Azide → Sinefungin

HRID 2273054Notes

Reaction equation

HRID 2273054 reaction equation
[NH2:1][C@@H:2]([CH2:6][CH2:7][C@H:8]([NH2:30])[CH2:9][C@@H:10]1[C@@H:14]([OH:15])[C@@H:13]([O:16]CC#C)[C@H:12]([N:20]2[CH:28]=[N:27][C:26]3[C:21]2=[N:22][CH:23]=[N:24][C:25]=3[NH2:29])[O:11]1)[C:3]([OH:5])=[O:4].O=C1O[C@H]([C@H](CO)O)C([O-])=C1O.[Na+].C(#N)C.O.C(O)(C(F)(F)F)=O.[N-]=[N+]=[N-]>O.CN(C=O)C.CO.S([O-])([O-])(=O)=O.[Cu+2].CO.O.CC(O)=O>[CH:23]1[N:24]=[C:25]([NH2:29])[C:26]2[N:27]=[CH:28][N:20]([C@@H:12]3[O:11][C@H:10]([CH2:9][C@@H:8]([NH2:30])[CH2:7][CH2:6][C@H:2]([NH2:1])[C:3]([OH:5])=[O:4])[C@@H:14]([OH:15])[C@H:13]3[OH:16])[C:21]=2[N:22]=1

Participants

10

Reactant 4

HCID 33558AzideHCID 22145617Water TFA acetonitrile2×HCID 23667548Sodium AscorbateHCID 86673677HCID 86673677, compound

Product 1

HCID 65482Sinefungin

Reagents, catalysts & solvents 5

HCID 24462Copper SulfateCatalystHCID 887MethanolSolventHCID 962WaterSolvent3×HCID 6228DimethylformamideSolventHCID 86607755Acetic acid methanol waterSolvent2×

Procedure

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).

Workup

  1. customone product mixture (12.3 min and 13 min, 40%)
  2. waitto continue stifling a second night
  3. customto purify the product which
  4. washeluted from the column at around 8-9 min
  5. concentrationThe product solution was concentrated
  6. customto remove the ACN
  7. customto afford the title detection analyte compound, which
  8. customat 100% B, 9.1-15 min
  9. customthat expected for pure title detection analyte compound regioisomer mixture (5.4 min, ESI+ m/z=1226/2=613)

Sources & evidence

ORD record
ord-a10496e54f1345a6993532757bf999f4
Source dataset
uspto-grants-2015_09
Patent number
US09120820B2

Reaction notes

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