The Skaggs Institute
for Chemical Biology
Chemistry and Drug Discovery
C.-H. Wong, C. Behrens,
C. Bennett, M. Best, A. Brik, S. Dean, M. Fujio, S. Hanson, Z. Hong, J. Hsu, T.-L. Hsu, C.-Y. Huang,
D.-R. Hwang, A. Krebs, J.-C. Lee, F.-S. Liang, H. Liu, L. Liu, M. Numa, T. Polat, M. Sawa, P. Schanen,
M. Sugiyama, D. Thayer, S.-K. Wang, L. Whalen, C.-Y. Wu, D. Wu, M. Wuchrer, Y.-Y. Yang
of our interest in developing new chemical and enzymatic methods for organic synthesis, we developed
new aldolases via directed evolution and structure-based directed mutagenesis and used the modified
enzymes in synthesis of iminocyclitol derivatives as glycosidase inhibitors. We also developed
a method to assemble saccharide and aminoglycoside microarrays in microtiter plates for high-throughput
screening. The arrays were then used in a high-throughput assay for discovery of glycosyltransferase
inhibitors as potential drug candidates for the treatment of inflammatory diseases, for dissection
of carbohydrate specificity, and for identification of selective binding of aminoglycosides
to certain RNA sequences.
A current major interest in our laboratory
is the development of new synthetic methods and strategies to tackle major problems associated
with carbohydrate-mediated biological recognition, such as bacterial and viral infection,
cancer metastasis, and immunoresponses. We developed the programmable 1-pot method for oligosaccharide
synthesis and covalent and noncovalent methods for arraying oligosaccharides for the high-throughput
analysis of sugar-protein interactions.
In our efforts to develop enzyme inhibitors
as potential therapeutic agents, we continue to develop new methods to rapidly identify novel
inhibitors. We use a well-designed core structure with acceptable binding affinity for an enzyme
of interest to mimic the reaction transition state of the enzymatic reaction, and we search for
a single group that can be added to the core to further improve the affinity and specificity of the
core structure for the enzyme. This additional group is identified from an array of readily available
and synthetic building blocks through a reaction with the core in microtiter plates without manipulation
to isolate or protect any groups and is followed by high-throughput screening in situ. We used this
strategy with the amide bondforming reaction, the triazole-forming reaction, the epoxide-opening
reaction with amines, and the tetrabutylammonium fluoridemediated N- and O-alkylation,
and we rapidly identified several molecules that inhibit, at low nanomolar concentrations, glycosidases,
glycosyltransferases, sulfotransferases, HIV protease, and the anthrax lethal factor.
In our work on glycoarrays, we developed
a new method to prepare arrays in microtiter plates covalently and showed the usefulness of this
method in the high-throughput analysis of the interactions of the polymannose moieties on HIV
with the broadly neutralizing antibody 2G12. We used the programmable 1-pot synthesis method
to prepare various oligomannoses and spotted them on the surface of microtiter plates containing
the succinimide ester groups, through a chemical or photocleavable linker. With this method,
we defined the epitope specificity of 2G12, and the optimal epitope identified has been used in
the design of vaccines for immunization. Our goal is to develop new carbohydrate-based vaccines
for the treatment of AIDS.
Brik, A., Alexandratos, J., Lin,
Y.-C., Elder, J.H., Olson, A.J., Wlodawer, A., Goodsell, D.S., Wong, C.-H. 1,2,3-Triazole
as a peptide surrogate in the rapid synthesis of HIV-1 protease inhibitors. Chembiochem 6:1167,
Brik, A., Wu, C.-Y., Best, M.D.,
Wong, C.-H. Tetrabutylammonium fluoride-assisted rapid
N9-alkylation on purine ring: application to combinatorial reactions in microtiter
plates for the discovery of potent sulfotransferase inhibitors in situ. Bioorg. Med. Chem. 13:4622,
Calarese, D.A., Lee, H.-K., Huang,
C.-Y., Best, M.D., Astronomo, R.D., Stanfield, R.L., Katinger, H., Burton, D.R., Wong, C.-H.,
Wilson, I.A. Dissection of the carbohydrate specificity
of the broadly neutralizing anti-HIV-1 antibody 2G12. Proc. Natl. Acad. Sci. U. S. A. 102:13372,
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.
Synthesis of α-galactosyl
ceramide and the related glycolipids for evaluation of their activities on mouse splenocytes.
Tetrahedron 61:1855, 2005.
Fridman, M., Belakhov, V., Lee,
L.V., Liang, F.-S., Wong, C.-H., Baasov, T. Dual effect of
synthetic aminoglycosides: antibacterial activity against Bacillus anthracis and
inhibition of anthrax lethal factor. Angew. Chem. Int. Ed. 44:447, 2005.
Hsu, C.-C., Hong, Z., Wada, M., Franke,
D., Wong, C.-H. Directed evolution of D-sialic acid
aldolase L-3-deoxy-manno-2-octulosonic acid (L-KDO) aldolases. Proc. Natl. Acad.
Sci. U. S. A. 102:9122, 2005.
Kinjo, Y., Wu, D., Kim, G., Xing,
G.-W., Poles, M.A., Ho, D.D., Tsuji, M., Kawahara, K., Wong, C.-H., Kronenberg, M.
Recognition of bacterial glycosphingolipids by natural killer T cells. Nature 434:520, 2005.
Klostermeier, D., Sears, P., Wong,
C.-H., Millar, D.P., Williamson, J.R. A three-fluorophore
FRET assay for high-throughput screening of small-molecule inhibitors of ribosome assembly.
Nucleic Acids Res. 32:2707, 2004.
Liang, F.-S., Wong C.-H.
Surface plasmon resonance study of HCV IRES RNA-aminoglycoside interactions. Methods Mol. Biol.,
Lin, H., Thayer, D.A., Wong, C.-H.,
Walsh, C.T. Macrolactamization of glycosylated peptide
thioesters by the thioesterase domain of tyrocidine synthetase. Chem. Biol. 11:1635, 2004.
Mocharla, V.P., Colasson, B., Lee,
L.V., Romper, S., Sharpless, K.B., Wong, C.-H., Kolb, H.C. In
situ click chemistry: enzyme-generated inhibitors of carbonic anhydrase II. Angew. Chem. Int.
Ed. 44:116, 2004.
Numa, M.M.D., Lee, L.V., Hsu, C.-C.,
Bower, K.E., Wong, C.-H. Identification of novel anthrax
lethal factor inhibitors generated by combinatorial Pictet-Spengler reaction followed by screening
in situ. Chembiochem 6:1002, 2005.
Shie, J.-J., Fang, J.-M., Kuo, C.-J.,
Kuo, T.-H., Liang, P.-H., Huang, H.-J., Yang, W.-B., Lin, C.-H., Chen, J.-L., Wu, Y.T., Wong, C.-H.
Discovery of potent anilide inhibitors against the severe
acute respiratory syndrome 3CL protease. J. Med. Chem. 48:4469, 2005.
Shie, J.-J., Fang, J.-M., Kuo, T.-H.,
Kuo, C.-J., Liang, P.-H., Huang, H.-J., Wu, Y.-T., Jan, J.-T., Cheng, Y.-S.E., Wong, C.-H. Inhibition
of the severe acute respiratory syndrome 3CL protease by peptidomimetic α,β-unsaturated
esters. Bioorg. Med. Chem. 13:5240, 2005.
Thayer, D.A., Yu, H.N., Galan, M.C.,
Wong, C.-H. A general strategy toward S-linked glycopeptides.
Angew. Chem. Int. Ed. 44:4596, 2005.
Tolbert, T.J., Franke, D., Wong,
C.-H. A new strategy for glycoprotein synthesis: ligation
of synthetic glycopeptides with truncated proteins expressed in E coli as TEV protease
cleavable fusion protein. Bioorg. Med. Chem. 13:909, 2005.
Tolbert, T.J., Wong, C.-H. Carbohydrate
chains: enzymatic and chemical synthesis. In: Encyclopedia of Biological Chemistry.
Lennarz, W.J., Lane, M.D. (Eds.). Academic Press, San Diego, 2005, Vol. 1, p. 307.
Wei, S.H., Rosen, H., Matheu, M.P.,
Sanna, M.G., Wang, S.-K., Wong, C.-H., Parker, I., Cahalan, M.D. Sphingosine
1-phosphate type 1 receptor agonism inhibits transendothelial migration of medullary T cells
to lymphatic sinuses. Nat. Immunol. 6:1228, 2005.
Wong, C.-H. Protein
glycosylation: new challenges and opportunities. J. Org. Chem. 70:4219, 2005.
Wu, C.-Y., Brik, A., Wang, S.-K.,
Chen, Y.-H., Wong, C.-H. Tetrabutylammonium fluoride-mediated
rapid alkylation reaction in microtiter plates for the discovery of enzyme inhibitors in situ.
Chembiochem. 6:2176, 2005.
Wu, C.-Y., King, K.-Y., Kuo, C.-J.,
Liang, P.-H., Wu, Y.-T., Ho, M.-Y., Liao, C.-L., Fang, J.-M., Wong, C.-H.
Discovery of stable benzotriazol esters as potent inhibitors of the severe acute respiratory
syndrome 3CL protease. Chem. Biol., in press.
Wu, D., Xing, G.-W., Poles, M., Kinjo,
Y., Sullivan, B., Bodmer-Narkevitch, V., Plettenburg, O., Kronenberg, M., Tsuji, M., Ho, D.D.,
Wong, C.-H. Bacterial glycolipids and analogs as antigens
for CD1d-restricted NKT cells. Proc. Natl. Acad. Sci. U. S. A. 102:1351, 2005.
Xing, G.-W., Wu, D., Poles, M.A.,
Horowitz, A., Tsuji, M., Ho, D.D., Wong, C.-H. Synthesis
and human NKT cell stimulating properties of 3-O-sulfo-α/β-galactosylceramides.
Bioorg. Med. Chem. 13:2907, 2005.