Acetic Acid → HCID 131719480, compound

Participants

Reaction Steps

2-((2-Amino-1,6-dihydro-6-oxo-purin-9-yl)methoxy)-1,3-propanediyl bis-(1-amino-cyclohexane-carboxylate) tris acetate, 98 mg (lyophilized sample contained 3.0 equivalents of acetic acid by NMR), 0.143 mmol (=a total of 0.43 mmol of acetic acid) is dissolved in de-ionized water, 0.4 mL, and 38 μL of a 0.015M ammonium hydroxide solution (=0.57 mmol) is added. The mixture is left at room temperature for 7 hrs., the vial is placed in a oil bath maintained at 48°-50° C. and left in the oil bath for 22 hrs. The sample is injected in two equal lots onto a preparative reverse phase HPLC column (YMC-Pack, ODS-AM DM-33-5, 20×250 mm; YMC Inc.). Separation is achieved with a solvent system of 15% methanol/85% 0.1M ammonium acetate buffered to pH 4 with acetic acid, flow rate: 9.5 mL/min and the detector set to 256 nm. The peak representing the mono ester is collected. The solvent is removed under high vacuum to about 2 mL and the residue is lyophilized three times from water containing acetic acid (0.1%) to remove the buffer. Excess acetic acid is then removed by drying at room temperature under high vacuum for 42 hrs. 30 mg (0.068 mmol=48%) of 2-(2-amino-1,6-dihydro-6-oxo purin-9-yl)methoxy)-3-hydroxy-1-propanyl (1-amino-cyclohexane-carboxylate) acetate, identical by HPLC and NMR with the product of Example 4B, is obtained.

Provenance & Evidence