4290
N. Veerapen et al. / Bioorg. Med. Chem. Lett. 19 (2009) 4288–4291
tine procedures, similar to those described above and exhibited
spectroscopic data consistent with the literature.24
We next tested the biological activity of
OAc
AcO
AcO
Si
a
-GalCer and
O
O
O
+
Gal(
cells in the presence of CD1d-expressing antigen-presenting cells
(APC) (Fig. 1a and b). -GalCer, but not Gal( 1?2GalCer), stimu-
lated iNKT cells in an APC-free CD1d-Fc fusion protein plate assay
(Fig. 1c). In fix/pulse, pulse/fix experiments -GalCer stimulated an
iNKT cell response under both conditions, whereas Gal( 1?2Gal-
Cer) resulted in cytokine release only under the pulse/fix condition
(Fig. 1d). Together, these data suggest that -GalCer and
Gal( 1?2GalCer) described here can stimulate human and mouse
iNKT cells. Furthermore, Gal( 1?2GalCer), in contrast to -GalCer,
a1?2GalCer). Both lipids stimulated human and mouse iNKT
N3 OBz
C14H29
O
HO
O
SPh
BzO
a
a
BzO
4
OBz
10
a
a
OAc
a
AcO
AcO
O
a
N3 OBz
C14H29
O
a
BzO
O
BzO
a
a
OBz
O
required uptake and processing to generate the biologically active
monoglycosyl ceramide.
O
O
Si
In conclusion, we have demonstrated the versatility of both
compounds 4 and 10 as crucial intermediates in practical and
11
high-yielding syntheses of
tant derivative, such as Gal(
a
a
-GalCer and other biologically impor-
b,c,d,e
1?2GalCer).26
OH
OH
HO
HO
OH
O
C25H51
OH
O
O
Acknowledgements
HN
OH
O
G.S.B. acknowledges support in the form of a Personal Research
Chair from Mr. James Badrick, Royal Society Wolfson Research
Merit Award, as a former Lister Institute-Jenner Research Fellow,
the Medical Council and The Wellcome Trust (084923/B/08/7).
C14H29
OH
O
OH
12
Scheme 4. Reagents and conditions: (a) NIS/TfOH, CH2Cl2, 67%; (b) TBAF, THF,
quant; (c) NaOMe/MeOH, quant; (d) H2, Pd, MeOH, 80%; (e) C25H51COCl, THF,
NaOAc, 78%.
References and notes
1. Kronenberg, M. Annu. Rev. Immunol. 2005, 23, 877.
2. Brutkiewicz, R. R. J. Immunol. 2006, 177, 769.
3. Bendelac, R.; Rivera, M. N.; Park, S. H.; Roark, J. H. Annu. Rev. Immunol. 1997, 15,
535.
4. Hong, S.; Scherer, D. C.; Mendiratta, S. K.; Serizawa, I.; Koezuka, Y.; Van, K. L.
Immunol. Rev. 1997, 169, 31.
5. Kawano, T.; Cui, J.; Koezuka, Y.; Toura, I.; Kaneko, Y.; Motoki, K.; Ueno, H.;
Nakagawa, R.; Sato, H.; Kondo, E.; Koseki, H.; Taniguchi, M. Science 1997, 278,
1626.
6. Crowe, N.; Uldrich, A. P.; Kyparissoudis, K.; Hammond, K. J. L.; Hayakawa, Y.;
Sidobre, S.; Keating, R.; Kronenberg, M.; Smyth, M. J.; Godfrey, D. I. J. Immunol.
2003, 171, 4020.
7. Burdin, N.; Brossay, L.; Kronenberg, M. Eur. J. Immunol. 1999, 29, 2014.
8. Carnaud, C.; Lee, D.; Donnars, O.; Park, S. H.; Beavis, A.; Koezuka, Y.; Bendelac,
A. J. Immunol. 1999, 163, 4647.
9. Taniguchi, M.; Harada, M.; Kojo, S.; Nakayama, T.; Wakao, H. Annu. Rev.
Immnunol. 2003, 21, 483.
10. Godfrey, D. I.; MacDonald, H. R.; Kronenberg, M.; Smyth, M. J.; Van, K. L. Nat.
Rev. Immunol. 2004, 4, 231.
11. Gonzalez-Aseguinolaza, G.; Van Kaer, L.; Bergmann, C. C.; Wilson, J. M.;
Schmieg, J.; Kronenberg, M.; Nakayama, T.; Taniguchi, M.; Koezuka, Y.; Tsuji, M.
J. Exp. Med. 2002, 195, 617.
12. Miyamoto, K.; Miyake, S.; Yamamura, T. Nature 2001, 413, 513.
13. Chiba, A.; Oki, S.; Miyamoto, K.; Hashimoto, H.; Yamamura, T.; Miyake, S.
Arthritis Rheum. 2004, 50, 305.
14. (a) Takikawa, H.; Muto, S. E.; Mori, K. Tetrahedron 1998, 54, 3141; (b) Figueroa-
Perez, S.; Schmidt, R. R. Carbohydr. Res. 2000, 328, 95; (c) Xia, C.; Yao, Q.;
Schuemann, J.; Rossy, E.; Chen, W.; Zhu, L.; Zhang, W.; De Libero, G.; Wang, P.
G. Bioorg. Med. Chem. Lett. 2006, 16, 2195; (d) Fan, G. T.; Pan, Y. S.; Lu, K. C.;
Cheng, Y. P.; Lin, W. C.; Lin, S.; Lin, C. H.; Wong, C. H.; Fang, J. M.; Lin, C. C.
Tetrahedron 2005, 61, 1855; (e) Ndonye, R. M.; Izmirian, D. P.; Dunn, M. F.; Yu,
K. O. A.; Porcelli, S. A.; Khurana, A.; Kronenberg, M.; Richardson, S. K.; Howell,
A. R. J. Org. Chem. 2005, 70, 10260; (f) Michieletti, M.; Bracci, A.; Compostella,
F.; De Libero, G.; Mori, L.; Fallarini, S.; Lombardi, G.; Panza, L. J. Org. Chem. 2008,
73, 9192; (g) Lee, A.; Farrand, K. J.; Dickgreber, N.; Hayman, C. M.; Juers, S.;
Hermans, I. F.; Painter, G. F. Carbohydr. Res. 2006, 341, 2785.
15. Kimura, A.; Imamura, A.; Ando, H.; Ishida, H.; Kiso, M. Synlett 2006, 15, 2379.
16. Akimoto, K.; Natori, T.; Morita, M. Tetrahedron Lett. 1993, 34, 5593.
17. Van den Berg, R. J. B. H. N.; Boltje, T. J.; Verhagen, C. P.; Litjens, R. E. J. N.; Van
der Marel, G. A.; Overkleeft, H. S. J. Org. Chem. 2006, 71, 836.
18. Imamura, A.; Ando, H.; Korogi, S.; Tanabe, G.; Muraoka, O.; Ishida, H.; Kiso, M.
Tetrahedron Lett. 2003, 44, 6725.
Figure 1.
a-GalCer and Gal(a1?2GalCer) stimulate CD1d-restricted iNKT cells. (a)
In vitro activation of 2.5 ꢁ 104 human iNKT cells (clone BM2a.3) in co-culture with
2.5 ꢁ 104 U937 cells and various concentrations of
a-GalCer (filled squares) and
Gal(a1?2GalCer) (open squares). After 16 h, cytokines were determined in culture
supernatants by ELISA; (b) in vitro activation of 5 ꢁ 104 mouse iNKT cells
(hybridoma DN32) in co-culture with 5 ꢁ 104 RAW cells transfected with CD1d
(filled symbols) or untransfected (open symbols) and various concentrations of a-
GalCer (squares) and Gal(
were determined in culture supernatants by ELISA; (c) Plate-bound murine
a1?2GalCer) (circles). After 16 h, cytokine concentrations
recombinant CD1d-Fc fusion proteins were loaded with
a-GalCer (filled squares)
or Gal(
a
1?2GalCer) (open squares) for 16 h, washed, and 5 ꢁ 104 iNKT cell
hybridomas were added per well. Cytokines were determined in culture superna-
tants by ELISA; (d) RAW cells transfected with CD1d were pulsed with 100 ng/ml of
19. Hada, N.; Oka, J.; Nishiyama, A.; Takeda, T. Tetrahedron Lett. 2006, 47, 6647.
20. (a) Stork, G.; Kim, G. J. Am. Chem. Soc. 1992, 114, 1087; (b) Bols, M. J. Chem. Soc.,
Chem. Commun. 1992, 12, 913; (c) Bols, M. J. Chem. Soc., Chem. Commun. 1993, 9,
791; (d) Bols, M. Acta Chem. Scand. 1993, 47, 829.
21. Bols, M. Tetrahedron 1993, 49, 10049.
22. Saito, S.; Ichinose, K.; Sasaki, Y.; Sumita, S. Chem. Pharm. Bull. 1992, 40, 3261.
a
-GalCer or Gal(a1?2GalCer) for 3 h and then washed and fixed with glutaralde-
hyde (filled bars), or fixed and then pulsed for 3 h (open bars). 105 APCs were co-
cultured with 105 iNKT cell hybridomas for 16 h and cytokines were determined in
culture supernatants by ELISA. Cytokine responses are expressed as percent of
maximal response. Methods are described elsewhere.25