Synthesis of β‐³H‐mitotane for use in a rapid assay for mitotane metabolism

Abstract

SUMMARYA 3H+-release method has been developed for the assay of P-hydroxylation of the adrenolytic drug mitotane. P-3H-rnitotane was synthesized by the reduction of 142-chlorophenyl)-l-(4-chlorophenyl)-2,2,2-trichloroethane by an aluminium-HgZC12 couple in the presence of 3Hz0. For P-hydroxylation of mitotane, the jH+-release assay is more efficient and sensitive than a method utilizing 14C-mitotane and chromatographic separation of metabolites by HPLC. The 3H+-release assay has been used to evaluate the ability of adrenal tumors to metabolize mitotane via the Phydroxylation route.

Development of Mitotane Lipid Nanocarriers and Enantiomers: Two-in-One Solution to Efficiently Treat Adreno-Cortical Carcinoma

“…Among the major mitotane metabo- [61], and o,p'-DDA (1,1-(o,p'-dichlorodiphenyl) acetic acid) [62] which has been identified through a proposed route involving the adrenal mitochondrial cytochrome P450-catalyzed hydroxylation of mitotane at the -carbon [63,64]. Subsequent dehydrochlorination of the hydroxylated product forms the corresponding acyl chloride that, in the presence of water, formed the acidic metabolite, o,p'-DDA, although it could alternatively bind to tissue nucleophiles [65]. Interestingly, the synthesis route of -3 H-mitotane has been reported few years ago for use in a assay for mitotane metabolism [65], and consisted in the reduction of 1-(2-chlorophenyl)-1-(4-chlorophenyl)-2,2,2-trichloroethane (o,p´-DDT) by an aluminium-Hg 2 Cl 2 couple in the presence of tritied water ( 3 H 2 0).…”

“…Interestingly, the synthesis route of -3 H-mitotane has been reported few years ago for use in a assay for mitotane metabolism [65], and consisted in the reduction of 1-(2-chlorophenyl)-1-(4-chlorophenyl)-2,2,2-trichloroethane (o,p´-DDT) by an aluminium-Hg 2 Cl 2 couple in the presence of tritied water ( 3 H 2 0). Thereby, the determination of the 3 H + , released to the aqueous media after organic solvent extraction, constituted a specific, faster (about 2-3 hours), and more sensitive assay for mitotane metabolic activation mediated by -hydroxylation than 14 Cmitotane-high-performance liquid chromatography (HPLC) [65]. Initial experiments in rats using 14 C-labeled mitotane along with thin-layer chromatography (TLC), gas-liquid chromatography (GLC) and MS, reported that most of metabolites (87.8%) was concentrated in the feces (e.g.…”

“…Subsequent dehydrochlorination of the hydroxylated product forms the corresponding acyl chloride that, in the presence of water, formed the acidic metabolite, o,p'-DDA, although it could alternatively bind to tissue nucleophiles [65]. Interestingly, the synthesis route of -3 H-mitotane has been reported few years ago for use in a assay for mitotane metabolism [65], and consisted in the reduction of 1-(2-chlorophenyl)-1-(4-chlorophenyl)-2,2,2-trichloroethane (o,p´-DDT) by an aluminium-Hg 2 Cl 2 couple in the presence of tritied water ( 3 H 2 0). Thereby, the determination of the 3 H + , released to the aqueous media after organic solvent extraction, constituted a specific, faster (about 2-3 hours), and more sensitive assay for mitotane metabolic activation mediated by -hydroxylation than 14 Cmitotane-high-performance liquid chromatography (HPLC) [65].…”

“…In fact, mitotane metabolism is being studied for almost four decades to better understand its pharmacokinetics and pharmacodynamics and so, its molecular activity, which would help in carrying out the treatment [61][62][63][64][65][66][67][68][69][70]. Several approaches, using chromatography and/or spectrometry, have been developed to quantitatively determine mitotane and its metabolites in body fluids (e.g.…”

Current and Emerging Therapies for Advanced Adrenocortical Carcinoma

“…Unfortunately, at present, there is no widely accepted method for predicting the response of an individual to mitotane therapy. A tritium-release assay has been developed and is reported to correlate with the ability of the tumor to metabolize mitotane [62,63].…”