Combining Hit Identification Strategies: Fragment-Based and in Silico Approaches to Orally Active 2-Aminothieno[2,3-<i>d</i>]pyrimidine Inhibitors of the Hsp90 Molecular Chaperone

Brough, Paul A.; Barril, Xavier; Borgognoni, Jenifer; Chêne, Patrick; Davies, Nicholas; Davis, Ben; Drysdale, Martin J.; Dymock, Brian; Eccles, Suzanne A.; García‐Echeverría, Carlos; Fromont, Christophe; Hayes, Angela; Hubbard, Roderick E.; Jordan, Allan M.; Jensen, Michael Rugaard; Massey, Andrew; Merrett, Angela; Padfield, Antony; Parsons, Rachel; Radimerski, Thomas; Raynaud, Florence I.; Robertson, Alan; Roughley, Stephen D.; Schoepfer, Joseph; Simmonite, Heather; Sharp, Swee Y.; Surgenor, A.E.; Valenti, Melanie; Walls, Steven B.; Webb, Paul; Wood, Mike; Workman, Paul; Wright, Lisa

doi:10.1021/jm900357y

Cited by 158 publications

(168 citation statements)

References 80 publications

(141 reference statements)

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“…Compared with NVP-AUY922, the novel, structurally distinct Hsp90 inhibitor NVP-BEP800 tested here has an improved oral bioavailability (Brough et al, 2009;Massey et al, 2010). In this study, we systematically applied a multitarget approach to explore the impact of NVP-AUY922 and NVP-BEP800 on the radiation response of tumour cells.…”

Section: Discussionmentioning

confidence: 99%

“…NVP-AUY922 and NVP-BEP800 (Brough et al, 2009) were kindly provided by Novartis Institutes for Biomedical Research (Basel, Switzerland). 17-Dimethylaminoethylamino-17-demethoxygeldanamycin (17-DMAG) was purchased from Sigma (Taufkirchen,Germany,.…”

Section: Drug Treatmentmentioning

confidence: 99%

“…Both requirements are met by a series of pyrazole resorcinol compounds that have proven to be stronger inhibitors Revised 3 March 2010; accepted 12 April 2010 of Hsp90 than geldanamycin derivatives. Currently, the isoxazole resorcinol NVP-AUY922 shows the highest affinity for the NH 2 -terminal nucleotide-binding site of Hsp90 Eccles et al, 2008;Gao et al, 2010), whereas NVP-BEP800 represents a novel fully synthetic, orally available 2-aminothieno [2,3-d]pyrimidine class Hsp90 inhibitor (Brough et al, 2009;Stühmer et al, 2009;Massey et al, 2010). Both compounds have good pharmaceutical and pharmacological properties.…”

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confidence: 99%

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Novel HSP90 inhibitors, NVP-AUY922 and NVP-BEP800, radiosensitise tumour cells through cell-cycle impairment, increased DNA damage and repair protraction

et al. 2010

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BACKGROUND: Heat-shock protein 90 (Hsp90) has a crucial role in both the stabilisation and regulation of various proteins, including those related to radioresistance. Inhibition of Hsp90 may therefore provide a strategy for enhancing the radiosensitivity of tumour cells. This study explores the responses of four tumour cell lines (A549, GaMG, HT 1080 and SNB19) to combined treatment with ionising radiation (IR) and two novel inhibitors of Hsp90, NVP-AUY922 and NVP-BEP800. The techniques used included cell and colony counts, expression of Hsp90, Hsp70, Akt, survivin, cleaved caspase 3, p53, cell-cycle progression and associated proteins. DNA damage was analysed by histone gH2AX and Comet assays. RESULTS: We found that NVP-AUY922 and NVP-BEP800 enhanced radiosensitivity in all tested cell lines. In contrast, only two cell lines (HT 1080 and GaMG) exhibited an increased rate of apoptosis after drug pretreatment, as revealed by western blot. In all tested cell lines, the expression of histone gH2AX, a marker of DNA double-strand breaks, after combined drug-IR treatment was higher and its decay rate was slower than those after each single treatment modality. Drug-IR treatment also resulted in impaired cell-cycle progression, as indicated by S-phase depletion and G2/M arrest. In addition, the cell cycle-associated proteins, Cdk1 and Cdk4, were downregulated after Hsp90 inhibition. INTERPRETATION: These findings show that the novel inhibitors of Hsp90 can radiosensitise tumour cell lines of different entities through destabilisation and depletion of several Hsp90 client proteins, thus causing the depletion of S phase and G2/M arrest, increased DNA damage and repair protraction and, to some extent, apoptosis. The results might have important implications for the radiotherapy of solid tumours.

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Section: Discussionmentioning

confidence: 99%

Section: Drug Treatmentmentioning

confidence: 99%

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Novel HSP90 inhibitors, NVP-AUY922 and NVP-BEP800, radiosensitise tumour cells through cell-cycle impairment, increased DNA damage and repair protraction

et al. 2010

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“…Cell proliferation was determined using the sulforhodamine B (SRB) assay. VER-155008 and VER-82160 were synthesized as previously described [22,23].…”

Section: Methodsmentioning

confidence: 99%

“…VER-82160 is a highly potent, orally bioavailable small molecule inhibitor of Hsp90 [22]. In a fluorescence polarization (FP) assay using N 6 -(6-amino)-hexyl-ATP-5-FAM as the FP probe, VER-155008 potently competed for the binding to Hsp70 with an IC 50 of 0.5 lM (Table 1).…”

Section: Ver-155008 Is a Potent Inhibitor Of The Hsp70 Family Of Chapmentioning

confidence: 99%

A novel, small molecule inhibitor of Hsc70/Hsp70 potentiates Hsp90 inhibitor induced apoptosis in HCT116 colon carcinoma cells

Massey¹,

Williamson²,

Browne³

et al. 2009

Cancer Chemother Pharmacol

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266

254

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Purpose The anti-apoptotic function of the 70 kDa family of heat shock proteins and their role in cancer is well documented. Dual targeting of Hsc70 and Hsp70 with siRNA induces proteasome-dependent degradation of Hsp90 client proteins and extensive tumor specific apoptosis as well as the potentiation of tumor cell apoptosis following pharmacological Hsp90 inhibition. Methods We have previously described the discovery and synthesis of novel adenosine-derived inhibitors of the 70 kDa family of heat shock proteins; the first inhibitors described to target the ATPase binding domain. The in vitro activity of VER-155008 was evaluated in HCT116, HT29, BT474 and MDA-MB-468 carcinoma cell lines. Cell proliferation, cell apoptosis and caspase 3/7 activity was determined for VER-155008 in the absence or presence of small molecule Hsp90 inhibitors. Results VER-155008 inhibited the proliferation of human breast and colon cancer cell lines with GI 50 s in the range 5.3-14.4 lM, and induced Hsp90 client protein degradation in both HCT116 and BT474 cells. As a single agent, VER-155008 induced caspase-3/7 dependent apoptosis in BT474 cells and non-caspase dependent cell death in HCT116 cells. VER-155008 potentiated the apoptotic potential of a small molecule Hsp90 inhibitor in HCT116 but not HT29 or MDA-MB-468 cells. In vivo, VER-155008 demonstrated rapid metabolism and clearance, along with tumor levels below the predicted pharmacologically active level. Conclusion These data suggest that small molecule inhibitors of Hsc70/Hsp70 phenotypically mimic the cellular mode of action of a small molecule Hsp90 inhibitor and can potentiate the apoptotic potential of a small molecule Hsp90 inhibitor in certain cell lines. The factors determining whether or not cells apoptose in response to Hsp90 inhibition or the combination of Hsp90 plus Hsc70/ Hsp70 inhibition remain to be determined.

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Fragment‐Based Lead Discovery

Hubbard

2021

Burger's Medicinal Chemistry and Drug Discovery

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Fragment‐based discovery is an effective approach to finding hit compounds that bind to a target. Fragments can be found that bind to most sites on most proteins, providing a rich source of chemical ideas for the medicinal chemist to develop into hits and leads. There are two distinctive activities in fragment‐based discovery: (i) finding fragments that bind and (ii) evolving the fragments to hit compounds which can then be progressed using conventional (structure‐guided) optimization. A wide variety of methods have been developed for screening, many based on biophysical approaches to identify and then characterize the binding of the relatively low affinity (but efficient) compounds – a separate section summarizes the key features of the methods. Perhaps the most difficult aspect of fragment‐based discovery is deciding which fragments to evolve – another section describes some recent projects to highlight the key strategies that are used. The article concludes with some summary statements on the main features and advantages of the approach and a discussion about future directions.

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Combining Hit Identification Strategies: Fragment-Based and in Silico Approaches to Orally Active 2-Aminothieno[2,3-d]pyrimidine Inhibitors of the Hsp90 Molecular Chaperone

Cited by 158 publications

References 80 publications

Novel HSP90 inhibitors, NVP-AUY922 and NVP-BEP800, radiosensitise tumour cells through cell-cycle impairment, increased DNA damage and repair protraction

Novel HSP90 inhibitors, NVP-AUY922 and NVP-BEP800, radiosensitise tumour cells through cell-cycle impairment, increased DNA damage and repair protraction

A novel, small molecule inhibitor of Hsc70/Hsp70 potentiates Hsp90 inhibitor induced apoptosis in HCT116 colon carcinoma cells

Fragment‐Based Lead Discovery

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