Non-competitive inhibition of human HinT1 using tryptamine 5'-adenosine phosphoramidate as substrate compound pre-incubated for 30 secs prior substrate addition measured for 2 mins by Dixon plot analysis
Stability in human HeLa cell lysate assessed as HINT-1-mediated phosphoramidase activity by measuring compound remaining at 1 mM after 22 hrs by RP-HPLC analysis relative to control
Stability in human K562 cell lysate assessed as HINT-1-mediated phosphoramidase activity by measuring compound remaining at 1 mM after 22 hrs by RP-HPLC analysis relative to control
Drug metabolism in human K562 cell lysate assessed as HINT-1-mediated gemcitabine formation at 1 mM after 22 hrs by RP-HPLC analysis in presence of phosphatase inhibitor relative to control
Drug metabolism in human K562 cell lysate assessed as HINT-1-mediated gemcitabine formation at 1 mM in absence of phosphatase inhibitor after 22 hrs by RP-HPLC analysis relative to control
Drug metabolism in human K562 cell lysate assessed as HINT-1-mediated compound formation by measuring retention time at 1 mM after 22 hrs by RP-HPLC analysis relative to control
Drug metabolism in human K562 cell lysate assessed as HINT-1-mediated compound dephosphorylation at 1 mM after 22 hrs by RP-HPLC analysis relative to control
Drug metabolism in human K562 cell lysate assessed as HINT-1-mediated gemcitabine formation at 1 mM after 22 hrs by RP-HPLC analysis relative to control
Drug metabolism in human K562 cell lysate assessed as HINT-1-mediated gemcitabine 5'-monophosphate formation at 1 mM after 22 hrs by RP-HPLC analysis relative to control
Drug level in human K562 cell lysate treated with 1 mM gemcitabine 5'-phosphoramidate assessed as HINT-1-mediated compound formation after 22 hrs by RP-HPLC analysis relative to control
Drug metabolism in human HuH7 cells assessed as HINT1-mediated ((2R,3R,4R,5R)-5-(4-butyramido-2-oxopyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methyl dihydrogen phosphate metabolite formation at 50 uM after 48 hrs by LC/MS/MS analysis
Prodrug activation in human Mino cells assessed as Hint1 mediated 2-amino-7-(2-(4-chlorophenoxy)ethyl)-9-((2R,3R,4S,5R)-3,4-dihydroxy-5-(phosphonooxymethyl)tetrahydrofuran-2-yl)-6-oxo-6,9-dihydro-1H-purin-7-ium formation per 5 million cells at 150 uM after 12 hrs by LC-MS/MS analysis
Prodrug activation in human Z138 cells assessed as Hint1 mediated 2-amino-7-(2-(4-chlorophenoxy)ethyl)-9-((2R,3R,4S,5R)-3,4-dihydroxy-5-(phosphonooxymethyl)tetrahydrofuran-2-yl)-6-oxo-6,9-dihydro-1H-purin-7-ium formation per 5 million cells at 150 uM after 12 hrs by LC-MS/MS analysis
Prodrug activation in human GRANTA-519 cells assessed as Hint1 mediated 2-amino-7-(2-(4-chlorophenoxy)ethyl)-9-((2R,3R,4S,5R)-3,4-dihydroxy-5-(phosphonooxymethyl)tetrahydrofuran-2-yl)-6-oxo-6,9-dihydro-1H-purin-7-ium formation per 5 million cells at 150 uM after 12 hrs by LC-MS/MS analysis
Prodrug activation in human Z138 cells assessed as hint1-mediated 2-amino-7-(2-(4-chlorophenoxy)ethyl)-9-((2R,3R,4S,5R)-3,4-dihydroxy-5-(phosphonooxymethyl)tetrahydrofuran-2-yl)-6-oxo-6,9-dihydro-1H-purin-7-ium formation per 5 million cells at 150 uM after 12 hrs by LC-MS/MS analysis relative to 9-((2R,3R,4S,5R)-5-(((((2-(1H-Indol-3-yl)ethyl)amino)-(hydroxy)phosphoryl)oxy)methyl)-3,4-dihydroxytetrahydrofuran-2-yl)-2-amino-7-(2-(4-chlorophenoxy)ethyl)-6-oxo-6,9-dihydro-1H-purin-7-ium
Prodrug activation in human GRANTA-519 cells assessed as hint1-mediated 2-amino-7-(2-(4-chlorophenoxy)ethyl)-9-((2R,3R,4S,5R)-3,4-dihydroxy-5-(phosphonooxymethyl)tetrahydrofuran-2-yl)-6-oxo-6,9-dihydro-1H-purin-7-ium formation per 5 million cells at 150 uM after 12 hrs by LC-MS/MS analysis relative to 9-((2R,3R,4S,5R)-5-(((((2-(1H-Indol-3-yl)ethyl)amino)-(hydroxy)phosphoryl)oxy)methyl)-3,4-dihydroxytetrahydrofuran-2-yl)-2-amino-7-(2-(4-chlorophenoxy)ethyl)-6-oxo-6,9-dihydro-1H-purin-7-ium
Inhibition of PKCi in human SKCO1 cells at 1 uM after 4 hrs using biotin labeled DLKLDNVLLDSEGHIK probe by mass-spectrometric analysis relative to control
Inhibition of His-tagged human full length HINT1 expressed in Rosetta2 pLysS cells using TrpAMP as substrate measured for 2 to 30 mins by fluorescence assay