2776 J ournal of Medicinal Chemistry, 1998, Vol. 41, No. 15
Yasunaga et al.
(57 mg, 0.48 mmol), and the crude salt was recrystallized from
EtOH-Et2O to give 34 as white crystals (378 mg, 84%): 1H
NMR (400 MHz, DMSO-d6) δ 7.10 (d, J ) 8.5 Hz, 2H), 6.82 (d,
J ) 8.5 Hz, 2H), 6.80 (dd, J ) 10.8, 2.9 Hz, 1H), 6.71 (dd, J )
8.8, 2.9 Hz, 1H), 6.45 (s, 1H), 4.57 (t, J ) 4.8 Hz, 1H), 4.15 (t,
J ) 5.2 Hz, 2H), 4.11 (t, J ) 5.6 Hz, 2H), 3.71 (s, 3H), 3.04 (t,
J ) 5.6 Hz, 2H), 2.78 (t, J ) 6.8 Hz, 2H), 2.51 (t, J ) 6.8 Hz,
2H), 2.0-1.8 (m, 2H), 1.6-1.5 (m, 4H); MS (FAB) m/z 390 (M+
+ 1); mp 139-141 °C. Anal. (C22H28FNO4‚0.5C4H4O4) C, H,
N, F.
4H); MS (EI) m/z 371 (M+); mp 139-141 °C. Anal. (C22H26
FNO3‚0.5C4H4O4) C, H, N, F.
-
6-F lu or o-8-[2-[2-(4-m eth oxyp h en yl)eth yla m in o]eth ox-
y]ch r om a n -4-ol fu m a r a te (2:1) (37): prepared from 32 by a
procedure similar to that described for 34 (72%); 1H NMR (400
MHz, DMSO-d6) δ 7.14 (d, J ) 8.0 Hz, 2H), 6.85 (d, J ) 8.0
Hz, 2H), 6.80 (dd, J ) 10.0, 2.8 Hz, 1H), 6.70 (dd, J ) 8.8, 2.8
Hz, 1H), 6.49 (s, 1H), 4.57 (t, J ) 4.8 Hz, 1H), 4.2-4.1 (m,
2H), 4.09 (t, J ) 5.6 Hz, 2H), 3.72 (s, 3H), 3.05 (t, J ) 5.6 Hz,
2H), 2.94 (t, J ) 7.6 Hz, 2H), 2.74 (t, J ) 7.6 Hz, 2H), 2.0-1.8
(m, 2H); MS (FAB) m/z 362 (M+ + 1); mp 153-155 °C (from
EtOH). Anal. (C20H24FNO4‚0.5C4H4O4) C, H, N, F.
6-Flu or o-8-[2-[N-m eth yl-4-(4-m eth oxyph en yl)bu tylam i-
n o]eth oxy]ch r om a n -4-on e Hyd r och lor id e (39). N-Methyl-
4-(4-methoxyphenyl)butylamine (38) was prepared by reduc-
tion of an N-ethoxycarbonyl derivative of 19a (LiAlH4, THF,
reflux): 1H NMR (90 MHz, CDCl3) δ 7.11 (d, J ) 8.8 Hz, 2H,
meta to OMe), 6.82 (d, J ) 8.8 Hz, 2H, ortho to OMe), 3.77 (s,
3H, OCH3), 2.7-2.4 (m, 4H, benzylic and NCH2), 2.41 (s, 3H,
NCH3), 1.84 (br s, 1H, NH), 1.8-1.4 (m, 4H); MS (EI) m/z 193
(M+).
This was coupled with 13 by a procedure similar to method
C to give 39 (48%): 1H NMR (500 MHz, CDCl3) δ 12.70 (br s,
1H), 7.23-7.21 (m, 1H), 7.07 (d, J ) 8.5 Hz, 2H), 6.92-6.89
(m, 1H), 6.82 (d, J ) 8.5 Hz, 2H), 4.61 (br s, 1H), 4.54 (br s,
1H), 4.53 (t, J ) 6.1 Hz, 2H), 3.78 (s, 3H), 3.56 (br s, 1H), 3.47
(br s, 1H), 3.28 (br s, 1H), 3.12 (br s, 1H), 2.92 (s, 3H), 2.80 (t,
J ) 6.1 Hz, 2H), 2.63 (t, J ) 7.3 Hz, 2H), 1.93 (br s, 2H), 1.70
(br s, 2H); MS (FAB) m/z 402 (M+ + 1); mp 126-128 °C (from
THF-Et2O). Anal. (C23H28FNO4‚HCl) C, H, N, Cl, F.
(+)-6-F lu or o-8-[2-[4-(4-m eth oxyp h en yl)bu tyla m in o]et-
h oxy]ch r om a n -4-ol [(+)-34] a n d (-)-6-F lu or o-8-[2-[4-(4-
m eth oxyp h en yl)bu tyla m in o]eth oxy]ch r om a n -4-ol [(-)-
34]. A CHIRALPAK AD column (i.d. ) 10 mm, l ) 250 mm),
packed with amylose tris(3,5-dimethylphenylcarbamate) sta-
tionary phase,29 was treated with 0.1% Et3N in EtOH prior to
use. The free base of 34 (100 mg) was dissolved in EtOH (10
mL), and the solution (0.2 mL) was injected to the column and
eluted with EtOH (1.2 mL/min) at ambient temperature. The
eluates were detected by UV absorption at λ ) 285 nm and
pooled into fractions; the fractions from 40 runs were thus
collected automatically. First was eluted (+)-34 (tR 16.1 min,
total amount 31.0 mg): 1H NMR (400 MHz, CDCl3) δ 7.08 (d,
J ) 8.8 Hz, 2H, meta to OMe), 6.81 (d, J ) 8.8 Hz, 2H, ortho
to OMe), 6.66 (dd, J ) 8.8, 2.9 Hz, 1H, chroman aromatic),
6.59 (dd, J ) 9.8, 2.9 Hz, 1H, chroman aromatic), 4.73 (t, J )
4.3 Hz, 1H, chroman C4-H), 4.3-4.2 (m, 2H, chroman C2-
H2), 4.06 (t, J ) 5.4 Hz, 2H, OCH2CH2N), 3.78 (s, 3H, OCH3),
3.01 (t, J ) 5.4 Hz, 2H, OCH2CH2N), 2.68 (t, J ) 7.1 Hz, 2H,
ArCH2), 2.57 (t, J ) 7.3 Hz, 2H, NCH2), 2.2-2.0 (m, 2H,
chroman C3-H2), 1.7-1.5 (m, 4H); MS (FAB) m/z 390 (M+
+
6-F lu or o-8-[2-[N-p r op yl-4-(1,3-b en zod ioxol-5-yl)b u -
1); [R]20 (c 0.15, CHCl3) +39°; optical and chemical purity in
D
tyla m in o]eth oxy]ch r om a n -4-on e Oxa la te (1:1) (40).
A
the same chromatography conditions, respectively 100.0% and
99.5% (peak area). Subsequently was eluted (-)-34 (tR 18.5
min, total amount 30.1 mg): [R]20D (c 0.15, CHCl3) -40°; optical
and chemical purity, respectively 99.6% and 99.4%. The 1H
NMR and mass spectra of (-)-34 were identical to those
observed for (+)-34.
mixture of the free base of 31n (600 mg, 1.50 mmol), n-C3H7-
Br (185 mg, 1.50 mmol), and K2CO3 (207 mg, 1.50 mmol) in
MeCN (15 mL) was heated to reflux for 2 h. Insoluble matter
was filtered off, and the filtrate was concentrated and chro-
matographed (CHCl3-MeOH, 98:2) to give the free base of 40
as a solid (600 mg, 1.35 mmol, 90%). This was treated with
oxalic acid (121 mg, 1.35 mmol), and the crude salt was
recrystallized from MeCN to give 40 as a white solid (420 mg,
53% from the free base of 31n ): 1H NMR (400 MHz, DMSO-
d6) δ 7.28 (dd, J ) 9.8, 2.9 Hz, 1H), 7.05 (dd, J ) 8.3, 2.9 Hz,
1H), 6.80 (d, J ) 7.8 Hz, 1H), 6.79 (s, 1H), 6.65 (d, J ) 7.8 Hz,
1H), 5.95 (s, 2H), 4.53 (t, J ) 6.3 Hz, 2H), 4.34 (t, J ) 4.8 Hz,
2H), 3.40 (br s, 2H), 3.10-2.95 (m, 4H), 2.80 (t, J ) 6.3 Hz,
2H), 2.53 (t, J ) 7.8 Hz, 2H), 1.7-1.5 (m, 6H), 0.90 (t, J ) 7.3
Hz, 3H); MS (FAB) m/z 444 (M+ + 1); mp 148-149 °C. Anal.
(C25H30FNO5‚C2H2O4) C, H, N, F.
P h a r m a cology. F SC Assa y w ith Wh ole CHO Cells
Exp r essin g th e Hu m a n 5-HT1A Recep tor s. CHO cells
(stable transfectant of the human 5-HT1A receptor genomic
gene,30 1.5 × 105 cells/well)16 were plated in a 24-well plate 48
h before the experiment. The cells were preincubated with
“reaction buffer” (1 mM 3-isobutyl-1-methylxanthine, 1 mM
MgCl2, and 0.85 mM CaCl2 in PBS; 250 µL) for 20 min at 37
°C. After aspiration was added the reaction buffer (250 µL)
containing 10 µM forskolin and the test drug; for the reversal
experiments (to assess antagonist activity), 0.3 µM 8-OH-
DPAT was also included in the reaction buffer. Cyclic AMP
was then allowed to accumulate for 8 min at 37 °C. The
reaction was terminated by the addition of 0.2 N aqueous HCl
(250 µL), and the mixture was incubated on ice for 1 h. Cyclic
AMP production was measured with a radioimmunoassay kit
(Yamasa Shoyu, Chiba, J apan). The FSC activity was ex-
pressed as percentage of the control level based on the response
that was induced by 0.3 µM 8-OH-DPAT.
N-[2-[(6-F lu or o-4-m eth oxych r om a n -8-yl)oxy]eth yl]-4-
(4-m eth oxyp h en yl)bu tyla m in e Oxa la te (1:1) (35). The
free base of 34 (200 mg, 0.510 mmol) was mixed with MeOH
(2 mL) and concentrated HCl (0.08 mL), and the mixture was
heated to reflux for 14 h. The mixture was made basic with 1
N NaOH and extracted with CH2Cl2. The organic phase was
dried, concentrated, and chromatographed (CHCl3-MeOH, 99:
1) to give the free base of 35 (124 mg, 0.310 mmol, 60%). This
was treated with oxalic acid (27 mg, 0.30 mmol) in MeOH to
give 35 as white crystals (140 mg, 54% from the free base of
34): 1H NMR (500 MHz, DMSO-d6) δ 8.0 (2-3H, br s), 7.11
(d, J ) 8.5 Hz, 2H), 6.91 (dd, J ) 10.3, 3.0 Hz, 1H), 6.84 (d, J
) 8.5 Hz, 2H), 6.75 (dd, J ) 9.1, 3.0 Hz, 1H), 4.28-4.20 (m,
4H), 4.06 (td, J ) 11.0, 2.4 Hz, 1H), 3.71 (s, 3H), 3.35 (s, 3H),
3.29 (t, J ) 5.2 Hz, 2H), 3.02 (t, J ) 7.6 Hz, 2H), 2.53 (t, J )
7.0 Hz, 2H), 2.06-1.91 (m, 2H), 1.7-1.6 (m, 4H); MS (EI) m/z
403 (M+); mp 188 °C. Anal. (C23H30FNO4‚C2H2O4) C, H, N,
F.
N-[2-[(6-F lu or o-2H-ch r om en -8-yl)oxy]eth yl]-4-(4-m eth -
oxyp h en yl)bu tyla m in e F u m a r a te (2:1) (36). A mixture of
the free base of 34 (625 mg, 1.60 mmol), p-TsOH‚H2O (366
mg, 1.93 mmol), 1,4-dioxane (10 mL), and toluene (5 mL) was
heated to reflux for 1 h under azeotropic dehydration condi-
tions. The mixture was diluted with EtOAc and washed
successively with aqueous NaHCO3, water, and brine. The
organic phase was dried, concentrated, and chromatographed
(CHCl3-MeOH, 99:1) to give the free base of 36 (450 mg, 1.21
mmol, 76%). This was treated with fumaric acid (66 mg, 0.59
mmol), and the crude salt was recrystallized from EtOH-Et2O
to give 36 as crystals (384 mg, 56% from the free base of 34):
1H NMR (500 MHz, DMSO-d6) δ 7.10 (d, J ) 8.5 Hz, 2H), 6.83
(d, J ) 8.5 Hz, 2H), 6.82 (dd, J ) 10.8, 3.1 Hz, 1H), 6.58 (dd,
J ) 8.5, 3.1 Hz, 1H), 6.46 (dt, J ) 9.8, 1.8 Hz, 1H), 6.45 (s,
1H), 5.98 (dt, J ) 9.8, 3.6 Hz, 1H), 4.71 (dd, J ) 3.6, 1.8 Hz,
2H), 4.10 (t, J ) 5.5 Hz), 3.71 (s, 3H), 3.00 (t, J ) 5.5 Hz, 2H),
2.74 (t, J ) 7.0 Hz, 2H), 2.51 (t, J ) 7.3 Hz, 2H), 1.6-1.5 (m,
Beh a vior a l Resp on ses. Male Wistar rats (J apan SLC)
were treated with reserpine 18 h prior to use. Fifteen minutes
after the administration of a test compound (or a vehicle),
8-OH-DPAT (0.25 mg/kg) was subcutaneously injected to rats
to induce the behavioral responses. Forepaw treading (FT)
and flat body posture (FBP)25 were scored every 3 min for 15
min beginning 3 min after the injection of 8-OH-DPAT,
according to a ranked intensity scale where 0 ) absent, 1 )