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In vitro cytotoxicity, pharmacokinetics, tissue distribution, and metabolism of small-molecule protein kinase D inhibitors, kb-NB142-70 and kb-NB165-09, in mice bearing human cancer xenografts

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Abstract

Purpose

Protein kinase D (PKD) mediates diverse biological responses including cell growth and survival. Therefore, PKD inhibitors may have therapeutic potential. We evaluated the in vitro cytotoxicity of two PKD inhibitors, kb-NB142-70 and its methoxy analogue, kb-NB165-09, and examined their in vivo efficacy and pharmacokinetics.

Methods

The in vitro cytotoxicities of kb-NB142-70 and kb-NB165-09 were evaluated by MTT assay against PC-3, androgen-independent prostate cancer cells, and CFPAC-1 and PANC-1, pancreatic cancer cells. Efficacy studies were conducted in mice bearing either PC-3 or CPFAC-1 xenografts. Tumor-bearing mice were euthanized between 5 and 1,440 min after iv dosing, and plasma and tissue concentrations were measured by HPLC–UV. Metabolites were characterized by LC–MS/MS.

Results

kb-NB142-70 and kb-NB165-09 inhibited cellular growth in the low–mid μM range. The compounds were inactive when administered to tumor-bearing mice. In mice treated with kb-NB142-70, the plasma C max was 36.9 nmol/mL, and the PC-3 tumor C max was 11.8 nmol/g. In mice dosed with kb-NB165-09, the plasma C max was 61.9 nmol/mL, while the PANC-1 tumor C max was 8.0 nmol/g. The plasma half-lives of kb-NB142-70 and kb-NB165-09 were 6 and 14 min, respectively. Both compounds underwent oxidation and glucuronidation.

Conclusions

kb-NB142-70 and kb-NB165-09 were rapidly metabolized, and concentrations in tumor were lower than those required for in vitro cytotoxicity. Replacement of the phenolic hydroxyl group with a methoxy group increased the plasma half-life of kb-NB165-09 2.3-fold over that of kb-NB142-70. Rapid metabolism in mice suggests that next-generation compounds will require further structural modifications to increase potency and/or metabolic stability.

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Acknowledgments

This study was supported in part by US Public Health Service National Institutes of Health grant CA78039, CA129127, and CA142580. This project used the UPCI Clinical Pharmacology Analytical Facility (CPAF) and was supported in part by award P01CA078039. We acknowledge Diane Mazzei and other members of DLAR for the excellent care of the animals used in these studies. We also acknowledge the UPCI Writing Group for their helpful comments.

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Authors and Affiliations

  1. Molecular Therapeutics and Drug Discovery, The University of Pittsburgh Cancer Institute, Hillman Cancer Center, 5117 Centre Ave, Room G27b, Pittsburgh, PA, 15213, USA

    Jianxia Guo, Dana M. Clausen, Jan H. Beumer, Robert A. Parise, Merrill J. Egorin, Peter Wipf & Julie L. Eiseman

  2. Department of Pharmacology and Chemical Biology, School of Medicine, University of Pittsburgh, Pittsburgh, PA, 15260, USA

    Jianxia Guo, Merrill J. Egorin, Qiming Jane Wang & Julie L. Eiseman

  3. Department of Pharmaceutical Sciences, School of Pharmacy, University of Pittsburgh, Pittsburgh, PA, 15260, USA

    Jan H. Beumer & Robert A. Parise

  4. Department of Medicine, School of Medicine, University of Pittsburgh, Pittsburgh, PA, 15260, USA

    Merrill J. Egorin

  5. Department of Chemistry, University of Pittsburgh, Pittsburgh, PA, 15260, USA

    Karla Bravo-Altamirano & Peter Wipf

  6. Department of Pharmacology, University of Virginia, Charlottesville, VA, 22908, USA

    Elizabeth R. Sharlow

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  1. Jianxia Guo
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Correspondence to Julie L. Eiseman.

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Guo, J., Clausen, D.M., Beumer, J.H. et al. In vitro cytotoxicity, pharmacokinetics, tissue distribution, and metabolism of small-molecule protein kinase D inhibitors, kb-NB142-70 and kb-NB165-09, in mice bearing human cancer xenografts. Cancer Chemother Pharmacol 71, 331–344 (2013). https://doi.org/10.1007/s00280-012-2010-z

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