1196 Journal of Medicinal Chemistry, 2006, Vol. 49, No. 3
BeliaeV et al.
2-[5-((tert-Butyldimethylsilanyl)oxy)-4-hydroxypentyl]isoin-
dole-1,3-dione 69. Prepared from diol 68 by general procedure for
Boc derivatives; yield 91%.
2-[5-((tert-Butyldimethylsilanyl)oxy)-4-oxopentyl]isoindole-
1,3-dione 70. Prepared from compound 69 by general procedure
for Boc derivatives; yield 92%.
(0.1 M), sodium octylsulfate (1 mM), EDTA (0.15 mM), dibutyl-
amine (1 mM), and methanol (5%) adjusted to pH 3.5 with
perchloric acid. The detection was carried out electrochemically
with a glassy carbon electrode, an Ag/AgCl reference electrode,
and an amperometric detector (Gilson model 142); the detector cell
was operated at 0.75 V. Current produced was monitored with the
Gilson Unipoint LC system software.
(S)-2-{3-[3-(5,7-Difluoro-1,2,3,4-tetrahydronaphthalen-2-yl)-
2-thioxo-2,3-dihydro-1H-imidazol-4-yl]propyl}isoindole-1,3-di-
one 71 (General Procedure). A stirred mixture of (S)-5,7-difluoro-
1,2,3,4-tetrahydronaphthalen-2-ylamine hydrochloride 10f (0.22 g,
1.0 mmol), 2-[5-((tert-butyldimethylsilanyl)oxy)-4-oxopentyl]iso-
indole-1,3-dione 70 (0.38 g, 1.05 mmol), potassium thiocyanate
(0.11 g, 1.10 mmol), water (0.18 g, 1.0 mmol), and acetic acid
(0.3 mL, 5.0 mmol) in ethyl acetate (3 mL) was refluxed for 7 h,
then cooled to room temperature, washed by sodium bicarbonate
solution, dried over anhydrous magnesium sulfate, and evaporated
in vacuo. The residue was purified by column chromatography on
silica, with ethyl acetate-petroleum ether mixture as eluent, to
afford a yellow oil (0.18 g, 40%).
(S)-5-(3-aminopropyl)-1-(5,7-difluoro-1,2,3,4-tetrahydronaph-
thalen-2-yl)-1,3-dihydroimidazole-2-thione Hydrochloride 73
(General Procedure). Compound 71 (0.16 g, 0.35 mmol) was
dissolved in a mixture of 2-propanol (5 mL) and THF (2 mL). Water
(0.8 mL) and sodium borohydride (0.066 g, 1.74 mmol) were added
at room temperature, and the mixture was stirred for 15 h. Acetic
acid (0.6 mL, 10 mmol) was added, and and the solution was
refluxed for 2 h and then evaporated in vacuo to dryness. The
residue was taken up into acetone, the solid was filtered off, and
the filtrate was acidified with 2 N HCl solution in ethyl acetate.
The precipitate was collected and washed with acetone to afford
beige crystals (0.076 g, 60%): δH (400 MHz, DMSO-d6) 12.1 (1H,
br s, NH), 8.40 (3H, br s, N+H), 7.05 (1H, t, C6H), 6.91 (1H, t,
C8H), 6.76 (1H, s, C4′H), 4.60 (2H, br s, C1H and C2H), 3.00-
2.75 (8H, m, C1H, C3H, C4H, C1′′H and C3′′H), 2.00 (1H, m,
C3H), 1.80 (2H, m, C2′′H); δC (100 MHz, DMSO-d6) 161.3 (CS),
160.2 (dd, C7), 160.2 (dd, C5), 139.4 (m, C8a), 127.8 (C5′), 119.1
(dd, C5a), 115.3 (C4′), 111.0 (dd, C5), 101.3 (t, C6), 52.4 (C2),
38.0 (C3′′), 31.9 (C1), 25.4 (C3), 25.1 (C1′′), 21.8 (C4), 21.2 (C2′′).
Anal. (C16H20ClF2N3S‚0.33CH3CO2C2H5) C, H, N, S.
Male NMRI mice or Wistar rats were obtained from Interfauna
Ibe´rica (Spain) and were kept 10 and 5 per cage, respectively, under
controlled environmental conditions (12 h light/dark cycle and room
temperature 22 ( 1 °C). Food and tap water were allowed ad
libitum, and experimentation was performed during daylight hours.
At time ) 0 h, animals were administered with either test
compounds at a given dose or vehicle (water) delivered orally via
gavage. At 2, 6, 9, 12, 18, and 24 h postdose, the animals were
sacrificed by decapitation and heart (left atrium and left ventricle)
and brain (frontal and parietal cortex) were isolated, weighed, and
stored in a volume of 0.2 M perchloric acid for 12 h at 4 °C in the
dark. Postincubation, the resulting supernatants were collected by
centrifuge filtration of incubates (0.2 µM, 10 min, ∼5000 rpm, 4
°C). Supernatants were stored frozen at -80 °C until analysis.
Quantification of dopamine and noradrenaline in supernatants was
performed by high-pressure liquid chromatography with electro-
chemical detection as described above. All animal interventions
were performed in accordance with European Directive 86/609 and
the rules of the National Institute of Health’s Guide for the Care
index.htm).
Supporting Information Available: Full NMR and combustion
analysis data for target compounds not included in Experimental
Section and NMR data for selected intermediate compounds (PDF).
This material is available free of charge via the Internet at http://
pubs.acs.org.
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Pharmacology. SK-N-SH cells (ATCC HTB-11, passages 55-
79) were maintained in a humidified atmosphere of 5% CO2-95%
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medium (Sigma-Aldrich) supplemented with 100 units/mL penicil-
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contained sodium acetate pH 5.0 (200 mM), N-ethylmaleimide (30
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pargyline hydrochloride (1 mM), sodium fumarate (10 mM),
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dopamine (50 mM prepared in 20 mM ascorbic acid). After a
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h, after which solutions were removed and filtered through Spin-X
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