Synthesis of Phosphatidylinositol Phosphates
J . Org. Chem., Vol. 61, No. 17, 1996 5909
L-r-P h osp h a tid yl-D-m yo-in ositol 3-P h osp h a te, Dip a lm -
itoyl [P td In s(3)P ]. A solution of (-)-14 (90 mg, 0.06 mmol)
and palladium black (72 mg) in aqueous 80% EtOH (2 mL)
was shaken under H2 (50 psi) for 24 h, filtered, and concen-
trated. The residual aqueous solution was lyophilized to
furnish PtdIns(3)P (52 mg, 98%): [R]23D ) -1.2° (c 0.4, CHCl3);
1H NMR (CDCl3) δ 0.89 (t, J ) 7.2 Hz, 6 H), 1.10-1.32 (m, 48
H), 1.49-1.66 (m, 4 H), 2.28-2.39 (m, 4 H), 3.40-3.80 (m, 6
H), 4.06-4.24 (m, 5 H); 31P NMR (CDCl3, H3PO4 as external
standard) δ -0.66, 1.65; MS (negative ion FAB) m/z (rel inten)
889.5 (M - H, 6), 651 (19), 255.2 (18), 183 (100).
(-)-21 (236 mg, 0.14 mmol) was subjected to hydrogenolysis,
as described for PtdIns(3)P, to afford PtdIns(3,4)P2 (lyophilized
powder, 133 mg, 98%): [R]23D ) +1.9° (c 0.5, CHCl3); 1H NMR
(CDCl3) δ 0.89 (t, J ) 7.2 Hz, 6 H), 1.13-1.35 (m, 48 H), 1.48-
1.62 (m, 4 H), 2.25-2.40 (m, 4 H), 3.42-3.52 (m, 1 H), 3.61-
3.70 (m, 2 H), 3.72-3.82 (m, 3 H), 4.02-4.20 (m, 3 H), 4.51
(m, 1 H), 5.32 (m, 1 H); 31P NMR (CDCl3, H3PO4 as external
reference) δ -1.402, -0.739, -0.327; MS (negative ion FAB)
m/z (rel inten) 969.3 (M - H, 4), 731.2 (6), 255.2 (100).
(+)-3,4,5-Tr i-O-allyl-6-O-ben zyl-1-O-(p-m eth oxyben zyl)-
m yo-in ositol (23). Regioselective protection at the 1-OH of
(-)-22 (140 mg, 0.3 mmol) with a p-methoxybenzyl group, as
(+)-3,4-Di-O-allyl-5,6-di-O-ben zyl-O-(p-m eth oxyben zyl)-
m yo-in ositol (16). Selective introduction of a p-methoxyben-
zyl function at the C-1 of (+)-15 (160 mg, 0.36 mmol), as
described for (+)-15, gave (+)-23 (syrup, 146 mg, 96%): [R]23
D
1
) +8.2° (c 0.8, CHCl3); H NMR (CDCl3) δ 2.44 (s, 1H), 3.12
described for (-)-3, yielded (+)-16 (syrup, 195 mg, 96%): [R]23
(dd, J ) 2.3, 9.9 Hz, 1 H), 3.21-3.30 (m, 2 H), 3.78-3.84 (m,
4 H), 3.91-3.97 (m, 2 H), 4.04-4.08 (m, 2 H), 4.25-4.35 (m, 4
H), 4.77-4.87 (m, 4 H), 5.12-5.17 (m, 2 H), 5.23-5.31 (m, 2
H), 5.83-6.02 (m, 2 H), 6.81-6.86 (m, 2 H), 7.21-7.43 (m, 7
H); MS (EI) m/z (rel inten) 509.4 [(M - H)+, 3], 469.4 (13),
419.4 (21), 389.2 (9), 91 (100).
D
) +14° (c 1.0, CHCl3); 1H NMR (CDCl3) δ 2.67 (d, J ) 4.2 Hz,
1 H), 3.51 (dd, J ) 3, 10 Hz, 1 H), 3.62-3.81 (m, 2 H), 4.08-
4.18 (m, 5 H), 4.20-4.30 (m, 1 H), 4.45-4.58 (m, 2 H), 4.61-
4.70 (m, 2 H), 4.90-5.01 (m, 2 H), 5.12-5.23 (m, 4 H), 5.40-
5.65 (m, 4 H), 6.21-6.42 (m, 2 H), 6.81-6.87 (m, 2 H), 7.24-
7.43 (m, 12 H); MS (EI) m/z (rel inten) 559.3 (M+ - H, 18),
469.3 (1.2), 439.2 (2.3), 121 (100).
(-)-3,4,5-Tr i-O-a llyl-2,6-d i-O-b en zyl-1-O-(p -m et h oxy-
ben zyl)-m yo-in ositol (24). Conventional benzylation of (+)-
23, as described for (+)-6, yielded (-)-24 (syrup, 140 mg,
98%): [R]23D ) -4° (c 0.5, CHCl3); 1H NMR (CDCl3) δ 3.14 (dd,
J ) 2.3, 9.4 Hz, 1 H), 3.21-3.30 (m, 2 H), 3.78-3.84 (m, 4 H),
3.91-3.97 (m, 2 H), 4.04-4.08 (m, 2 H), 4.25-4.35 (m, 4 H),
4.50-4.59 (m, 2 H), 4.77-4.87 (m, 4 H), 5.12-5.17 (m, 2 H),
5.23-5.31 (m, 2 H), 5.83-6.02 (m, 2 H), 6.81-6.86 (m, 2 H),
7.21-7.43 (m, 12 H); MS (EI) m/z (rel inten) 600 (M+, 18), 560
(29), 510 (28), 480 (17), 420 (9), 121 (100).
(+)-3,4-Di-O-a llyl-2,5,6-t r i-O-b en zyl-1-O-(p -m et h oxy-
ben zyl)-m yo-in ositol (17). Benzylation of (+)-16 (170 mg,
0.3 mmol), as described for (+)-6, gave (+)-17 (syrup, 193 mg,
1
98%): [R]23 ) +10° (c 1.2, CHCl3); H NMR (CDCl3) δ 3.24
D
(dd, J ) 3, 10 Hz, 1 H), 3.40-3.60 (m, 2 H), 3.80-4.01 (m, 4
H), 4.08-4.30 (m, 4 H), 4.36-4.40 (m, 2 H), 4.61-4.73 (m, 2
H), 4.91-5.12 (m, 6 H), 5.20-5.50 (m, 4 H), 5.97-6.18 (m, 2
H), 6.81-6.87 (m, 2 H), 7.23-7.58 (m, 17 H); MS (EI) m/z (rel
inten) 649.3 (M+ - H, 2.3), 559.3 (1.6), 529.3 (2.4), 121 (100).
(+)-2,5,6-Tr i-O-ben zyl-1-O-(p-m eth oxyben zyl)-myo-in os-
itol (18). Deallylation of (+)-17 (176 mg, 0.27 mmol) with
(-)-2,6-Di-O-ben zyl-1-O-(p-m eth oxyben zyl)-m yo-in osi-
tol (25). Deallylation of (-)-24 (120 mg, 0.20 mmol) with Pd/C
and PTSA, as described for (+)-10, gave (-)-25 (amorphous,
Pd/C and PTSA, as described for (+)-10, yielded (+)-18
79 mg, 82%): [R]23 ) -15° (c 0.5, CHCl3); 1H NMR (CDCl3) δ
D
1
(amorphous, 139 mg, 90%): [R]23 ) +5.4° (c 0.4, CHCl3); H
1.6 (br s, 1 H), 2.4 (br s, 1 H), 2.7 (br s, 1 H), 3.45 (t, J ) 9.2
Hz, 1 H), 3.54 (m, 1 H), 3.74-3.81 (m, 4 H), 3.89 (t, J ) 9.2
Hz, 1 H), 4.06 (t, J ) 2.7 Hz, 1 H), 4.65-4.87 (m, 5 H), 6.81-
6.86 (m, 2 H), 7.25-7.37 (m, 12 H); MS (EI) m/z (rel inten)
479.3 [M+ - H, 4], 389.3 (6), 359.1 (28), 299.6 (4), 121 (100).
Anal. Calcd for C28H32O7: C, 69.98; H, 6.71. Found: C,
69.90, H 6.46.
D
NMR (CDCl3) δ 2.21 (br s, 2 H), 3.42-3.51 (m, 3 H), 3.77-
3.83 (m, 4 H), 3.99-4.07 (m, 2 H), 4.62-5.01 (m, 8 H), 6.81-
6.87 (m, 2 H), 7.23-7.38 (m, 17 H); MS (FAB) m/z (rel inten)
593.3 (M + Na, 100), 121 (38).
Anal. Calcd for C35H38O7: C, 73.70; H, 6.67. Found: C,
73.00, H 6.54.
(-)-2,5,6-Tr i-O-ben zyl-1-O-(p-m eth oxyben zyl)-myo-in os-
itol 3,4-Bis(d iben zyl p h osp h a te) (19). Phosphorylation of
(+)-18 (150 mg, 0.26 mmol) via the phosphoramidite method,
as described for (-)-11, provided (-)-19 (syrup, 250 mg, 88%):
(-)-2,6-Di-O-ben zyl-1-O-(p-m eth oxyben zyl)-m yo-in osi-
tol 3,4,5-Tr is(d iben zyl p h osp h a te) (26). Phosphorylation
of (-)-25 (60 mg, 0.125 mmol) via the phosphoramidite method,
as described for (-)-11, provided (-)-26 (syrup, 150 mg, 95%):
1
[R]23 ) -7.3° (c 0.8, CHCl3); 1H NMR (CDCl3) δ 3.34-3.48
[R]23 ) -12.3° (c 0.4, CHCl3); H NMR (CDCl3) δ 3.38-3.49
D
D
(m, 3 H), 3.78-3.83 (m, 4 H), 3.98-4.07 (m, 1 H), 4.21-4.30
(m, 1 H), 4.41-4.54 (m, 2 H), 4.69-5.10 (m, 14 H), 6.81-6.86
(m, 2 H), 7.24-7.38 (m, 37 H); 31P NMR (CDCl3, H3PO4 as
external standard) δ -1.306, -0.856; MS (FAB) m/z (rel inten)
1090 (M, 100), 121 (34).
(m, 3 H), 3.78-3.84 (m, 4 H), 3.96-4.04 (m, 1 H), 4.19-4.28
(m, 1 H), 4.42-4.48 (m, 2 H), 4.65-5.10 (m, 16 H), 6.81-6.86
(m, 2 H), 7.24-7.37 (m, 42 H); 31P NMR (CDCl3, H3PO4 as
external reference) δ -2.23, -1.76, -1.56; MS (FAB) m/z (rel
inten) 1260 (M, 100), 1170 (40), 1140 (33), 1080 (19), 1050(8).
(-)-2,6-Tr i-O-b en zyl-m yo-in osit ol 3,4,5-Tr is(d ib en zyl
p h osp h a te) (27). Removal of the p-methoxybenzyl group of
(-)-26 (140 mg, 0.11 mmol) with TFA, as described for (-)-
(+)-2,5,6-Tr i-O-b en zyl-m yo-in osit ol 3,4-Bis(d ib en zyl
p h osp h a te) (20). Removal of the p-methoxybenzyl function
of (-)-19 (230 mg, 0.21 mmol) with TFA, as described for (-)-
12, gave (+)-20 (syrup, 190 mg, 93%): [R]23 ) +1.8° (c 1.2,
12, gave (-)-27 (syrup, 120 mg, 95%): [R]23 ) -8.2° (c 1,
D
D
1
1
CHCl3); H NMR (CDCl3) δ 1.80-2.41 (br s, 1 H), 3.38-3.50
CHCl3); H NMR (CDCl3) δ 1.95 (d, J ) 9.6 Hz, 1 H), 3.45-
(m, 2 H), 3.52-3.70 (m, 1 H), 3.75-3.88 (m, 1 H), 4.20-4.32
(m, 2 H), 4.63-4.74 (m, 2 H), 4.76-4.85 (m, 2 H), 4.86-5.12
(m, 10 H), 7.23-7.42 (m, 35 H); 31P NMR (CDCl3, H3PO4 as
external standard) δ -1.151, -0.699; MS (FAB) m/z (rel inten)
971.5 (M, 100), 881.4 (14), 791 (3).
3.55 (m, 1 H), 3.81 (t, J ) 9.6 Hz, 1 H), 4.76-4.86 (m, 2 H),
4.87-5.12 (m, 13 H), 7.09-7.43 (m, 40 H); 31P NMR (CDCl3,
H3PO4 as external reference) δ -2.12, -1.55, -1.43; MS (FAB)
m/z (rel inten) 1141.3 (M + H, 100), 1051.4 (50), 961.4 (16),
871 (2).
(-)-1-O-(1,2-Di-O-p a lm it oyl-sn -glycer ol-3-b en zyloxy-
ph osph or yl)-2,5,6-tr i-O-ben zyl-myo-in ositol 3,4-Bis(diben -
zyl p h osp h a te) (21). Coupling of (+)-20 (170 mg, 0.175
mmol) with the 1,2-di-O-palmitoyl-sn-glycerophosphoryl moi-
(-)-1-O-(1,2-Di-O-p a lm it oyl-sn -glycer ol-3-b en zyloxy-
ph osph or yl)-2,6-di-O-ben zyl-myo-in ositol 3,4,5-Tr is(diben -
zyl p h osp h a te) (28). Coupling of (-)-27 (102 mg, 0.09 mmol)
with the 1,2-di-O-palmitoyl-sn-glycerophosphoryl moiety, as
described for (-)-14, yielded (-)-28 (syrup, 143 mg, 86%):
[R]23D ) -2° (c 0.8, CHCl3); 1H NMR (CDCl3) δ 0.87 (t, J ) 6.2
Hz, 6 H), 1.21-1.29 (m, 48 H), 1.42-1.51 (m, 4 H), 2.11-2.22
(m, 4 H), 3.72-4.08 (m, 5 H), 4.24-4.69 (m, 3 H), 4.71-5.07
(m, 21 H), 6.85-7.42 (m, 45 H); 31P NMR (CDCl3, H3PO4 as
external reference) δ -2.28 (1P), -2.05 (0,25 P), -1.93 (0.5
P), -1.84 (0.25 P), -1.75 (1P), -1.52 (1 P); MS (FAB) m/z (rel
inten) 1862 (M + 2 H, 9), 1772 (5), 1681.6 (1), 1311 (8), 551
(100).
ety, as described for (-)-14, yielded (-)-21 (syrup, 254 mg,
1
86%): [R]23 ) -8.3° (c 1, CHCl3); H NMR (CDCl3) δ 0.89 (t,
D
J ) 7.2 Hz, 6 H), 1.18-1.40 (m, 48 H), 1.43-1.62 (m, 4 H),
2.26-2.40 (m, 4 H), 3.48-3.55 (m, 2 H), 3.63-3.75 (m, 1 H),
3.80-4.40 (m, 8 H), 4.60-5.15 (m, 16 H), 7.01-7.41 (m, 40
H); 31P NMR (CDCl3, H3PO4 as external reference) δ -1.367
(0.25 P), -1.337 (0.5 P), -1.145 (0.25 P), -0.995 (1 P), -0.704
(1 P); MS (negative ion FAB) m/z (rel inten) 969.3 (M - H, 4),
731.2 (6), 255.2 (100).
L-r-P h osp h a tid yl-D-m yo-in ositol 3,4-Bisp h osp h a te, Di-
p a lm itoyl [P td In s(3,4)P 2]. The perbenzylated derivative
L-r-P h osp h a tid yl-D-m yo-in ositol 3,4,5-tr isp h osp h a te,
Dip a lm itoyl [P td In s(3,4,5)P 3]. The perbenzylated deriva-