glycylprolylglycine-amide and glycine-amide

Alpha-hydroxy glycineamide (αHGA) is the active antiviral metabolite of tri-peptide glycyl-prolyl-glycine-amide (GPG-NH2 ). αHGA inhibits the replication of HIV-1 in vitro by interfering with the capsid formation. It has also an effect on viral gp160 envelope protein. Since drug transport is an important aspect of drug function, we investigated the mechanism of [(14) C] αHGA uptake by a human T cell line.. H9 cells were incubated with defined amounts of radiolabelled αHGA for definite time durations. After harvesting the cells and removal of radiolabelled material, the radioactivity associated with the cells was assayed. Experiments were also designed to address the effect of metabolic inhibitors, temperature and extra unlabelled compound as potential competitor on the cellular uptake of αHGA.. Uptake of αHGA into H9 cells was time- and dose-dependent. The uptake properties showed a low temperature dependency (Q10  < 2). Moreover the uptake was not inhibited by increasing concentrations of cold competitors. There was no effect on cellular uptake of αHGA by known metabolic inhibitors, NaN3 and NaF.. Kinetic analysis of compound uptake, metabolic inhibition studies, saturation studies and the Q10 value of αHGA uptake indicate that the compound enters H9 cells by a mechanism of passive diffusion.

Topics: Anti-HIV Agents; Diffusion; Glycine; HIV-1; Humans; Kinetics; Oligopeptides; T-Lymphocytes; Temperature; Virus Replication

The tripeptide amide glycyl-prolyl-glycinamide (GPG-amide) is a new antiretroviral drug candidate, but its absorption mechanism is unknown. In this investigation, the transport and metabolism of GPG-amide were studied in a model of the human intestinal epithelium, Caco-2 cell monolayers. The results show that when the tripeptide amide came into contact with the apical enterocyte membrane, it was degraded by CD26 (dipeptidyl peptidase IV) to glycylproline and the antiretrovirally active metabolite glycinamide. Glycinamide retained antiretroviral activity in vitro after transport through the Caco-2 cell monolayers. The transport of glycinamide across Caco-2 cell monolayers occurred via passive diffusion with an apparent permeability coefficient of about 2 x 10(-6) cm s(-1), which suggests that it is absorbed by the oral route in sufficient amounts to be considered for oral administration. In conclusion, the tripeptide GPG-amide acts as a prodrug that is activated by CD26 to release the orally active antiretroviral compound glycinamide.

Topics: Administration, Oral; Antiviral Agents; Biological Transport; Biotransformation; Caco-2 Cells; Dipeptidyl Peptidase 4; Glycine; Humans; Hydrogen-Ion Concentration; Intestinal Mucosa; Oligopeptides

The chemically modified tripeptide glycyl-prolyl-glycine-amide (GPG-NH(2)) inhibits replication of human immunodeficiency virus (HIV) type 1 (HIV-1) in vitro, probably by interfering with capsid formation. The aim of the present study was to determine whether the metabolites glycyl-proline (GP-OH), glycine (G-OH), prolyl-glycine-amide (PG-NH(2)), proline (P-OH), and glycine-amide (G-NH(2)) from proteolytic cleavage may inhibit the replication of HIV-1 in vitro. PG-NH(2) has previously been shown to have a modest effect on HIV-1 replication. In the present study we show that G-NH(2) exhibits a pronounced inhibitory effect on HIV-1. This effect was not due to a decrease in cell proliferation or viability and could not be shown for herpes simplex virus type 1. The G-NH(2) concentration that inhibited virus replication by 50% (IC(50)) was equimolar to that of GPG-NH(2) and ranged from 3 to 41 microM. Transmission electron microscopy revealed that the effect of G-NH(2) on HIV-1 morphology was equivalent to that of GPG-NH(2) and showed disarranged capsid structures, indicating interference with capsid formation. Serial passage of HIV-infected cells with G-NH(2) for more than 20 subcultivations did not decrease the susceptibility to the compound. The results from this study suggest that GPG-NH(2) might act as a prodrug and that G-NH(2) is an active antiretroviral metabolite.

Topics: Animals; Anti-HIV Agents; Cell Line; Cells, Cultured; Drug Resistance, Viral; Glycine; HIV-1; Humans; Leukocytes, Mononuclear; Microbial Sensitivity Tests; Oligopeptides; Serial Passage; Virus Assembly; Virus Replication