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Metabolic Profile of 3-Acetyl-11-Keto-β-Boswellic Acid and 11-Keto-β-Boswellic Acid in Human Preparations In Vitro, Species Differences, and Bioactivity Variation

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A Correction to this article was published on 14 August 2019

Abstract

3-Acetyl-11-keto-β-boswellic acid (AKBA) and 11-keto-β-boswellic acid (KBA) are widely used in the clinic as anti-inflammatory drugs. However, these drugs have the poor bioavailability, which may be caused by their extensive metabolism. In this study, we systemically characterized both phase I and II metabolism of AKBA and KBA in vitro. In total, four major metabolites were firstly biosynthesized and identified using 1D and 2D NMR spectroscopy. Among them, three metabolites were novel. The kinetic parameters (K m , V max , CL int, and K i ) were also analyzed systematically in various biological samples. Finally, the deacetylation of AKBA and hydroxylation of KBA were confirmed to be the major metabolic pathways based on their large CL int and the high amounts of KBA (46.7%) and hydroxylated KBA (50.8%) along with a low amount of AKBA (2.50%) in human primary hepatocytes. Carboxylesterase 2 (CE2) selectively catalyzed the deacetylation of AKBA to form KBA. Although CYP3A4, CYP3A5, and CYP3A7 catalyzed the metabolism of KBA, CYP3A4 played a predominant role in the hydroxylation reaction of KBA in human. Notably, deacetylation and regioselective hydroxylation exhibited considerable species differences. Deacetylation was only observed in human liver microsomes and primary human hepatocytes; 21- and 20-mono-hydroxylation of KBA were primarily observed in human, monkey, and dog; and 16- and 30-mono-hydroxylation were observed in other species. More importantly, all four mono-hydroxylation metabolites exhibited a moderate anti-inflammatory activity. The 21- and 20-hydroxylation metabolites inhibited the expression of iNOS, the LPS-induced activation of IkBα and p65 phosphorylation, and suppressed p65 nuclear translocation in RAW264.7 cells.

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Abbreviations

AKBA:

3-Acetyl-11-keto-β-boswellic acid

KBA:

11-Keto-β-boswellic acid

HLM:

Human liver microsomes

HIM:

Human intestine microsomes

CyLM:

Monkey liver microsomes

RLM:

SD rat liver microsomes

AChE:

Acetylcholinesterase

BuChE:

Butyrylcholinesterase

HSA:

Human serum albumin

RSA:

Rat serum albumin

PLM:

Pig liver microsomes

DLM:

Dog liver microsomes

MLM:

Mouse liver microsomes

RaLM:

Rabbit liver microsomes

GLM:

Guinea pig liver microsomes

CEs:

Carboxylesterases

NADPH:

β-Nicotinamide adenine dinucleotide phosphate disodium salt

M-2:

21β-Hydroxy-11-keto-β-boswellic acid

M-3:

16β-Hydroxy-11-keto-β-boswellic acid

M-4:

30-Hydroxy-11-keto-β-boswellic acid

M-5:

20β-Hydroxy-11-keto-β-boswellic acid

HA:

Huperzine A

iso-OMPA:

Tetraisopropylpyrophosphoramide

BNPP:

Bis-p-nitrophenyl phosphate

LPA:

Loperamide

LPS:

Lipopolysaccharide

TEPA:

Triethylenethiophosphoramide

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Acknowledgments

This study was supported by the National Natural Science Foundation of China (Nos. 81274047, 81473334, and 81503201), the Distinguished Professor of Liaoning Province, Dalian Outstanding Youth Science and Technology Talent program (2014J11JH132 and 2015J12JH201), and the Liaoning Bai Qian Wan Talent Program and Innovation Team of Dalian Medical University.

Author Contributions

Xiaochi Ma, Yonglei Cui, and Xiangge Tian designed the experiments. Yonglei Cui and Jing Ning performed the experiments. Chao Wang, Zhenlong Yu, and Yan Wang analyzed the data. Xiaokui Huo, Lingling Jin, Sa Deng, and Baojing Zhang prepared the figures. Yonglei Cui and Xiangge Tian wrote the main text. All authors reviewed the manuscript.

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    Authors and Affiliations

    1. College of Pharmacy, Academy of Integrative Medicine, Dalian Medical University, 9 West Section, Lvshun South Road, Lvshunkou District, Dalian, 116044, China

      Yonglei Cui, Jing Ning, Chao Wang, Zhenlong Yu, Yan Wang, Xiaokui Huo, Lingling Jin, Sa Deng, Baojing Zhang & Xiaochi Ma

    2. College of Basic Medical Science, Dalian Medical University, Dalian, China

      Xiangge Tian

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    1. Yonglei Cui
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    Cui, Y., Tian, X., Ning, J. et al. Metabolic Profile of 3-Acetyl-11-Keto-β-Boswellic Acid and 11-Keto-β-Boswellic Acid in Human Preparations In Vitro, Species Differences, and Bioactivity Variation. AAPS J 18, 1273–1288 (2016). https://doi.org/10.1208/s12248-016-9945-7

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