Formation Of Active Metabolite Research Articles

New insights into the pharmacokinetics and metabolism of (R,S)-ifosfamide in cancer patients using a population pharmacokinetic-metabolism model.

To describe the pharmacokinetics of R- and S-Ifosfamide (IFF), and their respective 2 and 3 N-dechloroethylated (DCE) metabolites (R2-, R3-, S2, S3-DCE-IFF) in cancer patients. (R,S)-IFF was administered (1.5 g/m2) daily for 5 days in 13 cancer patients. Plasma and urine samples were collected and analyzed using an enantioselective GC-MS method. An average of 97 observations per patient were simultaneously fitted using a pharmacokinetic-metabolism (PK-MB) model. A population PK analysis was performed using an iterative 2-stage method (IT2S). Auto-induction of IFF metabolism was observed over the 5 day period. Increases were seen in IFF clearance (R: 4 vs. 7 L/h; S: 5 vs. 10 L/h), and in the formation of DCE (R: 7 vs. 9%; S: 14 vs. 19%) and active metabolites (4-OHM-IFF; R: 71 vs. 77%; S: 67 vs. 71%). A novel finding of this analysis was that the renal excretion of the DCE metabolites was also induced. This population PK-MB model for (R,S)-IFF may be useful in the optimization of patient care, and gives new insight into the metabolism of (R,S)-IFF.

Antinociceptive effects of the neuroactive steroid, 3α-hydroxy-5α-pregnan-20-one and progesterone in the land snail, Cepaea nemoralis

Results of investigations with vertebrates have implicated neuroactive steroids and in particular 5α-reduced metabolites of progesterone such as 3α-hydroxy-5α-pregnan-20-one (3α,5α-THP/3A5P and originally allopregnanolone) in the rapid modulation of diverse functions including that of nociceptive sensitivity. These effects have been indicated to involve modulation of GABA receptors. Results of recent phylogenetic studies have revealed the presence of GABA receptors in invertebrates that may also be subject to modulation by steroids and neuroactive steroids. The present study examined the effects of the neuroactive steroid, 3α-hydroxy-5α-pregnan-20-one, as well as progesterone on aversive thermal (nociceptive) responses in a mollusc, the land snail, Cepaea nemoralis. 3α-Hydroxy-5α-pregnan-20-one had significant dose-related (0.01–1.0 μg) antinociceptive effects in Cepaea increasing the latency of response to a 40°C surface, with maximum effects being evident 15–30 min after administration. These effects of 3α-hydroxy-5α-pregnan-20-one were stereospecific, with the stereoisomer 3β-hydroxy-5α-pregnan-20-one (3B5P) failing to affect nociceptive responses. Progesterone also had significant dose-related (0.10–10 μg) antinociceptive effects that, however, were delayed in onset and relatively prolonged (60–120 min), suggestive of the formation of active metabolites. The presence of endogenous progesterone (12.36±0.17 ng/g tissue) was ascertained by a radioimmunoassay further supporting a functional role for steroids in Cepaea. The antinociceptive effects of 3α-hydroxy-5α-pregnan-20-one and progesterone were blocked by the GABA antagonists, bicuculline and picrotoxin, while being relatively insensitive to opioid and N-methyl- d-aspartate antagonists. These results suggest an early evolutionary development and phylogenetic continuity of neuroactive steroid and GABA involvement in the mediation of nociception.

Modulation of Drug‐Metabolising Enzyme Expression by Condensation Products of Indole‐3‐ylcarbinol, an Inducer in Cruciferous Vegetables

Indole-3-ylcarbinol (13C) is formed during processing of cruciferous vegetables and is suggested to be one of the modulators of drug-metabolising enzymes. Indole-3-ylcarbinol is a far less efficient inducer of hepatic enzymes after parenteral than after oral administration, due to formation of active metabolites in the gastrointestinal tract. As indole-3-ylcarbinol is unstable in weakly acidic aqueous solutions, non-active condensation products may be formed from indole-3-ylcarbinol, that cannot be transformed to the active products when reaching the stomach. The purpose of the present study was to test the ability of the condensation products formed at a pH corresponding to that of fresh vegetable juice to modulate the metabolism of xenobiotics. Indole-3-ylcarbinol was incubated in vitro at room temperature in the dark at pH 5.5 and samples taken at various times, for oral administration to rats and for chemical analysis. Indole-3-ylcarbinol was rapidly transformed into various oligomeric products. The 7-ethoxyresorufin O-deethylase activities (marker of cytochrome Cytochrome P450 1A enzymes, CYP1A) in liver, kidney and colon increased with the duration of the in vitro condensation period whereas the formation of 6beta-, 15beta- and and 2alpha-hydroxytestosterone was not affected significantly, indicating no effect on CYP2C11 or CYP3A enzymes. The hepatic metabolism of the food mutagen 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP). was increased by indole-3-ylcarbinol condensation products and the 4'-OH-PhIP/N-OH-PhIP ratio was decreased due to a significantly increased formation of the proximate genotoxic metabolite. N-OH-PhIP. The activities of DT-diaphorase and glutathione S-transferase were not changed significantly in the rat organs. These experiments clearly indicate that indole-3-ylcarbinol is not the definitive CYP1A inducer and that indole-3-ylcarbinol at near-neutral pH, is transformed to compounds that are inducers by themselves or may be further converted into inducing compounds in the rat stomach. Also, the enzyme inducing potency of indole-3-ylcarbinol containing vegetable juice is apparently enhanced by incubation in vitro before the intake.

Metabolism of tricyclic antidepressants.

1. Despite the considerable advances in the treatments available for mood disorders over the past generation, tricyclic antidepressants (TCAs) remain an important option for the pharmacotherapy of depression. 2. The pharmacokinetics of TCAs are characterized by substantial presystemic first-pass metabolism, a large volume of distribution, extensive protein binding, and an elimination half-life averaging about 1 day (up to 3 days for protriptyline). 3. Clearance of tricyclics is dependent primarily on hepatic cytochrome P450 (CYP) oxidative enzymes. Although the activities of some P450 isoenzymes are largely under genetic control, they may be influenced by external factors, such as the concomitant use of other medications or substances. Patient variables, such as ethnicity and age, also affect TCA metabolism. The impact of gender and related reproductive issues is coming under increased scrutiny. 4. Metabolism of TCAs, especially their hydroxylation, results in the formation of active metabolites, which contribute to both the therapeutic and the adverse effects of these compounds. 5. Renal clearance of the polar metabolites of TCAs is reduced by normal aging, accounting for much of the increased risk of toxicity in older patients. 6. Knowledge of factors affecting the metabolism of TCAs can further the development and understanding of newer antidepressant medications.

The efficiency concept in pharmacodynamics.

The classic approach to describe the pharmacological response to a drug is to analyse its concentration-effect relationship, using a variety of possible models such as maximum effect (Emax) models or sigmoid Emax models. The aim of this review is to discuss an alternative way of describing the pharmacological effect in terms of effect per unit of drug concentration, instead of simple effect. This variable is called efficiency, analogous with concepts used in other fields. The pharmacodynamic model for efficiency is derived from the sigmoid Emax model and is dependent on the same parameters. Since the sigmoid Emax model incorporates 'the law of diminishing returns', requiring ever higher concentrations to increase the effect by a given percentage, efficiency is bound to decrease with increasing concentrations. However, as a mathematical consequence of its derivation from the sigmoid Emax model, efficiency also has a maximum value, which can be expressed as a function of the slope factor (s) and drug concentration associated with half the maximum effect (C50), provided that the slope factor is greater than 1. The efficiency concept is potentially applicable to all drugs and particularly useful for those that follow concentration-effect relationships according to Emax or sigmoid Emax models. Most experience has been obtained with loop diuretics, especially with furosemide (frusemide). Slow administration of furosemide, leading to slow excretion of the drug, has been shown, in many studies, to significantly increase the total diuretic effect per amount of drug recovered in urine. In this review, some examples of the applicability of the efficiency concept to other drugs, such as antibacterials, opioids and antineoplastics, are discussed. In addition to pharmacodynamically varying efficiency, other saturable processes, such as the formation of active metabolites and saturable transport, may form a basis for the application of the efficiency concept. The efficiency of a drug dosage may also be influenced by tolerance and counter-regulation produced by the drug. All these factors contribute to schedule dependency. It is concluded that the shape of the time course of drug presentation to its site of action is an independent determinant of overall response. The possibility of adjusting the drug input profile to maximise therapeutic effect per dose and to separate cumulated therapeutic from cumulated adverse effects should be considered in designing administration schedules and in drug development.

Pharmacokinetics of (R)- and (S)-cyclophosphamide and their dechloroethylated metabolites in cancer patients.

The complete pharmacokinetics (PK) of (R)- and (S)-cyclophosphamide (CP) and their dechloroethylated (DCE) metabolites have not been reported to date. We collected plasma and urine samples from 12 cancer patients and determined concentrations of both enantiomers of CP and DCE-CP using a chiral GC-MS method. All concentrations of (R)-CP, (S)-CP, (R)-DCE-CP, and (S)-DCE-CP were simultaneously modeled using an enantiospecific compartmental PK model. A population PK analysis was performed. Enantiospecific differences between (R)- and (S)-CP were found for the formation clearance of CP to the DCE metabolites (Clf: 0.25 (R) vs. 0.14 (S) L/h). No difference was found between enantiomers for Cl40H, Cld, Cl(m)R, ClT, or T1/2. In contrast to the adolescent and adult group of patients, a child (6 years old) appeared to have a very different PK and metabolic profile (Bayesian control analysis). Proportions of the (R,S)-CP doses transformed to the (R)-DCE- and (S)-DCE-CP were much higher (R: 25 vs. 1.9%, and S: 38 vs. 3.6%), while formation of active metabolites was much lower (R: 42 vs. 74%, and S: 48 vs. 77%). CP appears to be enantioselectively metabolized to the DCE metabolites. This PK model can evaluate the proportion of a CP dose that is transformed to toxic or active metabolites. It may therefore be used to optimize CP treatment, to identify important drug interactions and/or patients with an abnormal metabolic profile.

Formation and clearance of active and inactive metabolites of opiates in humans.

The results of recent investigations of the analgesic and the nonanalgesic effects of opioid glucuronides are relevant to the research on drug abuse in forensic toxicology. As has been shown for heroin, knowledge of the state of distribution and elimination of active and inactive metabolites and glucuronides offers new possibilities in forensic interpretation of analytic results. Because of similar metabolic degradation, calculation of the time-dependent ratio of the concentration of morphine and its glucuronide metabolites in blood or serum allows a rough estimation of increased dosage and of time elapsed since the last application. Drug effects can be examined with respect to individual case histories, including overdose and survival time if the patient died. However, different methods of administration and the strong influence of different volumes or compartments of distribution of parent compounds and metabolites on concentrations in human body tissues require careful use of glucuronide concentration data. In Germany, dihydrocodeine (DHC) is prescribed as a heroin substitute, and relative overdoses are needed to be effective. DHC metabolism was studied in three patients who died from overdoses. All metabolites (dihydrocodeine-6-glucuronide [DHC6], nor-DHC [NDHC], dihydromorphine [DHM], nor-DHM [NDHM], and DHM-3- and 6-glucuronide [DHM3G, DHM6G]) were determined using HPLC and fluorescence detection. Concentrations of DHM (0.16 mg/L to 0.22 mg/L serum) were found. The DHM glucuronide ratios were similar to those of morphine. Receptor binding studies showed that the binding affinity of DHM to porcine mu-receptor was higher than that of morphine, and DHM6G's binding affinity was as high as that of morphine-6-glucuronide (M6G). Metabolites may play an important role in the effectiveness of DHC in substitution and toxicity. Because of enzyme polymorphism, the formation of DHC poses a risk for proper dosage in patients who are either poor or extensive metabolizers. The distribution of opioid glucuronides in cerebral spinal fluid in relation to transcellular transport in central nervous tissue is discussed with respect to the receptor binding of opiates and drug effect.

Opioid Drug-Drug Interactions: A Review

Opioid analgesics are among the most commonly prescribed medications. Frequently, they are combined with other therapeutic agents and pharmacodynamic or pharmacokinetic interactions may ensue. This review summarizes published case reports and studies of potential opioid drug interactions. A MED-LINE computer literature search (1966-1998) was undertaken to retrieve all pertinent case reports and studies of opioid drug interactions published in the English language. The results of the search indicate that numerous compounds from various therapeutic classes may participate in clinically significant pharmacodynamic and pharmacokinetic drug-drug interactions. Pharmacodynamic interactions usually involved additive central nervous system depression. Additionally, propoxyphene and tramadol can potentiate a hyperserotonergic state when coadministered with the SSRIs and MAOIs. Pharmacokinetic interactions typically involved inhibition or induction by specific hepatic cytochrome P-450 isoenzymes. Agents with enzyme inhibiting ability such as erythromycin, cimetidine, and selective serotonin reuptake inhibitors have been shown to potentiate the effects of certain opioid analgesics while codeine, which requires metabolic conversion via CYP 2D6 for pharmacological effectiveness, has reduced analgesic efficacy in the presence of inhibitors. The enzyme inducers rifampin and several anticonvulsants have been involved in the emergence of methadone withdrawal when added to existing methadone treatment. Additionally, enzyme inducers can increase the formation of the toxic metabolite of meperidine. Genetic polymorphism also potentially impacts the effectiveness of agents such as codeine since reduced active metabolite formation and analgesic efficacy has been demonstrated in individuals who lack CYP 2D6 activity.

Functional role of estrogen metabolism in target cells: review and perspectives.

Cytochrome P450 enzymes that metabolize estrogens are expressed in the mammary gland, uterus, brain and other target tissues for estrogen action, and this results in the formation of hydroxylated estrogens in these tissues. Estradiol metabolites formed in target tissues at or near estrogen receptors may either be inactive or have important biological effects, and changes in the activities of estrogen-metabolizing enzymes in target tissues may profoundly influence estrogen action. Although some active estrogen metabolites exert hormonal effects in target tissues by interaction with the classical estrogen receptor, other metabolites appear to elicit unique biological responses that are not associated with activation of this receptor. Therefore, some of the many actions of estradiol may not be caused by estradiol per se, but may result from the formation of active estrogen metabolite(s) which function as local mediators or may activate their own unique receptors or effectors. This is an important area in need of more research. The present paper represents a review of the literature and perspectives by the authors on the functional role of estrogen metabolism in target tissues.

The role of gemcitabine in the treatment of small cell lung cancer

Gemcitabine, a new anticancer agent with novel metabolic properties and mechanisms of action, is a nucleoside analogue, closely related to cytosine arabinoside. However, gemcitabine has demonstrated much greater activity against a variety of solid tumors than cytosine arabinoside, perhaps due to higher rates of accumulation of the active drug within cells, and a prolonged biphasic elimination leading to longer intracellular exposure to the drug. Gemcitabine is a pro-drug which must be metabolized intracellularly to its active diphosphate and triphosphate forms by the enzyme deoxycytidine kinase, the rate-limiting enzyme for active metabolite formation. Incorporation of gemcitabine triphosphate into DNA is the mechanism by which gemcitabine causes cell death. Following the gemcitabine nucleotide incorporation, one further deoxynucleotide is incorporated, but thereafter, the DNA polymerases are unable to proceed. Furthermore, proof-reading enzymes are unable to remove gemcitabine from this penultimate position which ultimately results in cell death. Gemcitabine has been investigated extensively in the treatment of advanced non-small cell lung cancer, and singleagent response rates greater than 20% have been reported in almost all studies. Experience with gemcitabine in the treatment of small cell lung cancer is much more limited. Only one phase II single-agent trial has been undertaken to date in chemotherapy-naive patients with small cell lung cancer. The results of this National Cancer Institute of Canada study are summarized in the accompanying table.

Subcutaneous versus Intravenous Epoetin in Patients with Renal Failure

The purpose of this overview was to compare the bioavailability of epoetin (recombinant human erythropoietin) with its activity after subcutaneous injection. Weighted means for pharmacokinetic parameters of epoetin were calculated from 15 studies with a total of 160 human subjects. After intravenous injection, total body clearance was 0.13 ml/min/kg, and terminal half-life 6.7 hours. After subcutaneous injection, maximum serum concentrations were found after 16.7 hours; the terminal half-life of 22.7 hours was determined by the rate of absorption from the subcutaneous depot. In patients with renal failure, the subcutaneous maintenance doses were 61% of the equipotent intravenous doses. The bioavailability after subcutaneous injection was 33.6%. The average steady-state serum concentrations of epoetin during intravenous therapy were 4.9 times higher than during subcutaneous treatment with equipotent doses. This discrepancy may partly be explained by the high initial concentrations after an injection that contribute little to the therapeutic effect, and partly by an underestimation of the bioavailability by the formation of active metabolites with low affinity for the antibody used in the radioimmunoassay. We conclude that the bioavailability of epoetin given via different routes of administration does not permit the estimation of equipotent doses.

Decreased levels of 2-amino-3-methylimidazo[4,5-f]quinoline-DNA adducts in rats treated with beta-carotene, alpha-tocopherol and freeze-dried aloe.

To assess mechanisms of chemoprevention of hepatocarcinogenesis by trans‐β‐carotene (β‐C), DL‐α‐tocopherol (α‐T), and freeze‐dried whole leaves of Kidachi aloe (Aloe), formation of 2‐amino‐3‐methylimidazo[4,5‐f]quinoline (IQ)‐DNA adducts was measured by 32P‐post‐labeling analysis, and CYP1A1 and CYP1A2 protein levels were analyzed by ELISA. Group 1 rats were fed diet containing 0.02%β‐C, 1.5%α‐T or 30% Aloe over an 8‐day period, while group 2 was given basal diet alone. On day 7, all animals were subjected to two‐thirds partial hepatectomy (PH). Twelve hours after PH, they received a single dose of the carcinogenic food pyrolysate IQ (100 mg/kg) intragastrically, to initiate hepatocarcinogenesis. Rats were killed 6, 12, 24 and 48 h after IQ administration. The levels of adducts, expressed as relative adduct labeling values in rats treated with β‐C, α‐T and Aloe, were decreased as compared with the control group at hour 24 (36 h after PH), with a significant difference in the case of the β‐C group (46.4% of the control value). Similarly, all showed a tendency for decrease at hour 48. Furthermore, the levels of CYP1A2, known to be responsible for activation of IQ, showed a significant reduction at hour 24. It is concluded that β‐C, and possibly also α‐T and Aloe, have the potential to reduce IQ‐DNA adduct formation, presumably as a result of decreased formation of active metabolites. The results may explain, at least in part, the previously observed inhibitory effects of these compounds on induction of preneoplastic hepatocellular lesions.

Pharmacokinetic-pharmacodynamic relationships for benzodiazepines.

This article reviews the literature on the plasma concentration-effect relationships for benzodiazepines, in humans and in experimental animals. Only literature that explicitly links pharmacokinetics to pharmacodynamics is included. The following questions are evaluated. Can concentration-effect relationships be demonstrated? If so, are these relations stable? Are the influences of specific factors such as age and disease on these relationships established? It is clear that, when studies are conducted and interpreted appropriately, relations can be found for a wide range of benzodiazepine effects. These include objective measures such as electroencephalography, semisubjective measures such as psychomotor performance, and subjective measures such as mood/sedation scales. A generally applicable model of the relationship which will allow prediction of effect is, however, not yet established. The relationship appears to be dependent on route and rate of administration, because of factors such as distributional delay, formation of active metabolites and, probably, acute tolerance. Furthermore, intra- and interindividual variability is considerable, probably due to varying experimental conditions and intrinsic interindividual differences. The limited data available on factors influencing the plasma concentration-effect relationships for benzodiazepines demonstrate clear changes in the pharmacodynamics after multiple doses, suggesting the development of tolerance, and a subsensitivity in patients with panic disorder. The influence of factors such as age, disease and drug interactions on the pharmacokinetic-pharmacodynamic relationship remains less clear.

Stimulation of microsomal spironolactone metabolism by reduced glutathione.

The first step in the conversion of spironolactone (SP) to its biologically active metabolites is deacetylation to 7 alpha-thiospirolactone (7 alpha-thio-SL). Studies were done to evaluate the effects of reduced glutathione (GSH) on SP deacetylation by adrenal microsomal preparations. In the absence of GSH, adrenal microsomes catalyzed the conversion of SP to 7 alpha-thio-SL at low rates. Addition of GSH to the incubation medium caused a concentration-dependent stimulation of SP deacetylation. At a concentration of 10 mM, GSH caused a 4- to 5-fold increase in the rate of 7 alpha-thio-SL production. The results suggest that GSH may have an important role in the overall disposition of SP, including the formation of active metabolites.

5β‐Reductase and other Androgen‐Metabolizing Enzymes in Primary Cultures of Developing Zebra Finch Telencephalon

Enzymes in the avian brain irreversibly catalyze the conversion of androgens into either active metabolites (aromatase and 5 alpha-reductase) or inactive metabolites (5 beta-reductase), 5 beta-reductase is thought to influence the formation of active metabolites by reducing the concentration of androgenic substrate available for these reactions. However, because these enzymes have different regional, cellular and subcellular distributions in brain, the traditional method to measure enzyme activity in brain homogenates may be inaccurate because of artificial mixing of enzymes. Recently, we have prepared primary cell cultures from the telencephalons of developing zebra finches. Cell cultures offer the advantage that enzyme activity can be measured in live cells in which the relative distribution of enzymes may more closely reflect that found in vivo. We have previously reported that aromatase is expressed at high levels in these cultures, and that it is active in both neurons and in glia. Here we report on the activities of 5 alpha- and 5 beta-reductase in these cell cultures. Along with aromatase, 5 beta-reductase was expressed at high levels in these mixed cell cultures, including cultures highly enriched in glia. This suggests that 5 beta-reductase is present in non-neuronal cells in brain, possibly co-localized with aromatase. However, despite the presence of 5 beta-reductase, aromatase was detected reliably in vitro even when the concentration of substrate was low. Thus, 5 beta-reductase does not prevent the synthesis of estrogen in the telencephalon of developing zebra finches. By contrast, 5 alpha-reductase activity was very low or absent in these cultures.(ABSTRACT TRUNCATED AT 250 WORDS)

Pharmacokinetic and pharmacodynamic studies of centrally acting drugs in rat: effect of pentobarbital and chlorpromazine on electroencephalogram in rat.

Electroencephalogram (EEG) alterations in rat after the i.v. administration of pentobarbital (PTB) and chlorpromazine (CPZ) were measured by power spectral analysis. The time courses of PTB concentrations in plasma, cerebrospinal fluid (CSF) and brain were determined after the i.v. administration of PTB (20, 40 mg/kg) by GC-MS. The PTB concentrations in plasma, CSF and brain could be described by a biexponential equation, a CSF model and a blood flow limited model, respectively. The relationship between the alteration of EEG and the PTB concentrations in the CSF or brain or the effect compartment were analyzed using the sigmoid Emax model. The alteration of EEG after PTB administration could be described by the PTB concentration in these compartments using the sigmoid Emax model. These results indicated that the site of action for the alteration of EEG after PTB administration is in instantaneous equilibrium with the CSF, the brain and the effect compartment. Thus, alterations in EEG after PTB administration can be predicted by monitoring the total PTB concentration in plasma. The alteration of EEG after i.v. administration of CPZ (4 mg/kg) showed a two-phase variation. Although the relationship between the alteration of EEG and the CPZ concentrations in CSF or the striatum or the effect compartment (total and free drug) were analyzed using the linear model, the Emax model or the sigmoid Emax model, the two-phase alteration of EEG after CPZ administration could not be described by any of these models. These results indicated that the pharmacokinetic and pharmacodynamic modeling of CPZ during the alteration of EEG may be complicated due to several pharmacokinetic and pharmacodynamic factors, such as an alteration of the free fraction of CPZ in the striatum, the formation of active metabolites, and two different intrinsic effects of CPZ on the EEG (one in an increase and the other in a decrease of the brain's electrical activity.

Thiopurine methyltransferase activity in American white subjects and black subjects.

Thiopurine methyltransferase (TPMT) is a cytoplasmic enzyme that preferentially catalyzes the S-methylation of aromatic and heterocyclic sulfhydryl compounds, including 6-mercaptopurine. TPMT exhibits genetic polymorphism in white populations, with 89% of individuals having high TPMT activity, 11% having intermediate activity, and one in 300 having extremely low or absent activity. TPMT activity is inversely correlated with formation of active 6-mercaptopurine metabolites (thioguanine nucleotides), thereby influencing 6-mercaptopurine toxicity and efficacy. To investigate ethnic and gender differences in TPMT, we measured erythrocyte TPMT activity in 209 white healthy subjects and 196 black healthy subjects (202 women and 303 men). The black population had lower TPMT activity than the white population (median, 14.4 versus 16.8 units/ml packed erythrocytes; p < 0.001). Maximum likelihood estimation of TPMT activity distribution identified 91.9% and 93.9% with high activity and 7.7% and 6.1% with intermediate activity in the white and black groups, respectively. These data indicate that TPMT activity is similarly polymorphic in American black subjects and white subjects, although median TPMT activity is approximately 17% lower in black subjects.

CYP2D6 phenotype determines the metabolic conversion of hydrocodone to hydromorphone

The contribution of cytochrome P450 2D6 (CYP2D6) to the formation of hydrocodone's active metabolite, hydromorphone, was examined in vitro and in vivo. Human liver microsomes prepared from an individual homozygous for the D6-B mutation of the CYP2D6 gene catalyzed this reaction at a negligible rate. Urinary metabolic ratios of hydrocodone/hydromorphone were highly correlated with O-demethylation ratios for dextromethorphan, an established marker drug of CYP2D6 activity (rs = 0.85; n = 18). The kinetics of hydrocodone after a single oral dose and its partial metabolic clearance to hydromorphone were investigated in five extensive metabolizers of dextromethorphan, six poor metabolizers, and four extensive metabolizers after pretreatment with quinidine, a selective inhibitor of CYP2D6 activity. The mean values for partial metabolic clearance by O-demethylation in the three groups were 28.1 +/- 10.3, 3.4 +/- 2.4, and 5.0 +/- 3.6 ml/hr/kg, respectively. No statistically significant phenotypic differences in physiologic measures were observed. However, over the first hour after dosing, the extensive metabolizers reported more "good opiate effects" and fewer "bad opiate effects" than poor metabolizers and extensive metabolizers in whom CYP2D6 was inhibited by quinidine. These data establish the importance of CYP2D6 in the formation of hydromorphone from hydrocodone and suggest that the activity of this enzyme may limit the abuse liability of hydrocodone.

Synthesis of cyclophosphamide analogs from aminotrideoxy sugars

Cyclophosphamide la is a highly effective and extensively used agent for the treatment of human cancers’. The metabolism, pharmacokinetics, and mechanism of action of la have been the subject of several recent reviews2-5. The activation process of la involves initial hydroxylation by the hepatic cytochrome P450 system to produce one or both isomers ‘9’ of 4-hydroxycyclophosphamide lb. Subsequent formation of aldophosphamide 2 leads to acrolein 3 and phosphoramide mustard’ 4 generally believed to be the ultimate cytotoxic agent that cross-links interstrand DNA’-‘“. Detoxification involves enzymic reactions at C-4 with formation of 4-ketocyclophosphamide 5 or carboxyphosphamide 6 by aldehyde dehydrogenasemediated oxidation. Thus, both formation of active metabolites and detoxification require enzymic reactions at C-4 of the six-membered cyclophosphamide ring. Recent data suggest that acrolein (3), produced by the decomposition of aldophosphamide, is toxic to cultured tumor cells , ‘I but does not play a significant role in the anticancer activity of 1. Acrolein may be responsible for the cardiac and pulmonary toxicities of cyclophosphamide’2 and also for cystitis and renal damage’3,‘4. Since la functions as a prodrug, such pre-activated analogs of la as benzo-annelated cyclophosphamide’5 or 4-hydroxy-(lb), and 4-hydroperoxy-cyclophosphamide”j (lc) have been synthesized both for possible enhancement of activity and for an understanding of the pathways of cyclophosphamide activation and metabolism. By analogy with the well-known strategy in the design of potential anticancer drugs with increased selectivity, attachment of a cyclophosphamide ring system to “carrier” molecules has been reported. This includes steroids for hormonally dependent neoplasm”, amino acids or peptides’*, nucleosides’9-22, and amino

Metabolism and activation of pancreas specific nitrosamines by pancreatic ductal cells in culture.

Metabolism of 14C labeled N-nitrosobis(2-oxopropyl)amine (BOP), N-nitroso(2-hydroxypropyl)(2-oxopropyl)amine (HPOP) and N-nitrosobis(2-hydroxypropyl)amine (BHP) by pancreatic duct cells in culture involves the following two pathways: reduction or oxidation reactions at the beta-carbon which result in the inter-conversion of these nitrosamines and activation reactions which result in the decomposition of the nitrosamine, the evolution of 14CO2 and the labeling of macromolecules. Reduction of BOP to HPOP seems to contribute significantly to the metabolism of the former nitrosamine by pancreatic duct cells, however, redox reactions at the beta-carbon of HPOP or BHP are not extensive. In terms of DNA damage, all three nitrosamines yield methyl and hydroxypropyl adducts. As expected, HPOP and BHP yield higher levels of O6-hydroxypropylguanine than BOP, while the latter yields higher levels of O6-methylguanine. There is no correlation between the ability of these nitrosamines to alkylate duct cell DNA in vitro and their carcinogenic potency in vivo. Concentrations of DNA adducts induced by pancreas specific nitrosamines (PSNs) in cultured duct cells at concentrations comparable to those found in the pancreatic juice of animals treated with BOP, are almost an order of magnitude lower than those induced in the pancreas of such animals. Discrepancies between in vitro and in vivo formation of active metabolites and DNA adducts may be attributed to the decline of the cells' ability to activate PSNs during culturing. In the same vein, the ductal cell may not be the main source of active metabolites targeting its DNA in the animal model.