Mode of action of DNA-competitive small molecule inhibitors of tyrosyl DNA phosphodiesterase 2

Peter  Hornyak; Trevor  Askwith; Sarah Walker; Emilia  Komulainen; Michael  Paradowski; Lewis E.  Pennicott; Edward  J. Bartlett; Nigel Brissett; Ali  Raoof; Mandy  Watson; Allan M.  Jordan; Donald J. Ogilvie; Simon E.  Ward; John R.  Atack; Laurence H.  Pearl; Keith W.  Caldecott; Antony W.  Oliver

doi:10.1042/bcj20160180

Mode of action of DNA-competitive small molecule inhibitors of tyrosyl DNA phosphodiesterase 2

Peter Hornyak, Trevor Askwith, Sarah Walker, Emilia Komulainen, Michael Paradowski, Lewis E. Pennicott, Edward J. Bartlett, Nigel Brissett, Ali Raoof, Mandy Watson, Allan M. Jordan, Donald J. Ogilvie, Simon E. Ward, John R. Atack, Laurence H. Pearl, Keith W. Caldecott, Antony W. Oliver

Research output: Contribution to journal › Article › peer-review

Abstract

Tyrosyl-DNA phosphodiesterase 2 (TDP2) is a 5'-tyrosyl DNA phosphodiesterase important for the repair of DNA adducts generated by non-productive (abortive) activity of topoisomerase II (TOP2). TDP2 facilitates therapeutic resistance to topoisomerase poisons, which are widely used in the treatment of a range of cancer types. Consequently, TDP2 is an interesting target for the development of small molecule inhibitors that could restore sensitivity to topoisomerase-directed therapies. Previous studies identified a class of deazaflavin-based molecules that showed inhibitory activity against TDP2 at therapeutically useful concentrations, but their mode of action was uncertain. We have confirmed that the deazaflavin series inhibits TDP2 enzyme activity in a fluorescence-based assay, suitable for high-throughput screen (HTS)-screening. We have gone on to determine crystal structures of these compounds bound to a 'humanized' form of murine TDP2. The structures reveal their novel mode of action as competitive ligands for the binding site of an incoming DNA substrate, and point the way to generating novel and potent inhibitors of TDP2.

Original language	English
Pages (from-to)	1869–1879
Journal	Biochemical Journal
Volume	473
Issue number	13
DOIs	https://doi.org/10.1042/bcj20160180
Publication status	Published - 20 Apr 2016

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Hornyak, P., Askwith, T., Walker, S., Komulainen, E., Paradowski, M., Pennicott, L. E., J. Bartlett, E., Brissett, N., Raoof, A., Watson, M., Jordan, A. M., Ogilvie, D. J., Ward, S. E., Atack, J. R., Pearl, L. H., Caldecott, K. W., & Oliver , A. W. (2016). Mode of action of DNA-competitive small molecule inhibitors of tyrosyl DNA phosphodiesterase 2. Biochemical Journal, 473(13), 1869–1879. https://doi.org/10.1042/bcj20160180

@article{af6f47f5e0fb4351ade651b850c5493e,

title = "Mode of action of DNA-competitive small molecule inhibitors of tyrosyl DNA phosphodiesterase 2",

abstract = "Tyrosyl-DNA phosphodiesterase 2 (TDP2) is a 5'-tyrosyl DNA phosphodiesterase important for the repair of DNA adducts generated by non-productive (abortive) activity of topoisomerase II (TOP2). TDP2 facilitates therapeutic resistance to topoisomerase poisons, which are widely used in the treatment of a range of cancer types. Consequently, TDP2 is an interesting target for the development of small molecule inhibitors that could restore sensitivity to topoisomerase-directed therapies. Previous studies identified a class of deazaflavin-based molecules that showed inhibitory activity against TDP2 at therapeutically useful concentrations, but their mode of action was uncertain. We have confirmed that the deazaflavin series inhibits TDP2 enzyme activity in a fluorescence-based assay, suitable for high-throughput screen (HTS)-screening. We have gone on to determine crystal structures of these compounds bound to a 'humanized' form of murine TDP2. The structures reveal their novel mode of action as competitive ligands for the binding site of an incoming DNA substrate, and point the way to generating novel and potent inhibitors of TDP2.",

author = "Peter Hornyak and Trevor Askwith and Sarah Walker and Emilia Komulainen and Michael Paradowski and Pennicott, {Lewis E.} and {J. Bartlett}, Edward and Nigel Brissett and Ali Raoof and Mandy Watson and Jordan, {Allan M.} and Ogilvie, {Donald J.} and Ward, {Simon E.} and Atack, {John R.} and Pearl, {Laurence H.} and Caldecott, {Keith W.} and Oliver, {Antony W.}",

year = "2016",

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day = "20",

doi = "10.1042/bcj20160180",

language = "English",

volume = "473",

pages = "1869–1879",

journal = "Biochemical Journal",

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Hornyak, P, Askwith, T, Walker, S, Komulainen, E, Paradowski, M, Pennicott, LE, J. Bartlett, E, Brissett, N, Raoof, A, Watson, M, Jordan, AM, Ogilvie, DJ, Ward, SE, Atack, JR, Pearl, LH, Caldecott, KW & Oliver , AW 2016, 'Mode of action of DNA-competitive small molecule inhibitors of tyrosyl DNA phosphodiesterase 2', Biochemical Journal, vol. 473, no. 13, pp. 1869–1879. https://doi.org/10.1042/bcj20160180

TY - JOUR

T1 - Mode of action of DNA-competitive small molecule inhibitors of tyrosyl DNA phosphodiesterase 2

AU - Hornyak, Peter

AU - Askwith, Trevor

AU - Walker, Sarah

AU - Komulainen, Emilia

AU - Paradowski, Michael

AU - Pennicott, Lewis E.

AU - J. Bartlett, Edward

AU - Brissett, Nigel

AU - Raoof, Ali

AU - Watson, Mandy

AU - Jordan, Allan M.

AU - Ogilvie, Donald J.

AU - Ward, Simon E.

AU - Atack, John R.

AU - Pearl, Laurence H.

AU - Caldecott, Keith W.

AU - Oliver , Antony W.

PY - 2016/4/20

Y1 - 2016/4/20

N2 - Tyrosyl-DNA phosphodiesterase 2 (TDP2) is a 5'-tyrosyl DNA phosphodiesterase important for the repair of DNA adducts generated by non-productive (abortive) activity of topoisomerase II (TOP2). TDP2 facilitates therapeutic resistance to topoisomerase poisons, which are widely used in the treatment of a range of cancer types. Consequently, TDP2 is an interesting target for the development of small molecule inhibitors that could restore sensitivity to topoisomerase-directed therapies. Previous studies identified a class of deazaflavin-based molecules that showed inhibitory activity against TDP2 at therapeutically useful concentrations, but their mode of action was uncertain. We have confirmed that the deazaflavin series inhibits TDP2 enzyme activity in a fluorescence-based assay, suitable for high-throughput screen (HTS)-screening. We have gone on to determine crystal structures of these compounds bound to a 'humanized' form of murine TDP2. The structures reveal their novel mode of action as competitive ligands for the binding site of an incoming DNA substrate, and point the way to generating novel and potent inhibitors of TDP2.

AB - Tyrosyl-DNA phosphodiesterase 2 (TDP2) is a 5'-tyrosyl DNA phosphodiesterase important for the repair of DNA adducts generated by non-productive (abortive) activity of topoisomerase II (TOP2). TDP2 facilitates therapeutic resistance to topoisomerase poisons, which are widely used in the treatment of a range of cancer types. Consequently, TDP2 is an interesting target for the development of small molecule inhibitors that could restore sensitivity to topoisomerase-directed therapies. Previous studies identified a class of deazaflavin-based molecules that showed inhibitory activity against TDP2 at therapeutically useful concentrations, but their mode of action was uncertain. We have confirmed that the deazaflavin series inhibits TDP2 enzyme activity in a fluorescence-based assay, suitable for high-throughput screen (HTS)-screening. We have gone on to determine crystal structures of these compounds bound to a 'humanized' form of murine TDP2. The structures reveal their novel mode of action as competitive ligands for the binding site of an incoming DNA substrate, and point the way to generating novel and potent inhibitors of TDP2.

UR - https://europepmc.org/articles/PMC4925160

U2 - 10.1042/bcj20160180

DO - 10.1042/bcj20160180

M3 - Article

C2 - 27099339

SN - 1470-8728

VL - 473

SP - 1869

EP - 1879

JO - Biochemical Journal

JF - Biochemical Journal

IS - 13

ER -

Mode of action of DNA-competitive small molecule inhibitors of tyrosyl DNA phosphodiesterase 2

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