2194 J ournal of Medicinal Chemistry, 2002, Vol. 45, No. 11
Szabo et al.
ments were performed in triplicate using an internal maleic
acid quantitation standard.
extractions were repeated g5 times until a white solid was
completely formed. The extraction sequence was then repeated
with 4:5:5 (v/v/v) 25 mM aqueous NH4HCO3/CH3CN/2-pro-
panol. The combined supernant solutions were concentrated
to near dryness by rotary evaporation. The solid was dissolved
in 25 mM NH4HCO3 (10 mL), and the solution was lyophilized
to dryness to give 3-aza FPP 6 (308 mg, 60% yield, 91% purity
by 31P NMR) as a white solid. 1H NMR (500 MHz, ND4OD/
D2O): δ 5.01 (br, 2 H), 4.18 (br, 2 H), 3.35 (br, 2 H), 3.03 (br,
2 H), 2.83 (s, 3 H), 2.39 (br, 2 H), 1.97 (q, 2 H, J ) 6.2), 1.90
(br, 2 H), 1.57 (s, 3 H), 1.55 (s, 3 H), 1.49 (s, 3 H). 13C NMR
(125 MHz, ND4OD/D2O): δ 139.3, 131.2, 124.2 (d, J ) 7.4),
118.0 (d, J ) 5.5), 59.5, 55.4 (d, J ) 17.5), 39.5 (d, J ) 8.3),
39.1, 38.4 (d, J ) 9.2), 26.0, 25.1 (d, J ) 6.4), 22.1, 17.0 (d, J
) 6.4), 15.4 (d, J ) 6.4). 31P NMR (202 MHz, ND4OD/D2O): δ
-6.34 (d, 1 P, J ) 19.8), -10.02 (d, 1 P, J ) 19.8). LRFABMS
N-Met h yl-4,8-d im et h yl-3E,7E-n on a d ien -1-a m in e (2b).
The procedure was based on the literature.15b,36 Lithiation of
N′-tert-butyl-N,N-dimethylformamidine (2.4 g, 18.8 mmol) in
dry THF (60 mL) with tert-butyllithium (13 mL, 1.7 M, 22.1
mmol) in pentane at -25 °C for 1 h was followed by alkylation
with geranyl chloride (2.9 g, 17 mmol) in THF (10 mL) at -78
°C to room temperature (2 h). Hydrolysis of the crude forma-
midine in 4:1 methanol-water (70 mL) containing KOH (4.8
g, 85 mmol) at reflux (ca. 12 h) afforded amine 2b (2.7 g, 95%)
as a colorless oil, which was used without further purification.
1
TLC Rf 0.12 (33% MeOH/CH2Cl2). H NMR, 13C NMR, and IR
data agreed with the literature values.15b
1,1-Dim eth yleth yl 2-[N-Meth yl-N-[4,8-d im eth yl-3E,7E-
n on a d ien yl]a m in o]eth a n oa te (3b). Alkylation of amine (2.7
g, 16.2 mmol) with tert-butyl bromoacetate (3.3 g, 17.0 mmol)
in the presence of triethylamine (1.8 g, 17.8 mmol) in dry THF
(18 mL) under N2 was carried out by a literature procedure.15b
Purification of the crude product by column chromatography
with 50% ether/pentane gave 4.1 g (90%) of the known amino
(H2O): m/z 386.2. HRFABMS (positive ion) calcd for C14H30
-
NO7P2, 386.1498; found, 386.1498. The 31P NMR spectrum
showed an extra peak attributed to inorganic pyrophosphate
[δP -7.49 (s, 0.09 P)].
1,1-Dim eth yleth yl 3-[N-Meth yl-N-[4,8-d im eth yl-3E,7E-
n on a d ien yl]a m in o]p r op a n oa te (9). The procedure was
1
ester 3b as a colorless oil. TLC Rf 0.63 (33% EA/hexane). H
based on a literature procedure for a different compound.38
A
NMR, 13C NMR, and IR data agreed with the literature
values.15b
solution of the N-methylamine 2b (715 mg, 4.3 mmol) in dry
MeOH (15 mL) under N2 was stirred at room temperature as
tert-butyl acrylate (1.1 mL, 8.5 mmol) was added. The solution
was refluxed for 1 day. After it was concentrated under
reduced pressure, the residue was subjected to column chro-
matography with 33% EA/hexane to give 913 mg (72%) of the
â-amino ester 9 as a colorless oil. TLC Rf 0.40 (33% EA/
hexane). 1H NMR (500 MHz, CDCl3): δ 5.09-5.03 (m, 2 H),
2.65 (t, 2 H, J ) 7.3), 2.35 (t, 2 H, J ) 7.7), 2.32 (t, 2 H, J )
7.5), 2.21 (s, 3 H), 2.12 (q, 2 H, J ) 7.9), 2.03 (q, 2 H, J ) 7.9),
1.94 (t, 2 H, J ) 8.1), 1.64 (d, 3 H, J ) 1.1), 1.58 (s, 3 H), 1.56
(s, 3 H), 1.41 (s, 9 H). 13C NMR (125 MHz, CDCl3): δ 172.0,
136.2, 131.3, 124.2, 121.7, 80.2, 57.2, 52.8, 41.8, 39.6, 33.6, 28.0,
26.6, 26.0, 25.6, 17.6, 16.0. IR (thin film): νmax 2975, 2927,
2-[N-Meth yl-N-[4,8-d im eth yl-3E,7E-n on a d ien yl]a m in o]-
eth a n ol (4b). The procedure was based on the literature for
a related compound.15b A suspension of LiAlH4 (742 mg, 19.5
mmol) in dry ether (50 mL) under N2 was stirred and cooled
at 0 °C as amino ester 3b (1.7 g, 5.9 mmol) in ether (15 mL)
was added dropwise. The mixture was stirred at room tem-
perature for 2 h and cooled to 0 °C. Water (0.8 mL), 15%
aqueous KOH (0.8 mL), and water (2.2 mL) were added in
succession.37 After it was stirred at room temperature for 1 h,
the insoluble salts were removed by filtration, and the filtrate
was concentrated. Purification by column chromatography
with 50% ether/pentane gave 1.3 g (94%) of amino alcohol 4b
1
as a colorless oil. TLC Rf 0.35 (33% MeOH/CH2Cl2). H NMR
2852, 2795, 1731, 1454, 1367, 1255, 1155, 1039, 848 cm-1
.
(500 MHz, CDCl3): δ 5.08-5.01 (m, 2 H), 3.53 (t, 2 H, J )
5.4), 3.22 (br. s, 1 H), 2.48 (t, 2 H, J ) 5.4), 2.37 (t, 2 H, J )
7.1), 2.22 (s, 3 H), 2.12 (q, 2 H, J ) 7.5), 2.01 (q, 2 H, J ) 7.5),
1.93 (t, 2 H, J ) 8.1), 1.62 (s, 3 H), 1.57 (s, 3 H), 1.55 (s, 3 H).
13C NMR (125 MHz, CDCl3): δ 136.3, 131.2, 124.1, 121.6, 58.6,
58.3, 57.4, 41.5, 39.6, 26.5, 25.8, 25.5, 17.5, 15.9. IR (thin
film): νmax 3421, 2966, 2923, 2854, 2798, 1668, 1452, 1376,
1041 cm-1. EIMS: m/z 226.2 [M + H]+ (1), 194.2 (2), 124.1
(1), 88.1 (100), 69.1 (9), 57.0 (5). HREIMS: m/z 224.2018 (calcd
for C14H26NO, [M - H]+ 224.2014).
EIMS: m/z 310.5 [M + H]+ (1), 254.2 (1), 236.2 (3), 194.2 (13),
172.1 (40), 116.1 (100), 88.1 (1), 74.1 (9), 57.1 (5). HREIMS:
m/z 308.2586 (calcd for C19H34NO2 [M - H]+ 308.2590).
3-[N-Meth yl-N-[4,8-d im eth yl-3E,7E-n on a d ien yl]a m in o]-
p r op a n ol (10b). The LiAlH4 reduction of 9 was carried out
as described above for 4b. Yield, 389 mg (91%). TLC Rf 0.35
(33% MeOH/CH2Cl2). 1H NMR (500 MHz, CDCl3): δ 5.09-
5.03 (m, 2 H), 3.76 (t, 2 H, J ) 5.1), 2.57 (t, 2 H, J ) 5.6), 2.35
(t, 2 H, J ) 7.3), 2.23 (s, 3 H), 2.15 (q, 2 H, J ) 7.7), 2.04 (q,
2 H, J ) 7.7), 1.95 (t, 2 H, J ) 7.9), 1.66 (quintet, 2 H, J )
5.1), 1.64 (s, 3 H), 1.59 (s, 3 H), 1.57 (s, 9 H). 13C NMR (125
MHz, CDCl3): δ 136.5, 131.2, 124.2, 121.3, 64.6, 58.3, 57.8,
41.8, 39.6, 27.6, 26.5, 25.8, 25.6, 17.6, 15.9. IR (thin film): νmax
3378 (-OH), 2925, 2852, 2800, 1668, 1452, 1376, 1128, 1051,
835 cm-1. EIMS: m/z 240.2 [M + H]+ (2), 194.2 (3), 172.1 (2),
116.1 (4), 102.1 (100), 88.1 (5), 69.1 (8), 58.1 (61). HREIMS:
m/z 238.2172 (calcd for C15H28NO [M - H]+ 238.2171).
3-[N-Meth yl-N-[4,8-d im eth yl-3E,7E-n on a d ien yl]a m in o]-
p r op yl Dip h osp h a te (3-a za h om oF P P , 12). The diphosphate
was prepared from amino alcohol 10b as described above for
4b except that the extraction buffer was 1:2:2 (v/v/v) 25 mM
aqueous NH4HCO3/CH3CN/2-propanol. Yield, 353 mg (80%,
93% purity by 31P NMR). 1H NMR (500 MHz, ND4OD/D2O):
δ 4.93 (br d, 2 H, J ) 6.9), 3.84 (t, 2 H, J ) 5.8, 5.6), 3.09 (t,
2 H, J ) 7.1), 2.90 (t, 2 H, J ) 7.5), 2.65 (s, 3 H), 2.26 (q, 2 H,
J ) 7.9), 1.92-1.86 (m, 4 H), 1.84 (q, 2 H, J ) 7.5), 1.47 (s, 6
H), 1.40 (s, 3 H). 13C NMR (125 MHz, ND4OD/D2O): δ 139.9,
132.6, 124.2, 118.1, 62.9, 54.7, 53.1, 39.7, 39.0, 38.5 (d, J )
3.7), 25.9, 25.1, 24.6 (d, J ) 8.3), 22.1, 17.1, 15.4. 31P NMR
(202 MHz, ND4OD/D2O): δ -7.66 (d, 1 P, J ) 19.8), -9.70 (d,
1 P, J ) 21.4). LRFABMS (H2O): m/z 400.2. HRFABMS
(positive ion) Calcd for C15H32NO7P2, 400.1654; found, 400.1652.
The 31P NMR spectrum showed an extra peak attributed to
inorganic monophosphate [δP 1.44 (s, 0.07 P)].
2-[N-Meth yl-N-[4,8-d im eth yl-3E,7E-n on a d ien yl]a m in o]-
eth yl Dip h osp h a te (3-a za F P P , 6). Literature methods20
were used with some modifications including omission of the
cellulose column chromatography step. A solution of amino
alcohol 4b (257 mg, 1.1 mmol) and triethylamine (173 mg, 173
mmol) in dry CH2Cl2 (11 mL) under N2 was stirred and cooled
at -15 °C as methanesulfonyl chloride (0.1 mL, 1.3 mmol) was
added dropwise. After 0.5 h, the product was extracted with
ice-cold CH2Cl2 (10 mL). The organic layer was washed with
ice-cold saturated NaHCO3 (2 × 10 mL), dried (MgSO4), and
concentrated to 5 mL. MeCN (15 mL) was added to the solution
of mesylate/chloride in CH2Cl2 (5 mL). The solution was
concentrated to 2 mL and stirred in an ice bath as tris(tetra-
n-butyl)ammonium hydrogen pyrophosphate (2.7 g, 2.8 mmol)
in MeCN (3 mL) was added. The suspension was stirred at
room temperature for 6 h. After it was washed with pentane
(3 × 10 mL) and concentrated, the residue was dissolved in 2
mL of ion exchange buffer (25 mM NH4HCO3 containing 2%
(v/v) 2-propanol). The resulting solution was passed through
a column (2 cm × 10 cm) of Dowex AG 50W-X8 (100-200
mesh) cation-exchange resin (ammonium form). The column
was eluted with 2 column volumes of ion exchange buffer, and
the eluant was lyophilized to dryness. The residual white solid
was partially dissolved in 20 mL of 3:5:5 (v/v/v) 25 mM aqueous
NH4HCO3-MeCN-2-propanol. The gellike mixture was vor-
texed, the suspension was centrifuged, and the supernatant
solution was decanted into another round-bottomed flask. The
N-Octyla m in om eth a n e-1,1-bisp h osp h on ic Acid (30).
This compound was prepared by reaction of n-octylamine with