Inhibition of the heat shock protein 90 molecular chaperone in vitro and in vivo by novel, synthetic, potent resorcinylic pyrazole/isoxazole amide analogues

Swee Y. Sharp; Chrisostomos Prodromou; Kathy Boxall; Marissa V. Powers; Joanna L. Holmes; Gary Box; Thomas P. Matthews; Kwai-Ming J. Cheung; Andrew Kalusa; Karen E. Ball; Angela Hayes; Anthea Hardcastle; Brian Dymock; Paul A. Brough; Xavier Barril; Julie E. Cansfield; Lisa Wright; Allan Surgenor; Nicolas Foloppe; Roderick E. Hubbard; Wynne Aherne; Laurence Pearl; Keith Jones; Edward McDonald; Florence Raynaud; Sue Eccles; Martin Drysdale; Paul Workman

doi:10.1158/1535-7163.MCT-07-0149

Inhibition of the heat shock protein 90 molecular chaperone in vitro and in vivo by novel, synthetic, potent resorcinylic pyrazole/isoxazole amide analogues

Swee Y. Sharp, Chrisostomos Prodromou, Kathy Boxall, Marissa V. Powers, Joanna L. Holmes, Gary Box, Thomas P. Matthews, Kwai-Ming J. Cheung, Andrew Kalusa, Karen E. Ball, Angela Hayes, Anthea Hardcastle, Brian Dymock, Paul A. Brough, Xavier Barril, Julie E. Cansfield, Lisa Wright, Allan Surgenor, Nicolas Foloppe, Roderick E. HubbardWynne Aherne, Laurence Pearl, Keith Jones, Edward McDonald, Florence Raynaud, Sue Eccles, Martin Drysdale, Paul Workman

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

Abstract

Although the heat shock protein 90 (HSP90) inhibitor 17-allylamino-17-demethoxygeldanamycin (17-AAG) shows clinical promise, potential limitations encourage development of alternative chemotypes. We discovered the 3,4-diarylpyrazole resorcinol CCT018159 by high-throughput screening and used structure-based design to generate more potent pyrazole amide analogues, exemplified by VER-49009. Here, we describe the detailed biological properties of VER-49009 and the corresponding isoxazole VER-50589. X-ray crystallography showed a virtually identical HSP90 binding mode. However, the dissociation constant (K_d) of VER-50589 was 4.5 +/- 2.2 nmol/L compared with 78.0 +/- 10.4 nmol/L for VER-49009, attributable to higher enthalpy for VER-50589 binding. A competitive binding assay gave a lower IC₅₀ of 21 +/- 4 nmol/L for VER-50589 compared with 47 +/- 9 nmol/L for VER-49009. Cellular uptake of VER-50589 was 4-fold greater than for VER-49009. Mean cellular antiproliferative GI₅₀ values for VER-50589 and VER-49009 for a human cancer cell line panel were 78 +/- 15 and 685 +/- 119 nmol/L, respectively, showing a 9-fold potency gain for the isoxazole. Unlike 17-AAG, but as with CCT018159, cellular potency of these analogues was independent of NAD(P)H:quinone oxidoreductase 1/DT-diaphorase and P-glycoprotein expression. Consistent with HSP90 inhibition, VER-50589 and VER-49009 caused induction of HSP72 and HSP27 alongside depletion of client proteins, including C-RAF, B-RAF, and survivin, and the protein arginine methyltransferase PRMT5. Both caused cell cycle arrest and apoptosis. Extent and duration of pharmacodynamic changes in an orthotopic human ovarian carcinoma model confirmed the superiority of VER-50589 over VER-49009. VER-50589 accumulated in HCT116 human colon cancer xenografts at levels above the cellular GI(50) for 24 h, resulting in 30% growth inhibition. The results indicate the therapeutic potential of the resorcinylic pyrazole/isoxazole amide analogues as HSP90 inhibitors.

Original language	English
Pages (from-to)	1198-1211
Journal	Molecular Cancer Therapeutics
Volume	6
Issue number	4
DOIs	https://doi.org/10.1158/1535-7163.MCT-07-0149
Publication status	Published - Apr 2007

Keywords

Adenosine Triphosphatases
Animals
Apoptosis
Biological Markers
Cell Cycle
Cell Proliferation
Crystallography, X-Ray
Drug Resistance, Neoplasm
Endothelial Cells
Female
HCT116 Cells
HSP90 Heat-Shock Proteins
HT29 Cells
Humans
Isoxazoles
Mice
Mice, Nude
NAD(P)H Dehydrogenase (Quinone)
Protein Binding
Pyrazoles
Treatment Outcome
Xenograft Model Antitumor Assays

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

Access to Document

10.1158/1535-7163.MCT-07-0149

Cite this

Sharp, S. Y., Prodromou, C., Boxall, K., Powers, M. V., Holmes, J. L., Box, G., Matthews, T. P., Cheung, K.-M. J., Kalusa, A., Ball, K. E., Hayes, A., Hardcastle, A., Dymock, B., Brough, P. A., Barril, X., Cansfield, J. E., Wright, L., Surgenor, A., Foloppe, N., ... Workman, P. (2007). Inhibition of the heat shock protein 90 molecular chaperone in vitro and in vivo by novel, synthetic, potent resorcinylic pyrazole/isoxazole amide analogues. Molecular Cancer Therapeutics, 6(4), 1198-1211. https://doi.org/10.1158/1535-7163.MCT-07-0149

@article{a5192f737b6c428485bce5781ece10b8,

title = "Inhibition of the heat shock protein 90 molecular chaperone in vitro and in vivo by novel, synthetic, potent resorcinylic pyrazole/isoxazole amide analogues",

abstract = "Although the heat shock protein 90 (HSP90) inhibitor 17-allylamino-17-demethoxygeldanamycin (17-AAG) shows clinical promise, potential limitations encourage development of alternative chemotypes. We discovered the 3,4-diarylpyrazole resorcinol CCT018159 by high-throughput screening and used structure-based design to generate more potent pyrazole amide analogues, exemplified by VER-49009. Here, we describe the detailed biological properties of VER-49009 and the corresponding isoxazole VER-50589. X-ray crystallography showed a virtually identical HSP90 binding mode. However, the dissociation constant (Kd) of VER-50589 was 4.5 +/- 2.2 nmol/L compared with 78.0 +/- 10.4 nmol/L for VER-49009, attributable to higher enthalpy for VER-50589 binding. A competitive binding assay gave a lower IC50 of 21 +/- 4 nmol/L for VER-50589 compared with 47 +/- 9 nmol/L for VER-49009. Cellular uptake of VER-50589 was 4-fold greater than for VER-49009. Mean cellular antiproliferative GI50 values for VER-50589 and VER-49009 for a human cancer cell line panel were 78 +/- 15 and 685 +/- 119 nmol/L, respectively, showing a 9-fold potency gain for the isoxazole. Unlike 17-AAG, but as with CCT018159, cellular potency of these analogues was independent of NAD(P)H:quinone oxidoreductase 1/DT-diaphorase and P-glycoprotein expression. Consistent with HSP90 inhibition, VER-50589 and VER-49009 caused induction of HSP72 and HSP27 alongside depletion of client proteins, including C-RAF, B-RAF, and survivin, and the protein arginine methyltransferase PRMT5. Both caused cell cycle arrest and apoptosis. Extent and duration of pharmacodynamic changes in an orthotopic human ovarian carcinoma model confirmed the superiority of VER-50589 over VER-49009. VER-50589 accumulated in HCT116 human colon cancer xenografts at levels above the cellular GI(50) for 24 h, resulting in 30% growth inhibition. The results indicate the therapeutic potential of the resorcinylic pyrazole/isoxazole amide analogues as HSP90 inhibitors.",

keywords = "Adenosine Triphosphatases, Animals, Apoptosis, Biological Markers, Cell Cycle, Cell Proliferation, Crystallography, X-Ray, Drug Resistance, Neoplasm, Endothelial Cells, Female, HCT116 Cells, HSP90 Heat-Shock Proteins, HT29 Cells, Humans, Isoxazoles, Mice, Mice, Nude, NAD(P)H Dehydrogenase (Quinone), Protein Binding, Pyrazoles, Treatment Outcome, Xenograft Model Antitumor Assays",

author = "Sharp, {Swee Y.} and Chrisostomos Prodromou and Kathy Boxall and Powers, {Marissa V.} and Holmes, {Joanna L.} and Gary Box and Matthews, {Thomas P.} and Cheung, {Kwai-Ming J.} and Andrew Kalusa and Ball, {Karen E.} and Angela Hayes and Anthea Hardcastle and Brian Dymock and Brough, {Paul A.} and Xavier Barril and Cansfield, {Julie E.} and Lisa Wright and Allan Surgenor and Nicolas Foloppe and Hubbard, {Roderick E.} and Wynne Aherne and Laurence Pearl and Keith Jones and Edward McDonald and Florence Raynaud and Sue Eccles and Martin Drysdale and Paul Workman",

year = "2007",

month = apr,

doi = "10.1158/1535-7163.MCT-07-0149",

language = "English",

volume = "6",

pages = "1198--1211",

journal = "Molecular Cancer Therapeutics",

issn = "1535-7163",

publisher = "American Association for Cancer Research Inc.",

number = "4",

}

Sharp, SY, Prodromou, C, Boxall, K, Powers, MV, Holmes, JL, Box, G, Matthews, TP, Cheung, K-MJ, Kalusa, A, Ball, KE, Hayes, A, Hardcastle, A, Dymock, B, Brough, PA, Barril, X, Cansfield, JE, Wright, L, Surgenor, A, Foloppe, N, Hubbard, RE, Aherne, W, Pearl, L, Jones, K, McDonald, E, Raynaud, F, Eccles, S, Drysdale, M & Workman, P 2007, 'Inhibition of the heat shock protein 90 molecular chaperone in vitro and in vivo by novel, synthetic, potent resorcinylic pyrazole/isoxazole amide analogues', Molecular Cancer Therapeutics, vol. 6, no. 4, pp. 1198-1211. https://doi.org/10.1158/1535-7163.MCT-07-0149

TY - JOUR

T1 - Inhibition of the heat shock protein 90 molecular chaperone in vitro and in vivo by novel, synthetic, potent resorcinylic pyrazole/isoxazole amide analogues

AU - Sharp, Swee Y.

AU - Prodromou, Chrisostomos

AU - Boxall, Kathy

AU - Powers, Marissa V.

AU - Holmes, Joanna L.

AU - Box, Gary

AU - Matthews, Thomas P.

AU - Cheung, Kwai-Ming J.

AU - Kalusa, Andrew

AU - Ball, Karen E.

AU - Hayes, Angela

AU - Hardcastle, Anthea

AU - Dymock, Brian

AU - Brough, Paul A.

AU - Barril, Xavier

AU - Cansfield, Julie E.

AU - Wright, Lisa

AU - Surgenor, Allan

AU - Foloppe, Nicolas

AU - Hubbard, Roderick E.

AU - Aherne, Wynne

AU - Pearl, Laurence

AU - Jones, Keith

AU - McDonald, Edward

AU - Raynaud, Florence

AU - Eccles, Sue

AU - Drysdale, Martin

AU - Workman, Paul

PY - 2007/4

Y1 - 2007/4

N2 - Although the heat shock protein 90 (HSP90) inhibitor 17-allylamino-17-demethoxygeldanamycin (17-AAG) shows clinical promise, potential limitations encourage development of alternative chemotypes. We discovered the 3,4-diarylpyrazole resorcinol CCT018159 by high-throughput screening and used structure-based design to generate more potent pyrazole amide analogues, exemplified by VER-49009. Here, we describe the detailed biological properties of VER-49009 and the corresponding isoxazole VER-50589. X-ray crystallography showed a virtually identical HSP90 binding mode. However, the dissociation constant (Kd) of VER-50589 was 4.5 +/- 2.2 nmol/L compared with 78.0 +/- 10.4 nmol/L for VER-49009, attributable to higher enthalpy for VER-50589 binding. A competitive binding assay gave a lower IC50 of 21 +/- 4 nmol/L for VER-50589 compared with 47 +/- 9 nmol/L for VER-49009. Cellular uptake of VER-50589 was 4-fold greater than for VER-49009. Mean cellular antiproliferative GI50 values for VER-50589 and VER-49009 for a human cancer cell line panel were 78 +/- 15 and 685 +/- 119 nmol/L, respectively, showing a 9-fold potency gain for the isoxazole. Unlike 17-AAG, but as with CCT018159, cellular potency of these analogues was independent of NAD(P)H:quinone oxidoreductase 1/DT-diaphorase and P-glycoprotein expression. Consistent with HSP90 inhibition, VER-50589 and VER-49009 caused induction of HSP72 and HSP27 alongside depletion of client proteins, including C-RAF, B-RAF, and survivin, and the protein arginine methyltransferase PRMT5. Both caused cell cycle arrest and apoptosis. Extent and duration of pharmacodynamic changes in an orthotopic human ovarian carcinoma model confirmed the superiority of VER-50589 over VER-49009. VER-50589 accumulated in HCT116 human colon cancer xenografts at levels above the cellular GI(50) for 24 h, resulting in 30% growth inhibition. The results indicate the therapeutic potential of the resorcinylic pyrazole/isoxazole amide analogues as HSP90 inhibitors.

AB - Although the heat shock protein 90 (HSP90) inhibitor 17-allylamino-17-demethoxygeldanamycin (17-AAG) shows clinical promise, potential limitations encourage development of alternative chemotypes. We discovered the 3,4-diarylpyrazole resorcinol CCT018159 by high-throughput screening and used structure-based design to generate more potent pyrazole amide analogues, exemplified by VER-49009. Here, we describe the detailed biological properties of VER-49009 and the corresponding isoxazole VER-50589. X-ray crystallography showed a virtually identical HSP90 binding mode. However, the dissociation constant (Kd) of VER-50589 was 4.5 +/- 2.2 nmol/L compared with 78.0 +/- 10.4 nmol/L for VER-49009, attributable to higher enthalpy for VER-50589 binding. A competitive binding assay gave a lower IC50 of 21 +/- 4 nmol/L for VER-50589 compared with 47 +/- 9 nmol/L for VER-49009. Cellular uptake of VER-50589 was 4-fold greater than for VER-49009. Mean cellular antiproliferative GI50 values for VER-50589 and VER-49009 for a human cancer cell line panel were 78 +/- 15 and 685 +/- 119 nmol/L, respectively, showing a 9-fold potency gain for the isoxazole. Unlike 17-AAG, but as with CCT018159, cellular potency of these analogues was independent of NAD(P)H:quinone oxidoreductase 1/DT-diaphorase and P-glycoprotein expression. Consistent with HSP90 inhibition, VER-50589 and VER-49009 caused induction of HSP72 and HSP27 alongside depletion of client proteins, including C-RAF, B-RAF, and survivin, and the protein arginine methyltransferase PRMT5. Both caused cell cycle arrest and apoptosis. Extent and duration of pharmacodynamic changes in an orthotopic human ovarian carcinoma model confirmed the superiority of VER-50589 over VER-49009. VER-50589 accumulated in HCT116 human colon cancer xenografts at levels above the cellular GI(50) for 24 h, resulting in 30% growth inhibition. The results indicate the therapeutic potential of the resorcinylic pyrazole/isoxazole amide analogues as HSP90 inhibitors.

KW - Adenosine Triphosphatases

KW - Animals

KW - Apoptosis

KW - Biological Markers

KW - Cell Cycle

KW - Cell Proliferation

KW - Crystallography, X-Ray

KW - Drug Resistance, Neoplasm

KW - Endothelial Cells

KW - Female

KW - HCT116 Cells

KW - HSP90 Heat-Shock Proteins

KW - HT29 Cells

KW - Humans

KW - Isoxazoles

KW - Mice

KW - Mice, Nude

KW - NAD(P)H Dehydrogenase (Quinone)

KW - Protein Binding

KW - Pyrazoles

KW - Treatment Outcome

KW - Xenograft Model Antitumor Assays

U2 - 10.1158/1535-7163.MCT-07-0149

DO - 10.1158/1535-7163.MCT-07-0149

M3 - Article

C2 - 17431102

SN - 1535-7163

VL - 6

SP - 1198

EP - 1211

JO - Molecular Cancer Therapeutics

JF - Molecular Cancer Therapeutics

IS - 4

ER -

Inhibition of the heat shock protein 90 molecular chaperone in vitro and in vivo by novel, synthetic, potent resorcinylic pyrazole/isoxazole amide analogues

Abstract

Keywords

UN SDGs

Access to Document

Fingerprint

Cite this