Triethylamine → HCID 20638914, compound

To a 500 mL round bottom flask, charged with 9.00 g (36.8 mmol) of biotin (Sigma Chemical Company), 7.93 g (40.5 mmol, 1.1 equiv.) of amine hydrochloride 12, and 200 mL of anhydrous dimethylformamide was added 17 mL of triethylamine (120 mmol) followed by 17.1 g (38.7 mmol, 1.05 equiv.) of benzotriazolyloxytris(dimethylamino)phosphonium hexaphosphonate (BOP, commercially available from Aldrich Chemical Company and Chem-Impex International, Wood Dale, Ill.) coupling agent. The reaction was stirred at room temperature for 13 h and then concentrated. The residue was diluted with 100 mL of 2-propanol and 300 mL of methylene chloride and the resulting mixture was washed with 2×150 mL aliquots of 1N aqueous HCl and then with 150 mL of de-ionized water. The organic phase was dried with anhydrous magnesium sulfate and vacuum filtered. The solids were rinsed with 100 mL of 25% 2-propanol/methylene chloride. The combined filtrates were concentrated. The residue was chromatographed on 9×22 cm of silica gel, eluting with 20% methanol/ethyl acetate. The fractions containing product (13) were combined and concentrated via rotary evaporation. The residue was chromatographed on 7×18 cm of RP-18 silica gel, eluting with 800 mL of 50:50 methanol/water, 1 liter of 55:45 methanol/water and 2 liters of 60:40 methanol/water. The fractions containing product (13) were combined and concentrated, first via rotary evaporation and finally under full vacuum pump pressure (<0.5 mm Hg) to afford 11.58 of the product (13) as a near colorless oil: 1H NMR (d6-DMSO) d 6.42 (1H, s), 6.33 (1H, s), 4.29 (1H, m), 4.12 (1H, m), 3.57 (3H, s), 3.22 (2H, t), 3.09 (1H, m), 2.91 and 2.77 (3H, 2 s), 2.81 (1H, dd), 2.57 (1H, d), 2.34-2.19 (4H, m) and 1.70-1.10 (12H, m) ppm; 1H NMR (CD3pk OD) d 4.48 (1H, dd), 4.29 (1H, dd), 3.63 and 3.62 (3H, 2 s), 3.34 (2H, t), 3.20 (1H, m), 3.02 and 2.88 (3H, 2 s), 2.91 (1H, dd), 2.68 (1H, d), 2.43-2.27 (4H, m) and 1.80-1.20 (12H, m) ppm; Thin Layer Chromatography (Visualization with p-aminocinnimaldehyde spray); Silica Gel, Rf=0.31 ({fraction (80/20)} ethyl acetate/methanol); RP-18 Silica Gel, Rf=0.29 ({fraction (60/40)} methanol/water).

1. concentrationconcentrated
2. additionThe residue was diluted with 100 mL of 2-propanol and 300 mL of methylene chloride
3. washthe resulting mixture was washed with 2×150 mL aliquots of 1N aqueous HCl
4. dry with materialThe organic phase was dried with anhydrous magnesium sulfate and vacuum
5. filtrationfiltered
6. washThe solids were rinsed with 100 mL of 25% 2-propanol/methylene chloride
7. concentrationThe combined filtrates were concentrated
8. customThe residue was chromatographed on 9×22 cm of silica gel
9. washeluting with 20% methanol/ethyl acetate
10. additionThe fractions containing product (13)
11. concentrationconcentrated via rotary evaporation
12. customThe residue was chromatographed on 7×18 cm of RP-18 silica gel
13. washeluting with 800 mL of 50:50 methanol/water, 1 liter of 55:45 methanol/water and 2 liters of 60:40 methanol/water
14. additionThe fractions containing product (13)
15. concentrationconcentrated, first via rotary evaporation and finally under full vacuum pump pressure (<0.5 mm Hg)

ORD record

ord-d395050729c149de81d3c329c18a3ae1

Source dataset

uspto-grants-2001_01

DOI

10.6084/m9.figshare.5104873.v1

Patent number

US06172045B2