6 -hydroxy- 1 -tetrahydrocannabinol synthesis and biological activity.

6beta-Hydroxy-delta(1)-tetrahydrocannabinol, a metabolite of delta(1)-tetrahydrocannabinol has been synthesized from delta(6)-tetrahydrocannabinol. It shows high tetrahydrocannabinol-type activity in rhesus monkeys. The implications of this funding are discussed.

Stereochemical requirements for cannabinoid activity.

Several pairs of cannabinoid isomers were synthesized and tested for psychotropic activity in rhesus monkeys. Two regularities were observed: (a) In the absence of the other substituents, the equatorial stereochemistry of the substituent at C-1 determines activity. (b) Two groups of THC-type cannabinoids which differ only in that the chemical groupings in one of them at C-1, C-2 are situated at C-1, C-6 in the other (but retain their stereochemistry) have almost equivalent psychotropic activity.

Monitoring theophylline therapy using citric acid-stimulated saliva in infants and children with asthma.

Saliva stimulation is required for measurement of drugs in saliva. Chewing on a piece of paraffin, which is the method usually used for saliva stimulation, requires cooperation of the patient and, thus, is inapplicable in infants and young children. To assess the value of determining theophylline concentrations from noninvasively obtained saliva in this age group, we studied the theophylline plasma to saliva concentration ratio in citric acid-stimulated saliva. Theophylline concentration was measured in 137 simultaneously obtained paired specimens of plasma and saliva from 68 patients 2 1/2 months to 14 years of age treated with theophylline for asthma (dosage 20.8 +/- 5.2 mg/kg/d, mean +/- SD). Saliva secretion was stimulated by placing citric acid crystals on the tongue. A strong and highly significant correlation was observed between both determinations (r = .96; P less than .01). The plasma to saliva ratio was 1.78 +/- 0.22 (mean +/- SD), with theophylline concentrations between 3.1 and 32.1 micrograms/mL of plasma. The ratio of estimated to actual plasma theophylline concentrations was 1.02 +/- 0.12 (mean +/- SD). Interindividual coefficient of variation of plasma to saliva theophylline concentrations ratios was 12.4%; mean intraindividual coefficient of variation was 5.3%. The use of citric acid for saliva stimulation is easily applicable to infants and young children. Compared with blood drawing, stimulation of saliva secretion by citric acid is painless and noninvasive, is more readily accepted to patients, is at least as clinically relevant for theophylline determination, and allows frequent measurements of drug levels for individualization of the dosage with samples taken at home.(ABSTRACT TRUNCATED AT 250 WORDS)

An evaluation of the initial treatment of acute asthma.

Two treatment regimens for the initial treatment of acute asthma in 50 patients between the ages of 12 and 20 years seen in the emergency room were evaluated. The treatments were randomized such that 26 patients received 2.5 mg of the beta 2-agonist fenoterol by nebulizer and 24 patients received 0.3 mg of epinephrine followed by 0.75 mg of Sus-Phrine. Clinical assessment and spirometry were performed over a two-hour period. Both groups responded within ten minutes and peak improvement was reached within one hour. Peak expiratory flow and clinical score were better following fenoterol treatment in the first hour (P less than .05). The one-second forced expiratory volume and the forced expiratory flow in the middle half of the vital capacity were greater at 20 minutes with fenoterol (P less than .05). Those with more severe obstruction (forced expiratory volume less than 30%) receiving aerosol therapy also had significantly greater improvement in the first 20 minutes compared with those who received injections. Four patients failed to respond to epinephrine whereas all patients showed improvement with fenoterol (P less than .05). These results demonstrated that an inhaled beta 2-agonist is effective in the initial treatment of acute asthma in children, regardless of severity, and avoids the need for injections.

Clonidine effects on disposition of xenobiotics in rats: inhibited elimination of flow-limited but not extraction-limited agents.

1. The alpha 2-adrenoceptor agonist, clonidine, reduces the hepatobiliary clearance of the anionic dye, sulphobromophthalein (BSP) in rodents. We now compare the effects of clonidine on BSP elimination with its effects on disposition of compounds which are metabolized by hepatic microsomal mixed function oxidases. 2. BSP, 100 mg kg-1 was administered i.v. to rats at 4 h after s.c. saline or clonidine, 0.2 mg kg-1. Thirty min later, plasma BSP levels were 121.4 +/- 2.25 micrograms ml-1 in saline-treated rats, while in clonidine-treated rats they were 631.5 +/- 141.0 micrograms ml-1. Clonidine raised hepatic BSP levels from 256.0 +/- 28.9 micrograms g-1 tissue to 568.5 +/- 86.5 micrograms g-1. 3. Acute administration of clonidine (0.2 mg kg-1 s.c.) or repeated clonidine dosing (0.2 mg kg-1, s.c. twice daily for 10 days) did not affect the disposition of intravenously administered [14C]-antipyrine (15 mg kg-1). 4. Activities of the P450 mixed function oxidase enzymes, aniline hydroxylase and aminopyrine N-demethylase, were identical in liver microsomes from saline-treated rats and in microsomes from rats given single or multiple s.c. doses of clonidine (0.2 mg kg-1). 5. Addition of clonidine or other 2-substituted imidazoles at concentrations up to 2 microM did not affect the activities of aniline hydroxylase or of aminopyrine N-demethylase in suspensions of rat liver microsomes. Other substituted imidazoles, including cimetidine, clotrimazole and metronidazole, at concentrations of 0.2 microM or higher, inhibited the activities of these microsomal enzymes. 6. Clonidine slowed BSP elimination, which is probably hepatic blood flow-limited, but not the extraction-limited elimination of antipyrine, which is metabolized by hepatic microsomal enzymes.

Changes in theophylline metabolism during postnatal development in rat liver slices.

The metabolism of theophylline was studied in liver slices of young and adult rats. Theophylline and six metabolite fractions were recognized in adult liver by thin-layer radiochromatography and high performance liquid chromatography: 1-methyluric acid; 1-methylxanthine; 1,3-dimethyluric acid and/or 3-methylxanthine; caffeine; a uracil derivative and two unknown polar compounds. Preincubation with caffeine or theobromine inhibited theophylline metabolism. Allopurinol decreased the formation of three metabolite fractions but markedly increased the production of 1-methylxanthine. SKF 525-A inhibited the overall metabolism of theophylline. The specific activity of the enzyme system was 3.2 +/- 0.4 nmoles X (g liver)-1 X hr-1 in the 4- to 5-day-old rat and increased to a peak of 25.7 +/- 1.7 in the 28-day-old; values for Km and Vmax in the 7- and 28-day-olds were 132.1 and 67.5 microM, and 23.9 and 52.1 nmoles X (g liver)-1 X hr-1 respectively. Theophylline and the same six metabolites were identified in young and adult rats, but the development pattern was not uniform. Peak age-related activity and involvement of mixed-function oxidase system are features which are common to theophylline and caffeine metabolism. Xanthine oxidase played a role in theophylline metabolism. Formation of caffeine from theophylline was not dependent on a lack of activity of other pathways.

Hepatic and extrahepatic metabolism of 14C-styrene oxide.

With 8-(14)C-styrene oxide as substrate, specific glutathione S-transferase and epoxide hydrase activities were determined in subcellular fractions of liver, lungs, kidney, and intestinal mucosa from rabbit, rat, and guinea pig. Liver had the highest enzyme activities in each species. Rat and guinea pig had higher glutathione S-transferase activity in both liver and kidney than rabbit. Rat testis also had appreciable glutathione S-transferase activity. The perinatal development of epoxide hydrase and glutathione S-transferase was followed in liver and several extrahepatic tissues of fetal and neonatal guinea pigs and rabbits. The rates at which enzyme activities reached adult levels in the extrahepatic tissues differed from the liver in both species. Epoxide hydrase and glutathione S-transferases developed at different rates in each organ, demonstrating that the relative importance of these two detoxifying pathways for styrene oxide may shift before and after birth. The effects of pretreating male and female rats with phenobarbital (PB), 1,2,3,4-dibenzanthracene (DBA), pregnenolone-16alpha-carbonitrile (PCN), or 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) on hepatic and extrahepatic epoxide hydrase and glutathione S-transferase activities toward styrene oxide were determined. PB increased both enzyme activities in liver of both sexes. PCN induced only glutathione S-transferase activity in female liver. Extrahepatic epoxide hydrase and glutathione S-transferase activities were unaffected except that TCDD doubled female renal epoxide hydrase activity and PB increased intestinal epoxide hydrase activity in both sexes. Styrene oxide biotransformation was studied in isolated, perfused rat liver and rabbit lung preparations. Conjugation with glutathione was a major metabolic pathway although significant amounts of diol were also formed in each instance. In rat liver, 27-40% of the administered styrene oxide was excreted via the bile mainly as S-(1-phenyl-2-hydroxyethyl)glutathione.

Effects of adrenoceptor agonists and antagonists on sulfobromophthalein disposition in mice.

The effects of alpha 2-adrenoceptor agonists on sulfobromophthalein (BSP) disposition in mice were studied. It was found that agents with both central and peripheral activities (clonidine, guanabenz, B-HT 920 and methyldopa) as well as peripherally acting alpha 2-adrenoceptor agonists (para amino-clonidine and St 91) inhibited sulfobromophthalein disposition in the mouse, and also caused substantial hypothermia. The effects of these agonists were inhibited by yohimbine, a specific alpha 2-antagonist, except for those of para amino-clonidine where only partial reversal was achieved. The effects of clonidine on BSP disposition were also reversed by piperoxan but not by the alpha 1-adrenoceptor antagonists, prazosin and phenoxybenzamine, nor by the beta-blocker, propranolol. These results suggest that, in mice, peripheral alpha 2-adrenoceptors are involved in the effects of the alpha-agonists on BSP disposition.

Tolerance to effects of clonidine and morphine on sulfobromophthalein disposition in mice.

Chronic treatment of mice with clonidine or morphine caused tolerance to the analgesic and thermoregulatory effects of these drugs. After chronic morphine, mice also became tolerant to the analgesic and thermoregulatory effects of clonidine. Cross tolerance to the hypothermic effect of morphine was demonstrated after chronic clonidine administration, but no diminution of morphine-induced analgesia could be shown. Morphine and clonidine acutely increased the retention of sulfobromophthalein (BSP) in plasma and liver. Chronic dosing with morphine or clonidine caused partial tolerance and cross-tolerance to the rise in hepatic BSP caused by an acute challenge with either agonist. However, both drugs elevated plasma BSP levels similarly in tolerant and non-tolerant mice. Thus, regimens which readily induced tolerance to the analgesic and hypothermic effects of morphine or clonidine were only partially effective in modifying the acute hepatobiliary effects of these drugs.

Loperamide effects on hepatobiliary function, intestinal transit and analgesia in mice.

Loperamide effects on hepatobiliary function, analgesia and gut transit were studied in mice. Varying doses of the antidiarrheal drug, loperamide, were administered to mice by intracerebroventricular, intravenous, subcutaneous and intragastric routes. Gut motility was determined by intestinal transit of India ink, analgesia by warm water tail flick latency, and hepatobiliary function by retention of the anionic dye, sulfobromophthalein in plasma and liver. When given by all routes at modest doses, loperamide slowed intestinal transit. Analgesia, a centrally mediated opiate effect, was only detected after intracerebroventricular or subcutaneous loperamide at high, near-toxic doses. Elevations of plasma and liver sulfobromophthalein were noted at routes and doses which slowed gut transit, well below those needed for analgesia. Intragastric loperamide at one fortieth its LD50 caused marked elevation of sulfobromophthalein levels and gut slowing, but no analgesia. Sulfobromophthalein elevation and gut slowing by intragastric loperamide were not affected by spinal cord transection but were reversed by naltrexone, an opiate antagonist. Non-toxic doses of loperamide slow gut transit and modify hepatobiliary function in mice by opiate actions at peripheral sites.

Albumin-dependent digoxin transfer in isolated perfused human placenta.

OBJECTIVE: To determine the effects of albumin (BSA) concentration in perfusion medium on digoxin transfer in isolated perfused human placental cotyledon. STUDY DESIGN: Isolated placental cotyledons from 13 normal human placentas were dually perfused after cannulating artery and vein of the chorionic plate and piercing 4 catheters through the corresponding basal plate with M199 medium enriched with BSA and glucose. Flow rates were 12 and 6 ml/min in the maternal and fetal circuits, respectively. Digoxin was added to the maternal reservoir at a final concentration of 5.51 +/- 1.00 ng/ml. BSA in maternal and fetal perfusate was kept at 3 concentrations: 1, 3 and 5 mg/ml (Groups I, II, III). Transplacental passage of digoxin was calculated from repeated fetal and maternal perfusate samples collected over 3 hours in the 3 groups. Digoxin levels were measured by FPIA (TDx, Abbott). RESULTS: There was no transfer of digoxin from the maternal to fetal compartment when the concentration of BSA was 1 mg/ml. Increasing the concentration of BSA led to a substantial increase in the transfer of digoxin to the fetal compartment. Steady state levels of digoxin in the fetal compartment were 0.61 +/- 0.19 ng/ml at 3 mg/ml of BSA. CONCLUSION: Maternal and fetal serum concentration of BSA affect digoxin transfer in isolated perfused human placentas. Three mg/ml are considered to be the optimal albumin concentration.

Effects of morphine and clonidine on sulphobromophthalein disposition in mice.

Levels of sulphobromophthalein (BSP) in plasma and liver were elevated by the opiate, morphine, and by the alpha 2-adrenoceptor agonist, clonidine. Neither morphine, 1 mg kg-1, nor clonidine, 0.01 mg kg-1, affected BSP levels significantly. When given together at these doses, they caused BSP levels in plasma and liver to be raised. At 20 mg kg-1, the effect of morphine on BSP levels was maximal, as was that of clonidine, 1.0 mg kg-1. However, the effect of these drugs given together on plasma BSP exceeded the maximal effect of either alone. Yohimbine, an alpha 2-adrenoceptor antagonist, did not affect BSP levels, nor did the opiate antagonist, naloxone. Each of these antagonists reversed the hepatobiliary effects of its respective agonist, as shown by return of BSP levels to those of saline-treated mice. Yohimbine did not reverse morphine, nor did naloxone reverse clonidine. The additive effects of morphine and clonidine and the specificities of their respective antagonists strongly suggest the involvement of discrete receptors mediating their essentially identical hepatobiliary effects.

Disposition of hexobarbitone and antipyrine in DOCA-hypertensive rats.

The disposition of antipyrine and hexobarbitone, and their effects on drug metabolizing hepatic enzymes have been investigated in DOCA-hypertensive rats. Antipyrine pharmacokinetic parameters were the same in hypertensive and control animals. Hexobarbitone sleeping time was longer in hypertensive rats compared with controls, while the activity of hepatic hexobarbitone hydroxylase was the same in both groups. Hepatic aminopyrine-N-demethylase activity was elevated in hypertensive rats while aniline hydroxylase and aryl hydrocarbon hydroxylase were lower. Glucuronyl transferase was the same in both groups. The sensitivity of the central nervous system of hypertensive rats to hexobarbitone was not altered, as determined by hexobarbitone concentration in blood and in brain. The total hepatic blood flow (arterial and portal) was significantly increased. Thus it is suggested that the difference in the disposition of the two drugs is probably not due to drug metabolizing enzyme activity. It is likely that the increase in total hepatic blood flow and rapid saturation of hepatic hexobarbitone metabolizing enzymes have significant roles in the slower metabolism and increased activity of hexobarbitone in hypertensive rats as compared with control rats.

Drug metabolizing enzymes in the ostrich (Struthio camelus): comparison with the chicken and the rat.

The activities of selected hepatic and renal drug-metabolizing enzymes of the ostrich, chicken and rats were compared. The concentration of glutathione in the liver and kidneys of the avian species was significantly lower than in the rat. The activity of ostrich hepatic glutathione S-transferase was 2-fold higher than that of the chicken and the rat and the renal glutathione S-transferase of the ostrich was 10 times higher than that of the rat. The activity of ostrich hepatic UDP-glucuronyl transferase was significantly lower than that of the rat. The activities of hepatic cytochrome P450 1A and 2B1/2 as measured by the dealkylation of ethoxy- and methoxyresorufin, respectively, were higher int he avian species than the rat; no difference was noticed in the activity of aniline hydroxylase. The results show that the activity of ostrich drug-metabolizing enzyme system is quantitatively different from the rat and in many cases also from the chicken.

Stridor caused by vocal cord malfunction associated with emotional factors.

We describe two adolescent patients in whom a disorder of the vocal cords associated with emotional factors resulted in acute episodes of stridor. Adduction of the vocal cords on inspiration and abduction on expiration was found on indirect laryngoscopy. The problem responded to either placebo treatment or psychotherapy. The similarity between vocal cord dysfunction presenting as stridor and that presenting as asthma is discussed. The importance of diagnosing these functional problems in children is emphasized in order to avoid unnecessary diagnostic procedures and hazardous treatment.

Effects of clonidine and IBMX on sulfobromophthalein disposition in rats.

Clonidine, an alpha 2-adrenoceptor agonist, inhibited the biliary excretion, reduced the plasma clearance and increased the hepatic retention of sulfobromophthalein (BSP) in a dose related fashion in rats. The maximal effects of clonidine on BSP disposition occurred about 4 h after pretreatment. The effects of clonidine on biliary excretion and hepatic retention of BSP were retained following laparotomy (with or without bile duct cannulation); however, the effect of clonidine on plasma disappearance profile was not retained following abdominal surgery. Isobutylmethylxanthine (IBMX) affected BSP disposition in a similar fashion as clonidine, in rats without bile duct cannulation only; no effect of IBMX could be observed in bile duct cannulation rats. Yohimbine, an alpha 2-adrenoceptor antagonist, reversed the effects of clonidine, but not of IBMX, on BSP disposition. It thus seems that clonidine and IBMX exert their effects on BSP disposition by different mechanisms and probably at different sites.

The effects of estrus cycle on drug metabolism in the rat.

The effect of the female rat estral cycle on microsomal drug metabolism in-vivo and in-vitro has been studied. Two microsomal enzymes, aminopyrine-N-demethylase and aniline hydroxylase showed a greater specific activity (p less than 0.01) in the diestrus phase of the estral cycle while the oxidative enzyme aryl hydrocarbon hydroxylase and the conjugative enzyme, glucuronyl transferase, were not affected. In vivo studies which included theophylline and antipyrine metabolism, and hexobarbital sleeping times showed no difference between the different phases of the estral cycle. Conflicting evidence about the effect of steroid sex hormones on hepatic drug metabolism is discussed.