Probenecid, its metabolites (including 14C labeled), and several analogs were synthesized. The uv spectral properties, p.Ka's, and partition coefficients were determined. The binding of probenecid and metabolites to human plasma, human albumin, and dog plasma was measured. Pmr parameters of probenecid, its metabolites, and 9 other analogs were obtained. There was no correlation between the nature of the alkyl side-chain substituents or the partition coefficient and the chemical shifts of the aromatic ring protons. In rats all the metabolites with a single exception were terminal metabolites; similar results were obtained in vitro. Propionic acid was identified as one of the metabolic products of probenecid. The effects on urate clearance of probenecid, the various metabolites, and the piperidyl analog were determined in Dalmatian and mongrel dogs. In Dalmatians the clearance decreased while in mongrels it increased. Our studies suggest that the metabolites of probenecid may very well play a significant role in the overall uricosuric effect of the parent drug.The structures of essentially all the metabolites of probenecid in rat bile1 and human urine2'4 have been elucidated.The major routes of biotransformation are oxidation of the side chain (Figure 1, Table I) and glucuronide conjugation. In man, formation of the acyl glucuronide accounts for the disposition of about 0.2 of the drug.2'3 Incubation of probenecid-^C with reinforced rat liver preparations leads to the same metabolites as found in vivo.5The purpose of the present study was (1) to synthesize the metabolites (labeled and unlabeled) of probenecid, (2) to learn whether they could contribute to the overall pharmacologic activity of the parent drug, and (3) to extend previous investigations of structure-activity relationships in this series.6 The latter studies included detailed analyses of high-resolution proton magnetic resonance (pmr) spectra, pAa determinations, and measurement of drug-protein binding. Experimental SectionA. Synthesis.Our general procedure for synthesis of probenecid analogs (adapted from Miller7) was: to an ice-cold soln of the appropriate secondary amine (0.12 mole) in 25 ml of anhyd MeOH 8.8 g (0.040 mole) of p-(chlorosulfonyl)benzoic acid (1)1 was added and the mixt stirred at room temp overnight. The MeOH was removed in vacuo and the oily residue taken up in 25 ml of H20. The pH was adjusted to 1 with coned HC1 and the resulting ppt collected by filtration, then dissolved in a slight excess of 0.1 N NaOH (pH >10). After one extn with Et20 and repptn with HC1, the compd was crystd from EtOH-H20, with prior charcoal treatment, and dried at 110°in vacuo. Purity of the analogs was detd by capillary mp in a Thomas-Hoover Uni-Melt, by known Rf values on tic,2 and by elemental analysis. The 14C compds# were synthesized by a modification of the procedure of Motoichi, et al.8 Ring 14C-labeled probenecid and some nonlabeled analogs were a gift from Dr.

ABSTRACT. Hypoxia influences many physiological processes, such as respiration, cardiovascular, neurophysiology, and digestion. Skeletal muscle is an important motor organ, which relies on oxygen of oxidation; however, the study of hypoxia in skeletal muscle is lacking. In order to understand the effect of hypoxia on skeletal muscle, we determined the expression level of four hypoxia-related genes (ADAM17, ARG2, MMP, and HIF1A) in two distinct skeletal muscle tissues from Tibetan pigs that live at different altitudes (500 and 3650 m). Consistent with the well-characterized role of four hypoxia-related genes in the adaptation to high altitude, we found that, compared with the plain pigs, the plateau pigs had higher mRNA abundances of the four genes and lower myofiber ratio in skeletal muscle. The negative correlation between the myofiber ratio and mRNA abundance of the four hypoxia-related genes highlights their critical roles in skeletal 11588 J. Zhang et al. ©FUNPEC-RP www.funpecrp.com.br Genetics and Molecular Research 14 (3): 11587-11593 (2015) muscle. These findings may be important for understanding skeletal muscle adaptation to high altitude and hypoxia-related muscle diseases in humans.

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