Rational Design and Simple Chemistry Yield a Superior, Neuroprotective HDAC6 Inhibitor, Tubastatin A

Butler, Kyle V.; Kalin, Jay H.; Brochier, Camille; Vistoli, Giulio; Langley, Brett; Kozikowski, Alan P.

doi:10.1021/ja102758v

Cited by 634 publications

(694 citation statements)

References 29 publications

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“…Table S1), which is also consistent with prior data. 19 In total, the single concentration screen of known inhibitors is consistent with the expected selectivities observed with SAHA, apicidin, MS-275, and tubastatin.…”

Section: Screening Of Known Hdac Inhibitorssupporting

confidence: 64%

“…1) was selected due to its HDAC6 selectivity. 19 As an initial screen, SAHA, apicidin, MS-275, and tubastatin were incubated at a single concentration (4.8 µM for MS-275 and 1 µM for all others), with each individual HDAC isoform affixed on the plate. The remaining deacetylase activity of the HDAC isoforms was then determined by adding the HDAC-Glo reagents.…”

Section: Screening Of Known Hdac Inhibitorsmentioning

confidence: 99%

“…Although no IC 50 values with mammalian cell-derived isoforms have been previously reported, baculovirus expressed HDAC1, 2, 3, and 6 proteins showed the same trend (16,400 ± 2600, >30,000, >30,000, and 15 ± 1 nM, respectively). 19 Interestingly, while baculovirus-derived isoforms displayed >1000-fold selectivity for HDAC6 compared with HDAC1, 19 the HDAC6 versus HDAC1 selectivity was only 87-fold with mammalian cell-derived proteins. The data with tubastatin also emphasize that the source of HDAC activity has a significant influence on the quantified selectivity of the inhibitor.…”

Section: Dose-dependent Inhibitionmentioning

confidence: 99%

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Development of an ELISA-Based HDAC Activity Assay for Characterization of Isoform-Selective Inhibitors

2015

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Histone deacetylase (HDAC) proteins are promising targets for cancer treatment, with several HDAC inhibitors used clinically as anticancer drugs. Most HDAC inhibitors nonspecifically interact with all or many of the 11 HDAC isoforms. Isoform-selective HDAC inhibitors would be useful tools to dissect the individual functions of HDAC proteins in cancer formation, in addition to potentially displaying effective anticancer properties. We report here a robust HDAC activity assay for screening selective HDAC inhibitors, which is inspired by the traditional enzyme-linked immunosorbent assay (ELISA). The key feature of the ELISA-based HDAC activity assay is use of mammalian cell-derived HDAC isoforms instead of recombinant proteins. Importantly, the assay was validated with several known HDAC inhibitors. The ELISAbased HDAC activity assay will facilitate the characterization of isoform-selective HDAC inhibitors against mammalian cell-derived HDAC proteins, which will enhance HDAC-centered cancer research and provide a foundation for anticancer drug development.

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Section: Screening Of Known Hdac Inhibitorssupporting

confidence: 64%

Section: Screening Of Known Hdac Inhibitorsmentioning

confidence: 99%

Section: Dose-dependent Inhibitionmentioning

confidence: 99%

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Development of an ELISA-Based HDAC Activity Assay for Characterization of Isoform-Selective Inhibitors

2015

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“…Mice were treated intraperitoneally and daily for 7 days with CAY10603 (CAY) or tubastatin A (TubA), two specific inhibitors of HDAC6 [21,22]. Analysis of motile cilia in the tracheal surface epithelium and primary cilia in the renal tubular epithelium revealed that these HDAC6 inhibitors partially rescued the ciliary length defect of CYLD knockout mice, but they did not have obvious effect on the percentage of ciliated cells ( Figure npg www.cell-research.com | Cell Research 6A-6F).…”

Section: Hdac6 Inhibitors Partially Rescue the Ciliary Length Defectsmentioning

confidence: 99%

CYLD mediates ciliogenesis in multiple organs by deubiquitinating Cep70 and inactivating HDAC6

et al. 2014

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Cilia are hair-like organelles extending from the cell surface with important sensory and motility functions. Ciliary defects can result in a wide range of human diseases known as ciliopathies. However, the molecular mechanisms controlling ciliogenesis remain poorly defined. Here we show that cylindromatosis (CYLD), a tumor suppressor protein harboring deubiquitinase activity, plays a critical role in the assembly of both primary and motile cilia in multiple organs. CYLD knockout mice exhibit polydactyly and various ciliary defects, such as failure in basal body anchorage and disorganization of basal bodies and axenomes. The ciliary function of CYLD is partially attributed to its deconjugation of the polyubiquitin chain from centrosomal protein of 70 kDa (Cep70), a requirement for Cep70 to interact with γ-tubulin and localize at the centrosome. In addition, CYLD-mediated inhibition of histone deacetylase 6 (HDAC6), which promotes tubulin acetylation, constitutes another mechanism for the ciliary function of CYLD. Small-molecule inhibitors of HDAC6 could partially rescue the ciliary defects in CYLD knockout mice. These findings highlight the importance of protein ubiquitination in the modulation of ciliogenesis, identify CYLD as a crucial regulator of this process, and suggest the involvement of CYLD deficiency in ciliopathies.

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“…Tubacin has non-drug-like qualities, high lipophilicity, and difficult synthesis and has proved to be more useful as a research tool rather than as a potential drug (18). We and others (12)(13)(14)(15)19) have developed HDAC6-selective inhibitors whose pharmacokinetics, toxicity, and efficacy make them potentially more useful than tubacin as therapeutic agents. ACY-1215, 2-(Diphenylamino)-N-(7-(hydroxyamino)-7-oxoheptyl)pyrimidine-5-carboxamide, a HDAC6-selective inhibitor, is currently being evaluated in clinical trials (http://clinicaltrials.gov).…”

mentioning

confidence: 99%

Development of a histone deacetylase 6 inhibitor and its biological effects

Lee

Mahendran

Yao

et al. 2013

Proc. Natl. Acad. Sci. U.S.A.

118

105

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Development of isoform-selective histone deacetylase (HDAC) inhibitors is important in elucidating the function of individual HDAC enzymes and their potential as therapeutic agents. Among the eleven zinc-dependent HDACs in humans, HDAC6 is structurally and functionally unique. Here, we show that a hydroxamic acid-based small-molecule N-hydroxy-4-(2-[(2-hydroxyethyl)(phenyl)amino]-2-oxoethyl)benzamide (HPOB) selectively inhibits HDAC6 catalytic activity in vivo and in vitro. HPOB causes growth inhibition of normal and transformed cells but does not induce cell death. HPOB enhances the effectiveness of DNA-damaging anticancer drugs in transformed cells but not normal cells. HPOB does not block the ubiquitin-binding activity of HDAC6. The HDAC6-selective inhibitor HPOB has therapeutic potential in combination therapy to enhance the potency of anticancer drugs.anticancer agents | epigenetics-based chemotherapy | drug discovery H istone deacetylase 6 (HDAC6) is unique among the eleven zinc-dependent HDACs in humans. HDAC6 is located in the cytoplasm, and it has two catalytic domains and an ubiquitinbinding domain at the C-terminal region (1-3). This study focused on the development of a HDAC6-selective inhibitor and its biological effects. The substrates of HDAC6 include nonhistone proteins such as α-tubulin, peroxiredoxin (PRX), cortactin, and heat shock protein 90 (Hsp90) but not histones (4-7). HDAC6 plays a key role in the regulation of microtubule dynamics including cell migration and cell-cell interactions. The reversible acetylation of Hsp90, a substrate of HDAC6, modulates its chaperone activity and, accordingly, the stability of survival and antiapoptotic factors, including epidermal growth factor receptor (EGFR), protein kinase AKT, proto-oncogene C-RAF, survivin, and other factors. HDAC6, through its ubiquitin-binding activity and interaction with other partner proteins, plays a role in the degradation of misfolded proteins by binding polyubiquitinated proteins and delivering them to the dynein and motor proteins for transport into aggresomes which are degraded by lysosomes (8-10). Thus, HDAC6 has multiple biological functions through deacetylasedependent and -independent mechanisms modulating many cellular pathways relevant to normal and tumor cell growth, migration, and death. HDAC6 is an attractive target for potential cancer treatment.There are several previous reports on the development of HDAC6-selective inhibitors (11)(12)(13)(14)(15). The most extensively studied is tubacin (16,17). Tubacin has non-drug-like qualities, high lipophilicity, and difficult synthesis and has proved to be more useful as a research tool rather than as a potential drug (18). We and others (12)(13)(14)(15)19) have developed HDAC6-selective inhibitors whose pharmacokinetics, toxicity, and efficacy make them potentially more useful than tubacin as therapeutic agents. ACY-1215, 2-(Diphenylamino)-N-(7-(hydroxyamino)-7-oxoheptyl)pyrimidine-5-carboxamide, a HDAC6-selective inhibitor, is currently being evaluated in clinical trial...

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Rational Design and Simple Chemistry Yield a Superior, Neuroprotective HDAC6 Inhibitor, Tubastatin A

Cited by 634 publications

References 29 publications

Development of an ELISA-Based HDAC Activity Assay for Characterization of Isoform-Selective Inhibitors

Development of an ELISA-Based HDAC Activity Assay for Characterization of Isoform-Selective Inhibitors

CYLD mediates ciliogenesis in multiple organs by deubiquitinating Cep70 and inactivating HDAC6

Development of a histone deacetylase 6 inhibitor and its biological effects

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