Inhibition of human sterol Δ7-reductase and other postlanosterol enzymes by LK-980, a novel inhibitor of cholesterol synthesis

Jure Ačimovič, Tina Korošec, Matej Seliškar, Ingemar Bjorkhem, K. Monostory, P. Szabó, Jean Marc Pascussi, Ales Belič, Uroš Urleb, Darko Kocjan, Damjana Rozman

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

Novel potential inhibitors of the postsqualene portion of cholesterol synthesis were screened in HepG2 cells. 2-(4-Phenethylpiperazin- 1-yl)-1-(pyridine-3-yl)ethanol (LK-980) was identified as a prospective compound and was characterized further in cultures of human primary hepatocytes from seven donors. In vitro kinetic measurements show that the half-life of LK-980 is at least 4.3 h. LK-980 does not induce CYP3A4 mRNA nor enzyme activity. Target prediction was performed by gas chromatography-mass spectrometry, allowing simultaneous separation and quantification of nine late cholesterol intermediates. Experiments indicated that human sterol Δ7- reductase (DHCR7) is the major target of LK-980 (34-fold increase of 7-dehydrocholesterol), whereas human sterol Δ14-reductase (DHCR14), human sterol Δ24-reductase (DHCR24), and human sterol C5-desaturase (SC5DL) represent minor targets. In the absence of purified enzymes, we used the mathematical model of cholesterol synthesis to evaluate whether indeed more than a single enzyme is inhibited. In silico inhibition of only DHCR7 modifies the flux of cholesterol intermediates, resulting in a sterol profile that does not support experimental data. Partial inhibition of the DHCR14, DHCR24, and SC5DL steps, in addition to DHCR7, supports the experimental sterol profile. In conclusion, we provide experimental and computational evidence that LK-980, a novel inhibitor from the late portion of cholesterol synthesis, inhibits primarily DHCR7 and to a lesser extent three other enzymes from this pathway.

Original languageEnglish
Pages (from-to)39-46
Number of pages8
JournalDrug Metabolism and Disposition
Volume39
Issue number1
DOIs
Publication statusPublished - Jan 2011

Fingerprint

Anticholesteremic Agents
Sterols
Oxidoreductases
Cholesterol
Enzymes
Cytochrome P-450 CYP3A
Hep G2 Cells
2-(4-phenethylpiperazin-1-yl)-1-(pyridine-3-yl)ethanol
Computer Simulation
Gas Chromatography-Mass Spectrometry
Half-Life
Hepatocytes
Theoretical Models
Messenger RNA

ASJC Scopus subject areas

  • Pharmacology
  • Pharmaceutical Science

Cite this

Inhibition of human sterol Δ7-reductase and other postlanosterol enzymes by LK-980, a novel inhibitor of cholesterol synthesis. / Ačimovič, Jure; Korošec, Tina; Seliškar, Matej; Bjorkhem, Ingemar; Monostory, K.; Szabó, P.; Pascussi, Jean Marc; Belič, Ales; Urleb, Uroš; Kocjan, Darko; Rozman, Damjana.

In: Drug Metabolism and Disposition, Vol. 39, No. 1, 01.2011, p. 39-46.

Research output: Contribution to journalArticle

Ačimovič, J, Korošec, T, Seliškar, M, Bjorkhem, I, Monostory, K, Szabó, P, Pascussi, JM, Belič, A, Urleb, U, Kocjan, D & Rozman, D 2011, 'Inhibition of human sterol Δ7-reductase and other postlanosterol enzymes by LK-980, a novel inhibitor of cholesterol synthesis', Drug Metabolism and Disposition, vol. 39, no. 1, pp. 39-46. https://doi.org/10.1124/dmd.110.035840
Ačimovič, Jure ; Korošec, Tina ; Seliškar, Matej ; Bjorkhem, Ingemar ; Monostory, K. ; Szabó, P. ; Pascussi, Jean Marc ; Belič, Ales ; Urleb, Uroš ; Kocjan, Darko ; Rozman, Damjana. / Inhibition of human sterol Δ7-reductase and other postlanosterol enzymes by LK-980, a novel inhibitor of cholesterol synthesis. In: Drug Metabolism and Disposition. 2011 ; Vol. 39, No. 1. pp. 39-46.
@article{e5b6c3a7640143bdb38339bc4af34da9,
title = "Inhibition of human sterol Δ7-reductase and other postlanosterol enzymes by LK-980, a novel inhibitor of cholesterol synthesis",
abstract = "Novel potential inhibitors of the postsqualene portion of cholesterol synthesis were screened in HepG2 cells. 2-(4-Phenethylpiperazin- 1-yl)-1-(pyridine-3-yl)ethanol (LK-980) was identified as a prospective compound and was characterized further in cultures of human primary hepatocytes from seven donors. In vitro kinetic measurements show that the half-life of LK-980 is at least 4.3 h. LK-980 does not induce CYP3A4 mRNA nor enzyme activity. Target prediction was performed by gas chromatography-mass spectrometry, allowing simultaneous separation and quantification of nine late cholesterol intermediates. Experiments indicated that human sterol Δ7- reductase (DHCR7) is the major target of LK-980 (34-fold increase of 7-dehydrocholesterol), whereas human sterol Δ14-reductase (DHCR14), human sterol Δ24-reductase (DHCR24), and human sterol C5-desaturase (SC5DL) represent minor targets. In the absence of purified enzymes, we used the mathematical model of cholesterol synthesis to evaluate whether indeed more than a single enzyme is inhibited. In silico inhibition of only DHCR7 modifies the flux of cholesterol intermediates, resulting in a sterol profile that does not support experimental data. Partial inhibition of the DHCR14, DHCR24, and SC5DL steps, in addition to DHCR7, supports the experimental sterol profile. In conclusion, we provide experimental and computational evidence that LK-980, a novel inhibitor from the late portion of cholesterol synthesis, inhibits primarily DHCR7 and to a lesser extent three other enzymes from this pathway.",
author = "Jure Ačimovič and Tina Korošec and Matej Seliškar and Ingemar Bjorkhem and K. Monostory and P. Szab{\'o} and Pascussi, {Jean Marc} and Ales Belič and Uroš Urleb and Darko Kocjan and Damjana Rozman",
year = "2011",
month = "1",
doi = "10.1124/dmd.110.035840",
language = "English",
volume = "39",
pages = "39--46",
journal = "Drug Metabolism and Disposition",
issn = "0090-9556",
publisher = "American Society for Pharmacology and Experimental Therapeutics",
number = "1",

}

TY - JOUR

T1 - Inhibition of human sterol Δ7-reductase and other postlanosterol enzymes by LK-980, a novel inhibitor of cholesterol synthesis

AU - Ačimovič, Jure

AU - Korošec, Tina

AU - Seliškar, Matej

AU - Bjorkhem, Ingemar

AU - Monostory, K.

AU - Szabó, P.

AU - Pascussi, Jean Marc

AU - Belič, Ales

AU - Urleb, Uroš

AU - Kocjan, Darko

AU - Rozman, Damjana

PY - 2011/1

Y1 - 2011/1

N2 - Novel potential inhibitors of the postsqualene portion of cholesterol synthesis were screened in HepG2 cells. 2-(4-Phenethylpiperazin- 1-yl)-1-(pyridine-3-yl)ethanol (LK-980) was identified as a prospective compound and was characterized further in cultures of human primary hepatocytes from seven donors. In vitro kinetic measurements show that the half-life of LK-980 is at least 4.3 h. LK-980 does not induce CYP3A4 mRNA nor enzyme activity. Target prediction was performed by gas chromatography-mass spectrometry, allowing simultaneous separation and quantification of nine late cholesterol intermediates. Experiments indicated that human sterol Δ7- reductase (DHCR7) is the major target of LK-980 (34-fold increase of 7-dehydrocholesterol), whereas human sterol Δ14-reductase (DHCR14), human sterol Δ24-reductase (DHCR24), and human sterol C5-desaturase (SC5DL) represent minor targets. In the absence of purified enzymes, we used the mathematical model of cholesterol synthesis to evaluate whether indeed more than a single enzyme is inhibited. In silico inhibition of only DHCR7 modifies the flux of cholesterol intermediates, resulting in a sterol profile that does not support experimental data. Partial inhibition of the DHCR14, DHCR24, and SC5DL steps, in addition to DHCR7, supports the experimental sterol profile. In conclusion, we provide experimental and computational evidence that LK-980, a novel inhibitor from the late portion of cholesterol synthesis, inhibits primarily DHCR7 and to a lesser extent three other enzymes from this pathway.

AB - Novel potential inhibitors of the postsqualene portion of cholesterol synthesis were screened in HepG2 cells. 2-(4-Phenethylpiperazin- 1-yl)-1-(pyridine-3-yl)ethanol (LK-980) was identified as a prospective compound and was characterized further in cultures of human primary hepatocytes from seven donors. In vitro kinetic measurements show that the half-life of LK-980 is at least 4.3 h. LK-980 does not induce CYP3A4 mRNA nor enzyme activity. Target prediction was performed by gas chromatography-mass spectrometry, allowing simultaneous separation and quantification of nine late cholesterol intermediates. Experiments indicated that human sterol Δ7- reductase (DHCR7) is the major target of LK-980 (34-fold increase of 7-dehydrocholesterol), whereas human sterol Δ14-reductase (DHCR14), human sterol Δ24-reductase (DHCR24), and human sterol C5-desaturase (SC5DL) represent minor targets. In the absence of purified enzymes, we used the mathematical model of cholesterol synthesis to evaluate whether indeed more than a single enzyme is inhibited. In silico inhibition of only DHCR7 modifies the flux of cholesterol intermediates, resulting in a sterol profile that does not support experimental data. Partial inhibition of the DHCR14, DHCR24, and SC5DL steps, in addition to DHCR7, supports the experimental sterol profile. In conclusion, we provide experimental and computational evidence that LK-980, a novel inhibitor from the late portion of cholesterol synthesis, inhibits primarily DHCR7 and to a lesser extent three other enzymes from this pathway.

UR - http://www.scopus.com/inward/record.url?scp=78650722521&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=78650722521&partnerID=8YFLogxK

U2 - 10.1124/dmd.110.035840

DO - 10.1124/dmd.110.035840

M3 - Article

C2 - 20952551

AN - SCOPUS:78650722521

VL - 39

SP - 39

EP - 46

JO - Drug Metabolism and Disposition

JF - Drug Metabolism and Disposition

SN - 0090-9556

IS - 1

ER -