In vitro studies on the deacetylation-reacetylation of arylamides and the transacetylation of arylamines by human and rat whole blood.
Human and rat whole blood were shown to metabolize the aromatic amides acetanilide and phenacetin by deacetylation followed by reacetylation in vitro. Derivatives of the parent compounds labelled with deuterium in the N-acetyl group produced non-labelled material after incubation. The reaction was monitored by capillary gas chromatographic-mass spectrometric (GC-MS) analysis. There was no significant difference in the acetyl group exchange of these substrates using blood samples donated by non-diabetic volunteers or Type 2 diabetic patients (respective mean +/- SEM values = 4.0 +/- 0.2% and 4.2 +/- 0.3% for trideuteroacetanilide, 6.2 +/- 0.6% and 6.1 +/- 0.3% for trideuterophenacetin). Increasing the glucose concentration in the incubation medium by 50 mmol/L significantly (P less than 0.01) increased deacetylation-reacetylation of trideuteroacetanilide in each group (4.6 +/- 0.2% and 4.7 +/- 0.2% for non-diabetic and diabetic subjects, respectively). In rat blood the amount of deacetylation-reacetylation was much higher: 7.2 +/- 0.6% and 8.3 +/- 0.7% for trideuteroacetanilide and trideuterophenacetin, respectively. Induction of experimental diabetes using streptozotocin did not significantly change the extent of deacetylation-reacetylation of either deuterated substrate (10.1 +/- 2.1% and 9.5 +/- 1.1%). Elevation of the incubation glucose concentration by 50 mmol/L produced an increase in acetyl group exchange (for trideuteroacetanilide) in diabetic (14.3 +/- 2.2%) and non-diabetic (10.6 +/- 1.0%) rats. The donation of acetyl groups (transacetylation) was observed after incubation of blood samples from both diabetic and non-diabetic human subjects and rats with trideuterophenacetin and a molar excess of aniline. This reaction significantly (P less than 0.001) decreased the acetyl group exchange of trideuterophenacetin (these values were 4.5 +/- 0.4% and 3.4 +/- 0.6% using samples from non-diabetic human subjects and rats, respectively) and demonstrated the ability of whole blood to catalyse transacetylation (acetyl-CoA-independent acetylation). There was correlation between the amount of (unlabelled) acetanilide produced by acetylation with acetyl-CoA and the percentage present as trideuteroacetanilide. The proportion of trideuteroacetanilide was higher using rat blood (e.g. the values for non-diabetic subjects were 25.5 +/- 1.7% vs 8.5 +/- 0.3%; P less than 0.001) although the total amount of acetanilide produced was lower (0.54 +/- 0.14 nmol vs 1.82 +/- 0.23 nmol; P less than 0.05) than that observed using human blood.
Lindsay
R M
RM
Department of Biochemical Medicine (University of Dundee), Ninewells Hospital and Medical School, U.K.
Fox
W R
WR
Baty
J D
JD
Willis
R G
RG
D000083:Acetanilides / Q000378:metabolism; D000107:Acetylation; D000814:Aniline Compounds / Q000494:pharmacology; D000818:Animals; D001191:Arylamine N-Acetyltransferase / Q000097:blood / Q000378:metabolism*; D001786:Blood Glucose / Q000032:analysis; D002851:Chromatography, High Pressure Liquid; D003903:Deuterium; D003920:Diabetes Mellitus / Q000097:blood; D003921:Diabetes Mellitus, Experimental / Q000097:blood; D008401:Gas Chromatography-Mass Spectrometry; D005947:Glucose / Q000494:pharmacology; D006801:Humans; D008297:Male; D010615:Phenacetin / Q000378:metabolism; D051381:Rats; D011919:Rats, Inbred Strains
D000083:Acetanilides; D000814:Aniline Compounds; D001786:Blood Glucose; D003903:Deuterium; D001191:Arylamine N-Acetyltransferase; D010615:Phenacetin; D005947:Glucose; C023650:aniline