Prototype trial design for rapid dose selection of antiretroviral drugs: an example using emtricitabine (Coviracil).

Antiretroviral monotherapy for initial drug characterization risks the selection of resistant virus, yet monotherapy is the only setting where many fundamental properties of a new drug can be reliably determined. Using data on viral replication kinetics and dynamics, we designed an accelerated (14 day) open-label study of single agent emtricitabine (formerly known as FTC)--a nucleoside reverse transcriptase inhibitor--to select a dosing regimen for further therapeutic study. Five regimens (25 mg bd, 100 mg od, 200 mg od, 100 mg bd and 200 mg bd) were evaluated in HIV-1-infected subjects over a 14 day dosing period to determine the optimal dose and pharmacokinetics. Serial blood samples for virological, pharmacokinetic and intracellular FTC-triphosphate measurements were drawn frequently. A dose-response relationship for the antiviral activity of emtricitabine was established, with total daily doses of 200 mg or more producing the greatest median HIV-1 viral load suppression: 1.72-1.92 log10. Based on virological outcomes, dose-response analysis and intracellular triphosphate levels, a once-daily dose of 200 mg was selected for further long-term clinical study. Adverse events possibly related to emtricitabine were unremarkable. The antiviral activity of emtricitabine correlated well with intracellular FTC-triphosphate concentrations. This study design is a safe, useful tool for early dose selection for drugs with potent antiretroviral activity and linear pharmacokinetics.

The influence of suckling interval on milk production of sows.

The objective of this study was to determine whether sow milk yield per gland could be increased by reducing the interval between suckles (suckling interval). Eighteen sows were allocated at their first farrowing to three treatments comprising litter sizes of 6 or 12 piglets or a cross-suckling treatment that was imposed to increase suckling frequency. The cross-suckled treatment comprised two groups of six piglets each. Each suckling group was allowed to suckle the sow during 30-min intervals each day between d 6 and d 28 of lactation. The suckling interval was shorter (P < .05) for cross-suckled sows than for sows suckling single litters of 6 or 12 piglets during early lactation (d 10 to 14) and late lactation (d 24 to 28). Average piglet growth rate between birth and 28 d of age was greatest (P < .05) for piglets in the single litters of six and lowest for piglets in the cross-suckled treatments. Single litters of 12 piglets had the highest (P < .05) litter growth rates, followed by the cross-suckled litters and then the single litters of six piglets. The concentration of lactose and fat in milk from sows remained relatively stable, although milk from the cross-suckled sows contained more protein in early lactation (P < .05). Milk yield of sows was not significantly increased (P > .05) by the cross-suckle treatment, although during early lactation, milk yield tended to be greater from sows in the cross-suckle treatment than from sows suckling single litters of six (8,920 g/d vs 7,819 g/d, P < .1). The concentration of DNA and total RNA and the RNA:DNA ratio in mammary glands was unaffected by treatment (P > .05). Sows with single litters of 12 piglets had the greatest total DNA in their udders (P < .05). However, individual gland weights were heavier (P < .05) in cross-suckled sows than in sows with single litters of 6 or 12 piglets. Increased suckling frequency seemed to play a role in increased mammary gland weight and milk production during lactation.

Protein synthesis during oxygen conformance and severe hypoxia in the mouse muscle cell line C2C12.

Oxygen conformance can be described as the ability to reduce energy demand, and hence oxygen consumption, in response to a decline in oxygen availability without a decrease in the concentration of ATP. It has been proposed that oxygen conformance may enhance cellular survival at low oxygen concentrations. We demonstrate that non-contracting C2C12 cells, a mouse skeletal muscle cell line, are capable of oxygen conformance. Typically, we found oxygen consumption to decline by 30-40% as the concentration of oxygen was reduced from 100 microM to 10 microM. Unexpectedly, the rate of protein synthesis, a major energy consumer in the cell, did not decrease significantly during oxygen conformance. Unlike oxygen conformance, severe hypoxia (<0.5 microM) caused a 36% decline in the concentration of PCr, and under these conditions of energy stress, the rate of protein synthesis declined by 43%. We conclude that there are two distinct metabolic responses to declines in oxygen concentration in non-contracting C2C12 cells.

A perifusion system to control oxygen concentration in cell suspensions.

A system is described for the perifusion of cells with a perifusate containing oxygen at concentrations defined by the user. The metabolic competency of cells in this system was assessed by measuring total ATP turnover for human platelets. Human platelets, which are small (radius of 1-2 micron) anucleate cells involved in blood clotting, maintained ATP turnover over a 4-h period at 37 degrees C. Oxygen concentration in this system was controlled by adjusting the proportions of air-saturated and nitrogen-saturated perifusates. Examples of oxygen consumption and lactate output as a function of oxygen concentration are described.

Duration of effects of isradipine during twice daily therapy in angina pectoris.

Isradipine, a 1,4 dihydropyridine calcium channel antagonist, is a potent coronary artery dilator that increases coronary blood flow with little effect on cardiac contractility. Isradipine is an approved antihypertensive agent, but its antianginal effects have not been well documented. In this placebo-controlled, double-blind, parallel-group design study we evaluated the duration of effects and safety of isradipine 10 mg bid in male patients with chronic stable angina pectoris. Seventy-two patients experiencing moderately severe angina between 3 and 7.5 minutes during a standard Bruce exercise test received placebo in a single-blind manner for 8-14 days. Sixty-one of these patients had reproducible treadmill exercise test results on three consecutive occasions and underwent further exercise tests at 3, 8, and 12 hours after a placebo period. Patients were then randomized (double blind) to either placebo or isradipine 10 mg bid for 2 weeks. Symptom-limited exercise tests were repeated predose and at 3, 8, and 12 hours after the 0800 hour dose dosing. Exercise duration increased significantly from baseline (last qualifying test during the single-blind placebo therapy, i.e., 0800 hours predose at visit 4) in the isradipine group compared to the placebo group prior to the administration of the 0800 hour dose (i.e., 12 hours after the 2000 hour dose) by 51 vs. 18 seconds, p = 0.04; and after the administration of the 0800 hour dose at 3 hours by 78 vs. 29 seconds, p = 0.005; and at 8 hours by 54 vs. 18 seconds, p = 0.04. Similarly, statistical significance was achieved when exercise data were analyzed using visit 4 (single-blind placebo therapy) corresponding time points as baseline. At 12 hours after the 0800 hour dose, exercise tolerance did not increase significantly after isradipine compared to placebo. Time to 1-mm ST-segment depression increased significantly after isradipine at 3 hours post 0800 hour dose compared to placebo (87 vs. 7 seconds, p < 0.01) but not at the 0, 8, or 12-hour postdose time points, regardless of which baseline was used. Isradipine therapy did not affect the rate-pressure double product. A significant correlation between the mean increase in total exercise time and mean plasma isradipine concentration was also present (p = 0.0295). During double-blind treatment, drug-related adverse events were experienced by four patients in the isradipine group and two patients in the placebo group. None of the patients experienced ischemic complications during the study.(ABSTRACT TRUNCATED AT 400 WORDS)