High-throughput screen identifies novel inhibitors of cancer biomarker α-methylacyl coenzyme A racemase (AMACR/P504S)

Brice A P Wilson, Haofan Wang, Benjamin A. Nacev, Ronnie Mease, Jun Liu, Martin Gilbert Pomper, William B Isaacs

Research output: Contribution to journalArticle

Abstract

α-methylacyl coenzyme A racemase (AMACR) is a metabolic enzyme whose overexpression has been shown to be a diagnostic indicator of prostatic adenocarcinoma and other solid tumors. Here, we confirm that attenuation of AMACR expression diminishes the growth of prostate cancer cell lines by using stably expressed short-hairpin RNA constructs. This observation strongly suggests that the AMACR enzyme may be a target for therapeutic inhibition in prostate cancer. To this end, we report here a novel assay capable of screening libraries of diverse small molecules for inhibitors of AMACR activity. This assay facilitated the screening of approximately 5,000 unique compounds and the discovery of 7 distinct chemical entities capable of inhibiting AMACR at low micromolar concentrations. The most potent inhibitor discovered is the selenoorganic compound ebselen oxide [inhibitory concentration (IC 50): 0.80 μmol/L]. The parent compound, ebselen (IC50: 2.79 μmol/L), is a covalent inactivator of AMACR (KI(inact): 24 μmol/L). Two of the AMACR inhibitors are selectively toxic to prostate cancer cell lines (LAPC4/LNCaP/PC3) that express AMACR compared to a normal prostate fibroblast cell line (WPMY1) that does not express the protein. This report shows the first high-throughput screen for the discovery of novel AMACR inhibitors, characterizes the first nonsubstrate-based inhibitors, and validates that AMACR is a viable chemotherapeutic target in vitro.

Original languageEnglish (US)
Pages (from-to)825-838
Number of pages14
JournalMolecular Cancer Therapeutics
Volume10
Issue number5
DOIs
StatePublished - May 2011

Fingerprint

Racemases and Epimerases
Coenzyme A
Tumor Biomarkers
Prostatic Neoplasms
Cell Line
Inhibitory Concentration 50
Small Molecule Libraries
Poisons
Enzymes
Oxides
Small Interfering RNA
Prostate
Adenocarcinoma
Fibroblasts
Growth
Neoplasms
Proteins
ebselen
Therapeutics

ASJC Scopus subject areas

  • Oncology
  • Cancer Research

Cite this

High-throughput screen identifies novel inhibitors of cancer biomarker α-methylacyl coenzyme A racemase (AMACR/P504S). / Wilson, Brice A P; Wang, Haofan; Nacev, Benjamin A.; Mease, Ronnie; Liu, Jun; Pomper, Martin Gilbert; Isaacs, William B.

In: Molecular Cancer Therapeutics, Vol. 10, No. 5, 05.2011, p. 825-838.

Research output: Contribution to journalArticle

@article{4c3438fb34ca4f338eb0c38c001eb104,
title = "High-throughput screen identifies novel inhibitors of cancer biomarker α-methylacyl coenzyme A racemase (AMACR/P504S)",
abstract = "α-methylacyl coenzyme A racemase (AMACR) is a metabolic enzyme whose overexpression has been shown to be a diagnostic indicator of prostatic adenocarcinoma and other solid tumors. Here, we confirm that attenuation of AMACR expression diminishes the growth of prostate cancer cell lines by using stably expressed short-hairpin RNA constructs. This observation strongly suggests that the AMACR enzyme may be a target for therapeutic inhibition in prostate cancer. To this end, we report here a novel assay capable of screening libraries of diverse small molecules for inhibitors of AMACR activity. This assay facilitated the screening of approximately 5,000 unique compounds and the discovery of 7 distinct chemical entities capable of inhibiting AMACR at low micromolar concentrations. The most potent inhibitor discovered is the selenoorganic compound ebselen oxide [inhibitory concentration (IC 50): 0.80 μmol/L]. The parent compound, ebselen (IC50: 2.79 μmol/L), is a covalent inactivator of AMACR (KI(inact): 24 μmol/L). Two of the AMACR inhibitors are selectively toxic to prostate cancer cell lines (LAPC4/LNCaP/PC3) that express AMACR compared to a normal prostate fibroblast cell line (WPMY1) that does not express the protein. This report shows the first high-throughput screen for the discovery of novel AMACR inhibitors, characterizes the first nonsubstrate-based inhibitors, and validates that AMACR is a viable chemotherapeutic target in vitro.",
author = "Wilson, {Brice A P} and Haofan Wang and Nacev, {Benjamin A.} and Ronnie Mease and Jun Liu and Pomper, {Martin Gilbert} and Isaacs, {William B}",
year = "2011",
month = "5",
doi = "10.1158/1535-7163.MCT-10-0902",
language = "English (US)",
volume = "10",
pages = "825--838",
journal = "Molecular Cancer Therapeutics",
issn = "1535-7163",
publisher = "American Association for Cancer Research Inc.",
number = "5",

}

TY - JOUR

T1 - High-throughput screen identifies novel inhibitors of cancer biomarker α-methylacyl coenzyme A racemase (AMACR/P504S)

AU - Wilson, Brice A P

AU - Wang, Haofan

AU - Nacev, Benjamin A.

AU - Mease, Ronnie

AU - Liu, Jun

AU - Pomper, Martin Gilbert

AU - Isaacs, William B

PY - 2011/5

Y1 - 2011/5

N2 - α-methylacyl coenzyme A racemase (AMACR) is a metabolic enzyme whose overexpression has been shown to be a diagnostic indicator of prostatic adenocarcinoma and other solid tumors. Here, we confirm that attenuation of AMACR expression diminishes the growth of prostate cancer cell lines by using stably expressed short-hairpin RNA constructs. This observation strongly suggests that the AMACR enzyme may be a target for therapeutic inhibition in prostate cancer. To this end, we report here a novel assay capable of screening libraries of diverse small molecules for inhibitors of AMACR activity. This assay facilitated the screening of approximately 5,000 unique compounds and the discovery of 7 distinct chemical entities capable of inhibiting AMACR at low micromolar concentrations. The most potent inhibitor discovered is the selenoorganic compound ebselen oxide [inhibitory concentration (IC 50): 0.80 μmol/L]. The parent compound, ebselen (IC50: 2.79 μmol/L), is a covalent inactivator of AMACR (KI(inact): 24 μmol/L). Two of the AMACR inhibitors are selectively toxic to prostate cancer cell lines (LAPC4/LNCaP/PC3) that express AMACR compared to a normal prostate fibroblast cell line (WPMY1) that does not express the protein. This report shows the first high-throughput screen for the discovery of novel AMACR inhibitors, characterizes the first nonsubstrate-based inhibitors, and validates that AMACR is a viable chemotherapeutic target in vitro.

AB - α-methylacyl coenzyme A racemase (AMACR) is a metabolic enzyme whose overexpression has been shown to be a diagnostic indicator of prostatic adenocarcinoma and other solid tumors. Here, we confirm that attenuation of AMACR expression diminishes the growth of prostate cancer cell lines by using stably expressed short-hairpin RNA constructs. This observation strongly suggests that the AMACR enzyme may be a target for therapeutic inhibition in prostate cancer. To this end, we report here a novel assay capable of screening libraries of diverse small molecules for inhibitors of AMACR activity. This assay facilitated the screening of approximately 5,000 unique compounds and the discovery of 7 distinct chemical entities capable of inhibiting AMACR at low micromolar concentrations. The most potent inhibitor discovered is the selenoorganic compound ebselen oxide [inhibitory concentration (IC 50): 0.80 μmol/L]. The parent compound, ebselen (IC50: 2.79 μmol/L), is a covalent inactivator of AMACR (KI(inact): 24 μmol/L). Two of the AMACR inhibitors are selectively toxic to prostate cancer cell lines (LAPC4/LNCaP/PC3) that express AMACR compared to a normal prostate fibroblast cell line (WPMY1) that does not express the protein. This report shows the first high-throughput screen for the discovery of novel AMACR inhibitors, characterizes the first nonsubstrate-based inhibitors, and validates that AMACR is a viable chemotherapeutic target in vitro.

UR - http://www.scopus.com/inward/record.url?scp=79955984121&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=79955984121&partnerID=8YFLogxK

U2 - 10.1158/1535-7163.MCT-10-0902

DO - 10.1158/1535-7163.MCT-10-0902

M3 - Article

VL - 10

SP - 825

EP - 838

JO - Molecular Cancer Therapeutics

JF - Molecular Cancer Therapeutics

SN - 1535-7163

IS - 5

ER -