Fluorous‐Based Small‐Molecule Microarrays for the Discovery of Histone Deacetylase Inhibitors

Vegas, Arturo J.; Bradner, James E.; Tang, Weiping; McPherson, Olivia M.; Greenberg, Edward; Koehler, Angela N.; Schreiber, Stuart L.

doi:10.1002/anie.200703198

Cited by 82 publications

(57 citation statements)

References 38 publications

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“…We undertook a chemical strategy to identify more drug-like HDAC6 inhibitors using a highly parallel approach to biased chemical library synthesis (17). We designed a hydroxamic acid building block, compound 1, possessing a reactive hydrazide separated by a six-carbon linear aliphatic linker that mimics the structure of SAHA and tubacin (Fig.…”

Section: Resultsmentioning

confidence: 99%

Discovery of selective small-molecule HDAC6 inhibitor for overcoming proteasome inhibitor resistance in multiple myeloma

Hideshima

Paranal

et al. 2016

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

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120

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Multiple myeloma (MM) has proven clinically susceptible to modulation of pathways of protein homeostasis. Blockade of proteasomal degradation of polyubiquitinated misfolded proteins by the proteasome inhibitor bortezomib (BTZ) achieves responses and prolongs survival in MM, but long-term treatment with BTZ leads to drug-resistant relapse in most patients. In a proof-of-concept study, we previously demonstrated that blocking aggresomal breakdown of polyubiquitinated misfolded proteins with the histone deacetylase 6 (HDAC6) inhibitor tubacin enhances BTZ-induced cytotoxicity in MM cells in vitro. However, these foundational studies were limited by the pharmacologic liabilities of tubacin as a chemical probe with only in vitro utility.Emerging from a focused library synthesis, a potent, selective, and bioavailable HDAC6 inhibitor, WT161, was created to study the mechanism of action of HDAC6 inhibition in MM alone and in combination with BTZ. WT161 in combination with BTZ triggers significant accumulation of polyubiquitinated proteins and cell stress, followed by caspase activation and apoptosis. More importantly, this combination treatment was effective in BTZ-resistant cells and in the presence of bone marrow stromal cells, which have been shown to mediate MM cell drug resistance. The activity of WT161 was confirmed in our human MM cell xenograft mouse model and established the framework for clinical trials of the combination treatment to improve patient outcomes in MM.histone deacetylase 6 | proteasome inhibitor | multiple myeloma | bortezomib-resistance | WT161

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Section: Resultsmentioning

confidence: 99%

Discovery of selective small-molecule HDAC6 inhibitor for overcoming proteasome inhibitor resistance in multiple myeloma

Hideshima

Paranal

et al. 2016

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

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120

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“…Fluorous-tagged molecules can be retained on solid fluorous-phase materials, a property that has been used to simplify organic synthesis and purification. In addition, it has been shown that fluorous-phase immobilization can be used to construct small-molecule microarrays (17), including carbohydrate microarrays (18), to screen inhibitors (19), and to enrich and detect fluorous peptides by using mass spectrometry (20).…”

mentioning

confidence: 99%

A nanostructure-initiator mass spectrometry-based enzyme activity assay

Northen

Lee

Hoang

et al. 2008

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

133

124

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We describe a Nanostructure-Initiator Mass Spectrometry (NIMS) enzymatic (Nimzyme) assay in which enzyme substrates are immobilized on the mass spectrometry surface by using fluorousphase interactions. This ''soft'' immobilization allows efficient desorption/ionization while also enabling the use of surfacewashing steps to reduce signal suppression from complex biological samples, which results from the preferential retention of the tagged products and reactants. The Nimzyme assay is sensitive to subpicogram levels of enzyme, detects both addition and cleavage reactions (sialyltransferase and galactosidase), is applicable over a wide range of pHs and temperatures, and can measure activity directly from crude cell lysates. The ability of the Nimzyme assay to analyze complex mixtures is illustrated by identifying and directly characterizing ␤-1,4-galactosidase activity from a thermophilic microbial community lysate. The optimal enzyme temperature and pH were found to be 65°C and 5.5, respectively, and the activity was inhibited by both phenylethyl-␤-D-thiogalactopyranoside and deoxygalactonojirimycin. Metagenomic analysis of the community suggests that the activity is from an uncultured, unsequenced ␥-proteobacterium. In general, this assay provides an efficient method for detection and characterization of enzymatic activities in complex biological mixtures prior to sequencing or cloning efforts. More generally, this approach may have important applications for screening both enzymatic and inhibitor libraries, constructing and screening glycan microarrays, and complementing fluorous-phase organic synthesis.fluorous phase ͉ microbial community ͉ carbohydrate

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“…1,2) Chemical arrays represent one of the most promising and high-throughput approaches to searching ligands against proteins of interest. [3][4][5][6] Several successful results from chemical arrays have been reported. 7,8) One of the key steps in the technology is immobilization of small molecules on a solid surface.…”

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confidence: 99%

Robust and Systematic Drug Screening Method Using Chemical Arrays and the Protein Library: Identification of Novel Inhibitors of Carbonic Anhydrase II

Miyazaki

Simizu

Ichimiya

et al. 2008

Bioscience, Biotechnology, and Biochemistry

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The identification of specific interactions between small molecules and human proteins of interest is a fundamental step in chemical biology and drug development. Here we describe an efficient method to obtain novel binding ligands of human proteins by a chemical array approach. Our method includes large-scale ligand screening with two libraries, proteins and chemicals, the use of cell lysates that express proteins of interest fused with red fluorescent protein, and high-throughput screening by merged display analysis, which removes false positive signals from array experiments. Using our systematic platform, we detected novel inhibitors of carbonic anhydrase II. It is suggested that our systematic platform is a rapid and robust approach to screen novel ligands for human proteins of interest.

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Fluorous‐Based Small‐Molecule Microarrays for the Discovery of Histone Deacetylase Inhibitors

Cited by 82 publications

References 38 publications

Discovery of selective small-molecule HDAC6 inhibitor for overcoming proteasome inhibitor resistance in multiple myeloma

Discovery of selective small-molecule HDAC6 inhibitor for overcoming proteasome inhibitor resistance in multiple myeloma

A nanostructure-initiator mass spectrometry-based enzyme activity assay

Robust and Systematic Drug Screening Method Using Chemical Arrays and the Protein Library: Identification of Novel Inhibitors of Carbonic Anhydrase II

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