HR22C16: A Potent Small‐Molecule Probe for the Dynamics of Cell Division

Hotha, Srinivas; Yarrow, Justin C.; Yang, Ji-Young; Garrett, Sarah; Renduchintala, Kishore V.; Mayer, Thomas; Kapoor, Tarun M.

doi:10.1002/anie.200351173

Cited by 140 publications

(76 citation statements)

References 29 publications

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“…Monastrol was obtained from AgScientific (M116) aliquoted to 10 mM in Me 2 SO and stored at Ϫ20°C. HR22C16, A1, and other analogs were previously synthesized and stored as described in Hotha et al (18).…”

Section: Methodsmentioning

confidence: 99%

“…The cell permeable small molecule inhibitor of cell division, HR22C16, was identified using a forward chemical genetic screen as previously described (18). Using in vitro assays it was previously shown that HR22C16 inhibits the mitotic kinesin, Eg5 with an IC 50 of 800 Ϯ 10 nM; however, it is not known if this compound binds and inhibits Eg5 specifically or if it also binds other microtubule motor proteins.…”

Section: Hr22c16 Interacts With Eg5 But Not Other Microtubulebased Momentioning

confidence: 99%

“…These cells are ϳ25-fold resistant to Taxol because of acquired mutations in the Taxol binding site (7). To test the efficacy of the Eg5 inhibitors, HR22C16 and two of its analogs (A1 and E1) (18) in these Taxol-resistant cell lines, we performed 72-h antiproliferative assays. Our results (Table I) show that all Eg5 inhibitors tested, including the first generation Eg5 inhibitor monastrol, are active against the Taxol-resistant cell lines, with relative resistance values ranging from 1.8 -3.2-fold, in contrast to 21-fold resistance to Taxol.…”

Section: Hr22c16 and Its Analogs Have Antiproliferative Activity Agaimentioning

confidence: 99%

“…One such compound, CK0106023, is a specific allosteric inhibitor of Eg5 and possesses antitumor activity in an ovarian cancer xenograft (17). Another Eg5 inhibitor, HR22C16 was discovered in a microscopy-based forward chemical genetics screen of ϳ16,000 compounds (18). This compound has antimitotic activity and inhibits the Eg5 motor function in vitro with an IC 50 of 800 Ϯ 10 nM.…”

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Mitotic Kinesin Inhibitors Induce Mitotic Arrest and Cell Death in Taxol-resistant and -sensitive Cancer Cells

Marcus

Peters

Thomas

et al. 2005

Journal of Biological Chemistry

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Taxanes are powerful chemotherapy agents that target the microtubule cytoskeleton, leading to mitotic arrest and cell death; however, their clinical efficacy has been hampered due to the development of drug resistance. Therefore, other proteins involved in spindle assembly are being examined as potential targets for anticancer therapy. The mitotic kinesin, Eg5 is critical for proper spindle assembly; as such, inhibition of Eg5 leads to mitotic arrest making it a potential anticancer target. We wanted to validate Eg5 as a therapeutic target and determine if Eg5 inhibitors retain activity in Taxol-resistant cells. Using affinity chromatography we first show that the compound HR22C16 is an Eg5 inhibitor and does not interact with other microtubule motor proteins tested. Furthermore, HR22C16 along with its analogs, inhibit cell survival in both Taxol-sensitive and -resistant ovarian cancer cells with at least 15-fold greater efficacy than monastrol, the first generation Eg5 inhibitor. Further analysis with HR22C16-A1, the most potent HR22C16 analog, showed that it retains efficacy in PgP-overexpressing cells, suggesting that it is not a PgP substrate. We further show that HR22C16-A1 induces cell death following mitotic arrest via the intrinsic apoptotic pathway. Interestingly, the combination of HR22C16-A1 with Taxol results in an antagonistic antiproliferative and antimitotic effect, possibly due to the abrogation of Taxol-induced mitotic spindles by HR22C16-A1. Taken together, our results show that Eg5 inhibitors have promising anticancer activity and can be potentially used to overcome Taxol resistance in the clinical setting.

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Section: Methodsmentioning

confidence: 99%

Section: Hr22c16 Interacts With Eg5 But Not Other Microtubulebased Momentioning

confidence: 99%

Section: Hr22c16 and Its Analogs Have Antiproliferative Activity Agaimentioning

confidence: 99%

mentioning

confidence: 99%

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Mitotic Kinesin Inhibitors Induce Mitotic Arrest and Cell Death in Taxol-resistant and -sensitive Cancer Cells

Marcus

Peters

Thomas

et al. 2005

Journal of Biological Chemistry

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158

123

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“…In blast crisis chronic myeloid leukemia and prostate cancer cells, in which Eg5 is highly expressed, inhibition of Eg5 causes cell cycle arrest and significantly suppresses cell proliferation [5,6] . Over the last decade, the effects of various Eg5 inhibitors on the proliferation of cancer cells have been investigated, and the mechanisms of action of several Eg5 inhibitors have been studied [7][8][9][10][11] . Dimethylenastron is a cell-permeable quinazoline-thione compound that acts as a potent inhibitor of Eg5 [12] .…”

Section: Introductionmentioning

confidence: 99%

Dimethylenastron suppresses human pancreatic cancer cell migration and invasion in vitro via allosteric inhibition of mitotic kinesin Eg5

et al. 2011

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Aim:The mitotic kinesin Eg5 plays a critical role in bipolar spindle assembly, and its inhibitors have shown impressive anticancer activity in preclinical studies. This study was undertaken to investigate the effect of dimethylenastron, a specific inhibitor of Eg5, on the migration and invasion of pancreatic cancer cells. Methods: Human pancreatic cancer cell lines PANC1, EPP85, BxPC3, CFPAC1, and AsPAC1 were used. Eg5 expression was examined using immunofluorescence microscopy. Cell migration and invasion were analyzed with wound healing and transwell assays. Cell proliferation was examined using sulforhodamine B and MTT assays. The binding of dimethylenastron to Eg5 was analyzed with a molecular modeling study, and the ADP release rate was examined with the MANT-ADP reagent. Results: Eg5 expression was 9-16-fold up-regulated in the 5 pancreatic cancer cell lines. Treatment of PANC1 pancreatic cancer cells with dimethylenastron (3 and 10 μmol/L) for 24 h suppressed the migratory ability of the cancer cells in a concentration-dependent manner. The invasion ability of the cancer cells was also reduced by the treatment. However, treatment of PANC1 cells with dimethylenastron (3 and 10 μmol/L) for 24 h had no detectable effect on their proliferation, which was inhibited when the cancer cells were treated with the drug for 72 h. Molecular modeling study showed that dimethylenastron could allosterically inhibit the motor domain ATPase of Eg5 by decreasing the rate of ADP release. Conclusion: Dimethylenastron inhibits the migration and invasion of PANC1 pancreatic cancer cells, independent of suppressing the cell proliferation. The findings provide a novel insight into the mechanisms of targeting Eg5 for pancreatic cancer chemotherapy.

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Forward Chemical Genetics

Haggarty¹,

Schreiber²

2007

Chemical Biology

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HR22C16: A Potent Small‐Molecule Probe for the Dynamics of Cell Division

Cited by 140 publications

References 29 publications

Mitotic Kinesin Inhibitors Induce Mitotic Arrest and Cell Death in Taxol-resistant and -sensitive Cancer Cells

Mitotic Kinesin Inhibitors Induce Mitotic Arrest and Cell Death in Taxol-resistant and -sensitive Cancer Cells

Dimethylenastron suppresses human pancreatic cancer cell migration and invasion in vitro via allosteric inhibition of mitotic kinesin Eg5

Forward Chemical Genetics

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