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W Lenk

Publications and source records attributed to W Lenk.

At least 19 recordsLinked to original sources
PubMed1997

Evidence for two psoriasis susceptibility loci (HLA and 17q) and two novel candidate regions (16q and 20p) by genome-wide scan.

In a 12.5 cM genome-wide scan for psoriasis susceptibility loci, recombination-based tests revealed linkage to the HLA region (Zmax = 3.52), as well as suggestive linkage to two novel regions: chromosome 16q (60-83.1 cM from pter, Zmax = 2.50), and chromosome 20p (7.5-25 cM from pter, Zmax = 2.62). All three regions yielded P values < or = 0.01 by non-parametric analysis. Recombination-based and allele sharing methods also confirmed a previous report of a dominant susceptibility locus on distal chromosome 17q (108.2 cM from pter, Zmax = 2.09, GENEHUNTER P = 0.0056). We could not confirm a previously reported locus on distal chromosome 4q; however, a broad region of unclear significance was identified proximal to this proposed locus (153.6-178.4 cM from pter, Zmax = 1.01). Taken together with our recent results demonstrating linkage to HLA-B and -C, this genome-wide scan identifies a psoriasis susceptibility locus at HLA, confirms linkage to 17q, and recommends two novel genomic regions for further scrutiny. One of these regions (16q) overlaps with a recently-identified susceptibility locus for Crohn's disease. Psoriasis is much more common in patients with Crohn's disease than in controls, suggesting that an immunomodulatory locus capable of influencing both diseases may reside in this region.

Adult↗
PubMed1993

Toxicokinetics of diethylene glycol (DEG) in the rat.

Oral doses of 1 and 5 ml/kg 14C-diethylene glycol (DEG) given to rats were rapidly and almost completely absorbed, the invasion constants being 2.95 h-1 and 4.24 h-1. The kinetics of invasion were determined with the method of residuals (Rowland and Tozer 1989) and by reconstruction of the invasion curves according to Kübler (1970). 14C-DEG was rapidly distributed from the blood into the organs and tissues in the order kidneys > brain > spleen > liver > muscle > fat, i.e. the same order as the blood flow. The relative volume of distribution, app. VD, was determined at 298 ml, indicating distribution over the whole body. After oral doses of 1, 5, and 10 ml 14C-DEG/kg 64, 87, and 91% of 14C activity in rat blood disappeared in 12-16 h with a half-life of 3.4 h and the remaining 9, 5, and 4% with half-lives of 39 h, 45 h, and 49 h. A total of 73-96% of 14C activity in blood was excreted with the urine and 0.7-2.2% with the faeces. From the cumulative urinary excretion kinetics half-lives of 6 h were determined for doses of 1 and 5 ml/kg and 10 h for the dose of 10 ml/kg. After doses of 5 ml/kg and 10 ml/kg 14C-DEG semi-logarithmic plots of elimination rate versus time were constant for 5 and 9 h, respectively, indicating that DEG accelerated its renal elimination by inducing osmotic diuresis. Thereafter urinary excretion followed first order kinetics with elimination half-lives of 3.6 h. After oral doses of 5 ml/kg 14C-DEG given to rats of 336 g body weight with an app. VD of 297 ml, the total clearance of 14C activity was determined at 63 ml/h, and the renal clearance of unmetabolized DEG was 66 ml/h. The ratio of ClDEG to Cl(inulin) = 0.64 indicated that DEG and its metabolite 2-hydroxyethoxyacetate (2-HEAA) were reabsorbed from the tubuli into the blood capillaries. DEG produced metabolic acidosis, which was completely balanced after doses of 1 and 5 ml/kg, but doses greater than 10 ml/kg produced non-compensated metabolic acidosis, hydropic degeneration of the tubuli, oliguria, anuria, accumulation of urea-N, and death in uraemic coma.

Administration, Oral↗
PubMed1993

Metabolism of 2-acetylaminofluorene. I. Metabolism in vitro of 2-acetylaminofluorene and 2-acetylaminofluoren-9-one by hepatic enzymes.

1. 2-Acetylaminofluorene (AAF) was converted by rat liver microsomal and cytosolic enzymes to 2-aminofluorene (AF), 2-glycoloylaminofluorene (GAF), 2-acetylaminofluoren-3-, -7-, and -9-ol (3-, 7-, 9-hydroxy-AAF), and 2-acetylaminofluoren-9-one (AAF-9-one). In addition, a new metabolite MX1 was detected. 2. AAF was converted by rabbit liver microsomal and cytosolic enzymes to N-hydroxy-AAF, GAF, 5-, 7-, and 9-hydroxy-AAF, AAF-9-one, 5- and 7-hydroxy-AAF-9-one (new compounds), and AF, indicating species differences in the N- and ring-hydroxylation of AAF and secondary oxygenation of AAF. In addition, an unknown metabolite MX2 was detected. 3. AAF-9-one was converted by rat liver microsomal and cytosolic enzymes to optically active 9-hydroxy-AAF and 7-hydroxy-AAF-9-one; in addition MX1 was found. 4. Rabbit liver microsomal and cytosolic enzymes converted AAF-9-one to 2-aminofluoren-9-one (AF-9-one), 9-hydroxy-AAF, N-hydroxy-AAF-9-one, GAF-9-one, 7-hydroxy-AAF-9-one, and 7,9-dihydroxy-AAF. In addition, metabolite MX1 and its dihydro-dihydroxy derivative were found. 5. These results indicate that AAF and AAF-9-one have common metabolic pathways, as AAF after primary oxygenation to 9-hydroxy-AAF and partial dehydrogenation to AAF-9-one, undergoes secondary oxygenation to 7-hydroxy-AAF-one and MX1 as well as the corresponding dihydro-dihydroxy derivatives.

2-Acetylaminofluorene↗
PubMed1991

The metabolism of 4-aminobiphenyl in rat. I. Reaction of N-hydroxy-4-aminobiphenyl with rat blood in vivo.

1. 3H-4-Aminobiphenyl (ABP, 5 mg) given i.p. to rat had elimination half-lives of 15.6, 17 and 17 h, respectively, for urinary, faecal and total 3H elimination. 14C-ABP administered orally to rats at 100 mg/kg gave elimination half-lives of 31, 36.7 and 34 h, respectively, for urinary, faecal and total 14C elimination. 2. Semi-log plots of percentage dose remaining in the body versus time indicated that: (i) 82% of 3H activity was excreted in 36 h with a half-life of 14.4 h and 18% with a half-life of 46.2 h, and (ii) 77% of 14C activity was excreted in 48 h with a half-life of 15 h and 23% with a half-life of 180 h. 3. After i.p. injection of 10 mg/kg 14C-ABP to rats, ferrihaemoglobin (HbFe3+) concn increased to 60% in 2 h, accompanied by accumulation of 14C activity in erythrocytes, indicating that the active metabolite, N-hydroxy-4-aminobiphenyl (N-hydroxy-ABP) had oxidized haemoglobin-Fe2+ (HbFe2+) and was bound to the erythrocyte. 4. ABP given i.p. to rats at 0.24 mmol/kg rapidly appeared in blood, disappeared with a half-life of 30 min, and blood concn plateaued at 30 nmol/ml. The concn of 4-acetyl-aminobiphenyl (AABP) plateaued at 17 nmol/ml after 15 min, indicating a dynamic equilibrium between N-acetylation of ABP and N-deacetylation of AABP. The concn of 4'-hydroxy-4-acetylaminobiphenyl (4'-hydroxy-AABP) increased slowly at 1.65 nmol/h. 5. AABP given i.p. to rats at 0.88 mmol/kg slowly appeared in the blood, accompanied by the appearance of ABP and 4'-hydroxy-AABP and formation of HbFe3+. After 4 h the concn of AABP and ABP was 27-35 mmol/ml, indicating a dynamic equilibrium between N-deacetylation of AABP and acetylation of ABP. Neither N-hydroxy-ABP nor N-hydroxy-4-acetylaminobiphenyl (N-hydroxy-AABP) were found.

Aminobiphenyl Compounds↗
PubMed1991

The metabolism of 4-aminobiphenyl in rat. III. Urinary metabolites of 4-aminobiphenyl.

1. T.l.c. of the 24-48 h urine of rats dosed with 4-aminobiphenyl (ABP) showed that 4-acetylaminobiphenyl (AABP), 4'-hydroxy-4-aminobiphenyl(4'-hydroxy-ABP), 2'-hydroxy-4-acetylaminobiphenyl(2'-hydroxy-AABP), 4'-hydroxy-4-acetylaminobiphenyl(4'-hydroxy-AABP), 3'-hydroxy, 4'-methoxy-4-acetylaminobiphenyl (3'-hydroxy-4'methoxy-AABP), 4'-hydroxy, 3'-methoxy-4-acetylaminobiphenyl (4'-hydroxy-3'-methoxy-AABP), and 3',4'-dihydroxy-4-acetylaminobiphenyl(3',4'-dihydroxy-AABP) are urinary metabolites. Neither 4-nitrosobiphenyl(nitroso-BP) nor N-hydroxy-4-acetylaminobiphenyl(N-hydroxy-AABP) were detected. 2. Radiochromatography of the 48-h urine of rats dosed with 14C-ABP gave three fractions, U1, U2 and U3 containing 34.6%, 38.8% and 20.4%, respectively, of the total 14C. The conjugated metabolites of ABP were found in U1 and U2, and the unconjugated metabolites in U3, indicating that 80% of the 14C activity in urine was in conjugated, and only 20% in unconjugated metabolites. 3. Rechromatography of U3 gave six radioactive bands from which the following metabolites were isolated and identified as being 4'-hydroxy-AABP, 3'-hydroxy-4'-methoxy-AABP, 4'-hydroxy-3'-methoxy-AABP, 3',4'-dihydroxy-AABP, AABP, ABP, 4,4'-bisazoxybiphenyl (BABP) and 4-(4-aminophenyl)-1,2-benzoquinone. Neither nitroso-BP nor N-hydroxy-AABP were detected.

Aminobiphenyl Compounds↗
PubMed1991

The metabolism of 4-aminobiphenyl in rat. II. Reaction of N-hydroxy-4-aminobiphenyl with rat blood in vitro.

1. N-Hydroxy-4-aminobiphenyl (N-hydroxy-ABP) reacts with HbFe2+ of rat blood in vitro at a molar ratio of 1:47 to produce 20% HbFe3+ within 1 min; N-hydroxy-ABP oxidized 9.4 equiv. of HbFe2+. N-hydroxy-ABP rapidly disappeared and HbFe3+ was reduced at a rate of 44 microM/min. 2. On titration of rat blood in vitro with N-hydroxy-ABP up to 0.81 mM, 4-nitrosobiphenyl (nitroso-BP) disappeared within 5 min; with concn of N-hydroxy-ABP greater than 0.81 mM, N-hydroxy-ABP was present also as nitroso-BP, indicating saturation of reactive binding sites. When N-hydroxy-ABP reacted with HbFe2+ at a molar ratio of 1:103 to 1:1.9, 13 to 1.3 equiv. of HbFe3+ were formed per mol of N-hydroxy-ABP in 5 min, indicating that with increasing N-hydroxy-ABP concn side-reactions increased. 3. After incubation of N-hydroxy-ABP (1.72 mM) with rat Hb (7.66 mM HbFe2+), nitroso-BP disappeared with a half-life of 1 min, maximal HbFe3+ of 72% occurred at 47 min, and the concn of 4-aminobiphenyl (ABP) increased at a rate of 51 nmol/ml per h. 4. In rats injected with 0.24 mmol/kg ABP, HbFe3+ concn plateaued at 56% after 75 min, indicating an equilibrium between HbFe3+ formation and HbFe3+ reduction. Such equilibrium was simulated by titrating rat blood in vitro with N-hydroxy-ABP for 1 h. 5. The long-lasting HbFe3+ formation by ABP in rat results from a cycle of activation of ABP to N-hydroxy-ABP, its rapid co-oxidation with HbFe2+ to form HbFe3+ and nitroso-BP, and binding of nitroso-BP to erythrocyte thiol groups. ABP is released from the Hb adduct and enters a new cycle of activation and inactivation, until terminated by ring-hydroxylation.

Aminobiphenyl Compounds↗
PubMed1991

The metabolism of 4-aminobiphenyl in rat. IV. Ferrihaemoglobin formation by 4-aminobiphenyl metabolites.

1. Rats dosed with nitrosobenzene (56 mumol/kg), 4-chloronitrosobenzene (53 mumol/kg), 3,4-dichloronitrosobenzene (53 mumol/kg), 4-ethoxynitrosobenzene (86 mumol/kg), 4-nitrosobiphenyl(nitroso-BP, 55 mumol/kg) or 2-nitrosofluorene (256 mumol/kg) had maximal ferrihaemoglobin (HbFe3+) concn of 69, 68, 69, 67, 55 and 42% after 15, 25, 48, 35, 80 and 115 min, respectively, indicating differences in solubility of the nitrosoarenes in body fluids. 2. Nitroso-BP and 3-hydroxy-4-aminobiphenyl (3-hydroxy-ABP) catalytically oxidized HbFe2+ in bovine erythrocytes in vitro; nitroso-BP was three times as active as 3-hydroxy-ABP. 3',4'-Dihydroxy-4-aminobiphenyl (3',4'dihydroxy-ABP) showed only low catalytic activity, and seven other ABP metabolites exhibited only marginal activity. 3. Nitroso-BP was inactive in solutions of purified human Hb, but 3-hydroxy-ABP catalytically oxidized HbFe2+, indicating that nitrosoarenes oxidize HbFe2+ in erythrocytes in vitro and in vivo by a mechanism different from that of o-aminophenols. The second-order rate constant for HbFe2+ oxidation by 3-hydroxy-ABP at 37 degrees C was k2 = 19.1 +/- 1.31/mol per s.

Aminobiphenyl Compounds↗
PubMed1990

Evaluation of experimental combined toxicity by use of dose-frequency curves: comparison with theoretical additivity as well as independence.

Dose-frequency curves of toxic effects of a substance A were evaluated in the absence and in the presence of a fixed dose of a second substance B. Data were fitted by the curve-fitting program ALLFIT. Observed combined frequencies of A + B were compared statistically with the expected frequencies of additivity and (or) independence by the phi 2-square goodness-of-fit test. The theoretical dose-frequency curves expected for an additive response were obtained by a solely graphical procedure and the theoretical curves for independent effects were calculated from the effects of B and A at certain doses. In rotarod tests with trained mice, the combined deteriorating effect of ethanol and benzodiazepines were significantly over-additive. However, their lethal interaction appeared underadditive in mice. The lethal underadditive interaction of ethanol and phencyclidine (PCP) can be ascribed largely to independent actions of these compounds. Loss of righting reflex was additively enhanced by PCP, whereas PCP overadditively enhanced the effect of ethanol. The insecticidal action of the cholinesterase inhibitors malathion and parathion appeared additive and significantly different from independent interaction. A comparison of results from dose-response curves with isoboles showed good agreement. The method appears as an attractive alternative or as a complementary procedure to the isobolographic analysis. Combination experiments as described can be carried out and evaluated rather simply, with a minimum of expenditure and a maximum of information.

Animals↗
PubMed1989

N-hydroxy-N-arylacetamides. V. Differences in the mechanism of haemoglobin oxidation in vitro by N-hydroxy-4-chloroacetanilide and N-hydroxy-4-chloroaniline.

1. Autoxidation of N-hydroxy-4-chloroaniline(I) in buffer pH 7.4 was rapid and yielded 4,4'-azoxybischlorobenzene, 4-chloronitrosobenzene, 4-chloronitrobenzene, and 4-chlorophenyl nitroxide. In contrast, autoxidation of N-hydroxy-4-chloroacetanilide(II) was very slow, since in ether and water 78 and 92%, respectively, had decomposed in six months. 2. Haemoglobin(HbO2)-catalysed autoxidation of (I) occurred at a molar ratio of haemoglobin-Fe2+ to (I) of less than 0.25 and was accompanied by ferrihaemoglobin(HbFe3+)-formation and oxygen consumption. Coupled oxidation of HbO2 with (I) occurred at a molar ratio of greater than 0.2 and was accompanied by liberation of oxygen and the formation of HbFe3+, haemoglobin-4-chloronitrosobenzene complex, HbO2, desoxyhaemoglobin, 4-chloronitrosobenzene, 4-chloronitrobenzene, 4-chloroaniline, 4,4'-azoxybischlorobenzene, and 4-chlorophenyl nitroxide. At an equimolar ratio of 10(-3) M haemoglobin-Fe2+ to (I), 96% HbO2 was converted into HbFe3+ (50%) and haemoglobin-4-chloronitrosobenzene complex in the initial fast phase of the reaction, but only 34% of the bound oxygen was liberated, the rest was sequentially reduced to water. (I) completely disappeared, and 4-chloronitrosobenzene was the major metabolite, mainly bound to haemoglobin. 3. Chemical oxidation of (II) by PbO2 in benzene produced acetyl 4-chlorophenyl nitroxide, whose spontaneous decomposition gave 38% 4-chloronitrosobenzene, 33% N-acetoxy-4-chloroacetanilide, 10% 4-chloroacetanilide, and 8% 4-chloronitrobenzene. Its spontaneous decomposition in water also followed second order kinetics, K = 350 l mol-1 sec-1 and yielded N-(2-acetylamino-5-chlorophenyl)-p-benzo-quinoneimine-N-oxide in addition. 4. In the coupled oxidation of 10(-3) M haemoglobin-Fe2+ with 10(-3) M (II), 75% HbFe3+ was formed after 1 h, but only one third of the equivalent of oxygen was released, and two thirds were reduced to water. Concentration of (II) decreased by 5% only, indicating that one mol of (II) had catalysed the oxidation of 15 equivalents of haemoglobin-Fe2+. The identity of the product pattern formed with HbO2 with that produced by chemical one-electron oxidation indicated that oxygen bound to haemoglobin also functions as an acceptor for electrons from (II) as from (I), but the different redox potentials can explain why the secondary aromatic nitroxide was catalytically active and the primary nitroxide was not.

Acetanilides↗
PubMed1989

Pharmacokinetics and biotransformation of diethylene glycol and ethylene glycol in the rat.

1. 14C-Diethylene glycol (DEG), administered orally to rats at 1, 5, and 10 ml/kg, gave elimination half-lives of 6, 6, and 10 h, respectively, from urinary excretion data. Half-logarithmic plots of urinary 14C excretion rates versus time indicated zero-order elimination for the first 9 and 18 h after oral doses of 5 and 10 ml of 14C-DEG/kg, respectively. 14C-DEG urinary elimination kinetics changed into first-order 6, 9, and 18 h after oral doses of 1, 5, and 10 ml/kg, with a half-life of 3 h. 2. After oral doses of 3 and 5 ml ethylene glycol (EG)/kg, half-lives of 4.5 and 4.1 h were estimated from cumulative urinary excretion data for non-metabolized EG. A half-life of 2 h was determined from half-logarithmic plots of urinary excretion rates of non-metabolized EG after the same oral doses of EG. 3. The urinary concentrations of non-metabolized DEG and its metabolite, 2-hydroxyethoxyacetic acid (2-HEAA), determined by high-resolution n.m.r. spectroscopy in the urine of rats doses with DEG were 61-68% and 16-31% dose, respectively. 4. Urinary concentrations of non-metabolized EG and its metabolite, glycolic acid (GA), determined by n.m.r., gave 62-67% for non-metabolized EG and 28.7% for GA following oral doses of EG. 5. Oxidation of DEG and EG in rats was accompanied by a change of urinary pH, reflecting metabolic acidosis. 6. Comparison of the KM for DEG oxidation in vitro by ADH with that of ethanol oxidation, showed a 680-fold difference in substrate affinity. DEG inhibited ethanol oxidation non-competitively, the Ki being 0.44 M.

Acidosis↗
PubMed1987

N-hydroxy-N-arylacetamides--IV. Differences in the mechanism of haemoglobin oxidation in vitro between N-hydroxy-N-arylacetamides and arylhydroxylamines.

In solutions of purified human haemoglobin N-hydroxy-4-chloroacetanilide (N-hydroxy-4ClAA] was one of the most active compounds and N-hydroxy-acetanilide (N-hydroxy-AA) was the least active compound among the six N-hydroxy-N-arylacetamides tested for ferrihaemoglobin (HbFe3+)-forming activity. Co-oxidation of haemoglobin by N-hydroxy-4-chloroacetanilide was compared with that of N-hydroxy-4-chloroaniline(N-hydroxy-4ClA) and found to differ in the kinetics of HbFe2+-oxidation, in the catalytic activity of the two compounds, in the activation energy, and in the product pattern, indicating that the mechanism by which N-hydroxy-N-arylacetamides oxidize oxyhaemoglobin in vitro is different from that of arylhydroxylamines. Attempts have failed to detect by EPR spectroscopy acetyl 4-chlorophenyl nitroxide radical, the postulated catalytically-active oxidation product of N-hydroxy-4-chloroacetanilide.

Acetanilides↗
PubMed1987

N-hydroxy-N-arylacetamides. III: Mechanism of haemoglobin oxidation by N-hydroxy-4-chloroacetanilide in erythrocytes in vitro.

N-Hydroxy-4-chloroacetanilide(N-hydroxy-4C1AA) was the most active, and N-hydroxy-2-acetylaminofluorene(N-hydroxy-2AAF) the least active compound among six N-hydroxy-N-arylacetamides, in forming ferrihaemoglobin(HbFe3+) in bovine erythrocytes in the presence of 11 mM glucose. N-Hydroxy-4C1AA oxidized 25 equiv. of HbFe2+, both in the presence and absence of glucose or lactate. Therefore, its catalytic properties did not depend on metabolic regeneration by the NADPH- or NADH-dependent erythrocyte reductases. In contrast, N-hydroxy-4-chloroaniline(N-hydroxy-4C1A) oxidized 760 equiv. of HbFe2+ in the presence of glucose, but only 81 equiv. of HbFe2+ in the presence of lactate. These results indicate that the catalytic activity depended on the metabolic regeneration from 4-chloronitrosobenzene(4-C1NOB) by NADPH-dependent erythrocyte reductases. A relationship was established between HbFe3+ concn. and the concn. of N-hydroxy-4C1A and 4-C1NOB(determined together), 4-chloroacetanilide(4-C1AA) and 4-chloroaniline(4-C1A), indicating co-oxidation of N-hydroxy-4C1AA and oxyhaemoglobin in erythrocytes and partial reduction of the newly formed 4-C1NOB to 4-C1A. In rat blood in vitro incubated with N-hydroxy-4C1AA, 4-C1NOB concn. increased with increasing HbFe3+ concn., indicating that 4-C1NOB was formed by co-oxidation of oxyhaemoglobin and N-hydroxy-4C1AA, and not by enzymic N-deacetylation.

Acetanilides↗
PubMed1986

N-hydroxy-N-arylacetamides. II: Molecular aspects of ferrihaemoglobin formation by N-hydroxy-N-arylacetamides and arylhydroxylamines in the rat.

Ferrihaemoglobin (HbFe3+) formation in rats after i.p. injection of 6 N-hydroxy-N-arylacetamides has shown that N-hydroxy-4-chloroacetanilide(N-hydroxy-4ClAA) was the most active and N-hydroxy-2-acetylaminofluorene(N-hydroxy-2AAF) the least active compound tested. As N-hydroxy-N-arylacetamides were thought to produce HbFe3+ only after enzymic N-deacetylation, the corresponding arylhydroxylamines were also tested for HbFe3+-forming activity and were found to be more active, N-hydroxy-4-chloroaniline(N-hydroxy-4ClA) being one of the most active and N-hydroxy-2-aminofluorene(N-hydroxy-2AF) the least active compound tested. N-Hydroxy-4-chloroacetanilide given i.p. to rats more rapidly invaded the blood and produced larger amounts of ferrihaemoglobin than did N-hydroxy-2-acetylaminofluorene, due to differences in their availability in plasma. Injection of 50 mg/kg of N-hydroxy-4-chloroacetanilide gave similar concn of HbFe3+ and 4-chloronitrosobenzene(4-CINOB) as injections of 8 mg/kg of N-hydroxy-4-chloroaniline, indicating that the arylhydroxylamine, after N-deacetylation, was the active molecule in vivo. The concn of 4-chloronitrosobenzene declined faster than HbFe3+ concn. 4-Chloronitrosobenzene therefore is a further example of a 'hit-and-run' chemical. Inhibition by the microsomal carboxylesterase inhibitor, bis(4-nitrophenyl)phosphate(BNPP), indicated that ferrihaemoglobin formation by 4-chloroacetanilide, but not by N-hydroxy-4-chloroacetanilide, depends on the enzymic activity of hepatic microsomal carboxylesterases.

Acetamides↗
PubMed1985

The metabolism of N-hydroxyphenacetin in vitro and in vivo.

N-Hydroxyphenacetin (100 mg/kg) injected i.p. into rats rapidly appeared in the blood and disappeared with a t1/2 of 14 min; phenacetin and 4-acetamidophenol were major metabolites in blood. Ferrihaemoglobin was formed, but 4-nitrosophenetole was not detected in blood. N-Hydroxyphenacetin injected i.p. into rats was excreted in the urine unchanged (partly conjugated 2.1% of the dose, 2% was excreted as phenacetin, 19% as 4-acetamidophenol) and 1.8% as 2-hydroxyphenacetin. In addition, small amounts of 3-hydroxyphenacetin (0.4%) and traces of N-[4-(2-hydroxyethoxy)phenyl]acetamide (beta-HAP) (0.05%) were found. Time-course kinetics have shown that N-hydroxyphenacetin is metabolized in vitro to phenacetin, 2- and 3-hydroxyphenacetin, and 4-acetamidophenol by microsomal and cytosolic preparations of rat and rabbit liver. However, after the initial reaction, the formation of phenacetin and 2- and 3-hydroxyphenacetin did not continue with time, indicating that these products were not formed enzymically. N-Hydroxyphenacetin incubated with rat erythrocytes formed ferrihaemoglobin; the relationship between ferrihaemoglobin, phenacetin and 4-nitrosophenetole concn indicated that N-hydroxyphenacetin was oxidized by oxyhaemoglobin to acetyl 4-ethoxyphenyl nitroxide, which yielded phenacetin and 4-nitrosophenetole spontaneously.

Acetaminophen↗
PubMed1985

Additional routes in the metabolism of phenacetin.

omega-Hydroxylation of the ethyl moiety of phenacetin by rabbit-liver microsomal preparations was slow, but was increased 10-fold by pretreatment of the animals with phenobarbitone (PB), and was decreased 2.8-fold by treatment with 3-methylcholanthrene (3-MC) or beta-naphthoflavone (beta-NF). N-[4-(2-hydroxyethoxy)phenyl]acetamide (beta-HAP), the omega-hydroxylation product, which was detected in trace amounts only in the urine of rabbits injected with phenacetin, was converted into [4-(acetylamino)phenoxy]acetic acid (4-APA) by the microsomal and cytosolic fraction of liver homogenate and NADP+ or NAD+. Rabbits excreted 56% of a dose of beta-HAP as 4-APA in the 48 h urine. Phenacetin, injected i.p. into rabbits previously treated with PB, was excreted in the urine as 4-APA (12.2% of dose). beta-HAP formed endogenously or added as substrate in vitro was recovered as the O-acetyl derivative, when ethyl acetate was used for extraction of metabolites from microsomal incubation mixtures. (omega-1)-Hydroxylation of the ethyl moiety of phenacetin, which gave 4-acetamido-phenol, occurred rapidly with rabbit-liver microsomal preparations, and was not increased significantly after pretreatment of animals with either PB or 3-MC. omega-Hydroxylation of the acetic moiety of phenacetin by rabbit-liver preparations to give N-(4-ethoxyphenyl)glycolamide (4-GAP) was slow, but was increased three-fold after pretreatment of animals with 3-MC or beta-NF, whereas PB had no effect. 4-GAP was detected in trace amounts only in the urine of rabbits injected i.p. with phenacetin. N-Hydroxylation of phenacetin by rabbit-liver microsomal preparations was slow, but increased three-fold after treatment of animals with 3-MC, and was unchanged by PB. N-Hydroxylation of phenacetin by hepatic microsomes from 3-MC-treated rabbits was 26 times slower than that of 2-acetylaminofluorene; no N-hydroxy derivatives of N-(4-chlorophenyl)acetamide and propanil were detected in vitro.

Acetylation↗
PubMed1984

N-Hydroxy-N-arylacetamides. I. Toxicity of certain polycyclic and monocyclic N-hydroxy-N-arylacetamides in rats.

Of the two carcinogenic N-hydroxy-N-arylacetamides tested, N-hydroxy-4-acetylaminobiphenyl was as active as the monocyclic analogs in the oxidation of hemoglobin, whereas N-hydroxy-2-acetylaminofluorene produced less ferrihemoglobin after IP injection into female and male rats. Monocyclic N-hydroxy-N-arylacetamides, such as N-hydroxy-4-chloroacetanilide or N-hydroxyphenacetin, were more toxic than the parent N-arylacetamides, LD50 in mice being 190 mg/kg for N-hydroxy-4-chloroacetanilide vs 755 mg/kg for 4-chloroacetanilide, and 702 mg/kg for N-hydroxyphenacetin versus 1,220 mg/kg for phenacetin. The higher acute toxicities are probably due, at least in part, to the production of more ferrihemoglobin by the N-hydroxy-N-arylacetamides. Chronic toxicity of N-hydroxy-4-chloroacetanilide was tested on 10 male and 10 female Sprague Dawley rats after IP or SC injection of 20 mg (0.11 mmol)/kg twice weekly for 16 weeks into two groups of 10 animals each (five males, five females, total dose: 3.5 mmol/kg). The experiment, which was terminated after 2 years, did not yield any hint that N-hydroxy-4-chloroacetanilide was carcinogenic in the rat. Subchronic toxicity of N-hydroxyphenacetin was tested in two experiments on male and female Sprague Dawley rats after IP or SC injection of 50 or 100 mg (0.26 or 0.51 mmol)/kg. In the first experiment, two groups of 15 rats each (seven males, eight females) were injected either IP or SC with 50 and 100 mg/kg twice weekly for 29 weeks, and in the second experiment groups of 10 males and 10 females were injected SC with 100 mg/kg twice daily on 5 days a week for 12 weeks. The experiments, which were terminated after 29 weeks and 12 weeks treatment, respectively, did not provide evidence for chronic interstitial nephritis or tumor growth in the kidney. N-Hydroxy-N-arylacetamides were found to be inferior to the corresponding arylhydroxylamines in their ferrihemoglobin-forming capabilities in female rats. Large differences in activity of the arylhydroxylamines and no close relation to the number of rings was observed, N-hydroxy-2-acetylaminofluorene being the least active and N-hydroxy-4-acetylaminobiphenyl being as active as the monocyclic compounds, and exceeding all in the duration of its activity.(ABSTRACT TRUNCATED AT 400 WORDS)

Acetamides↗
PubMed1984

Peroxidase activity of oxyhaemoglobin in vitro.

In bovine erythrocyte suspensions incubated with 16 mM aniline, 4-phenetidine, 4-chloro- or 3,4-dichloroaniline for three hours at 37 degrees C, HbFe3+ concentrations of 10, 35, 77 and 93%, respectively, were found. N- and C-oxygenation products of aniline, 4-chloro-, and 3,4-dichloroaniline were formed, which can explain the oxidation of HbFe3+, indicative of peroxygenase activity of oxyhaemoglobin. The same N- and C-oxygenated derivatives of 4-chloro- and 3,4-dichloroaniline were also formed by hepatic microsomes, although at a 25- to 5000-fold higher rate. HbFe3+ was formed more readily on incubation of either bovine erythrocytes or purified human Hb with various N-arylacetohydroxamic acids. The metabolites of N-(4-chlorophenyl)-N-hydroxyacetamide are the same as the products of chemical oxidation of NOH-4ClAA by PbO2 or KMnO4, indicating the peroxidase activity of oxyhaemoglobin.

Aniline Compounds↗