Advanced

Characterization of drug-protein binding process by employing equilibrium sampling through hollow-fiber supported liquid membrane and Bjerrum and Scatchard plots.

Barri, Thaer LU ; Trtić-Petrović, Tatjana; Karlsson, Michael and Jönsson, Jan Åke LU (2008) In Journal of Pharmaceutical and Biomedical Analysis 48(1). p.49-56
Abstract
The technique equilibrium sampling through membrane (ESTM) was extended to measuring the free drug concentration in solutions of drug and protein. Bjerrum and Scatchard plots were employed for characterizing individual drug binding to pure human blood proteins. Four drugs were investigated as a model system: fluvoxamine and ropivacaine which dominantly bind to alpha-acid glycoprotein (AGP), and R,S-ibuprofen and S-ketoprofen which highly bind to human serum albumin (HSA). The level of drug binding to AGP and HSA relied on drug and protein concentrations. Bjerrum and Scatchard plots revealed high affinity constants (K(a)) at low protein concentration. Both Bjerrum and Scatchard plots of fluvoxamine and ropivacaine binding to AGP showed one... (More)
The technique equilibrium sampling through membrane (ESTM) was extended to measuring the free drug concentration in solutions of drug and protein. Bjerrum and Scatchard plots were employed for characterizing individual drug binding to pure human blood proteins. Four drugs were investigated as a model system: fluvoxamine and ropivacaine which dominantly bind to alpha-acid glycoprotein (AGP), and R,S-ibuprofen and S-ketoprofen which highly bind to human serum albumin (HSA). The level of drug binding to AGP and HSA relied on drug and protein concentrations. Bjerrum and Scatchard plots revealed high affinity constants (K(a)) at low protein concentration. Both Bjerrum and Scatchard plots of fluvoxamine and ropivacaine binding to AGP showed one specific binding site (n(1)=1) with ropivacaine K(a) value close to 5 times higher than the K(a) of fluvoxamine at 22.9muM AGP concentration. Bjerrum plots of ketoprofen and ibuprofen gave total number of binding sites or bound molecules of 6-7, which did not depend on the drug or protein concentration. Scatchard plots of ketoprofen and ibuprofen exhibited two binding sites (n(1) and n(2)) at 0.15muM and 0.75muM HSA concentrations. On one hand, at 0.15muM HSA, ketoprofen and ibuprofen were bound to site I at n(1)=1.2 and n(1)=1.0, respectively. However, at 0.75muM HSA, ketoprofen and ibuprofen were bound to site I at n(1)=1.2 and n(1)=1.9, respectively. On the other hand, site II, at 0.15muM HSA, interacted with ketoprofen and ibuprofen at n(2)=5.6 and 6.7, respectively. However, at 0.75muM HSA, site II interacted with ketoprofen at n(2)=7.4 and ibuprofen at n(2)=6.2. It would be concluded that, upon mixing ketoprofen and ibuprofen in a HSA solution, a ketoprofen-ibuprofen interaction would most likely occur at site II in HSA. (Less)
Please use this url to cite or link to this publication:
author
organization
publishing date
type
Contribution to journal
publication status
published
subject
in
Journal of Pharmaceutical and Biomedical Analysis
volume
48
issue
1
pages
49 - 56
publisher
Elsevier
external identifiers
  • wos:000259339800007
  • pmid:18565712
  • scopus:49849105510
ISSN
0731-7085
DOI
10.1016/j.jpba.2008.04.030
language
English
LU publication?
yes
id
f9c00858-4a78-4b07-bc53-7d850620b45c (old id 1168677)
date added to LUP
2009-03-02 09:36:00
date last changed
2017-10-01 03:44:11
@article{f9c00858-4a78-4b07-bc53-7d850620b45c,
  abstract     = {The technique equilibrium sampling through membrane (ESTM) was extended to measuring the free drug concentration in solutions of drug and protein. Bjerrum and Scatchard plots were employed for characterizing individual drug binding to pure human blood proteins. Four drugs were investigated as a model system: fluvoxamine and ropivacaine which dominantly bind to alpha-acid glycoprotein (AGP), and R,S-ibuprofen and S-ketoprofen which highly bind to human serum albumin (HSA). The level of drug binding to AGP and HSA relied on drug and protein concentrations. Bjerrum and Scatchard plots revealed high affinity constants (K(a)) at low protein concentration. Both Bjerrum and Scatchard plots of fluvoxamine and ropivacaine binding to AGP showed one specific binding site (n(1)=1) with ropivacaine K(a) value close to 5 times higher than the K(a) of fluvoxamine at 22.9muM AGP concentration. Bjerrum plots of ketoprofen and ibuprofen gave total number of binding sites or bound molecules of 6-7, which did not depend on the drug or protein concentration. Scatchard plots of ketoprofen and ibuprofen exhibited two binding sites (n(1) and n(2)) at 0.15muM and 0.75muM HSA concentrations. On one hand, at 0.15muM HSA, ketoprofen and ibuprofen were bound to site I at n(1)=1.2 and n(1)=1.0, respectively. However, at 0.75muM HSA, ketoprofen and ibuprofen were bound to site I at n(1)=1.2 and n(1)=1.9, respectively. On the other hand, site II, at 0.15muM HSA, interacted with ketoprofen and ibuprofen at n(2)=5.6 and 6.7, respectively. However, at 0.75muM HSA, site II interacted with ketoprofen at n(2)=7.4 and ibuprofen at n(2)=6.2. It would be concluded that, upon mixing ketoprofen and ibuprofen in a HSA solution, a ketoprofen-ibuprofen interaction would most likely occur at site II in HSA.},
  author       = {Barri, Thaer and Trtić-Petrović, Tatjana and Karlsson, Michael and Jönsson, Jan Åke},
  issn         = {0731-7085},
  language     = {eng},
  number       = {1},
  pages        = {49--56},
  publisher    = {Elsevier},
  series       = {Journal of Pharmaceutical and Biomedical Analysis},
  title        = {Characterization of drug-protein binding process by employing equilibrium sampling through hollow-fiber supported liquid membrane and Bjerrum and Scatchard plots.},
  url          = {http://dx.doi.org/10.1016/j.jpba.2008.04.030},
  volume       = {48},
  year         = {2008},
}