Azide → Sinefungin

Participants

Reaction Steps

To a mixture consisting of Compound 7 (˜1 mg, 0.0016 mmol) in water (100 μL) with sodium ascorbate (1 mg, 0.005 mmol) in water (200 μL) was added copper(II) sulfate (500 μg, 0.0031 mmol) in water (100 μL) and Azide-Fluor585 (Click Chemistry Tools, Catalog #AZ110, 1.3 mg, 0.0016 mmol) in DMF (600 μL). The reaction mixture was stirred at room temperature overnight. HPLC (Gemini C18, 110 {acute over (Å)}, 5μ, 250×4.6 mm, 1 mL/min, 5/95/0.05→90/10/0.05 ACN/H2O/TFA) showed mainly one product mixture (12.3 min and 13 min, 40%) and starting fluorophore azide mixture (18.7 min and 20.5 min, 50%). The reaction mixture was allowed to continue stifling a second night. Prep HPLC (Gemini C18, 110 {acute over (Å)}, 10μ, 250×21.2 mm, 20 mL/min, 15/85/0.05→90/10/0.05 ACN/H2O/TFA) was done to purify the product which eluted from the column at around 8-9 min. The product solution was concentrated to remove the ACN and then lyophilized to afford the title detection analyte compound, which was taken up in methanol (500 μL) and quantified by UV (895 μg/mL, or 448 μg); LC/MS (Gemini C18, 3μ, 2.0×50 mm, 400 μL/min, A: 90/10/0.01 H2O/MeOH/HOAc, B 90/10/0.01 MeOH/H2O/HOAc 0-6 min 0-100% B, 6-9 hold at 100% B, 9.1-15 min re-equilibrate at 0% B) consistent with that expected for pure title detection analyte compound regioisomer mixture (5.4 min, ESI+ m/z=1226/2=613).

Provenance & Evidence