High-throughput microplate assay for the determination of drug partition coefficients

• Type: Article in International Scientific Journal
• Date: 2010

Authors

Luis M Magalhaes (Author) Other; Claudia Nunes (Author) Other; Marlene Lucio (Author) FEUP; Marcela A Segundo (Author) FFUP; Salette Reis (Author) FFUP; Jose L F C Lima (Author) FFUP

Journal

Title: Nature Protocols

Vol. 5 No. 11

Pages: 1823-1830

ISSN: 1754-2189

Publisher: Springer Nature

Indexing

ISI Web of Knowledge - 34 Citations

ISI Web of Science

Scopus - 49 Citations

Pubmed / Medline

Scientific classification

FOS: Natural sciences > Biological sciences

CORDIS: Health sciences > Pharmacological sciences > Pharmacy

Other information

Authenticus ID: P-003-C2T

DOI: 10.1038/nprot.2010.137

Resumo (PT): Partition coefficients (Kp) of drugs between the phospholipid bilayer and the aqueous phase provide useful information in quantitative structure-activity relationship studies. Hexadecylphosphocholine (HePC) micelles, composed of a zwitterionic hydrophilic surface and a hydrophobic core, mimic the biomembranes and have several advantages over other lipid structures to assess Kp values. Their preparation is easy, fast and avoids the use of toxic organic solvents, and the output has fewer spectroscopic interferences. Here, we describe a high-throughput microplate protocol for assessing the Kp of drugs using HePC micelles as membrane models and derivative spectrophotometry as the detection technique. Moreover, the time-consuming data treatment to assess Kp values is easily performed by a dedicated Excel routine developed here and described in detail. The Kp values of nonsteroidal anti-inflammatory drugs (acemetacin, clonixin, diclofenac and indomethacin) were determined to show the simplicity of the method and to validate this protocol, which provides Kp values (n = 3) of two drugs in ~2 h. <br> <br>

Abstract (EN): Partition coefficients (K(p)) of drugs between the phospholipid bilayer and the aqueous phase provide useful information in quantitative structure-activity relationship studies. Hexadecylphosphocholine (HePC) micelles, composed of a zwitterionic hydrophilic surface and a hydrophobic core, mimic the biomembranes and have several advantages over other lipid structures to assess K(p) values. Their preparation is easy, fast and avoids the use of toxic organic solvents, and the output has fewer spectroscopic interferences. Here, we describe a high-throughput microplate protocol for assessing the K(p) of drugs using HePC micelles as membrane models and derivative spectrophotometry as the detection technique. Moreover, the time-consuming data treatment to assess K(p) values is easily performed by a dedicated Excel routine developed here and described in detail. The K(p) values of nonsteroidal anti-inflammatory drugs (acemetacin, clonixin, diclofenac and indomethacin) were determined to show the simplicity of the method and to validate this protocol, which provides K(p) values (n = 3) of two drugs in similar to 2 h.

Language: English

Type (Professor's evaluation): Scientific

No. of pages: 8