Metabolic stereoselectivity of cytochrome P450 3A4 towards deoxypodophyllotoxin: In silico predictions and experimental validation

Mattijs K. Julsing; Nikolay P. Vasilev; Dina Schneidman-Duhovny; Remco Muntendam; Herman J. Woerdenbag; Wim J. Quax; Haim J. Wolfson; Iliana Ionkova; Oliver Kayser

doi:10.1016/j.ejmech.2007.09.005

Metabolic stereoselectivity of cytochrome P450 3A4 towards deoxypodophyllotoxin: In silico predictions and experimental validation

Mattijs K. Julsing, Nikolay P. Vasilev, Dina Schneidman-Duhovny, Remco Muntendam, Herman J. Woerdenbag, Wim J. Quax, Haim J. Wolfson, Iliana Ionkova, Oliver Kayser^*

^*Corresponding author for this work

Research output: Contribution to journal › Article › peer-review

26 Scopus citations

Abstract

Deoxypodophyllotoxin is stereoselectively converted into epipodophyllotoxin by recombinant human cytochrome P450 3A4 (CYP3A4). Further kinetic analysis revealed that the Michaelis-Menten K_m and V_max for hydroxylation of deoxypodophyllotoxin by CYP3A4 at C7 position were 1.93 μM and 1.48 nmol/min/nmol, respectively. Deoxypodophyllotoxin was subjected to automated docking analysis in order to get better knowledge of the interaction between the CYP3A4 enzyme and the substrate, using the PatchDock algorithm with distance constraints. Automated docking showed that the β-hydrogen atom at C7 position is in the most appropriate binding orientation at the site of oxidation. The docking results are consistent with the experimental data for the bioconversion of deoxypodophyllotoxin into epipodophyllotoxin by CYP3A4. In addition, the effects of five lignans, deoxypodophyllotoxin, epipodophyllotoxin, podophyllotoxin, demethylenedeoxypodophyllotoxin, and demethylenepodophyllotoxin, on CYP3A4 were compared in order to investigate the influence of the methylenedioxy group on the biotransformation process, to give insight into the mode of metabolization and to explain inhibitory activity of lignans.

Original language	English
Pages (from-to)	1171-1179
Number of pages	9
Journal	European Journal of Medicinal Chemistry
Volume	43
Issue number	6
DOIs	https://doi.org/10.1016/j.ejmech.2007.09.005
State	Published - Jun 2008
Externally published	Yes

Bibliographical note

Funding Information:
The authors would like to express their gratitude to F.P. Guengerich for providing us with the plasmids containing the genes encoding the human CYP3A4 and NADPH-P450 reductase and M.A. Castro for providing us with the lignan compounds. The research of H.J. Wolfson has been supported in part by the Israel Science Foundation (grant no. 281/05) and Hermann Minkowski Minerva Center for Geometry at Tel Aviv University. Financial support by the Huygens Program to N.P. Vasilev and Miiggenburg – Foundation is gratefully acknowledged.

Keywords

Automated docking
Cytochrome P450 3A4 (EC 1.14.14.1)
Drug metabolism
Epipodophyllotoxin
Lignans

Access to Document

10.1016/j.ejmech.2007.09.005

Cite this

Julsing, M. K., Vasilev, N. P., Schneidman-Duhovny, D., Muntendam, R., Woerdenbag, H. J., Quax, W. J., Wolfson, H. J., Ionkova, I., & Kayser, O. (2008). Metabolic stereoselectivity of cytochrome P450 3A4 towards deoxypodophyllotoxin: In silico predictions and experimental validation. European Journal of Medicinal Chemistry, 43(6), 1171-1179. https://doi.org/10.1016/j.ejmech.2007.09.005

@article{606ab4e235b24cb6932535726efe8aa7,

title = "Metabolic stereoselectivity of cytochrome P450 3A4 towards deoxypodophyllotoxin: In silico predictions and experimental validation",

abstract = "Deoxypodophyllotoxin is stereoselectively converted into epipodophyllotoxin by recombinant human cytochrome P450 3A4 (CYP3A4). Further kinetic analysis revealed that the Michaelis-Menten Km and Vmax for hydroxylation of deoxypodophyllotoxin by CYP3A4 at C7 position were 1.93 μM and 1.48 nmol/min/nmol, respectively. Deoxypodophyllotoxin was subjected to automated docking analysis in order to get better knowledge of the interaction between the CYP3A4 enzyme and the substrate, using the PatchDock algorithm with distance constraints. Automated docking showed that the β-hydrogen atom at C7 position is in the most appropriate binding orientation at the site of oxidation. The docking results are consistent with the experimental data for the bioconversion of deoxypodophyllotoxin into epipodophyllotoxin by CYP3A4. In addition, the effects of five lignans, deoxypodophyllotoxin, epipodophyllotoxin, podophyllotoxin, demethylenedeoxypodophyllotoxin, and demethylenepodophyllotoxin, on CYP3A4 were compared in order to investigate the influence of the methylenedioxy group on the biotransformation process, to give insight into the mode of metabolization and to explain inhibitory activity of lignans.",

keywords = "Automated docking, Cytochrome P450 3A4 (EC 1.14.14.1), Drug metabolism, Epipodophyllotoxin, Lignans",

author = "Julsing, {Mattijs K.} and Vasilev, {Nikolay P.} and Dina Schneidman-Duhovny and Remco Muntendam and Woerdenbag, {Herman J.} and Quax, {Wim J.} and Wolfson, {Haim J.} and Iliana Ionkova and Oliver Kayser",

note = "Funding Information: The authors would like to express their gratitude to F.P. Guengerich for providing us with the plasmids containing the genes encoding the human CYP3A4 and NADPH-P450 reductase and M.A. Castro for providing us with the lignan compounds. The research of H.J. Wolfson has been supported in part by the Israel Science Foundation (grant no. 281/05) and Hermann Minkowski Minerva Center for Geometry at Tel Aviv University. Financial support by the Huygens Program to N.P. Vasilev and Miiggenburg – Foundation is gratefully acknowledged.",

year = "2008",

month = jun,

doi = "10.1016/j.ejmech.2007.09.005",

language = "אנגלית",

volume = "43",

pages = "1171--1179",

journal = "European Journal of Medicinal Chemistry",

issn = "0223-5234",

publisher = "Elsevier Masson s.r.l.",

number = "6",

}

Julsing, MK, Vasilev, NP, Schneidman-Duhovny, D, Muntendam, R, Woerdenbag, HJ, Quax, WJ, Wolfson, HJ, Ionkova, I & Kayser, O 2008, 'Metabolic stereoselectivity of cytochrome P450 3A4 towards deoxypodophyllotoxin: In silico predictions and experimental validation', European Journal of Medicinal Chemistry, vol. 43, no. 6, pp. 1171-1179. https://doi.org/10.1016/j.ejmech.2007.09.005

TY - JOUR

T1 - Metabolic stereoselectivity of cytochrome P450 3A4 towards deoxypodophyllotoxin

T2 - In silico predictions and experimental validation

AU - Julsing, Mattijs K.

AU - Vasilev, Nikolay P.

AU - Schneidman-Duhovny, Dina

AU - Muntendam, Remco

AU - Woerdenbag, Herman J.

AU - Quax, Wim J.

AU - Wolfson, Haim J.

AU - Ionkova, Iliana

AU - Kayser, Oliver

N1 - Funding Information: The authors would like to express their gratitude to F.P. Guengerich for providing us with the plasmids containing the genes encoding the human CYP3A4 and NADPH-P450 reductase and M.A. Castro for providing us with the lignan compounds. The research of H.J. Wolfson has been supported in part by the Israel Science Foundation (grant no. 281/05) and Hermann Minkowski Minerva Center for Geometry at Tel Aviv University. Financial support by the Huygens Program to N.P. Vasilev and Miiggenburg – Foundation is gratefully acknowledged.

PY - 2008/6

Y1 - 2008/6

N2 - Deoxypodophyllotoxin is stereoselectively converted into epipodophyllotoxin by recombinant human cytochrome P450 3A4 (CYP3A4). Further kinetic analysis revealed that the Michaelis-Menten Km and Vmax for hydroxylation of deoxypodophyllotoxin by CYP3A4 at C7 position were 1.93 μM and 1.48 nmol/min/nmol, respectively. Deoxypodophyllotoxin was subjected to automated docking analysis in order to get better knowledge of the interaction between the CYP3A4 enzyme and the substrate, using the PatchDock algorithm with distance constraints. Automated docking showed that the β-hydrogen atom at C7 position is in the most appropriate binding orientation at the site of oxidation. The docking results are consistent with the experimental data for the bioconversion of deoxypodophyllotoxin into epipodophyllotoxin by CYP3A4. In addition, the effects of five lignans, deoxypodophyllotoxin, epipodophyllotoxin, podophyllotoxin, demethylenedeoxypodophyllotoxin, and demethylenepodophyllotoxin, on CYP3A4 were compared in order to investigate the influence of the methylenedioxy group on the biotransformation process, to give insight into the mode of metabolization and to explain inhibitory activity of lignans.

AB - Deoxypodophyllotoxin is stereoselectively converted into epipodophyllotoxin by recombinant human cytochrome P450 3A4 (CYP3A4). Further kinetic analysis revealed that the Michaelis-Menten Km and Vmax for hydroxylation of deoxypodophyllotoxin by CYP3A4 at C7 position were 1.93 μM and 1.48 nmol/min/nmol, respectively. Deoxypodophyllotoxin was subjected to automated docking analysis in order to get better knowledge of the interaction between the CYP3A4 enzyme and the substrate, using the PatchDock algorithm with distance constraints. Automated docking showed that the β-hydrogen atom at C7 position is in the most appropriate binding orientation at the site of oxidation. The docking results are consistent with the experimental data for the bioconversion of deoxypodophyllotoxin into epipodophyllotoxin by CYP3A4. In addition, the effects of five lignans, deoxypodophyllotoxin, epipodophyllotoxin, podophyllotoxin, demethylenedeoxypodophyllotoxin, and demethylenepodophyllotoxin, on CYP3A4 were compared in order to investigate the influence of the methylenedioxy group on the biotransformation process, to give insight into the mode of metabolization and to explain inhibitory activity of lignans.

KW - Automated docking

KW - Cytochrome P450 3A4 (EC 1.14.14.1)

KW - Drug metabolism

KW - Epipodophyllotoxin

KW - Lignans

UR - http://www.scopus.com/inward/record.url?scp=44349130284&partnerID=8YFLogxK

U2 - 10.1016/j.ejmech.2007.09.005

DO - 10.1016/j.ejmech.2007.09.005

M3 - ???researchoutput.researchoutputtypes.contributiontojournal.article???

C2 - 17998150

AN - SCOPUS:44349130284

SN - 0223-5234

VL - 43

SP - 1171

EP - 1179

JO - European Journal of Medicinal Chemistry

JF - European Journal of Medicinal Chemistry

IS - 6

ER -

Metabolic stereoselectivity of cytochrome P450 3A4 towards deoxypodophyllotoxin: In silico predictions and experimental validation

Abstract

Bibliographical note

Keywords

Access to Document

Other files and links

Fingerprint

Cite this