Improvement of cyclophosphamide activation by CYP2B6 mutants: from in silico to ex vivo

Molecular Pharmacology
Thien-An NguyenIsabelle de Waziers

Abstract

Cyclophosphamide (CPA) is a chemotherapeutic agent that is primarily activated in the liver by cytochrome P4502B6 (CYP2B6) and then transported to the tumor via blood flow. To prevent deleterious secondary effects, P450-based gene-directed enzyme prodrug therapy (GDEPT) consists of expressing CYP2B6 in tumor cells before CPA treatment. Given the relatively low affinity of CYP2B6 for CPA, the aim of our work was to modify CYP2B6 to increase its catalytic efficiency (V(max)/K(m)) to metabolize CPA into 4'-OH CPA. A molecular model of CYP2B6 was built, and four residues in close contact with the substrate were subjected to mutagenesis. Canine CYP2B11 exhibiting a particularly low K(m) to CPA, the amino acids exclusively present in the CYP2B11 substrate recognition sequences were substituted in human CYP2B6. All mutants (n = 26) were expressed in Saccharomyces cerevisiae and their enzymatic constants (K(m), V(max)) evaluated using CPA as substrate. Five mutants exhibited a 2- to 3-fold higher catalytic efficiency than wild-type CYP2B6. A double mutant, comprising the two most effective mutations, showed a 4-fold increase in K(m)/V(max). Molecular dynamic simulations of several mutants were found to be consistent with the observed m...Continue Reading

References

Jun 1, 1992·Journal of Clinical Oncology : Official Journal of the American Society of Clinical Oncology·L J AyashE Frei
Mar 1, 1972·European Journal of Clinical Pharmacology·B SchoeneH F von Oldershausen
Dec 5, 1993·Journal of Molecular Biology·A Sali, T L Blundell
Jun 1, 1993·Journal of Clinical Oncology : Official Journal of the American Society of Clinical Oncology·W P PetersE Shpall
Aug 1, 1996·Methods : a Companion to Methods in Enzymology·H LussA K Nussler
May 21, 1999·Drug Metabolism Reviews·D J WaxmanY Jounaidi
Sep 2, 1999·Pharmacogenetics·L GervotI de Waziers
Jul 27, 2001·Current Drug Metabolism·T L Domanski, J R Halpert
Dec 17, 2002·Journal of Molecular Modeling·Christine BatheltJürgen Pleiss
Dec 20, 2002·The Journal of Pharmacology and Experimental Therapeutics·Margit SpatzeneggerJames R Halpert
Apr 30, 2003·Protein Science : a Publication of the Protein Society·Björn Wallner, Arne Elofsson
Oct 18, 2003·Proceedings of the National Academy of Sciences of the United States of America·Emily E ScottC David Stout
Feb 1, 2005·Archives of Biochemistry and Biophysics·Wataru HonmaJames R Halpert
Mar 5, 2005·Clinical Cancer Research : an Official Journal of the American Association for Cancer Research·Jeremy P BraybrookeAdrian L Harris
Jun 24, 2006·BMC Bioinformatics·Martin PetrekJirí Damborský
Aug 22, 2006·Biochimica Et Biophysica Acta·Vlad CojocaruRebecca C Wade
Aug 29, 2006·Biochimica Et Biophysica Acta·Yonghong Zhao, James R Halpert

❮ Previous
Next ❯

Citations

Mar 26, 2010·Drug Metabolism and Disposition : the Biological Fate of Chemicals·Qingbiao Huang, Grazyna D Szklarz
Jan 31, 2013·Future Medicinal Chemistry·Paul R Ortiz de Montellano
Mar 27, 2016·Biochimica Et Biophysica Acta·Thomas LautierGilles Truan
Feb 22, 2012·Biotechnology Advances·Jan BrezovskyJiri Damborsky
Aug 28, 2016·Journal of Controlled Release : Official Journal of the Controlled Release Society·Ikrame AmaraIsabelle de Waziers
Dec 14, 2016·Medicinal Research Reviews·Sergio M MarquesJiri Damborsky
Feb 12, 2017·Journal of Analytical Toxicology·Taha AhmadGary O Rankin
Nov 2, 2017·Cardiovascular and Interventional Radiology·Olivier PellerinIsabelle de Waziers
Jun 1, 2018·International Journal of Molecular Sciences·Philippe UrbanGilles Truan
Sep 8, 2018·Journal of Cellular Physiology·Faroogh MarofiMajid Farshdousti Hagh
May 28, 2009·Journal of Separation Science·Daniel I RanieriOliver Hofstetter
Jul 14, 2017·Journal of Cellular Physiology·Rana Moradian TehraniHamed Mirzaei
Apr 2, 2019·Current Topics in Medicinal Chemistry·Benjamin Diethelm-VarelaFengtian Xue
Jun 20, 2018·International Journal of Molecular Sciences·Sonia Nava-SalazarAmerica Vanoye-Carlo
Nov 20, 2019·Oncoimmunology·Benjamin NayagomIsabelle de Waziers
May 13, 2009·Journal of Chemical Information and Modeling·Jamel Eddine MeslamaniMichel Petitjean
Jun 3, 2021·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Zeinab Breijyeh, Rafik Karaman

❮ Previous
Next ❯

Related Concepts

Trending Feeds

COVID-19

Coronaviruses encompass a large family of viruses that cause the common cold as well as more serious diseases, such as the ongoing outbreak of coronavirus disease 2019 (COVID-19; formally known as 2019-nCoV). Coronaviruses can spread from animals to humans; symptoms include fever, cough, shortness of breath, and breathing difficulties; in more severe cases, infection can lead to death. This feed covers recent research on COVID-19.

Blastomycosis

Blastomycosis fungal infections spread through inhaling Blastomyces dermatitidis spores. Discover the latest research on blastomycosis fungal infections here.

Nuclear Pore Complex in ALS/FTD

Alterations in nucleocytoplasmic transport, controlled by the nuclear pore complex, may be involved in the pathomechanism underlying multiple neurodegenerative diseases including Amyotrophic Lateral Sclerosis and Frontotemporal Dementia. Here is the latest research on the nuclear pore complex in ALS and FTD.

Applications of Molecular Barcoding

The concept of molecular barcoding is that each original DNA or RNA molecule is attached to a unique sequence barcode. Sequence reads having different barcodes represent different original molecules, while sequence reads having the same barcode are results of PCR duplication from one original molecule. Discover the latest research on molecular barcoding here.

Chronic Fatigue Syndrome

Chronic fatigue syndrome is a disease characterized by unexplained disabling fatigue; the pathology of which is incompletely understood. Discover the latest research on chronic fatigue syndrome here.

Evolution of Pluripotency

Pluripotency refers to the ability of a cell to develop into three primary germ cell layers of the embryo. This feed focuses on the mechanisms that underlie the evolution of pluripotency. Here is the latest research.

Position Effect Variegation

Position Effect Variagation occurs when a gene is inactivated due to its positioning near heterochromatic regions within a chromosome. Discover the latest research on Position Effect Variagation here.

STING Receptor Agonists

Stimulator of IFN genes (STING) are a group of transmembrane proteins that are involved in the induction of type I interferon that is important in the innate immune response. The stimulation of STING has been an active area of research in the treatment of cancer and infectious diseases. Here is the latest research on STING receptor agonists.

Microbicide

Microbicides are products that can be applied to vaginal or rectal mucosal surfaces with the goal of preventing, or at least significantly reducing, the transmission of sexually transmitted infections. Here is the latest research on microbicides.