Target validation using in-cell small molecule clickable imaging probes

Abstract

The application of click chemistry to the visualization of chemical probes in in-cell chemical biology experiments is reviewed and the influence this research has had on target validation and molecular mode of action studies is also highlighted.

Synthesis and Structure–Activity Relationship Study of Chemical Probes as Hypoxia Induced Factor-1α/Malate Dehydrogenase 2 Inhibitors

“…The combined extracts were dried on anhydrous MgSO 4 , filtered, and concentrated to give a brown oil (0.3 g, 70.4% yield). 1 The compound 7a (0.06 g, 0.08 mmol) was dissolved in the 2 mL mixture of TFA and CH 2 Cl 2 (1:3), and the solution was stirred at room temperature for 1 h. Then, the solvents were removed and coevaporated with toluene three times to obtain crude free amine product 8a. The amine product 8a without further purification was dissolved in 1 mL of DMF, to which 0.5 mL of TEA was added.…”

“…The resulting residue was purified by silica gel column chromatography (n-hexane:EtOAc = 4:6) to form N-(2-hydroxyphenyl)-2-phenoxyacetamide as a yellow solid (0.364 g, 76.0% yield). 1 03 mmol) and triethylamine (0.56 g, 4.06 mmol) in methanol (4.0 mL) was added di-tert-butyl dicarbonate (0.55 mL, 2.43 mmol). The reaction mixture was stirred at room temperature overnight, and the methanol and TEA were removed in vacuo to yield oily residue, which was dissolved in CH 2 Cl 2 and washed with a solution of sodium carbonate.…”

“…The resulting residue was purified by silica gel column chromatography (hexane:EtOAc = 9:1) to give methyl 4-(4-methoxybenzyloxy)-3-nitrobenzoate as a brown solid (2.56 g, 88.5% yield). 1 4-(4-Methoxybenzyloxy)-3-nitrobenzoic Acid (12). A solution of methyl 4-(4-methoxybenzyloxy)-3-nitrobenzoate (11) (2.0 g, 6.3 mmol) in THF/H 2 O (3:1 20 mL) was treated with lithium hydroxide monohydrate (1.05 g, 25.2 mmol) and stirred at room temperature until the reaction was complete as judged by TLC.…”

“…The resulting residue was purified by silica gel column chromatography (n-hexane:EtOAc = 7:3) to form prop-2-ynyl 4-hydroxy-3-(2-phenoxyacetamido) benzoate as a white solid (0.16 g, 76.2% yield). 1 Phenyl Propargyl Ether (18). A suspension of phenol (16) (1.0 g, 10.6 mmol), anhydrous potassium carbonate (4.4 g, 31.8 mmol), and propargyl bromide (17) (1.6 mL, 21.2 mmol) in DMF (10 mL) was stirred overnight at room temperature.…”

“…The resulting residue was purified by silica gel column chromatography (hexane:EtOAc = 4:6) to give prop-2-ynyl 3-amino-4-hydroxybenzoate as a yellow solid (0.61 g, 88.4% yield). 1 (4-Adamantan-1-yl-phenoxy)acetic Acid (5a). This synthetic procedure followed our previous method.…”