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Nucleic Acids Research, 1993, Vol. 21, No. 16 3671-3675
© 1993

CHEMISTRY

A quantitative assay to measure the relative DNA-binding affinity of pyrrolo[2,1-c][1,4]benzodiazepine (PBD) antitumour antibiotics based on the inhibition of restriction endonuclease BamhI

Madhu S. Puvvada, John A. Hartley1, Terence C. Jenkins2 and David E. Thurston*

School of Pharmacy and Biomedical Sciences, University of Portsmouth Park Building, King Henry 1st Street, Portsmouth PO1 2DZ 1University College London Medical School 91 Riding House Street, london W1P 8BT 2CRC Biomolecular Structure Unit, The Institute of Cancer Research Sutton, Surrey SM2 5NG, UK

*To whom correspondence should be addressed

Received May 14, 1993. Revised July 9, 1993. Accepted July 9, 1993.

An assay has been developed (restriction endonuclease digestion assay-RED100) based on inhibition of the restriction endonuclease BamHI that is capable of quantitative evaluation of the relative DNA-binding affinity of pyrrolo[2,1-c][1,4]benzodlazeplne (PBD) antitumour antibiotics. This method provides comparable results to those obtained from thermal denaturation and ethidium bromide displacement assays but is much more sensitive, discriminating between molecules of similar structure such as DC-81, Iso-DC-81 and neothramycin. The results reveal a trend between relative DNA-binding affinity and in vitro cytotoxlclty for the PBDs in two tumour cell lines studied.


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