Separation of electronic and hydrophobic effects for the papain hydrolysis of substituted N-benzoylglycine esters.

The role of hydrophobic and electronic effects on the kinetic constants kcat and Km for the papain hydrolysis of a series of 22 substituted N-benzoylglycine pyridyl esters was investigated. The series studied comprises a wide variety of substituents on the N-benzoyl ring, with about a 300,000-fold range in their hydrophobicities, and 2.1-fold range in their electronic Hammet constants (sigma). It was found that the variation in the log kcat and log 1/Km constants could be explained by the following quantitative-structure activity relationships (QSAR): log 1/Km = 0.40 pi 4 + 4.40 and log 1/kcat = 0.45 sigma + 0.18. The substituent constant, pi 4, is the hydrophobic parameter for the 4-N-benzoyl substituents. QSAR analysis of two smaller sets of glycine phenyl and methyl esters produced similar results. A clear separation of the substituent effects indicates that in the case of these particular esters, acylation appears to be the rate limiting catalytic step.

Effects of the respiratory tract on inhaled materials.

Inhalation exposure is often compared to intravenous or oral routes of administration with regard to the biological fate of inhaled materials. Such comparisons, however, overlook the contribution of respiratory tract enzymes to the metabolic fate and toxicity of inhaled materials. The effect of respiratory tract metabolism on the toxicity of inhaled materials is thought to be substantial for many compounds for the following reasons. (1) High concentrations of xenobiotic metabolizing enzymes occur in the nose and substantial concentrations occur in the lung. (2) The respiratory tract tissues are the first exposed to inhaled materials and are exposed to the highest concentrations (barring tissue specific uptake). (3) The products of respiratory tract metabolism may have different toxicities from those of hepatic metabolism. (4) Tissues at risk to toxic metabolites formed in the respiratory tract are different from those formed in the liver. These four reasons for concluding that respiratory tract metabolism may influence the toxicity of inhaled materials are backed by a solid body of expanding experimental data. Therefore, a complete assessment of the fate of inhaled materials should include assessment of potential contributions of respiratory tract metabolism.

In vivo metabolism of nasally instilled benzo[a]pyrene in dogs and monkeys.

Metabolism of inhaled materials deposited in the nasal cavity potentially influences their biological fate and toxicity. Metabolic enzymes, including cytochrome P-450-dependent monooxygenases, are not evenly distributed throughout the nasal cavity. The purpose of this study was to determine whether benzo[a]pyrene (BaP) deposited in the nasal cavity could be metabolized and cleared by the nasal tissue in the ethmoid and maxillary turbinate regions of Beagle dogs and cynomolgus monkeys. Nasopharyngeal mucus was collected at frequent intervals during periodic nasal instillations of BaP (and for dogs 24 h after instillation) for analysis of BaP and its metabolites. During and up to 48 h after nasal instillation of [14C]BaP, blood, urine and feces were collected to determine BaP clearance from the nose. High pressure liquid chromatographic analysis of organic phase extracts of nasopharyngeal mucus demonstrated that [14C]BaP instilled in either turbinate region was metabolized to dihydrodiols, quinones, phenols and tetrols in both species. Phenols were the major metabolic product, although all treated animals produced trans-7,8-dihydrobenzo[a]pyrene-7,8-diol. The dog mucus sampled at 24 h had no detectable radioactivity. The excreta from both species contained only small amounts of the instilled radioactivity. There was no distinctive pattern of metabolite production based on instillation site.

Inhibition of nasal and liver cytochrome P-450 mono-oxygenases by dioxolanes.

1. A series of six alkyl-substituted dioxolanes were studied for their inhibitory effects on mono-oxygenase activities in vitro with nasal and hepatic microsomes from rats and rabbits. 2. Carbon monoxide binding and hexamethylphosphoramide (HMPA) N-demethylase activity were most susceptible to inhibition by the test compounds. 3. Inhibition of HMPA N-demethylase activity in both nasal and liver microsomes increased with lipophilicity of the inhibiting compound. In olfactory mucosa, the bulk of the substituent at the 4-position also seemed to have an effect on inhibition. 4. Mono-oxygenase activity in the nasal mucosa was inhibited more readily than that in the liver.

Carboxylesterases in the respiratory tracts of rabbits, rats and Syrian hamsters.

Esters are a widespread class of organic compounds found both in industry and the environment. Because esters are often volatile and, therefore, readily inhaled, the capacity of respiratory tract tissues as well as liver S-9 homogenates from rats, rabbits, and Syrian hamsters to hydrolyze a variety of esters was investigated. A new technique to determine hydrolysis rates by measuring carboxylic acid residues using ion chromatography was proven effective. The results indicated that esters, including potentially carcinogenic beta-lactones, are readily hydrolyzed by respiratory tract enzymes. Species and tissue differences were apparent. The nasal ethmoturbinates had especially high levels of esterase activity with tissue weight-normalized activities from rabbits and hamsters for most substrates exceeding all other tissues tested, including liver. Phenyl acetate was the most rapidly hydrolyzed by ethmoturbinate tissue of the esters tested. Among straight chain aliphatic alcohol acetates, hydrolysis rates increased with carbon number up to pentyl alcohol and then decreased. Branched 4-carbon alcohol acetates were less rapidly hydrolyzed than n-butyl acetate. Correlation of hydrophobicity constants with hydrolysis rates indicated that, for the straight chain aliphatic acetates, a bilinear model best fit the data.

In vivo metabolism of nasally instilled dihydrosafrole [1-(3,4-methylenedioxyphenyl)propane] in dogs and monkeys.

Nasal metabolism of inhaled material may influence its biological fate and toxicity. The purpose of this study was to investigate, in a noninvasive and qualitative manner, the in vivo nasal metabolic activity towards 1-(3,4-methylenedioxyphenyl)propane (dihydrosafrole). Dihydrosafrole was the compound of choice as a representative of the methylenedioxyphenyl compounds. Methylenedioxyphenyl compounds, inhaled as essences or insecticide synergists, have complex interactions with cytochrome P-450-dependent monooxygenases, causing both inhibition and induction. Clearance of dihydrosafrole and its metabolites from both the ethmoid (olfactory) and maxillary (respiratory) turbinate regions of Beagle dogs and Cynomolgus monkeys was examined. Nasopharyngeal mucus was collected at frequent intervals during periodic instillation of dihydrosafrole (and, for the dogs, 24 h after instillation). Blood, urine and feces were collected to examine dihydrosafrole clearance from the nose during instillations and up to 48 h after completion of the nasal instillations of [3H]dihydrosafrole. Analysis of mucus for dihydrosafrole metabolites was by HPLC. Most of the recovered radioactivity was in urine and blood samples over the first 24 h. Radioactivity was recovered from the nasopharyngeal mucus in both organic extractable and water soluble forms. HPLC of the organic extracts demonstrated that [3H]dihydrosafrole instilled in either turbinate region was metabolized to 2-methoxy-4-propylphenol, 2-methoxy-4-propenylphenol and 1-(3,4-methylenedioxyphenyl)propan-1-ol. A number of minor metabolites were produced in both species. One mucus sample from an ethmoid-instilled dog contained 1-(3,4-methylenedioxyphenyl)propene (isosafrole) as a metabolite. Results from this study indicate that interspecies, inter-individual, and inter-regional differences occur in the metabolism of nasally deposited dihydrosafrole in monkeys and dogs.