Discovery and structure-activity relationship of auriculatone: A potent hepatoprotective agent against acetaminophen-induced liver injury

doi:10.1016/j.bmcl.2017.07.028

Bioorganic & Medicinal Chemistry Letters

Volume 27, Issue 16, 15 August 2017, Pages 3636-3642

https://doi.org/10.1016/j.bmcl.2017.07.028 Get rights and content

Abstract

Acetaminophen (APAP, paracetamol) overdose has been the most frequent cause of drug-induced liver failure. APAP-induced liver toxicity can be fatal in many cases even with treatment of the clinically used N-acetylcysteine (NAC), and the need for novel therapeutic agents is apparent. Through evaluating the hepatoprotective effects of the co-occurring substances present in oleanolic acid tablets which have been used in China for decades as an adjuvant therapy for acute and chronic hepatitis, auriculatone was found to protect HL-7702 cells from APAP-induced liver injury comparable to NAC at the concentration of 10 μM. Structure activity relationship on auriculatone and its analogs showed that absence of the C17 carboxyl group of auriculatone was essential to achieve good hepatoprotective activity, and that the C3-OH, C16 carbonyl and C12-C13 olefinic group were critical for retaining the exceptional activity of auriculatone. Any modifications in the current investigation were all detrimental to the hepatoprotective activity. Docking and drug-metabolizing activity studies demonstrated that CYP3A4 was likely the main target of auriculatone, and that auriculatone elicited the hepatoprotective effect possibly through inhibiting CYP3A4’s metabolism of APAP to the toxic metabolite NAPQI. The work may pave the way for the use of auriculatone in the treatment of APAP-induced liver injury.

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Acknowledgements

This work was supported by the National Natural Science Foundation of China (81260506), Joint Foundation of Guizhou Province Department of Science and Technology and Guizhou Medical University ([2015]7357), Foundation for Doctor of Guiyang Medical University ([2014]008), Excellent Scientific and Technological Yong Talent of Guizhou Province ([2015]32), Natural Science Foundation of Education Department of Guizhou Province ([2015]441), and the Innovation Team of Natural Science Foundation of

References (37)

S.S. Lee et al.
Role of CYP2E1 in the hepatotoxicity of acetaminophen
J Biol Chem
(1996)
K.E. Thummel et al.
Oxidation of acetaminophen to N-acetyl-p-aminobenzoquinone imine by human CYP3A4
Biochem Pharmacol
(1993)
L.F. Prescott et al.
Treatment of paracetamol (acetaminophen) poisoning with N-acetylcysteine
Lancet
(1977)
Y. Xie et al.
Mechanisms of acetaminophen-induced cell death in primary human hepatocytes
Toxicol Appl Pharmacol
(2014)
M.E. Mullins et al.
What is the rate of adverse events with intravenous versus oral N-acetylcysteine in pediatric patients?
Ann Emerg Med
(2004)
A.F. Pizon et al.
Adverse reaction from use of intravenous N-acetylcysteine
J Emerg Med
(2006)
H. Wang et al.
Elucidation of the pharmacophore of echinocystic acid, a new lead for blocking HCV entry
Eur J Med Chem
(2013)
M.T. Periasamy
M. Methods of enhancement of reactivity and selectivity of sodium borohydride for applications in organic synthesis
J Organomet Chem
(2000)
M.J. Hodgman et al.
A review of acetaminophen poisoning
Crit Care Clin
(2012)
G. Branden et al.
Structure-based ligand design to overcome CYP inhibition in drug discovery projects
Drug Discov Today
(2014)

E.S. Bjornsson

Drug-induced liver injury: an overview over the most critical compounds

Arch Toxicol

(2015)

A.M. Larson et al.

Acetaminophen-induced acute liver failure: results of a United States multicenter, prospective study

Hepatology

(2005)

S.D. Nelson

Molecular mechanisms of the hepatotoxicity caused by acetaminophen

Semin Liver Dis

(1990)

D.J. Jollow et al.

Acetaminophen-induced hepatic necrosis. II. Role of covalent binding in vivo

J Pharmacol Exp Ther

(1973)

Z. Lu et al.

SIRT3-dependent deacetylation exacerbates acetaminophen hepatotoxicity

EMBO Rep

(2011)

W.Z. Potter et al.

Acetaminophen-induced hepatic necrosis. 3. Cytochrome P-450-mediated covalent binding in vitro

J Pharmacol Exp Ther

(1973)

C. Cheung et al.

The cyp2e1-humanized transgenic mouse: role of cyp2e1 in acetaminophen hepatotoxicity

Drug Metab Dispos

(2005)

C.J. Patten et al.

Cytochrome P450 enzymes involved in acetaminophen activation by rat and human liver microsomes and their kinetics

Chem Res Toxicol

(1993)

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Although therapeutic doses of APAP are effective and safe, administration of an overdose can lead to acute hepatic necrosis, liver failure, and even death (Yuan et al., 2010; Zimmerman, 1981). At present, although the metformin (Kim et al., 2015), anti-TNF-α agents (Gao and Bataller, 2011) and N-acetylcysteine (NAC) (Zhou et al., 2017) were used to cure liver injury induced by APAP, these drugs were not recommended for patients because of the potential severe hepatotoxicity and adverse effect. Some studies began to found new agents from other sources, such as the traditional medicinal plants with some active properties, which could relieve the liver injury induced by APAP and possess huge promise for being developed into drugs for treatment of the liver diseases caused by APAP. (
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^e: These authors contributed equally to this work.

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Discovery and structure-activity relationship of auriculatone: A potent hepatoprotective agent against acetaminophen-induced liver injury

Abstract

Graphical abstract

Section snippets

Acknowledgements

J Biol Chem

Biochem Pharmacol

Lancet

Toxicol Appl Pharmacol

Ann Emerg Med

J Emerg Med

Eur J Med Chem

J Organomet Chem

Crit Care Clin

Drug Discov Today

Drug-induced liver injury: an overview over the most critical compounds

Arch Toxicol

Acetaminophen-induced acute liver failure: results of a United States multicenter, prospective study

Hepatology

Molecular mechanisms of the hepatotoxicity caused by acetaminophen

Semin Liver Dis

Acetaminophen-induced hepatic necrosis. II. Role of covalent binding in vivo

J Pharmacol Exp Ther

SIRT3-dependent deacetylation exacerbates acetaminophen hepatotoxicity

EMBO Rep

Acetaminophen-induced hepatic necrosis. 3. Cytochrome P-450-mediated covalent binding in vitro

J Pharmacol Exp Ther

The cyp2e1-humanized transgenic mouse: role of cyp2e1 in acetaminophen hepatotoxicity

Drug Metab Dispos

Cytochrome P450 enzymes involved in acetaminophen activation by rat and human liver microsomes and their kinetics

Chem Res Toxicol