High-Throughput Screening System for Identifying Phototoxic Potential of Drug Candidates Based on Derivatives of Reactive Oxygen Metabolites

Onoue, Satomi; Ochi, Mitsukazu; Gandy, Graham; Seto, Yoshiki; Igarashi, Naoko; Yamauchi, Yukiko; Yamada, Shizuo

doi:10.1007/s11095-010-0161-3

Cited by 20 publications

(2 citation statements)

References 32 publications

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“…These generated ROS result in local oxidative stress, causing breakage in genomic DNA and oxidative modifications in proteins and lipids within cell membranes . In order to avoid the undesirable effects of photoreactive compounds, efforts have been made to establish a model system for estimation of photosensitive potential through analytical and biochemical methods .…”

Section: Introductionmentioning

confidence: 99%

Naproxen intercalates with DNA and causes photocleavage through ROS generation

Husain

Yaseen

Rehman

et al. 2013

FEBS J

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Naproxen is an important non-steroidal anti-inflammatory drug with many pharmacological and biological properties. In this study, we have attempted to ascertain the mode of action and the mechanism of binding of naproxen to DNA. We have also demonstrated that, upon irradiation with white light, naproxen generates reactive oxygen species, causing DNA cleavage. Generation of reactive oxygen species from photo-irradiated naproxen as determined spectrophotometrically was found to lead to nicking of plasmid DNA as analyzed by agarose gel electrophoresis. Without photo-irradiation, naproxen binds to DNA and forms drug-DNA complexes as revealed by spectroscopic techniques. Several experiments such as determination of the effect of urea, iodide-induced quenching and a competitive binding assay with ethidium bromide showed that naproxen binds to DNA primarily in an intercalative manner. These observations were further supported by CD analysis, viscosity measurements and molecular docking. Using DNA as a template, fluorescence resonance energy transfer between naproxen and ethidium bromide was also observed, further strengthening the evidence for intercalative binding of naproxen with DNA.

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Section: Introductionmentioning

confidence: 99%

Naproxen intercalates with DNA and causes photocleavage through ROS generation

Husain

Yaseen

Rehman

et al. 2013

FEBS J

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“…Chemicals for the validation study were selected by chemical management group of the VMT in cooperation with Dr Manfred Liebsch (ZEBET), the Europe Center for the Validation of Alternative Methods (ECVAM), the Interagency Coordinating Committee on the Validation of Alternative Methods (ICCVAM) and the Korean Center for the Validation of Alternative Methods (KoCVAM). According to the reported in vitro/in vivo photosafety information and clinical observations (Durbize et al, 2003;Moore, 2002;Motley and Reynolds, 1989;Onoue et al, 2010;Peters and Holzhutter, 2002;Portes et al, 2002;Spielmann, 1994;Spielmann et al, 1994aSpielmann et al, , 1994bSpielmann et al, , 1998aSpielmann et al, , b, 1995Trevisi et al, 1994) and in-house assay results Spielmann (1994); Spielmann et al (1994aSpielmann et al ( , 1994bSpielmann et al ( , 1995 Phototoxic chemicals Validation study on ROS assay for photosafety assessment from in vitro 3 T3 neutral red uptake phototoxicity testing (3 T3 NRU PT), two standard chemicals and 42 test chemicals, including 23 phototoxins and 19 nonphototoxic drugs/chemicals, were selected for the intra-/inter-laboratory validation study (Table 1).…”

Section: Chemicals and Reagentsmentioning

confidence: 99%

Establishment and intra‐/inter‐laboratory validation of a standard protocol of reactive oxygen species assay for chemical photosafety evaluation

Onoue

Hosoi

Wakuri

et al. 2012

J of Applied Toxicology

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A reactive oxygen species (ROS) assay was previously developed for photosafety evaluation of pharmaceuticals, and the present multi-center study aimed to establish and validate a standard protocol for ROS assay. In three participating laboratories, two standards and 42 coded chemicals, including 23 phototoxins and 19 nonphototoxic drugs/chemicals, were assessed by the ROS assay according to the standardized protocol. Most phototoxins tended to generate singlet oxygen and/or superoxide under UV-vis exposure, but nonphototoxic chemicals were less photoreactive. In the ROS assay on quinine (200 µm), a typical phototoxic drug, the intra- and inter-day precisions (coefficient of variation; CV) were found to be 1.5-7.4% and 1.7-9.3%, respectively. The inter-laboratory CV for quinine averaged 15.4% for singlet oxygen and 17.0% for superoxide. The ROS assay on 42 coded chemicals (200 µm) provided no false negative predictions upon previously defined criteria as compared with the in vitro/in vivo phototoxicity, although several false positives appeared. Outcomes from the validation study were indicative of satisfactory transferability, intra- and inter-laboratory variability, and predictive capacity of the ROS assay.

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