Identification and characterization of human metabolites of CAI [5-amino-1-1(4'-chlorobenzoyl-3,5-dichlorobenzyl)-1,2,3-triazole- 4-carboxamide).
The calcium influx inhibitor and cytostatic agent, 5-amino-1-1(4'-chlorobenzoyl-3,5-dichlorobenzyl)-1,2,3-triazole-4-carboxamide (CAI), is in phase I clinical trial for patients with refractory cancer. Additional chromatography peaks were observed during HPLC analysis of patient samples. Identification and characterization of physiological metabolites were undertaken using HPLC techniques developed for their purification from blood, pleural fluid, and urine samples. A hydrophobic metabolite, M1, was purified and functionally characterized. Structural analysis of the purified compound indicated that it is a 3,5-dichloro-4(p-chlorobenzoyl)-benzoic acid. Quantitative analysis of M1 concentration during CAI administration indicated that the rise in M1 concentration lagged behind that of CAI and persisted after CAI was no longer detectable. No clear relationship between CAI or M1 and either toxicity or efficacy was observed. Chromatography of patient blood and urine samples under conditions favoring hydrophilic metabolite detection suggested the presence of a glucuronide compound; this was also indicated by sample treatment with beta-glucuronidase. Attempts at purification did not yield a compound stable for structural analysis. The benzophenone metabolite, M1, was nonfunctional in assays of calcium influx inhibition or proliferation. No pharmacodynamic associations were observed for these metabolites, nor was there pharmacological activity of the M1 as an individual agent. These data suggest that CAI is processed into triazole and benzophenone moieties by phase I metabolism, and these metabolites or the parent compound may be conjugated for excretion by glucuronidation.
Soltis
M J
MJ
Signal Transduction and Prevention Unit, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892, USA.
Yeh
H J
HJ
Cole
K A
KA
Whittaker
N
N
Wersto
R P
RP
Kohn
E C
EC
D000970:Antineoplastic Agents / Q000493:pharmacokinetics*; D016718:Arachidonic Acid / Q000378:metabolism; D001202:Ascitic Fluid / Q000378:metabolism; D001565:Benzoates / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology; D002118:Calcium / Q000378:metabolism; D002455:Cell Division / Q000187:drug effects; D002460:Cell Line; D002851:Chromatography, High Pressure Liquid; D005122:Exudates and Transudates / Q000378:metabolism; D005965:Glucuronates / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D005966:Glucuronidase / Q000737:chemistry; D006801:Humans; D008168:Lung / Q000378:metabolism; D014230:Triazoles / Q000097:blood / Q000737:chemistry* / Q000493:pharmacokinetics*; D014407:Tumor Cells, Cultured / Q000187:drug effects
D000970:Antineoplastic Agents; D001565:Benzoates; D005965:Glucuronates; D014230:Triazoles; C104827:3,5-dichloro-4-(4-chlorobenzoyl)benzoic acid; D016718:Arachidonic Acid; C062058:carboxyamido-triazole; D005966:Glucuronidase; D002118:Calcium