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Synthesis of D-erythro-sphingomyelin and of D-erythro-ceramide-1-phosphoinositol
Teeahectron Lencrs.Vol.34.No.43. PP.6881-6884.1993 Printed inGreatBritain
CW40-4039193 $6.00+ .Wl Pergamon Rcss Ltd
Synthesis of D-erythro-Sphingomyelin and of D-erythro-Ceramide-1-Phosphoinositoll Bemd Kratzer, Thomas G. Mayer, and Richard R. Schmidt* Fakulttit Chemie, Universitit Konstanz, Postfach 5560 M 725, D-78434 Konstanz, Germany
Abstract: 3-O-Silyl-protected axidosphingceine3. readilyavailablefromD-q&o-azidosphin~ine, is tmnsformed into ceramidylphosphitederivative5 which is a versatilebuildingblock for the synthesisof ceramide-l-O-phosphate andderivatives.This is exhibitedfor the synthesisof the tide compounds1 and2.
Phosphosphingolipids
play an important role as membrane constituents. For instance, sphingomyelin
(Scheme 1.1) is found in a variety of different cell types2. Scheme 1
Several syntheses for this compound have been reporteds. Also, D-eryrhro-ceramide-1-phosphoinositol2
and
structural variants have been recently detected in plants, yeasts, and fungi? some of these compounds have been recognized to serve in glycophosphoinositol mediated anchoring of proteins (GPI anchors)s. Recent investigations towards the synthesis of 2 resulted in a diastereomeric mixturee. Furthermore. phosphorylated metabolites of sphingosine, for instance, sphingosine-l-phosphate,
sphingosine-phosphorylcholine,
and ceramide-l-
phosphate were found to participate in cell regulation and transmembrane signalling? Sphingosine-l-phosphate has become synthetically available s-9. We would like to report on a convenient and versatile synthesis of this type of natural products, as shown for 1 and 2 to, based on 3-0-tert-butyldimethylsilyl-protected sine 3s.“. which is readily available from D-eryrhro-azidosphingosinett
azidosphingo-
(Scheme 2).
Azide 3 was converted to the amine with triphenylphosphine (2 eq) and an excess of water in pyridine following a general procedure described by Dong and Butcher Jr. 38.Reaction with stearic acid anhydride in the presence of triethylamine in THF afforded the ceramide derivative in high yield which was condensed with the bifunctional
phosphitylation
reagent
bis(diisopropylamino)-cyanoethoxyphosphi
(4)‘2 in dichlomme-
thane/acetonitrile (1:l) under a nitrogen atmosphere in the presence of diisopropylammonium
tetrazolide. TLC-
analysis of the reaction mixture after 30 min revealed completion of the reaction. Treatment with aq. NaHC03
6881
6882
and silica gel chromatography (petroleum ether/ethyl acetate/triethylaminene, 7O:lO:l) provided veffatile intermediate V in pure form. Scheme 2
1. PPhJ. H& 2. (c,,H,&Q#. 3. 4. ‘R,NH
CP~~N~H,~&CN
pvridine (61 %I NEto (66 %I . Tetrazole: NaHcO,.
I+0
(SU)
4
1. ll.TeImzda w
%I
2 12,&O,
pY&he
3.33 % HNh$.
2. t-6uO2H
EtOH
(46 %I
3. 33 % HNMe2_ EtOH
6
I
TBAF. THF; I.E.
(68 %)
I1
7
pTsOH.
I
WH
(41 %)
cl 2
Reaction of 5 with choline tosylate in the presence of freshly sublimed tetrazole (1.0 eq) led to replacement of the diisopxopylamine group by the choline residue. The phosphite intermediate was oxidized in situ by successive addition of iodine (0.4 M solution in pyridine/dichlaromethane/water, 3:l:l) to afford the corresponding phosphotriester, treatment with a solution of dimethylamine (33% in dry ethanol) at room temperagave silyl-protected sphingomyelin 6 in 62% overall yield Final &protection was accomplished by using tetrabutylammonium
fluoride (TBAF, 1.2 eq) in THF at 35oC. Tnatment
of the product with mixed bed ion
6883
exchange resin (II+, OH-) furnished sphingomyelin (1)inhighoverall yield. It had physical data in agreement with mported valuests*t4. For the synthesis of 2. a l-O-unprotected D-isomer of m@nositol
was required. Though various me-
thods have been mported for the resolution of chiral my0-inositol derivatives, we employed a mod&d
proce+
dure to atrive at masonable amounts of the desired compound in an economical way (Scheme 3). scheme3
8a
sb I pTsOH. M-F/l-l20
(-)-Mnt-OH -
(521)
OH P A
10: R - (-)Mnl-OcO ,,: R_H I
ob lKi?w+OwY
Qa
(45w
To this end, myo-inositol was transformed as describedt5J6 into the 1,2:5&di-O-cyclohexylidene derivative, then treatment with bis(tributyltin)oxide MntOcoQl
afkded
in toluene and with (-)-menthyloxycsrbonyl
(u 2,3:4,5-) chloride [(-)-
mgioselectively the l-O-carbonates 8a.b (diastuwmeric xnixtuxe).Decyclohexylidena-
tion with p-toluenesulfonic acid (p-TsOH) in MeOH/l’HF/H~O (521) afforded known 9a,b17,which could be separated by crystallization of 9a with ethyl acetate/ethanol (2: 1)t7. Reaction of 9a with methoxymethyl chloride (MOM-Cl) in the presence of Hitnig’s base at Ooc and then slowly raising the temperature to 20% furnished O-MOM-protected 10,which was fully chsracteked13.
Treatment witb K$%JIvIeOH
atforded the
desired l-O-unprotected D-isomer 11. Reaction of 5 with 11inthepresence of tetmxole and then oxidation with t-BuOaH gave the corresponding phosphouiester which with dimethylamine in ethanol furnished O-protected 7. Treatment with p-TsOH in MeOH at 4ooc led to removal of all protective gmups, thus providing the desired target molecule 2 which was obtained by silica gel chromatography (CHCls/IvleOHL2 N NH3,65:35:8) as pure stereoisomerts. In conclusion, transformation of 3 into 5 provides a versatile intermediate for the synthesis of phospho sphmgolipids with different ester moieties. The generality of this approach has been shown for phosphorous ester formation with choline and inositol.
6884
References and Notes 1.
This work was supported by the Fonds der Chemischen
2.
Barenholz, Y.; Thompson, T.E.. Biochim. Biophys. Acru 1980,604. 129-158; Barenholz, Y.; Gatt, S. in PhosphoZipids (Hawthorne, J.N.; Ansell, G.B.; Eds.). Elsevier Biomedical Press, Amsterdam 1982, pp. 129-177.
3.
Syntheses
of optically pure Sphingomyelins:
Industrie.
a) Dong, Z.; Butcher Jr., J.A., Tetrahedron Lett. 1991, c) Shapiro, D.; Flowers,
32,5291-5294; b) Bruzik, KS.. J. Chem. Sot. Perkin Trans I 1988,423-431; H.M., J. Am. Chem. Sot. 1%2,84,1047-1050. 4.
Lhomme,
0.; Bruneteau.
M.; Costello, C.E.; Mas, P.; Molot, P.M.; Dell, A.; Tiller, P.; Michel, G.,
Eur. J. Biochem. lPPO,191,203-209, and references therein. 5.
Conzelmann. therein.
6.
Frantova,
A.; Puoti, A.; Lester, R.L.; Desponds,
A.Yu.; Stepanov,
A.E.; Bushnev,
C., EiUBO J. 1992, II, 457-466; and references
A.S.; Zvonkova,
E.N.; Shvets, V.I., Tetrahedron L.&t.
1992,33,3539-3542. 7.
Olivera, A.; Spiegel, S., Glycoconjugare
8.
Kratzer, B.; Schmidt, R.R., Tetrahedron Len. 1993.34, 1761-1764.
9. 10.
J. 1992.9, 110-I 17.
Ruan, F.; Sadahira, Y.; Hakomori, S.-i.; Igarashi, Y., Bioorg. Med. Chem. Len. lPP2,2,973-978. R.R. Schmidt, B. Kratzer, VIIth European Carbohydrate be presented.
Symposium,
Cracow, Poland, Aug. 1993, to
P.; Schmidt, R.R., Liebigs Ann. Chem. 1988,663-667.
11.
Zimmermann,
12.
Bannwarth,
13.
1: [a]Dm = + 5.8 (c = 1, chloroform/methanol, l:l)[ref.sc: [alo= = + 6.1 (chloroform/methanol, l:l)] m.p. 213-215OC (ref.? m.p. 213-214oC). iH NMR (250 MHz, CDCl CD OD 1:l) 6 = 0.86-0.91 (t. J = 6.3 Hz, 6 H), 1.27 (m, 50 H), 1.58 (m, 2 H), 1.98-2.04 (m, 2 H). 2. 1/5-2.31 (t, J = 6.8 Hz, 2 H), 3.22 (s. 9 H), 3.62 (t, J = 5.4 Hz, 2 H), 3.92-4.0 (m. 2 H), 4.03-4.09 (t, J = 7.7 Hz, 1 H), 4.14-4.2 (m, 1 H), 4.27 (m, 2 H), 5.4-5.5 (dd. J = 7.4 Hz, J = 15.3 Hz, 1 H), 5.66-5.77 (dt, J = 6.5 Hz, J = 15.3 Hz, 1 H).
W.; Trzeciak, Helv. Chim. Actu lP87,70. 175-186.
2: rH NMR (250 MHz, CDCls/CD30D/D~0 2:1:0.2) 6 = 0.81-0.86 (t, J = 6.3 Hz, 6 H), 1.22-1.40 (m, 50 H), 1.52 (m, 2 H), 1.97 (m, 2 H), 2.1-216 (t, J = 7.9 Hz, 2 H). 3.16-3.23 (t, J = 9.2 Hz, 1 H), 3.353.4 (dd, J = 2.8 Hz, J = 9.5 Hz, 1 H), 3.56-3.63 (t. J = 9.5 Hz. 1 H), 3.68-3.75 (t, J = 9.5 Hz, 1 H). 3.82-3.88 (m, 3 H), 4.01-4.07 (t, J = 7.8 Hz, 1 H), 4.15 (t, J = 2.8 Hz, 1 H), 4.22-4.59 (m, 1 H, overlapped by broad signals of water), 5.33-5.43 (dd, J = 7.8 Hz, J = 15.3 Hz, 1 H), 5.61-5.72 (dt, J = 6.7 Hz, J = 15.3 Hz, 1 H). s1P NMR (161 MHz, CDCl&!D3OD&O 2:1:0.2) $, = 1.3 ppm. 5: rH NMR (250 MHz, CDC13) 6 = 0.01, 0.02 (2 s, 6 H), 0.82-0.93 (m, 15 H) 1.12-1.29 (m, 62 H), 1.53-1.63 (m, 2 H), 1.93-2.01 (m, 2 H), 2.06-2.14 (m, 2 H), 2.56-2.62 (m, 2 H), 3.58-3.86 (m, 6 H), 4.01-4.09 (m, 1 H), 4.16-4.21 (t. J = 6.3 Hz, 1 H), 5.33-5.43 (dd, J = 6.9 Hz, J = 15.3 Hz, 1 H), 5.535.65 (m, 2 H). 3rP NMR (161 MHz, CDC13) $, = 148.7, 148.8 ppm. 10: lH NMR (250 MHz, CDCls) 6 = 0.74-2.05 (m, 18 H), 3.35-3.37 (3 s, 9 I-I), 3.40 - 3.47 (m, 7 H), 3.50-3.59 (dd, J = 2.4 Hz, J = 10 Hz, 1 H), 3.87-3.99 (m, 2 H), 4.14-4.16 (t, J = 2.4 Hz, 1 H), 4.434.59 (m, 2 H), 4.62-4.87 (m, 10 H). 14.
See ref. 3a, c
15.
S. Skrypski, Diplomarbeit, Universitat Konstanz, 1986.
16.
Massy, D.J.R.; Wyss. P., Helv. Chim. Actu 1990, 73, 1037-1057; Garegg, P-J.; Iversen, T.; Johansson, R.; Lmdberg, B., Carbohydr. Res. 1984,130,322-326.
17.
Aguil6, A.; Martin-Lomas,
M.; Penades, S., Tetrahedron L&t. 1992.33,401-404.
(Received in Germany 18 August 1993; accepted 6 September
1993)
CW40-4039193 $6.00+ .Wl Pergamon Rcss Ltd
Synthesis of D-erythro-Sphingomyelin and of D-erythro-Ceramide-1-Phosphoinositoll Bemd Kratzer, Thomas G. Mayer, and Richard R. Schmidt* Fakulttit Chemie, Universitit Konstanz, Postfach 5560 M 725, D-78434 Konstanz, Germany
Abstract: 3-O-Silyl-protected axidosphingceine3. readilyavailablefromD-q&o-azidosphin~ine, is tmnsformed into ceramidylphosphitederivative5 which is a versatilebuildingblock for the synthesisof ceramide-l-O-phosphate andderivatives.This is exhibitedfor the synthesisof the tide compounds1 and2.
Phosphosphingolipids
play an important role as membrane constituents. For instance, sphingomyelin
(Scheme 1.1) is found in a variety of different cell types2. Scheme 1
Several syntheses for this compound have been reporteds. Also, D-eryrhro-ceramide-1-phosphoinositol2
and
structural variants have been recently detected in plants, yeasts, and fungi? some of these compounds have been recognized to serve in glycophosphoinositol mediated anchoring of proteins (GPI anchors)s. Recent investigations towards the synthesis of 2 resulted in a diastereomeric mixturee. Furthermore. phosphorylated metabolites of sphingosine, for instance, sphingosine-l-phosphate,
sphingosine-phosphorylcholine,
and ceramide-l-
phosphate were found to participate in cell regulation and transmembrane signalling? Sphingosine-l-phosphate has become synthetically available s-9. We would like to report on a convenient and versatile synthesis of this type of natural products, as shown for 1 and 2 to, based on 3-0-tert-butyldimethylsilyl-protected sine 3s.“. which is readily available from D-eryrhro-azidosphingosinett
azidosphingo-
(Scheme 2).
Azide 3 was converted to the amine with triphenylphosphine (2 eq) and an excess of water in pyridine following a general procedure described by Dong and Butcher Jr. 38.Reaction with stearic acid anhydride in the presence of triethylamine in THF afforded the ceramide derivative in high yield which was condensed with the bifunctional
phosphitylation
reagent
bis(diisopropylamino)-cyanoethoxyphosphi
(4)‘2 in dichlomme-
thane/acetonitrile (1:l) under a nitrogen atmosphere in the presence of diisopropylammonium
tetrazolide. TLC-
analysis of the reaction mixture after 30 min revealed completion of the reaction. Treatment with aq. NaHC03
6881
6882
and silica gel chromatography (petroleum ether/ethyl acetate/triethylaminene, 7O:lO:l) provided veffatile intermediate V in pure form. Scheme 2
1. PPhJ. H& 2. (c,,H,&Q#. 3. 4. ‘R,NH
CP~~N~H,~&CN
pvridine (61 %I NEto (66 %I . Tetrazole: NaHcO,.
I+0
(SU)
4
1. ll.TeImzda w
%I
2 12,&O,
pY&he
3.33 % HNh$.
2. t-6uO2H
EtOH
(46 %I
3. 33 % HNMe2_ EtOH
6
I
TBAF. THF; I.E.
(68 %)
I1
7
pTsOH.
I
WH
(41 %)
cl 2
Reaction of 5 with choline tosylate in the presence of freshly sublimed tetrazole (1.0 eq) led to replacement of the diisopxopylamine group by the choline residue. The phosphite intermediate was oxidized in situ by successive addition of iodine (0.4 M solution in pyridine/dichlaromethane/water, 3:l:l) to afford the corresponding phosphotriester, treatment with a solution of dimethylamine (33% in dry ethanol) at room temperagave silyl-protected sphingomyelin 6 in 62% overall yield Final &protection was accomplished by using tetrabutylammonium
fluoride (TBAF, 1.2 eq) in THF at 35oC. Tnatment
of the product with mixed bed ion
6883
exchange resin (II+, OH-) furnished sphingomyelin (1)inhighoverall yield. It had physical data in agreement with mported valuests*t4. For the synthesis of 2. a l-O-unprotected D-isomer of m@nositol
was required. Though various me-
thods have been mported for the resolution of chiral my0-inositol derivatives, we employed a mod&d
proce+
dure to atrive at masonable amounts of the desired compound in an economical way (Scheme 3). scheme3
8a
sb I pTsOH. M-F/l-l20
(-)-Mnt-OH -
(521)
OH P A
10: R - (-)Mnl-OcO ,,: R_H I
ob lKi?w+OwY
Qa
(45w
To this end, myo-inositol was transformed as describedt5J6 into the 1,2:5&di-O-cyclohexylidene derivative, then treatment with bis(tributyltin)oxide MntOcoQl
afkded
in toluene and with (-)-menthyloxycsrbonyl
(u 2,3:4,5-) chloride [(-)-
mgioselectively the l-O-carbonates 8a.b (diastuwmeric xnixtuxe).Decyclohexylidena-
tion with p-toluenesulfonic acid (p-TsOH) in MeOH/l’HF/H~O (521) afforded known 9a,b17,which could be separated by crystallization of 9a with ethyl acetate/ethanol (2: 1)t7. Reaction of 9a with methoxymethyl chloride (MOM-Cl) in the presence of Hitnig’s base at Ooc and then slowly raising the temperature to 20% furnished O-MOM-protected 10,which was fully chsracteked13.
Treatment witb K$%JIvIeOH
atforded the
desired l-O-unprotected D-isomer 11. Reaction of 5 with 11inthepresence of tetmxole and then oxidation with t-BuOaH gave the corresponding phosphouiester which with dimethylamine in ethanol furnished O-protected 7. Treatment with p-TsOH in MeOH at 4ooc led to removal of all protective gmups, thus providing the desired target molecule 2 which was obtained by silica gel chromatography (CHCls/IvleOHL2 N NH3,65:35:8) as pure stereoisomerts. In conclusion, transformation of 3 into 5 provides a versatile intermediate for the synthesis of phospho sphmgolipids with different ester moieties. The generality of this approach has been shown for phosphorous ester formation with choline and inositol.
6884
References and Notes 1.
This work was supported by the Fonds der Chemischen
2.
Barenholz, Y.; Thompson, T.E.. Biochim. Biophys. Acru 1980,604. 129-158; Barenholz, Y.; Gatt, S. in PhosphoZipids (Hawthorne, J.N.; Ansell, G.B.; Eds.). Elsevier Biomedical Press, Amsterdam 1982, pp. 129-177.
3.
Syntheses
of optically pure Sphingomyelins:
Industrie.
a) Dong, Z.; Butcher Jr., J.A., Tetrahedron Lett. 1991, c) Shapiro, D.; Flowers,
32,5291-5294; b) Bruzik, KS.. J. Chem. Sot. Perkin Trans I 1988,423-431; H.M., J. Am. Chem. Sot. 1%2,84,1047-1050. 4.
Lhomme,
0.; Bruneteau.
M.; Costello, C.E.; Mas, P.; Molot, P.M.; Dell, A.; Tiller, P.; Michel, G.,
Eur. J. Biochem. lPPO,191,203-209, and references therein. 5.
Conzelmann. therein.
6.
Frantova,
A.; Puoti, A.; Lester, R.L.; Desponds,
A.Yu.; Stepanov,
A.E.; Bushnev,
C., EiUBO J. 1992, II, 457-466; and references
A.S.; Zvonkova,
E.N.; Shvets, V.I., Tetrahedron L.&t.
1992,33,3539-3542. 7.
Olivera, A.; Spiegel, S., Glycoconjugare
8.
Kratzer, B.; Schmidt, R.R., Tetrahedron Len. 1993.34, 1761-1764.
9. 10.
J. 1992.9, 110-I 17.
Ruan, F.; Sadahira, Y.; Hakomori, S.-i.; Igarashi, Y., Bioorg. Med. Chem. Len. lPP2,2,973-978. R.R. Schmidt, B. Kratzer, VIIth European Carbohydrate be presented.
Symposium,
Cracow, Poland, Aug. 1993, to
P.; Schmidt, R.R., Liebigs Ann. Chem. 1988,663-667.
11.
Zimmermann,
12.
Bannwarth,
13.
1: [a]Dm = + 5.8 (c = 1, chloroform/methanol, l:l)[ref.sc: [alo= = + 6.1 (chloroform/methanol, l:l)] m.p. 213-215OC (ref.? m.p. 213-214oC). iH NMR (250 MHz, CDCl CD OD 1:l) 6 = 0.86-0.91 (t. J = 6.3 Hz, 6 H), 1.27 (m, 50 H), 1.58 (m, 2 H), 1.98-2.04 (m, 2 H). 2. 1/5-2.31 (t, J = 6.8 Hz, 2 H), 3.22 (s. 9 H), 3.62 (t, J = 5.4 Hz, 2 H), 3.92-4.0 (m. 2 H), 4.03-4.09 (t, J = 7.7 Hz, 1 H), 4.14-4.2 (m, 1 H), 4.27 (m, 2 H), 5.4-5.5 (dd. J = 7.4 Hz, J = 15.3 Hz, 1 H), 5.66-5.77 (dt, J = 6.5 Hz, J = 15.3 Hz, 1 H).
W.; Trzeciak, Helv. Chim. Actu lP87,70. 175-186.
2: rH NMR (250 MHz, CDCls/CD30D/D~0 2:1:0.2) 6 = 0.81-0.86 (t, J = 6.3 Hz, 6 H), 1.22-1.40 (m, 50 H), 1.52 (m, 2 H), 1.97 (m, 2 H), 2.1-216 (t, J = 7.9 Hz, 2 H). 3.16-3.23 (t, J = 9.2 Hz, 1 H), 3.353.4 (dd, J = 2.8 Hz, J = 9.5 Hz, 1 H), 3.56-3.63 (t. J = 9.5 Hz. 1 H), 3.68-3.75 (t, J = 9.5 Hz, 1 H). 3.82-3.88 (m, 3 H), 4.01-4.07 (t, J = 7.8 Hz, 1 H), 4.15 (t, J = 2.8 Hz, 1 H), 4.22-4.59 (m, 1 H, overlapped by broad signals of water), 5.33-5.43 (dd, J = 7.8 Hz, J = 15.3 Hz, 1 H), 5.61-5.72 (dt, J = 6.7 Hz, J = 15.3 Hz, 1 H). s1P NMR (161 MHz, CDCl&!D3OD&O 2:1:0.2) $, = 1.3 ppm. 5: rH NMR (250 MHz, CDC13) 6 = 0.01, 0.02 (2 s, 6 H), 0.82-0.93 (m, 15 H) 1.12-1.29 (m, 62 H), 1.53-1.63 (m, 2 H), 1.93-2.01 (m, 2 H), 2.06-2.14 (m, 2 H), 2.56-2.62 (m, 2 H), 3.58-3.86 (m, 6 H), 4.01-4.09 (m, 1 H), 4.16-4.21 (t. J = 6.3 Hz, 1 H), 5.33-5.43 (dd, J = 6.9 Hz, J = 15.3 Hz, 1 H), 5.535.65 (m, 2 H). 3rP NMR (161 MHz, CDC13) $, = 148.7, 148.8 ppm. 10: lH NMR (250 MHz, CDCls) 6 = 0.74-2.05 (m, 18 H), 3.35-3.37 (3 s, 9 I-I), 3.40 - 3.47 (m, 7 H), 3.50-3.59 (dd, J = 2.4 Hz, J = 10 Hz, 1 H), 3.87-3.99 (m, 2 H), 4.14-4.16 (t, J = 2.4 Hz, 1 H), 4.434.59 (m, 2 H), 4.62-4.87 (m, 10 H). 14.
See ref. 3a, c
15.
S. Skrypski, Diplomarbeit, Universitat Konstanz, 1986.
16.
Massy, D.J.R.; Wyss. P., Helv. Chim. Actu 1990, 73, 1037-1057; Garegg, P-J.; Iversen, T.; Johansson, R.; Lmdberg, B., Carbohydr. Res. 1984,130,322-326.
17.
Aguil6, A.; Martin-Lomas,
M.; Penades, S., Tetrahedron L&t. 1992.33,401-404.
(Received in Germany 18 August 1993; accepted 6 September
1993)