Design and evaluation of 'Linkerless' hydroxamic acids as selective HDAC8 inhibitors

University of California, San Francisco, San Francisco, California, United States
Bioorganic & Medicinal Chemistry Letters (Impact Factor: 2.33). 05/2007; 17(10):2874-8. DOI: 10.1016/j.bmcl.2007.02.064
Source: PubMed

ABSTRACT In this report, we describe new HDAC inhibitors designed to exploit a unique sub-pocket in the HDAC8 active site. These compounds were based on inspection of the available HDAC8 crystal structures bound to various inhibitors, which collectively show that the HDAC8 active site is unusually malleable and can accommodate inhibitor structures that are distinct from the canonical 'zinc binding group-linker-cap group' structures of SAHA, TSA, and similar HDAC inhibitors. Some inhibitors based on this new scaffold are >100-fold selective for HDAC8 over other class I and class II HDACs with IC(50) values <1microM against HDAC8. Furthermore, treatment of human cells with the inhibitors described here shows a unique pattern of hyperacetylated proteins compared with the broad-spectrum HDAC inhibitor TSA.

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    • "High potassium solution (KPSS) with composition as per PSS with an isosmotic substitution of NaCl) with KCl (60 mmol/L final concentration ). Phenylephrine (PE) hydrochloride and acetylcholine chloride stocks were made up daily in distilled water; TSA stock was made up with ethanol and compound 2 (manufactured as described previously Krennhrubec et al. 2008) and plumbagin in dimethylsulfoxide (DMSO). "
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    ABSTRACT: There is increasing interest in establishing the roles that lysine acetylation of non nuclear proteins may exert in modulating cell function. Lysine deacetylase 8 (KDAC8), for example, has been suggested to interact with α-actin and control the differentiation of smooth muscle cells. However, a direct role of smooth muscle non nuclear protein acetylation in regulating tone is unresolved. We sought to define the actions of two separate KDAC inhibitors on arterial tone and identify filament-interacting protein targets of acetylation and association with KDAC8. Compound 2 (a specific KDAC8 inhibitor) or Trichostatin A (TSA, a broad-spectrum KDAC inhibitor) inhibited rat arterial contractions induced by phenylephrine (PE) or high potassium solution. In contrast to the predominantly nuclear localization of KDAC1 and KDAC2, KDAC8 was positioned in extranuclear areas of native vascular smooth muscle cells. Several filament-associated proteins identified as putative acetylation targets colocalized with KDAC8 by immunoprecipitation (IP): cortactin, α-actin, tropomyosin, HSPB1 (Hsp27) and HSPB6 (Hsp20). Use of anti-acetylated lysine antibodies showed that KDAC inhibition increased acetylation of each protein. A custom-made antibody targeting the C-terminal acetylated lysine of human HSPB6 identified this as a novel target of acetylation that was increased by KDAC inhibition. HSPB6 phosphorylation, a known vasodilatory modification, was concomitantly increased. Interrogation of publicly available mass spectrometry data identified 50 other proteins with an acetylated C-terminal lysine. These novel data, in alliance with other recent studies, alert us to the importance of elucidating the mechanistic links between changes in myofilament-associated protein acetylation, in conjunction with other posttranslational modifications, and the regulation of arterial tone.
    11/2013; 1(6):e00127. DOI:10.1002/phy2.127
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    • "It is therefore clear that HDAC8 fulfils critical functions in cancer cells and that these are associated with its overexpression. This has stimulated interest in the development of HDAC8-specific inhibitors and treatment of neuroblastoma cells with a " linkerless " hydroxamic acid, selective HDAC8 inhibitor [100], inhibited cell proliferation and induced differentiation, reproducing the siRNAinduced phenotype [98]. Interestingly, knockdown of HDAC8 failed to affect global histone H4 acetylation, suggesting that the principal targets of this enzyme are either non-histone proteins or specific gene promoters. "
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    ABSTRACT: The histone modifying enzymes (HME) represent particularly promising targets for the development of alternatives to praziquantel, the only currently available drug to combat schistosomiasis. The inhibition of these enzymes frequently arrests the cell cycle or induces apoptosis in cancer cells, but not in normal cells and numerous HME inhibitors are under investigation as potential anticancer agents. The recent resolution of the genome sequences of Schistosoma mansoni and Schistosoma japonicum has allowed us to identify all the schistosome genes encoding histone acetyltransferases, deacetylases, methyltransferases and demethylases. We have chosen a strategy using phylogenetic screening with inhibitors of HME classes, screening of individual HME targets by both high-throughput and reasoned (in silico docking using resolved crystal structures) approaches in a project funded by the European Community, named SEtTReND (Schistosome Epigenetics: Targets, Regulation, New Drugs). The initial focus is on the class I histone deacetylase (HDAC) 8 since the comparison of the catalytic site of the schistosome and human enzymes shows crucial differences, rendering possible the development of inhibitors specific for SmHDAC8. However, phenotypic screening shows that inhibitors of all HME classes tested were able to induce apoptosis and death in parasites in vitro, indicating that other enzymes may prove to be viable targets.
    Current pharmaceutical design 05/2012; 18(24):3567-78. · 3.29 Impact Factor
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    • "However, attempts to improve selectivity by exploiting subtle differences in active site components, including the capping group, linker region, and metal-binding domains, have had some success (Bieliauskas and Pflum 2008). Selective compounds targeting exclusively HDAC1 or HDAC8 in the class I category and HDAC4 or HDAC6 in the class II category have been developed (Perez-Balado et al. 2007; Kozikowski et al. 2008; Krennhrubec et al. 2007; Bieliauskas and Pflum 2008). "
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    ABSTRACT: Epigenetic mechanisms have a central role in regulating gene expression and are capable of influencing complex cognitive functions. In particular, acetylation of histone proteins is an epigenetic modification involved in mediating synaptic plasticity, learning, and memory. Emerging evidence indicates that increased histone acetylation through the inhibition of histone deacetylases (HDACs) can facilitate the formation of long-term memories in preclinical studies. Moreover, HDAC inhibitors have been reported to ameliorate cognitive deficits in animal models relevant to neurodegenerative and neurodevelopmental disorders. HDAC inhibitors have also been found to enhance the extinction of learned behaviors, including drug-seeking behaviors. Consequently, HDAC inhibition may be a useful approach in the treatment of a wide range of disorders characterized by cognitive dysfunction. Future HDAC-based pharmacotherapies will benefit from a greater understanding of different HDAC isoforms and the molecular pathways specifically involved in inducing cognitive enhancement. KeywordsDrug addiction-Histone deacetylase (HDAC) inhibitors-Memory-Neurodegenerative disorders-Neurodevelopmental disorders-Synaptic plasticity
    05/2011: pages 245-266;
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