Oncolytic immunotherapies represent a new promising strategy in the treatment of cancer. In our efforts to develop oncolytic peptides, we identified a series of chemically modified 9-mer cationic peptides that were highly effective against both drug-resistant and drug-sensitive cancer cells and with lower toxicity toward normal cells. Among these peptides, LTX-315 displayed superior anticancer activity and was selected as a lead candidate. This peptide showed relative high plasma protein binding abilities and a human plasma half-life of 160 min, resulting in formation of nontoxic metabolites. In addition, the lead candidate demonstrated relatively low ability to inhibit CYP450 enzymes. Collectively these data indicated that this peptide has potential to be developed as a new anticancer agent for intratumoral administration and is currently being evaluated in a phase I/IIa study.

In vitro rates of metabolism and Michaelis-Menten constants were determined for 25 different C6 to C10 hydrocarbons using rat liver slices in a vial head-space equilibration system. The rates of metabolism were compared with steady-state levels obtained in vivo in the same strains of rats after inhalation. Aromates were metabolized at a higher rate than naphthenes n-alkanes, isoalkanes and 1-alkenes. The aromates showed, in contrast to the other hydrocarbons investigated, increased metabolism with increasing number of carbon atoms up to C8 (o-xylene, the most extensively metabolized compound). The in vivo steady-state concentrations of the aromates in blood were inversely related to the in vitro efficiency of their metabolism. This explains the pattern of blood levels observed for the C6 to C10 aromates in the rat after inhalation, with o-xylene demonstrating the lowest concentration. In general, the extent of tissue metabolism of the investigated hydrocarbons might be of greater importance for their body distribution than their lipophilicity, especially for the highly metabolized compounds. The high in vitro intrinsic liver clearances found for the aromates indicate a flow-dependent metabolism of these hydrocarbons in vivo. The head-space liver slice equilibration system seems to work adequately for metabolic studies of hydrocarbons with different volatility and water solubility.

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Ahstruct:In vitro systems of high biological organization. e.g. containing intact hepatocytes. have been considered as more reliable for metabolic studies and in vivo predictions of toxicokinetics than subcellular systems. For this reason, the kinetics and metabolism of low-molecular-weight volatile chemicals, i.e. head space elimination of substrate and formation of metabolites, were compared in liver S9 and in liver slices. Two substrates. toluene and n-hexane, were used as they represent differences in metabolic pathways and physical-chemical properties. The two systems responded similarly to diethyldithiocarbamate inhibition and acetone inhibitiodinduction of cytochrome P-450 mediated metabolism. In contrast to liver S9. liver slices did not respond adequately to phenobarbital induction raising the question of substrate availability for P-450 enzymes in liver slices. Liver slices appeared to be superior to liver S9 when specific metabolic pathways involving phase I1 enzymes were investigated. However, liver SY seemed to be at least as good as liver slices for estimation of total metabolic rate constants (K,,,, V,,,) as a basis for in vivo predictions. As the head space liver S9 system is faster and easier to operate than liver slices, it is a promising screening tool for the metabolism of volatile compounds and metabolic interactions.

The elimination of ethanol, diazepam and oxazepam which are metabolised by different enzymes, has been studied for 30, 60, 90 and 120 min at 37, 27, 17 and 7 degrees C in rat liver slice incubations. Ethanol elimination followed zero-order kinetics at all temperatures, while the benzodiazepines consistently displayed first-order kinetics. No sign of phase transition was observed in the respective Arrhenius-plots. Ethanol elimination was more temperature dependent than the elimination of diazepam, while the elimination of oxazepam was little influenced by temperature. This is shown by the temperature ratios (Q10) and energies of activation (Ea) of 1.76, 1.56, 1.24 and 40.5, 31.9, 15.2 for ethanol, diazepam and oxazepam, respectively. This means that ethanol, diazepam and oxazepam elimination was reduced by 25, 22 and 14%, respectively, for each 10 degrees C of temperature reduction, which is considerably lower than the commonly observed 50% reduction of enzyme activity. We conclude that observations made for one drug on temperature dependent elimination may not apply to other drugs.