Journal: Journal of Medicinal Chemistry

Article Title: Discovery of Pyrazole-Based Positron Emission Tomography Agent that Maps Histone Deacetylase 6 (HDAC6) in the Nonhuman Primate Brain

doi: 10.1021/acs.jmedchem.5c02216

Figure Lengend Snippet: Representative HDAC6 PET tracers with IC 50 values for HDAC6.

Article Snippet: The binding assay using AS-MS was performed at rt in a final volume of 10 μL using the recombinant HDAC6 (HDAC6, Active, H88-30G-10, Lot:B2010-9; SignalChem Biotech Inc., Richmond, BC, Canada).

Techniques:

Journal: Journal of Medicinal Chemistry

Article Title: Discovery of Pyrazole-Based Positron Emission Tomography Agent that Maps Histone Deacetylase 6 (HDAC6) in the Nonhuman Primate Brain

doi: 10.1021/acs.jmedchem.5c02216

Figure Lengend Snippet: Affinity selection (AS)–MS binding study of HDAC6 with 16a and bavarostat. Data points are the mean ± standard error (SE) (four technical replicates). (A) Saturation binding curve. (B) Time course for the dissociation. (C) Binding parameters calculated from AS–MS data. Functional IC 50 values are shown for comparison.

Article Snippet: The binding assay using AS-MS was performed at rt in a final volume of 10 μL using the recombinant HDAC6 (HDAC6, Active, H88-30G-10, Lot:B2010-9; SignalChem Biotech Inc., Richmond, BC, Canada).

Techniques: Selection, Binding Assay, Functional Assay, Comparison

Journal: Journal of Medicinal Chemistry

Article Title: Discovery of Pyrazole-Based Positron Emission Tomography Agent that Maps Histone Deacetylase 6 (HDAC6) in the Nonhuman Primate Brain

doi: 10.1021/acs.jmedchem.5c02216

Figure Lengend Snippet: Brain and plasma concentration of 16a in wild-type (WT) and HDAC6 knockout (KO) mice. Compound 16a (1 mg/kg) was intravenously administered to WT and HDAC6 KO mice. Two hours after administration, the hippocampus and plasma were collected and the concentration of 16a in the brain (A) and plasma (B) was measured using liquid chromatography with tandem mass spectrometry (LC–MS/MS). The data are expressed in percentage of injected dose per gram of brain tissue (%ID/g) or per milliliter of plasma (%ID/mL), respectively, as mean ± standard error (SE), n = 4. ** P < 0.01, n.s.: not significant.

Article Snippet: The binding assay using AS-MS was performed at rt in a final volume of 10 μL using the recombinant HDAC6 (HDAC6, Active, H88-30G-10, Lot:B2010-9; SignalChem Biotech Inc., Richmond, BC, Canada).

Techniques: Clinical Proteomics, Concentration Assay, Knock-Out, Liquid Chromatography, Mass Spectrometry, Liquid Chromatography with Mass Spectroscopy, Injection

Journal: Journal of Medicinal Chemistry

Article Title: Discovery of Pyrazole-Based Positron Emission Tomography Agent that Maps Histone Deacetylase 6 (HDAC6) in the Nonhuman Primate Brain

doi: 10.1021/acs.jmedchem.5c02216

Figure Lengend Snippet: Comparison of HDAC6 expression levels in a mouse and monkey using Western blotting. Hippocampus (Hip) was obtained from two mice, and HDAC6 expression was examined. Recombinant HDAC6 conjugated with GST (0.03, 0.1, 0.3, and 1 ng) was also used for the positive controls (A). Lysates of the hippocampus and corpus callosum (CC) obtained from a monkey were applied to Western blotting in duplicate. The same controls were used for comparison (B).

Article Snippet: The binding assay using AS-MS was performed at rt in a final volume of 10 μL using the recombinant HDAC6 (HDAC6, Active, H88-30G-10, Lot:B2010-9; SignalChem Biotech Inc., Richmond, BC, Canada).

Techniques: Comparison, Expressing, Western Blot, Recombinant

Journal: Journal of Medicinal Chemistry

Article Title: Discovery of Pyrazole-Based Positron Emission Tomography Agent that Maps Histone Deacetylase 6 (HDAC6) in the Nonhuman Primate Brain

doi: 10.1021/acs.jmedchem.5c02216

Figure Lengend Snippet: Immunohistochemical (IHC) staining of HDAC6 protein in a monkey brain. Coronal brain sections, including the areas below were prepared from a monkey (a, putamen; b, caudate nucleus; c, corpus callosum; d, white matter; e, anterior cingulate cortex; f, thalamus; g, hippocampus; h, cerebellum; i, brain stem [pons]). They were immunostained with HDAC6 antibody.

Article Snippet: The binding assay using AS-MS was performed at rt in a final volume of 10 μL using the recombinant HDAC6 (HDAC6, Active, H88-30G-10, Lot:B2010-9; SignalChem Biotech Inc., Richmond, BC, Canada).

Techniques: Immunohistochemical staining, Immunohistochemistry