DU145 cells and once tumors were established the mice were divided in two groups with ten mice in each. Mice were treated orally with 40 mg/kg GPA512 daily five times/week for four weeks. Tumor growth was measured by caliper and at the end of the study tumors were harvested for subsequent analyses using immunohistochemistry and mRNA expression analysis.
Results: In vitro studies showed that the prodrug GPA512 is rapidly converted to GL in plasma and that GPA512 is stable in buffer solution and has similar inhibitory effects on proliferation as GL on DU145 cells. The pharmacokinetics of GPA512 following a single oral dose indicated that the compound was rapidly absorbed and converted to GL with a tmax of 15 min. Oral administration of GPA512 in mice increased the plasma exposure (AUC) of the active parent compound 20-fold compared to when GL was dosed orally. GPA512 treated mice bearing subcutaneous DU145 tumors had significantly smaller tumors compared to mice treated with vehicle. No adverse effects or weight loss were observed. Analysis of tumors showed decreased cell proliferation and increased amount of apoptotic cells in GPA512 treated mice compared to control. The mRNA expression of STAT3 regulated anti-apoptotic gene MCl-1 was significantly reduced by GPA512 treatment.
Conclusions: The drug-like properties and safety profile of the prodrug GPA512 and galiellalactone's direct inhibition of STAT3, warrant further studies of GPA512 as a drug candidate for treatment of patients with CRPC.
Citation Format: Rebecka Hellsten, Zilma Escobar, Anders Bjartell, Giacomo Canesin, Susan Evans Axelsson, Olov Sterner, Martin Johansson. Preclinical characterization of GPA512: A prodrug of a direct STAT3 inhibitor for the treatment of prostate cancer. [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference: Molecular Targets and Cancer Therapeutics; 2015 Nov 5-9; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2015;14(12 Suppl 2):Abstract nr C179.