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Pharmaceutical Research

, Volume 30, Issue 9, pp 2290–2302 | Cite as

Benzylmorpholine Analogs as Selective Inhibitors of Lung Cytochrome P450 2A13 for the Chemoprevention of Lung Cancer in Tobacco Users

  • Linda C. Blake
  • Anuradha Roy
  • David Neul
  • Frank J. Schoenen
  • Jeffrey Aubé
  • Emily E. Scott
Research Paper

ABSTRACT

Purpose

4-(Methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK), one of the most prevalent and procarcinogenic compounds in tobacco, is bioactivated by respiratory cytochrome P450 (CYP) 2A13, forming DNA adducts and initiating lung cancer. CYP2A13 inhibition offers a novel strategy for chemoprevention of tobacco-associated lung cancer.

Methods

Twenty-four analogs of a 4-benzylmorpholine scaffold identified by high throughput screening were evaluated for binding and inhibition of both functional human CYP2A enzymes, CYP2A13 and the 94%-identical hepatic CYP2A6, whose inhibition is undesirable. Thus, selectivity is a major challenge in compound design.

Results

A key feature resulting in CYP2A13-selective binding and inhibition was substitution at the benzyl ortho position, with three analogs being >25-fold selective for CYP2A13 over CYP2A6.

Conclusions

Two such analogs were negative for genetic and hERG toxicities and metabolically stable in human lung microsomes, but displayed rapid metabolism in human liver and in mouse and rat lung and liver microsomes, likely due to CYP2B-mediated degradation. A specialized knockout mouse mimicking the human lung demonstrates compound persistence in lung and provides an appropriate test model. Compound delivered by inhalation may be effective in the lung but rapidly cleared otherwise, limiting systemic exposure.

KEY WORDS

chemopreventative cytochrome P450 2A13 cytochrome P450 2A6 lung cancer tobacco 

ABBREVIATIONS

CYP

Cytochrome P450

DMSO

Dimethyl sulfoxide

hERG

Human ether-a-go-go-related gene

Kd

Equilibrium dissociation constant

Ki

Inhibition constant

NNK

4-(Methylnitrosamino)-1-(3-pyridyl)-1-butanone

PEITC

Phenethyl isothiocyanate

POR

NADPH-cytochrome P450 oxidoreductase

SAR

Structure activity relationship

Notes

ACKNOWLEDGMENTS AND DISCLOSURES

Thanks are due to Kelin Li, who provided synthetic training to LB. We are grateful for purity analysis by Patrick Porubsky and Ben Neuenswander at The University of Kansas Center for Chemical Methodologies and Library Design. Toxicity studies were undertaken at Cerep. Metabolic stability in microsomes was undertaken at Xenotech, LLC. Mike Wester facilitated metabolic screening with the major human xenobiotic metabolizing CYP enzymes. Transgenic microsomes were a gift from Xinxin Ding at the Wadsworth Center, New York State Department of Health and State University of New York at Albany. This work was supported by the Kansas Masonic Cancer Research Institute, the University of Kansas General Research Fund, NIH grant NIGMS GM076343, and the Institute for Advancing Medical Innovation.

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Copyright information

© Springer Science+Business Media New York 2013

Authors and Affiliations

  • Linda C. Blake
    • 1
  • Anuradha Roy
    • 2
  • David Neul
    • 3
  • Frank J. Schoenen
    • 4
  • Jeffrey Aubé
    • 1
    • 4
  • Emily E. Scott
    • 1
  1. 1.Department of Medicinal ChemistryUniversity of KansasLawrenceUSA
  2. 2.High Throughput Screening LaboratoryUniversity of KansasLawrenceUSA
  3. 3.Pharmacokinetics, Dynamics and MetabolismPfizer Inc.La JollaUSA
  4. 4.University of Kansas Specialized Chemistry CenterUniversity of KansasLawrenceUSA

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