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

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ABSTRACT

Purpose

The present study aimed to develop a high-throughput screening strategy for predicting the phototoxic potential of pharmaceutical substances, using a derivatives-of-reactive-oxygen-metabolites (D-ROM) assay.

Methods

The assay conditions of the D-ROM assay were optimized with a focus on screening run time, sensitivity, solvent system, and reproducibility. The phototoxic potentials of 25 model compounds were assessed by the D-ROM assay, as well as by other screening systems for comparison, including the reactive oxygen species (ROS) assay, the DNA-photocleavage assay, and the 3T3 neutral red uptake phototoxicity test (3T3 NRU PT).

Results

Some phototoxic drugs tended to yield D-ROM when exposed to simulated sunlight (250 W/m2), whereas D-ROM generation was negligible for non-phototoxic chemicals. Compared with the ROS assay, the assay procedure for the D-ROM assay was highly simplified with a marked reduction in screening run time. Comparative experiments also demonstrated that D-ROM data were related to the outcomes of the DNA-photocleavage assay and the 3T3 NRU PT, with prediction accuracies of 76 and 72%, respectively.

Conclusion

The D-ROM assay has potential for identifying the phototoxic potential of a large number of new drugs as a 1st screening system in the early stages of drug discovery.

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Abbreviations

3T3 NRU PT:

3T3 neutral red uptake phototoxicity test

5-FU:

5-fluorouracil

8-MOP:

8-methoxypsoralen

AFM:

atomic force microscopy

AGE:

agarose gel electrophoresis

CD:

circular dichroism

DEPPD:

N,N-diethyl-p-phenylenediamine

DMEM:

Dulbecco’s Modified Eagle Medium

D-ROM:

derivatives of reactive oxygen metabolites

EBSS:

Earle’s Balanced Salt Solution

EtBr:

ethidium bromide

ECVAM:

Europe Center for the Validation of Alternative Methods

OC:

open circular

OECD:

Organisation for Economic Co-operation and Development

PIF:

photoirritation factor

ROS:

reactive oxygen species

SC:

supercoiled

UV:

ultraviolet

VIS light:

visible light

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ACKNOWLEDGEMENTS

This work was supported in part by a Grant-in-Aid from the Food Safety Commission, Japan (No. 0807).

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Authors and Affiliations

  1. Department of Pharmacokinetics and Pharmacodynamics and Global Center of Excellence (COE) Program, School of Pharmaceutical Sciences, University of Shizuoka, 52-1 Yada, Suruga-ku, Shizuoka, 422-8526, Japan

    Satomi Onoue, Masanori Ochi, Yoshiki Seto & Shizuo Yamada

  2. Drug Safety Research and Development, Pfizer Global Research and Development, Ramsgate Rd., Sandwich, CT13 9NJ, UK

    Graham Gandy

  3. Graduate School of Pharmaceutical Sciences, Chiba University, 1-8-1 Inohana, Chuo-ku, Chiba, 260-8675, Japan

    Naoko Igarashi

  4. Department of Pharmaceutical Physical Chemistry College of Pharmaceutical Sciences, Matsuyama University, 4-2, Bunkyo, Matsuyama, Ehime, 790-8578, Japan

    Yukinori Yamauchi

Authors
  1. Satomi Onoue
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  2. Masanori Ochi
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  3. Graham Gandy
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  4. Yoshiki Seto
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  5. Naoko Igarashi
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  6. Yukinori Yamauchi
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  7. Shizuo Yamada
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Corresponding author

Correspondence to Satomi Onoue.

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Onoue, S., Ochi, M., Gandy, G. et al. High-Throughput Screening System for Identifying Phototoxic Potential of Drug Candidates Based on Derivatives of Reactive Oxygen Metabolites. Pharm Res 27, 1610–1619 (2010). https://doi.org/10.1007/s11095-010-0161-3

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