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Flavonoids as inhibitors of human carbonyl reductase 1

Carlquist, Magnus LU ; Frejd, Torbjörn LU and Gorwa-Grauslund, Marie-Francoise LU (2008) In Chemico-Biological Interactions 174(2). p.98-108
Abstract
Human carbonyl reductase 1 (CBR1), that is one of the enzymes responsible for the reduced efficiency of treatments by the antineoplastic agents anthracyclines, was functionally expressed in Saccharomyces cerevisiae. CBR1 was purified and kinetically characterised using daunorubicin as substrate. CBR1-catalysed reduction of daunorubicin followed an apparent Michaelis-Menten kinetics with K-M = 85.2 +/- 26.7 mu M and V-max =3490 +/- 220 mu mol/(min g protein). The type of inhibition for the flavonoid compound rutin was determined by studying initial reaction rates in the presence of rutin. The inhibition kinetics was found to follow an apparent mixed inhibition with K-ic = 1.8 +/- 1.2 mu M and K-iu = 2.8 +/- 1.6 mu M. IC50-values were also... (More)
Human carbonyl reductase 1 (CBR1), that is one of the enzymes responsible for the reduced efficiency of treatments by the antineoplastic agents anthracyclines, was functionally expressed in Saccharomyces cerevisiae. CBR1 was purified and kinetically characterised using daunorubicin as substrate. CBR1-catalysed reduction of daunorubicin followed an apparent Michaelis-Menten kinetics with K-M = 85.2 +/- 26.7 mu M and V-max =3490 +/- 220 mu mol/(min g protein). The type of inhibition for the flavonoid compound rutin was determined by studying initial reaction rates in the presence of rutin. The inhibition kinetics was found to follow an apparent mixed inhibition with K-ic = 1.8 +/- 1.2 mu M and K-iu = 2.8 +/- 1.6 mu M. IC50-values were also determined for a set of flavonoids in order to identify essential structure for inhibition activity. Computational docking experiments of the four best inhibitors to the catalytic site of CBR1 showed that the flavonoid skeleton structure was the binding part of the molecule. The presence of a sugar moiety in I and 2, or a sugar mimicking part in 9. directed the orientation of the flavonoid so that the sugars were pointing outwards, giving rise to a stabilising effect to the binding. Finally, additional binding epitopes that interacted with various parts of the flavonoid ligand were identified and could potentially be targeted for further improvement of inhibition activity. These included; hydrogen-binding sites surrounding Ser139 and Cys226, Met234 and Tyr193 or Trp229; aromatic-aromatic interaction with Tyr193, Trp229 or NADPH; van der Waals interactions with IIe140. (C) 2008 Elsevier Ireland Ltd. All rights reserved. (Less)
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author
organization
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Contribution to journal
publication status
published
subject
keywords
flavonoids, Saccharomyces cerevisiae, adriamycin, doxorubicin, daunorubicin, anthracyclines, human carbonyl reductase 1, CBR1, mixed, inhibition, rutin, IC50
in
Chemico-Biological Interactions
volume
174
issue
2
pages
98 - 108
publisher
Elsevier
external identifiers
  • wos:000258057500003
  • scopus:46049109896
ISSN
1872-7786
DOI
10.1016/j.cbi.2008.05.021
language
English
LU publication?
yes
id
a1c0efc8-9bae-4316-9d55-ad434869389e (old id 1253790)
date added to LUP
2008-10-28 14:18:40
date last changed
2017-11-12 03:39:31
@article{a1c0efc8-9bae-4316-9d55-ad434869389e,
  abstract     = {Human carbonyl reductase 1 (CBR1), that is one of the enzymes responsible for the reduced efficiency of treatments by the antineoplastic agents anthracyclines, was functionally expressed in Saccharomyces cerevisiae. CBR1 was purified and kinetically characterised using daunorubicin as substrate. CBR1-catalysed reduction of daunorubicin followed an apparent Michaelis-Menten kinetics with K-M = 85.2 +/- 26.7 mu M and V-max =3490 +/- 220 mu mol/(min g protein). The type of inhibition for the flavonoid compound rutin was determined by studying initial reaction rates in the presence of rutin. The inhibition kinetics was found to follow an apparent mixed inhibition with K-ic = 1.8 +/- 1.2 mu M and K-iu = 2.8 +/- 1.6 mu M. IC50-values were also determined for a set of flavonoids in order to identify essential structure for inhibition activity. Computational docking experiments of the four best inhibitors to the catalytic site of CBR1 showed that the flavonoid skeleton structure was the binding part of the molecule. The presence of a sugar moiety in I and 2, or a sugar mimicking part in 9. directed the orientation of the flavonoid so that the sugars were pointing outwards, giving rise to a stabilising effect to the binding. Finally, additional binding epitopes that interacted with various parts of the flavonoid ligand were identified and could potentially be targeted for further improvement of inhibition activity. These included; hydrogen-binding sites surrounding Ser139 and Cys226, Met234 and Tyr193 or Trp229; aromatic-aromatic interaction with Tyr193, Trp229 or NADPH; van der Waals interactions with IIe140. (C) 2008 Elsevier Ireland Ltd. All rights reserved.},
  author       = {Carlquist, Magnus and Frejd, Torbjörn and Gorwa-Grauslund, Marie-Francoise},
  issn         = {1872-7786},
  keyword      = {flavonoids,Saccharomyces cerevisiae,adriamycin,doxorubicin,daunorubicin,anthracyclines,human carbonyl reductase 1,CBR1,mixed,inhibition,rutin,IC50},
  language     = {eng},
  number       = {2},
  pages        = {98--108},
  publisher    = {Elsevier},
  series       = {Chemico-Biological Interactions},
  title        = {Flavonoids as inhibitors of human carbonyl reductase 1},
  url          = {http://dx.doi.org/10.1016/j.cbi.2008.05.021},
  volume       = {174},
  year         = {2008},
}