The elimination of MTC-220, a novel anti-tumor agent of conjugate of paclitaxel and muramyl dipeptide analogue, in rats.

PURPOSE: MTC-220, a conjugate of paclitaxel and muramyl dipeptide analogue, was reported to exhibit anti-tumor ability and anti-metastatic effect. The aim of present study was to investigate the elimination of MTC-220 and the related mechanisms in rats. METHODS: The excretion of MTC-220 and its metabolites in bile and urine were determined in rats after intravenous administration at 4 mg/kg. Caco-2 cell monolayer, in situ liver perfusion model and in vivo pharmacokinetics with selected inhibitors in rats were used to confirm the involvement of hepatic transporters in the elimination of MTC-220. The metabolic stability of MTC-220 was assessed by the incubation with rat liver microsomes and plasma. RESULTS: Approximately 72 % of MTC-220 was excreted into bile and less than 0.02 % into urine after administration in rats. The Caco-2 cell monolayer was impermeable to MTC-220. In in situ liver perfusion model, the hepatic extraction ratio of MTC-220 was reduced to 40 % of control in the presence of rifampicin, an Oatps inhibitor, and the cumulative biliary excretion rates of MTC-220 were reduced to 52.9, 71.5 and 62.9 % of control when concomitant perfusion with probenecid, novobiocin and verapamil, the inhibitors of Mrp2, Bcrp and P-gp, respectively. Co-administration of rifampicin, probenecid, novobiocin and verapamil with MTC-220 increased the AUC0-t and decreased the CL of MTC-220 in certain extents in rats. MTC-220 remained metabolically intact in rat liver microsomes, but less stable in plasma incubation. CONCLUSIONS: In summary, the elimination of MTC-220 was mainly through the biliary excretion in unchanged form in rats. Liver transporters including Oatps, Mrp2, Bcrp and P-gp might be all involved in the hepatic elimination of MTC-220. MTC-220 exhibited the high metabolic stability in liver microsomes, but less stable in plasma. The esterases might involve in the metabolism of MTC-220 in plasma.

[Studies on the isolation of endogenous sleep factors from Tupaia Belangeri Chinensis (TBC) after sleep deprivation].

The present study was undertaken to explore endogenous sleep factors isolated from 48-72 h sleep deprived (SD) male Tupaia belangeri chinensis (TBC). Only drink ad libitum (10% glucose) was available within 24 h before collection of urine. Controlled "clean" urinary samples were pooled and stored at 20 C. Fraction I-V from the urine were determined after ultrafiltration and Sephadex-G15. Amino-acid analysis of each fraction was automatically done by a 835 Amino-acid Analyzer, respectively. Bioassay was performed in 40 adult rabbits weighing 2.5-3.5 kg of either sex. Experiments were undertaken via the mesodiencephalic intraventricular infusion. Results show that S2C (Fraction-III) (50 micrograms/rabbit, i.c.v.) exhibited significant delta-enhancing effect compared to the controls. Further purification was done with Sephadex G-25 and Sephadex LH-20. The more purified S4B (50 micrograms/rabbit, i.c.v.) also exhibited significant delta-enhancing effect compared to the controls. The amino-acid analysis of Fraction-III revealed that the compositional contents of S2C and S4B are different from what have been known with Factor S, DSIP and SPS.

Somnogenic muramyl peptides.

Sleep-promoting materials isolated from human urine and rabbit brain are muramyl peptides (MPs). The most active component of the urinary material is N-acetylglucosaminyl-1,6-anhydro-N-acetylmuramyl-Ala-Glu-diaminopimel yl-Ala; 1 pmol, infused into a lateral cerebral ventricle of rabbits, induced excess slow-wave sleep (SWS) for several hours. MP-induced sleep is normal in that it is similar to the deep sleep that follows sleep deprivation. Other biological actions of MPs (e.g., pyrogenicity and immunomodulatory activity) could be dissociated, but only in part, from somnogenic actions. Interleukin 1, a substance thought to mediate many MP activities, is somnogenic, and thus may be involved in MP-induced sleep. That MPs and other immunologically active substances can greatly enhance SWS suggests that some immunological mechanisms integrate sleep in their actions.

Muramyl peptides in mammalian tissues and their effects at the cellular level.

Muramyl peptides (MPs), presumably breakdown products of bacterial cell walls, have been found in the brain, liver, and kidney of the rat. They exert multiple physiological effects on higher animals as immunoadjuvants, activators of macrophages, pyrogens, antitumor agents, inducers of contractility of smooth muscle, and promoters of slow-wave sleep, as well as nonspecific protectors of animals against infection. Structure-function relationships of these substances have been extensively studied, especially with respect to somnogenicity. In the role an intact muramyl ring is required, and the 1,6-anhydro form is active. The presence of free carboxyls or amides on the glutamyl and diaminopimelyl entities have important effects. The stereochemistry is crucial: the alanine adjacent to the N-acetylmuramyl entity must be L, and the glutamate must be D. Studies were carried out with murine macrophages to establish mechanisms of action of these glycopeptides. There are two populations of binding sites for MPs on those cells. When compounds of different structure are compared, binding ability correlates with pyrogenic and somnogenic activity. Serotonin competes with these agents for binding sites. Binding of that substance induces at least one macrophage response characteristic of the binding of MP.

Pharmacokinetics and metabolism of muramyl dipeptide and nor-muramyl dipeptide [3H-labelled] in the mouse.

The fates of 3H-muramyl dipeptide (MDP) and 3H-nor-MDP have been investigated after intravenous (i.v.), intraperitoneal, and subcutaneous injection of a range of doses in the mouse. After i.v. injection both compounds were cleared rapidly from the circulation, distributed initially to the tissues, and finally excreted largely intact in the urine. Most of the tissues contained intact material at 2 min after injection, but the much lower levels of radioactivity persisting at 1 h had undergone considerable metabolism (except in intestine, where some intact material persisted for as long as 24 h). Some accumulation of radioactivity was observed in liver and kidney and there were quantitative and qualitative differences between the two compounds. Characterisation of some of the metabolites in these tissues was undertaken, and the deamidated muramyl dipeptide was tentatively identified which is known to have some biological activity. The mechanism of the biological effects, which may be expressed over a relatively long time period, remains to be explained in view of the rapid excretion of most of the dose.