Protease-mediated enzymatic hydrolysis and activation of aryl phosphoramidate derivatives of stavudine.
Several proteases are capable of hydrolyzing the aryl substituted phosphoramidate derivatives of stavudine resulting in the formation of the active metabolite, alaninyl d4T monophosphate. Subtilisin Protease A, Subtilisin Griseus, Subtilisin Carlsberg, Papaya, Bacillus were amongst the most effective proteases in hydrolyzing stavudine derivatives and specificity of their activity was confirmed using several protease inhibitors to block the hydrolysis of these phosphoramidate derivatives. We found that these proteases exhibit chiral selectivity at the phosphorus center of stavudine derivatives. Our results indicate that cellular proteases may be responsible for the activation of these phosphoramidate derivatives. In addition, we show that the enzymatic hydrolysis takes place at the carboxymethyl ester side chain of these pro-drugs and the direct attack on the phosphorus center by these enzymes does not occur. Finally, we describe a novel activation pathway hitherto unknown for the activation and viral inhibitory characteristic shown by these phosphoramidate derivatives of stavudine.
Venkatachalam
T K
TK
Department of Chemistry, Parker Hughes Institute and Paradigm Pharmaceuticals, 2699, Patton Road, Roseville, MN 55113, USA. jhumphrey@ih.org
Samuel
P
P
Qazi
S
S
Uckun
F M
FM
D000577:Amides / Q000138:chemical synthesis* / Q000737:chemistry / Q000493:pharmacokinetics; D001711:Biotransformation; D054306:Dideoxynucleotides; D009682:Magnetic Resonance Spectroscopy; D010447:Peptide Hydrolases / Q000378:metabolism*; D010756:Phosphoric Acids / Q000138:chemical synthesis* / Q000737:chemistry / Q000493:pharmacokinetics; D011355:Prodrugs / Q000493:pharmacokinetics*; D013056:Spectrophotometry, Ultraviolet; D018119:Stavudine / Q000031:analogs & derivatives* / Q000138:chemical synthesis / Q000378:metabolism / Q000493:pharmacokinetics; D013237:Stereoisomerism; D013938:Thymidine Monophosphate / Q000031:analogs & derivatives* / Q000378:metabolism
D000577:Amides; D054306:Dideoxynucleotides; D010756:Phosphoric Acids; D011355:Prodrugs; C472384:stampidine; D013938:Thymidine Monophosphate; C011067:phosphoramidic acid; D018119:Stavudine; D010447:Peptide Hydrolases