Biomedical subjects

R G Dickinson

Publications and source records attributed to R G Dickinson.

At least 19 recordsLinked to original sources

PubMed1998

Hepatobiliary transport of diflunisal conjugates and taurocholate by the perfused rat liver: the effect of chronic exposure of rats to diflunisal.

Acyl glucuronides are reactive electrophilic metabolites of carboxylate drugs which can form covalent adducts with endogenous macromolecules such as serum albumin and hepatic proteins. Such adducts have been suggested as initiating factors in certain immune and toxic responses to acidic drugs. In the present study, pretreatment of rats with high daily doses (50 mg/kg orally) of the non-steroidal anti-inflammatory drug (NSAID) diflunisal (DF) for 35 days, followed by perfusion of the isolated liver with 3 mg DF for 3 hr, resulted in appreciable concentrations of covalent adducts of DF with hepatic tissue (3.68 microg DF/g liver). Immunoblotting using a rabbit polyclonal DF antiserum showed the major DF-modified bands at about 110, 140 and 200 kDa. A vehicle-pretreated control group achieved adduct concentrations of only 0.37 microg DF/g liver, with the 200 kDa band not detectable in immunoblots. Elimination of DF from perfusate of the isolated perfused rat liver (IPRL) preparation was the same (t1/2 about 3.4 hr) in both DF- and vehicle-pretreated groups. Appearance of the sulfate (DS) conjugate, the major metabolite in perfusate, was also similar. However, higher concentrations of the acyl glucuronide (DAG) and phenolic glucuronide (DPG) conjugates were found in perfusate at later times, though a statistically significant difference in area under the concentration-time curve was found only in the case of DAG. At 3 hr, recoveries of dose as DAG and DPG were significantly higher in perfusate, but not in bile. No significant differences in uptake and biliary excretion of taurocholate were found between the two groups. The finding of higher perfusate concentrations of DAG and DPG could signal a minor compromise to biliary excretion processes for the glucuronides, though whether such a result is simply coincident with or attributable to DAG-derived covalent DF-protein adducts in liver remains indeterminate.

Animals↗

PubMed1998

Disposition and covalent binding of diflunisal and diflunisal acyl glucuronide in the isolated perfused rat liver.

Acyl glucuronides are intrinsically reactive metabolites of carboxylate drugs, capable of undergoing hydrolysis, intramolecular rearrangement (isomerization via acyl migration), and intermolecular transacylation reactions. Transacylation with nucleophilic groups located on protein molecules leads to covalent drug-protein adducts. Protein adducts can also form from the rearrangement isomers via a glycation mechanism. In this study, the isolated perfused rat liver preparation was used to separately trace the dispositions of the nonsteroidal anti-inflammatory drug diflunisal (DF), its reactive acyl glucuronide metabolite (DAG), and a mixture of DAG rearrangement isomers (iso-DAG), each administered at 30-microg DF equivalents/ml perfusate (four recirculating perfusions each group). After administration of DF, the drug was eliminated in a log linear manner over 3 hr, with apparent elimination half-life (t1/2) of 2.6 +/- 0.4 hr. The sulfate conjugate (DS), excreted almost exclusively into perfusate, accounted for 14.2% of the dose, with the phenolic glucuronide (DPG) and DAG (11.1 and 7.9% of dose, respectively) excreted primarily in bile. Only a small portion (2.3%) of the dose was recovered as novel "diglucuronides" (D-2G, arising from phenolic glucuronidation of iso-DAG), excreted exclusively in bile. Covalent DF-protein adducts were found in both perfusate (0.98%) and liver (0. 14%). After administration of DAG, rapid hydrolysis occurred (initial DAG t1/2 17.3 +/- 4.2 min). At 3 hr, recoveries (in comparison to DF-dosed perfusions) were similar for DF (51.7%) and DAG (8.3%), significantly decreased for DS (10.6%) and DPG (6.4%), and significantly increased for iso-DAG (0.8%), D-2G (9.1%), and covalent adducts in perfusate (1.49%) and liver (0.30%). After administration of iso-DAG, elimination from perfusate was slower (t1/2 55 +/- 15 min), and hydrolysis to DF was modest by comparison with DAG-dosed perfusions. Recoveries as iso-DAG and D-2G in bile were greatly enhanced (8.2 and 36.4%, respectively). Adduct formation was higher in liver (0.76% of dose) but not in perfusate (1.03%). Immunoblots of liver homogenates revealed drug-modified proteins at ca. 110 and 120 kDa. The results show that (a) DAG undergoes avid systemic deconjugation-conjugation cycling and isomerization to iso-DAG; (b) iso-DAG is more resistant to hydrolysis, is readily taken up by hepatocytes and undergoes novel metabolism (phenolic glucuronidation); and (c) the glycation pathway (i.e. using iso-DAG as substrate) plays a major role in formation of covalent DF-protein adducts in liver.

Animals↗

PubMed1997

Bioactivation of phenytoin by human cytochrome P450: characterization of the mechanism and targets of covalent adduct formation.

The cytochrome P450-dependent covalent binding of radiolabel derived from phenytoin (DPH) and its phenol and catechol metabolites, 5-(4'-hydroxyphenyl)-5-phenylhydantoin (HPPH) and 5-(3',4'-dihydroxyphenyl)-5-phenylhydantoin (CAT), was examined in liver microsomes. Radiolabeled HPPH and CAT and unlabeled CAT were obtained from microsomal incubations and isolated by preparative HPLC. NADPH-dependent covalent binding was demonstrated in incubations of human liver microsomes with HPPH. When CAT was used as substrate, covalent adduct formation was independent of NADPH, was enhanced in the presence of systems generating reactive oxygen species, and was diminished under anaerobic conditions or in the presence of cytoprotective reducing agents. Fluorographic analysis showed that radiolabel derived from DPH and HPPH was selectively associated with proteins migrating with approximate relative molecular weights of 57-59 kDa and at the dye front (molecular weights < 23 kDa) on denaturing gels. Lower levels of radiolabel were distributed throughout the molecular weight range. In contrast, little selectivity was seen in covalent adducts formed from CAT. HPPH was shown to be a mechanism-based inactivator of P450, supporting the contention that a cytochrome P450 is one target of covalent binding. These results suggest that covalent binding of radiolabel derived from DPH in rat and human liver microsomes occurs via initial P450-dependent catechol formation followed by spontaneous oxidation to quinone and semiquinone derivatives that ultimately react with microsomal protein. Targets for covalent binding may include P450s, though the catechol appears to be sufficiently stable to migrate out of the P450 active site to form adducts with other proteins. In conclusion, we have demonstrated that DPH can be bioactivated in human liver to metabolites capable of covalently binding to proteins. The relationship of adduct formation to DPH-induced hypersensitivity reactions remains to be clarified.

Animals↗

PubMed1997

The mechanism of the carbamazepine-valproate interaction in humans.

AIMS: The study investigated the mechanism of the interaction between valproate and carbamazepine which causes raised plasma carbamazepine-10,11-epoxide concentrations with unchanged plasma carbamazepine concentrations. This interaction has usually been attributed to valproate inhibiting epoxide hydrolase, the enzyme that catalyses the biotransformation of carbamazepine-10,11-epoxide to carbamazepine-10,11-trans-diol. METHODS: Clearances of plasma carbamazepine, carbamazepine-epoxide and carbamazepine-diol to relevant carbamazepine metabolites present in urine were measured under steady-state conditions in 17 adults receiving carbamazepine as anticonvulsant monotherapy, and in 10 adults taking the drug together with valproate. RESULTS: Plasma carbamazepine-epoxide concentrations were higher, relative to carbamazepine dose, in the co-medicated patients. Plasma apparent clearances of carbamazepine, relative to drug dose, were similar whether or not valproate was taken. Formation clearances of carbamazepine-10,11-trans-diol conjugate, and probably of carbamazepine-10,11-trans-diol, were lower in subjects co-medicated with valproate, and a higher proportion of the carbamazepine dose was excreted in urine as carbamazepine-10,11-epoxide. CONCLUSIONS: Valproate appears to inhibit the glucuronidation of carbamazepine-10,11-trans-diol, and probably also inhibits the conversion of carbamazepine-10,11-epoxide to this trans-diol derivative, rather than simply inhibiting the latter reaction only.

Adult↗

PubMed1997

Effects of pregnancy on various pathways of human antiepileptic drug metabolism.

Ratios of phenytoin and carbamazepine doses to steady-state plasma concentrations of the drugs (apparent clearances) increase in pregnant women. Mean phenytoin clearance to urinary unconjugated p-hydroxyphenytoin increased from 0.28 +/- SD 0.18 to 0.74 +/- SD 0.37 L/day in 13 pregnant women; mean clearance to p-hydroxyphenytoin glucuronide increased proportionately less (15.25 +/- SD 5.43 to 31.94 +/- SD 16.30 L/day), the proportion of the metabolite that was conjugated falling from 98.4 +/- SD 0.72% to 97.65 +/- SD 0.67%. Mean clearances to urinary phenytoin and phenytoin-dihydrodiol did not increase. In 10 epileptic women, mean clearances of carbamazepine to urinary (a) carbamazepine-10,11-epoxide (1.66 +/- SD 1.2 to 3.70 +/- SD 2.09 L/day), (b) unconjugated carbamazepine-10,11-trans-diol (33.93 +/- SD 10.21 to 47.01 +/- SD 19.58 L/day). (c) unconjugated carbamazepine-acridan (0.24 +/- SD 0.12 to 0.47 +/- SD 0.34 L/day), and (d) unconjugated 2-hydroxy-carbamazepine (0.08 +/- SD 0.09 to 0.66 +/- SD 1.14 L/day) all increased during pregnancy. Mean clearance to unconjugated 3-hydroxy-carbamazepine decreased (0.53 +/- SD 0.25 to 0.18 +/- SD 0.23 L/day). In contrast, mean clearances of carbamazepine to the glucuronides of its first stage metabolites (carbamazepine-diol, 2- and 3-hydroxy-carbamazepine and carbamazepine-acridan, respectively) did not increase in pregnancy. The conversion of carbamazepine to carbamazepine-epoxide increased proportionately more than the conversion of carbamazepine-epoxide to carbamazepine-diol. Pregnancy was thus associated with increased microsomal oxidations of phenytoin and carbamazepine, without proportionate increases in the subsequent hydrolysis of carbamazepine-10,11-epoxide and in the O-glucuronidations of the earlier stage metabolites.

Adult↗

PubMed1997

Anticonvulsant therapy in aged patients. Clinical pharmacokinetic considerations.

Alterations in drug disposition that occur with aging are now becoming widely recognised, and there is an increasing number of drugs for which the approach to therapy in elderly patients can be based on pharmacokinetic data. Both healthy aging and comorbid disease can alter the responsiveness of the body to drugs and to their absorption, distribution and elimination. Altered absorption in the elderly has not been documented after oral ingestion of any anticonvulsant drug. Increased adipose tissue in the elderly may raise the apparent volume of distribution (Vd) of lipid-soluble drugs. An increased Vd in the elderly has been shown for diazepam and clobazam, but not midazolam. The data are inconclusive for phenytoin and valproic acid (sodium valproate). The decreased plasma protein binding that often occurs in the elderly has few clinical consequences. The reduced liver function that to occur with aging seems to affect the elimination of drugs that are mainly cleared by oxidative metabolism [e.g. carbamazepine, phenytoin and phenobarbital (phenobarbitone)]. Reduced clearances for methylphenobarbital (methylphenobarbitone), diazepam, midazolam and clobazam occur in elderly men, but not in women. The reduced renal function that is seen in old age affects the disposition of drugs that are eliminated mainly by direct renal excretion. Thus. the clearances of vigabatrin and gabapentin correlate with creatinine clearance. Such considerations may help guide anticonvulsant dosage in the elderly.

Absorption↗

PubMed1996

Dose-dependent metabolism of carbamazepine in humans.

48-h steady-state metabolic balance studies were carried out in 17 adults receiving long-term anticonvulsant monotherapy. With increasing carbamazepine dosage (1) carbamazepine overall plasma apparent clearance (CL/F), (2) plasma clearance of carbamazepine to urinary carbamazepine-10,11-epoxide, (3) plasma clearance of carbamazepine-10,11-epoxide to urinary unconjugated carbamazepine-10,11-trans-diol and (4) plasma clearances of carbamazepine to urinary 2- and 3-hydroxy carbamazepine all increased. However, with increasing carbamazepine dose there was no increase in the clearance of carbamazepine to (5) its acridan derivative in urine or of (6) the diol, phenolic or acridan metabolites to their metabolically subsequent conjugates excreted in urine. These findings are consistent with ongoing dose-dependent autoinduction of carbamazepine metabolism along the first two stages, but not the final stage, of the epoxide-diol pathway and, to a lesser extent, along pathways yielding phenolic metabolites. However, conjugations of the various plasma phase I metabolites of carbamazepine are not dose-dependent. Plasma concentration ratios of substances involved in consecutive stages of the epoxide-diol pathway, as in previous published studies, suggested apparent dose dependence of the epoxide-->unconjugated diol stage only. Presumably, increased flux along the first two stages of the full epoxide-diol pathway reduces plasma carbamazepine and carbamazepine-10,11-epoxide concentrations largely in parallel, concealing the dose dependence of the conversion of carbamazepine to its epoxide.

Adult↗

PubMed1996

The utility of the bile-exteriorized rat as a source of reactive acyl glucuronides: studies with zomepirac.

Acyl glucuronide conjugates of acidic drugs are chemically unstable metabolites, able to undergo a number of reactions including covalent binding interactions with proteins. The question of whether any toxicological or immunological responses result from such covalent modification of native proteins in vivo is topical. Study of acyl glucuronide reactivity thus requires a convenient source of these metabolites. The utility of the bile-exteriorized rat for this purpose is highlighted herein using the formerly marketed nonsteroidal antiinflammatory agent zomepirac. Zomepirac was injected i.v. at 60 mg/kg four times into bile-exteriorized rats at 6-h intervals. The 24-h bile samples contained ca. 24% of zomepirac doses as zomepirac acyl glucuronide (ZAG). Purification was achieved by washing of the acidified bile with etherhexane, extraction into ethyl acetate, semipreparative HPLC, and crystallization. Overall recovery through the purification procedure was ca. 50%. Identity as ZAG was confirmed by mass spectrometry. The approach takes advantage of the robust glucuronidation capacity of the rat, especially at higher drug doses, and of its ability to preferentially excrete hepatically formed drug glucuronides into bile rather than into urine via blood. Prior to this work, ZAG was presumed to be only a minor metabolite of zomepirac in rats, based on early urinary recovery studies. Thus, measurement of urinary acyl glucuronide conjugates in the rat may severely underestimate their true formation in this species.

Animals↗

PubMed1996

Vesico-hepato-renal cycling of acidic drugs via their reactive acyl glucuronide metabolites? Studies with diflunisal in rats.

1. Deconjugation-reconjugation cycling of acidic drugs is known to occur in vivo via the hydrolysis of their reactive acyl glucuronide metabolites during their circulation in the blood (systemic cycling) or during their passage through the gut after biliary excretion (enterohepatic cycling). Whether such cycling occurs after renal excretion via hydrolysis in the urinary bladder followed by absorption of liberated drug (vesico-hepato-renal cycling) was investigated in rats using diflunisal (DF) and its acyl glucuronide (DFAG) as model compounds. 2. After administration of DF (1 mg/0.5 mL buffer, pH 7) into the bladder of anaesthetized bile-exteriorized rats, DF appeared rapidly in plasma, achieving peak concentrations of 7 micrograms/mL at 1 h. At 4 h, 30% of the dose was recovered as metabolites, mainly DFAG and DF phenolic glucuronide (DFPG) in bile, while 30% was recovered as unchanged DF from the bladder. 3. By contrast, after intravesical administration of an equimolar amount of DFAG at pH 7 or 5, DFAG itself was not detectable in plasma. Plasma concentrations of DF were barely detectable, with only approximately 1% of the administered dose recovered as metabolites in bile. 4. The data thus show that, although DF itself undergoes facile absorption from the urinary bladder of healthy rats, vesico-hepato-renal cycling of DF via DFAG appears to be of only minor quantitative importance.

Administration, Intravesical↗

PubMed1996

Effect of felbamate on valproic acid disposition in healthy volunteers: inhibition of beta-oxidation.

We assessed the effects of felbamate (FBM) on the disposition of valpr oic acid (VPA) in healthy volunteer men. Eighteen subjects received sodium VPA, 400 mg/day for 21 days. Plasma and urine samples were taken on day 7 to document the steady-state disposition of VPA alone. From day 8 to day 21, subjects received placebo or FBM at the following doses (mg/day): 1,200, 2,400, 3,000, or 3,600 (n = 2-4 per group). Many adverse events (AE) occurred from about day 10; 2 subjects dropped out and 1 continued on a reduced FBM dose. Pharmacokinetic studies were repeated on day 21 for the 16 subjects who completed the study. FBM was measured in plasma and urine by high-performance liquid chromatography (HPLC). VPA and its 2-en, 4-en, and 3-oxo metabolites in plasma, and VPA (nonconjugated and total), and its 3-oxo and 4-hydroxy metabolites in urine were measured by gas chromatography/mass spectrometry (GC/MS). Mean plasma FBM trough concentrations on day 21 ranged from 26.9 mu g/ml (1,200 mg dose) to 76.8 mu g/ml (3,600-mg dose). Mean plasma VPA C max values were 32-42 mu g/ml in the various subgroups when VPA only was administered. Higher plasma VPA levels were observed when FBM was administered concurrently (55.4-63.8 mu g/ml). The excretion of 3-oxo-VPA in urine was significantly lower on day 21 than on day 7, whereas VPA-glucuronide was significantly increased. The effects of FBM on VPA disposition were dose dependent and were maximal at approximately 2400 mg/day. FBM has caused significant inhibition of the beta-oxidation pathway for VPA metabolic clearance, and this had been largely compensated by increased VPA glucuronidation.

Adult↗

PubMed1995

Studies on the reactivity of acyl glucuronides--VIII. Generation of an antiserum for the detection of diflunisal-modified proteins in diflunisal-dosed rats.

Acyl glucuronide metabolites of carboxylic drugs such as the salicylate derivative diflunisal (DF) have been shown to react with proteins to produce covalent adducts. To aid in the study of the formation and distribution of these adducts in both humans and rats, we raised an antiserum against human serum albumin modified by covalent attachment of DF via an amide bond, using a carbodiimide reagent. This antiserum had wide reactivity, reacting with all types of DF-modified proteins tested and with free DF (albeit at a lower affinity). It did not cross-react with other salicylates or other non-steroidal anti-inflammatory drugs. The antiserum has been used in immunoblotting to detect proteins covalently modified by DF in the plasma and livers of rats treated with the drug for 7 days. Although some cross-reactivity was apparent on the blots, a series of DF-modified proteins was found in cytosolic, mitochondrial and mixed membrane fractions of hepatocytes, with molecular weights ranging from 28 to 130 kDa.

Animals↗

PubMed1995

Rat serum albumin modified by diflunisal acyl glucuronide is immunogenic in rats.

Acyl glucuronide metabolites of carboxylic acid drugs such as the salicylate derivative diflunisal (DF) have been shown to react with proteins in vitro and in vivo to produce covalent adducts. Such attachment of foreign compounds to endogenous molecules could be associated with toxic and/or immune consequences. In this study we have injected rats with rat serum albumin (RSA) modified (a) by DF using a carbodiimide reagent (-->DF-RSA-I, 4.9 micrograms DF/mg RSA) and (b) by incubation with DF acyl glucuronide (DAG) and its rearrangement isomers (iso-DAG) (-->DF-RSA-II, 0.34 micrograms DF/mg RSA). All of the six rats injected with DF-RSA-I produced antibodies reactive with DF-modified keyhole limpet hemocyanin (KLH), the coating protein used in the ELISA. Three out of six animals injected with DF-RSA-II generated similar antibodies. Cross-reactivity with other non-steroidal anti-inflammatory drugs (NSAIDs) such as naproxen and ketoprofen (as the free drugs) was not observed. This study shows that a self protein covalently modified by incubation with DAG and iso-DAG is immunogenic in rats. The data thus support the hypothesis that covalent modification of macromolecules by acyl glucuronide metabolites of acidic drugs in vivo can lead to the production of circulating antibodies which may be involved in aberrant immune responses such as drug hypersensitivity.

Animals↗

PubMed1995

Metabolism of carbamazepine and co-administered anticonvulsants during pregnancy.

Urinary excretions of carbamazepine, carbamazepine-10,11-epoxide, carbamazepine-10,11-trans-diol, 9-hydroxyacridan and 2- and 3-hydroxycarbamazepine were measured at various stages of pregnancy, and in the post-natal period, in ten epileptic women, six of whom took no other enzyme-inducing anticonvulsant and four of whom took such co-medication. Mean plasma carbamazepine apparent clearance was increased in pregnancy, but only by virtue of the increased clearance in the anticonvulsant co-medicated women. Alterations in the proportions of the carbamazepine dose cleared via the various excretion pathways studied were quantitatively minor, but there was evidence consistent with impaired conversion of carbamazepine-10,11-epoxide to carbamazepine-10,11-trans-diol during all pregnancies studied. Clearances of carbamazepine to the various excretory products studied were consistent with there being (i) increased urinary excretion of unmetabolised drug in pregnancy, possibly related to the increased glomerular filtration rate, (ii) increased formation of oxidative metabolites of the drug, particularly in women co-medicated with enzyme-inducing anticonvulsants, this effect being offset, in full (in non-co-medicated women) or in part (in co-medicated women) by (iii) inhibition of the epoxide-diol pathway in pregnancy, an inhibition to which folate intake may have contributed.

Adult↗

PubMed1995

Characterization and formation of the glutathione conjugate of clofibric acid.

The incubation of 1-O-clofibryl glucuronide (1-O-CAG), a metabolite of clofibrate, with glutathione (GSH) resulted in the appearance of a new peak when analyzed by HPLC. The use of HPLC coupled to electrospray-MS permitted the identification of the peak as S-(p-chlorophenoxy-2-methylpropanoyl)glutathione (CA-SG), formed by nucleophilic displacement of the glucuronide moiety from 1-O-CAG. Conjugate formation was enhanced 8-fold by rat liver glutathione S-transferases (GSTs). GSH was unreactive with isomers of 1-O-CAG formed by acyl migration, indicating that 1-O-CAG itself was the preferred substrate. Rearrangement of 1-O-CAG to its isomers in vitro, was found to be decreased in the presence of GSH. In vivo studies indicated that, following an intravenous infusion of clofibric acid to rats (75 mg/kg), the concentration of CA-SG excreted in bile over 4 hr, was approximately 0.1% of the concurrent CAG concentrations. Although these results indicate a minor role for GST-catalyzed reactions in clofibrate metabolism in vivo, they do define 1-O-acyl-linked glucuronides as a new class of substrates for GSTs.

Animals↗

PubMed1994

Studies on the reactivity of acyl glucuronides--VI. Modulation of reversible and covalent interaction of diflunisal acyl glucuronide and its isomers with human plasma protein in vitro.

Acyl glucuronide conjugates are chemically reactive metabolites which can undergo hydrolysis, rearrangement (isomerization via acyl migration) and covalent binding reactions with protein. The present study was undertaken to identify factors modulating the reactivity of diflunisal acyl glucuronide (DAG) with human serum albumin (HSA) in vitro, by comprehensively evaluating the interplay of the three pathways above when DAG and a mixture of its 2-, 3- and 4-isomers (iso-DAG) were incubated with protein. Buffer, plasma, fraction V HSA, fatty acid-free HSA, globulin-free HSA and fatty acid- and globulin-free HSA were investigated at pH 7.4 and 37 degrees, each in the absence and presence of warfarin, diazepam and diflunisal (DF) as reversible binding competitors. DAG and iso-DAG were highly reversibly bound (ca. 98-99.5%) in plasma and HSA solutions. The binding was primarily at the benzodiazepine site, since displacement occurred in the presence of diazepam and fatty acids but not warfarin. DAG degradation, via rearrangement, hydrolysis and covalent adduct formation (in that order of quantitative importance), was retarded in plasma and HSA solutions compared to buffer. The protective effect of protein was afforded by the high reversible binding to the (non-catalytic) benzodiazepine site. The warfarin site appeared to be catalytic for DAG hydrolysis, whereas rearrangement appeared to be hydroxide ion-catalysed only. In contrast to DAG, iso-DAG degradation was greatly accelerated in the presence of protein, through both covalent binding and catalysis of hydrolysis. Covalent binding via DAG was increased in the presence of warfarin but decreased in the presence of diazepam, DF and fatty acids. The opposite effects were found for covalent binding via iso-DAG. The data suggest that covalent binding of DF to HSA via DAG and iso-DAG occurs by different mechanisms (presumably transacylation and glycation, respectively) at different sites (benzodiazepine and warfarin, respectively) whereas reversible binding occurs primarily at the same site (benzodiazepine).

Diflunisal↗

PubMed1994

Studies on the reactivity of acyl glucuronides--VII. Salicyl acyl glucuronide reactivity in vitro and covalent binding of salicylic acid to plasma protein of humans taking aspirin.

Salicyl acyl glucuronide (SAG) is a significant metabolite of salicylic acid (SA) and aspirin. We have shown that, under physiological conditions in vitro, SAG undergoes rearrangement in a manner consistent with acyl migration to its 2-, 3- and 4-O-acyl positional isomers as the predominant pathway (T1/2 values were 1.4-1.7 hr in buffer at pH 7.4 and 37 degrees). Incubation of SAG or a mixture of its rearrangement isomers (iso-SAG) (each at approximately 50 micrograms SA equivalents/mL) with human serum albumin (HSA, at approximately 40 mg/mL) revealed the formation of covalent adducts with the protein, with peak concentrations of 1-2 micrograms SA equivalents/mL. The data support a role for the rearrangement/glycation mechanism of adduct formation. Covalent adducts of SA were also detected in the plasma of humans taking aspirin (at > or = 1200 mg/day), but the concentrations were low (<< 100 ng SA equivalents/mL). Reactivity of SAG thus provides a mechanism (though of uncertain quantitative importance) of covalent attachment of the salicyl moiety of aspirin to tissue macromolecules, which is in addition to its well-known acetylating capacity.

Animals↗

PubMed1994

Early stage autoinduction of carbamazepine metabolism in humans.

Six healthy young adult male volunteers were given two 600 mg (2540 mu moles) oral doses of carbamazepine (CBZ) 5 days apart. Serial concentrations of CBZ and its 10,11-epoxy (CBZ-epoxide) and 10,11-dihydro-10,11-trans-dihydroxy (CBZ-diol) metabolites in plasma, and daily excretions of these substances and the 2-hydroxy (2-OH-CBZ), 3-hydroxy (3-OH-CBZ) and 9-hydroxymethyl-10-carbamoylacridan (acridan) metabolites in urine were followed for 5 days after each dose. Pharmacokinetic analysis showed that autoinduction of CBZ metabolism was present within 6-10 days of the initial drug dose. The mean oral clearance of CBZ increased from 1.48 to 1.74 l.h-1 (difference 0.26 l.h-1, 95% confidence interval 0.11 to 0.41 l.h-1) and the mean percentage urinary recovery of the amount of CBZ eliminated increased from 41.8% to 44.6% (difference 2.8%, 95% confidence interval 0.5 to 5%) between the two studies 5 days apart. The data for daily clearance to metabolite and the time-courses of the plasma CBZ-epoxide to CBZ and CBZ-diol to CBZ concentration ratios suggested that autoinduction had begun by the second day after CBZ intake, and involved not only the epoxide-diol pathway but, to a lesser extent, the oxidations to phenolic derivatives.

Acridines↗

PubMed1994

Urinary excretion of phenobarbitone and its metabolites in chronically treated patients.

The elimination of phenobarbitone (PB) was studied in 14 chronically treated epileptic patients under steady state conditions. PB, [S]-PB-N-glucoside ([S]-PB-N-G) and p-hydroxy-PB (p-OH-PB) were assayed in urine by a HPLC method. Some 57% of the daily dose was recovered in urine, 14% as [S]-PB-N-G, 16% as p-OH-PB (conjugated plus non-conjugated) and 27% as unaltered PB. Thus PB-N-G formation contributed significantly to the elimination of PB during long-term administration of the drug, and there was reason to suspect that some of the PB-N-G formed may have already been degraded to untraced products before excretion from the body.

Adult↗