Application of oral bioavailability surrogates in the design of orally active inhibitors of rhinovirus replication.

Previous studies in rats and humans demonstrated poor oral bioavailability of potent in vitro 2-aminobenzimidazole inhibitors of rhinovirus replication due to significant first-pass elimination and possibly also to poor aqueous solubility. Estimations of aqueous solubility, as well as measurements of caco-2 permeability and NADPH dependent compound loss in rat liver microsomal incubations were employed alongside traditional in vivo experiments in rats to guide subsequent chemistry efforts. Retention of activity upon replacement of the metabolically labile vinyl oxime in the lead molecule with a vinyl carboxamide was a major breakthrough; however, oral bioavailability among the latter compounds was variable. Based on the ability to independently measure solubility, permeability, and metabolic stability of new compounds, variable solubility across the series (ranging from approximately 1 to 10 microg/mL) was identified as the cause of the inconsistent performance. Subsequent efforts to improve solubility led to the discovery of highly soluble (>10 mg/mL) and potent dessulfonyl vinyl carboxamide benzimidazoles. Determination of the metabolic stability of these compounds as a surrogate of the extent of their first-pass elimination supported a prediction of excellent oral bioavailability. In comparison to the sulfonyl-containing vinyl carboxamides, caco-2 permeabilities were reduced 5 to 10-fold; however, these were considered to be in the range of well-absorbed compounds based on comparison to a series of reference compounds of known percentage absorption in humans. Subsequent experiments in the rat verified the oral bioavailability of these N-alkyl compounds, with one compound (368177) having an absolute oral bioavailability of 89.4%. The application of solubility and caco-2 permeability as surrogates for oral absorption potential, in conjunction with the use of microsomal incubations as a surrogate for first-pass metabolism, was shown to augment a rational chemistry approach to discover orally bioavailable inhibitors of rhinovirus replication. Future expanded use of these surrogates is planned.

Serologic incidence of some diseases in Kansas wild turkeys.

Wild turkeys (Meleagris gallopavo, n = 1164) were tested for Mycoplasma gallisepticum, Mycoplasma meleagridis, Mycoplasma synoviae, and Salmonella pullorum from 1990 to 1997. Although 3.3% of the turkeys were suspect for one or more diseases, only 0.9% were serologically positive for M. gallisepticum. These 11 positives were all from one country in south-central Kansas.

Correlation of biocatalytic activity in an organic-aqueous two-liquid phase system with solvent concentration in the cell membrane.

Results presented here show that loss of progesterone 11 alpha-hydroxylase activity in Rhizopus nigricans in aqueous-organic two-liquid phase and cosolvent systems correlates well with the concentration of solvent in the cell membranes. Rhizopus nigricans is shown to retain full 11 alpha-hydroxylase activity at saturating aqueous phase concentrations of hexane and the higher primary alcohols. This reflects their inability to attain a critical concentration in the cell membranes. The relationship between our own findings and the previously described correlation of the logarithm of the partition coefficient with activity retention is explained and design parameters are proposed that may be used to select solvents for future biocatalytic systems.