346
Avramova, Danchev, Buyukliev, and Bogoslovova
water. The mixture was heated under reflux for 10 h. The solvents were
evaporated on a rotary evaporator. To the crude product was added dilute
HCl (1:1) to pH 3–4. The non-basic products were extracted with ether. The
ethereal solution was dried and the ether removed. The crude product was
recrystallized from acetone. Yield of 5a – 0.41 g (27%); mp 142–145 °C
(Table 2).
Biochemical Evaluations
Evaluation of the Degree of MAO Inhibition
Rat brain mitochondria preparations and MAO assay procedure were
performed according to Meyerson et al. [11]. The ammonia formed was
measured according to fluorimetric assay, proposed by Taylor et al. [12]
.
Substrate concentrations in the MAO inhibition assay were 1 mmol/l
serotonine (type A), 0.1 mmol/l betaphenylethylamine (type B), 1 mmol/l
tyrosine (type A+B). Enzyme preparation were preincubated with the inhibi-
tors for 15 min at 37 °C. IC50 values represent mean values of determination
in 3 to 5 rat brain preparations. Inhibitor concentrations over the range 0.01
–100 mmol/l were used .
Preparation of 4-[3-Phenyl)-3-hydroxypropyl]-2-(4-phenyl)-3-methylmor-
pholine 5b
From 3.45 g (10 mmol) 4e, 0.56 g (10 mmol) KOH, and 0.74 g (20 mmol)
NaBH4 in 20 ml methanol, after refluxing for 10 h was obtained 1.89 g (61%)
of the product 5b, recrystallized from acetone. Mp 80–82 °C (Table 2).
The MAO inhibition activity in the absence of the tested compounds
amounted to 6.12 ± 0.55 (MAO-A) 2.62 ± 0.25 (ΜΑΟ−Β), 6.98 ± 0.30
(MAO-A+B) mmol/l NH4+/min/mg of protein. Protein concentrations were
determined by the method of Lowry et al. [13]
.
The results of pharmacological and biochemical experiments underwent
statistical processing by the Student-Fisher t-test at level of significance
p ≤ 0.05.
Pharmacology
Materials and Methods
References
The experiments were performed on 368 male white mice with body
weight 18–22 g and 5 male Wistar rats (160–200 g body weight). Acute
toxicity (LD50) of the studied compounds was assessed by dissolving them
in saline (0.9% NaCl), and administering to mice by the intraperitoneal (ip)
route in 4 or 5 different doses (6 animals in group for each dose) and
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of LD50 (the same volume – 0.1 ml/10 g b/w of the solvent – 0.9% NaCl, was
administered to the controls). The solution of hexobarbital sodium (dose 80
mg/kg body weight) was administered ip to the animals 30 min after the
administration of the compounds. Sleeping time was measured in minutes by
observing the righting reflex recovery.
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A control group of 6 animals was put in an actometer (Activity Cage, Ugo
Basille, Italy) and the locomotor activity was determined in arbitrary units
at 10 min intervals for 90 min. The tested compounds in dose 1/10 of LD50
were administered to the animals and they were tested under analogous
conditions. The total locomotor activity was compared to that of the control
(vehicle-treated) group.
The antidepressive activity of the compounds was examined using the
screeningtest “behaviourdespair” [10], calculating the time of immobilisation
of the mice in seconds over a 5 min observation.
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Received: June 2, 1998 [FP307]
Arch. Pharm. Pharm. Med. Chem. 331, 342–346 (1998)