Modulation of alpha-interferon's antiviral and clinical effects by aspirin, acetaminophen, and prednisone in healthy volunteers.

The magnitude and duration of the antiviral and clinical effect of alpha-interferon was measured in healthy volunteers. A single 3 million unit intramuscular dose of interferon was given either alone (controls) or after 72 h of concomitant medications. These medications included either aspirin (650 mg every 4 h), acetaminophen (650 mg every 4 h), or prednisone (40 mg per day). Peripheral blood mononuclear cells were assayed for resistance to vesicular stomatitis virus infection and induction of 2'-5'-oligoadenylate synthetase activity as evidence of interferon's antiviral effect. Co-administration of acetaminophen increased both antiviral parameters by more than 70% (P < 0.05) and reduced symptoms after interferon dosing, compared to controls. Aspirin and prednisone did not demonstrate any significant differences from controls in antiviral effect. As a group, acetaminophen, aspirin, and prednisone reduced the clinical symptoms by 47% compared to controls (P = 0.03) after interferon dosing, although individual drug comparisons failed to reach statistical significance. Independent of treatment group, the changes in antiviral markers after interferon dosing correlated closely with each other (r = 0.72, P < 0.001), but neither correlated with symptoms or fever (r < 0.30, P > 0.05). Acetaminophen enhances the antiviral effects of a single intramuscular dose of alpha-interferon, considering the parameters measured in these healthy volunteers.

Aminopyrine infusion breath test for the determination of changes in P450 metabolism in vivo.

1. An osmotic mini-pump was used to maintain a constant infusion of radiolabelled [N-dimethyl-14C] aminopyrine into a rat. After implanting the mini-pump, 14CO2 expiration rate was constant within 12 h, and this rate was maintained for 192 h. 2. Treatment with 2-diethylaminoethyl-2,2-diphenylvalerate HCl (SKF 525-A) or cimetidine, inhibitors of P450-dependent metabolism, resulted in both dose- and time-dependent inhibition of the expiration of 14CO2.

Effects of interferon, polyriboinosinic acid--polyribocytidilic acid and steroids on the cytochrome P450 system of cultured primary mouse hepatocytes.

An earlier study from this laboratory showed that the hepatic murine cytochrome P450 (P450) system was depressed by interferon in vivo but induced in cultured primary hepatocytes. The current investigation attempted to resolve this contradiction. The P450 content of the cells used in the earlier study fell precipitously during the first 24 hr of culture and remained at the same low level throughout another 48 hr of incubation. This failure to maintain the P450 level suggested that the cells may not have been sufficiently viable to support the mechanisms involved in the depressant activity of interferon. Accordingly, a chemically defined medium containing hydrocortisone was devised which supported an acceptable level and function of the P450 system throughout a 72 hr incubation period. Functionality of the P450 system was evaluated by measuring aminopyrine N-demethylase and benzo[a]pyrene hydroxylase activities. When this steroid supplemented medium was used, interferon depressed both activities by about 25%; however, neither activity was affected significantly by poly IC. On the other hand, benzo[a]pyrene hydroxylase activity was depressed by both poly IC and interferon in hepatocytes induced with dexamethasone or with dexamethasone plus 3-methylcholanthrene. These studies emphasize the necessity of maintaining an acceptable level of homeostasis in cultured hepatocytes if one is to derive meaningful interpretations of certain biological events.

Intracellular zidovudine (ZDV) and ZDV phosphates as measured by a validated combined high-pressure liquid chromatography-radioimmunoassay procedure.

In vitro studies of zidovudine (ZDV) phosphorylation may not accurately reflect the in vivo dose-response relationship, which is crucial to determining the relationship between ZDV exposure, efficacy, and toxicity. However, measurement of ZDV phosphorylated anabolites in peripheral blood mononuclear cells (PBMCs) from ZDV-treated human immunodeficiency virus (HIV)-infected patients would be extremely useful in the more appropriate utilization of ZDV in the treatment of HIV infection. We developed a specific and sensitive combined high-pressure liquid chromatography (HPLC)-radioimmunoassay (RIA) procedure for the determination of ZDV, ZDV-monophosphate, ZDV-diphosphate, and ZDV-triphosphate in PBMCs taken from ZDV-treated HIV-infected patients. ZDV and its anabolites were extracted from washed, Ficoll-Paque-isolated PBMCs and then separated by HPLC using a strong anion-exchange column. The anabolites were then hydrolyzed to ZDV with acid phosphatase. ZDV was then measured by using a modified commercially available RIA protocol. Our method was validated by measuring [3H]ZDV anabolites generated in Molt-4 cells radioisotopically and simultaneously by the combined HPLC-RIA procedure. The ZDV determinations correlated well (r2 = 0.97) over the range of 0.037 to 5.2 pmol (10 to 1,400 pg) per assay tube. Furthermore, we defined the stability of ZDV anabolites during ficoll isolation and the recovery after extraction and cleanup. We then measured intracellular parent ZDV and its phosphorylated anabolites in PBMCs from six ZDV-treated HIV-infected patients (PBMCs were taken 2 h after a 300-mg oral dose). The mean concentrations ( +/- standard deviations) of parent and of mono-, di-, and triphosphates were 0.15 +/- 0.08, 1.4 +/-, 0.082 +/- 0.02, and 0.081 +/- 0.03 pmol/10(6) PBMC, respectively (one pmol/10(6) PBMC represents a concentration of approximately 1 microm). Concurrent serum ZDV concentrations were between 1.3 and 7.1 microm. This method should provide a useful tool for evaluating in vivo pharmacokinetics of ZDV anabolites in PBMCs and possibly other cell types, even at the low doses of ZDV currently administered therapeutically.

Ionic iodinated contrast medium and amobarbital sodium mixtures: potential for precipitation.

When a small amount of diatrizoate meglumine, an ionic iodinated contrast medium, is left in a catheter system before injection of amobarbital sodium or when the contrast medium is intentionally mixed with amobarbital sodium, a potentially dangerous situation occurs. The authors showed in vitro that a dense precipitate forms in this situation. This is due to an acid-base reaction between the relatively acidic contrast medium and basic barbiturate and the subsequent formation of insoluble amobarbital.

Pharmacokinetic screening of o-naphthoquinone 5-lipoxygenase inhibitors.

The o-naphthoquinone derivative, CGS 8515 (I), is a potent inhibitor (IC50, 0.1 microM) of 5-lipoxygenase, but its therapeutic potential is compromised by a short plasma half-life (22 min) and extremely poor oral bioavailability (less than 2%). Poor biopharmaceutical properties of CGS 8515 were attributed to poor aqueous solubility and rapid in vivo hydrolysis of its methyl ester function to an inactive metabolite (IC50, 100 microM). An active amide analogue (II) was synthesized to prevent rapid hydrolysis. While analogue II appeared to be stable in vivo, its plasma half-life was also short (10 min), possibly because of rapid tissue distribution rather than metabolic elimination. Therefore, three potent analogues with increased aqueous solubilities were synthesized and compared with respect to their pharmacokinetic properties. The analogue with the highest aqueous solubility (V) demonstrated a plasma concentration vs time profile with the largest area under the curve (AUC) and the smallest distribution (alpha) phase of all the analogues studied. The percentage AUC of the terminal phase (beta) for three analogs paralleled their aqueous solubilities. The oral bioavailability of V was improved to 27%, compared to 2% for the parent compound, CGS 8515.

Variations in demethylation of N-methylnaltrexone in mice, rats, dogs, and humans.

1. Rats and mice have a greater capacity than dogs or humans to N-demethylate the quaternary ammonium compound, N-methylnaltrexone. 2. In dogs, following the i.v. administration of N-[14C-methyl]methylnaltrexone, 50% of the radioactivity was excreted in the urine and an additional 30% in the faeces within 120 h. 3. In humans following the i.v. administration of 14C-N-methylnaltrexone, 40-60% of the radioactivity was excreted in the urine within the first 24 h. The plasma radioactivity-time curves indicated a biphasic decay and a short distribution phase between 6 and 9 min. with a longer elimination phase between 238 and 1320 min.