Formation of N-methylnicotinamide in the brain from a dihydropyridine-type prodrug: effect on brain choline.

The enhancement of brain choline levels is a possible therapeutic option in neurodegenerative diseases; however, brain choline levels are held within narrow limits by homeostatic mechanisms including the rapid clearance of excess choline from the brain. The present study tests whether N-methylnicotinamide (NMN), an inhibitor of the outward transport of choline from the brain, can elevate brain choline levels in vivo. As NMN does not cross the blood-brain barrier, we synthesized and administered the brain-permeable prodrug, 1,4-dihydro-N-methyl-nicotinamide (DNMN), and tested its effect on the levels of NMN and choline in brain extracellular fluid, using the microdialysis procedure. Administration of DNMN (1 mmol/kg s.c.) caused a 4- and 9-fold increase in plasma and liver NMN levels, respectively, as determined by HPLC. Concomitantly, the brain tissue levels of NMN were increased by a factor of twenty. In brain extracellular fluid, the injection of DNMN (1-3 mmol/kg s.c.) elevated NMN levels by 3- to 10-fold to maximum levels of >10 microM. In spite of these enhanced NMN levels, the choline concentrations in the brain extracellular fluid and in the cerebrospinal fluid (4.7 microM) remained unchanged or were even slightly decreased. Microsomal incubations of DNMN indicated that cytochrome P-450 3A isoforms may be involved in NMN formation in the liver, but not in the brain. We conclude that DNMN, a brain-permeable prodrug of NMN, is efficiently oxidized to NMN in the brain, but a 10-fold increase in extracellular NMN levels is not sufficient to reduce the clearance of choline from the brain.

Enhancement of brain choline levels by nicotinamide: mechanism of action.

Following the subcutaneous (s.c.) administration of nicotinamide (10 mmol/kg), the brain and CSF levels of nicotinamide were increased to millimolar concentrations, but the concentrations of N-methylnicotinamide (NMN) in the CSF, and of NMN and NAD+ in brain tissue were not significantly altered. Concomitantly, nicotinamide caused increases of the choline levels in the venous brain blood. In hippocampal slices, nicotinamide (1-10 mM) induced choline release in a calcium- and mepacrine-sensitive manner and, in [3H]choline-labelled slices, increased the levels of [3H]lyso-phosphatidylcholine and [3H]glycerophosphocholine. We conclude that nicotinamide enhances brain choline concentrations by mobilising choline from choline-containing phospholipids, presumably via activation of phospholipase A2, while the formation of NMN does not contribute to this effect.

Further assignment of resonances in 1H NMR spectra of cerebrospinal fluid (CSF).

A number of previously unidentified 1H NMR signals detected in CSF spectra of patients with various neurological and metabolic diseases are assigned to metabolites, drugs and drug excipients. Two-dimensional 1H NMR spectroscopy (COSY and J-resolved) is employed to resolve resonances which are hidden by superimposed peaks in one-dimensional spectra. Assignments obtained by making use of 2-D techniques, and of a 1-D 1H NMR data base created for ca. 150 authentic compounds, enable us to clarify the nature of complex signal patterns found in crowded spectral regions of CSF such as the aliphatic methyl region at ca. 1.0 ppm.

[Studies on the biotransformation and anti-ulcer effects of 2-(nicotinoylaminoethanesulfonylamino)pyridine].

Investigations were made of 2-(nicotinoylaminoethanesulfonylamino)pyridine (NTP), a new compound with anti-ulcer effects, to clarify the relationship between its biotransformation and pharmacological effects. The effect of NTP was investigated by using two types of experimental gastric ulcer models in rats. The anti-ulcer effect was revealed to be an action of NTP itself. In order to study the biotransformation of NTP, a quantitative analysis method by HPLC was established. When NTP was orally administered to rabbits, 2-(aminoethanesulfonylamino)pyridine (TP), which is a metabolite of NTP, appeared in plasma and lymph. When NTP was orally administered to rabbits to investigate its distribution in organs, a massive amount of NTP and TP appeared in various organs within 7 hr after administration. We conclude that the pharmacological effect of NTP appears to be associated mainly with the unchanged substance.