ISMSC 2007 - Università degli Studi di Pavia

Supra-Biomolecular Tandem Assays – Application Examples
Andreas Hennig, Hüseyin Bakirci, and Werner M. Nau
Jacobs University Bremen, School of Engineering and Science, Campus Ring 1, D-28759
Bremen, Germany, E-mail: w.nau@jacobs-university.de
PSA 73
The chemical and pharmaceutical community has great interest in simple, fast and reliable
screening methods to determine the activity of catalysts and enzymes. 1 Established methods
require antibodies which need to be raised for a specific analyte by time-consuming and
expensive methods. 2,3 In addition, the laborious chemical derivatization of substrates, which
presents an alternative approach, is often accompanied by a change (or even loss) in activity. 4,5
We present a novel label-free supramolecular method, in which macrocyclic hosts mimic the
role of antibodies for assaying enzymatic activity. A simple and versatile concept has been
developed based on the dynamic equilibria of a fluorescent dye, a substrate and a product as
well as their respective host-guest complexes. Accordingly, as the enzymatic reaction proceeds,
the conversion of the substrate to the product is coupled with a change in the host-guest
complexation equilibrium of the fluorescent dye with the macrocycle (supra-biomolecular
tandem). The latter can be detected continuously with high sensitivity in enzyme assays. We will
present application examples for enzymatic decarboxylation and hydrolysis as well as an
inhibitor screening.
[1] Reymond, J. L., Enzyme Assays. Wiley-VCH: Weinheim, 2005.
[2] Tawfik, D. S.; Green, B. S.; Chap, R.; Sela, M.; Eshhar, Z., Proc. Natl. Acad. Sci. USA 1993,
90, 373-377.
[3] MacBeath, G.; Hilvert, D., J. Am. Chem. Soc. 1994, 116, 6101-6106.
[4] Hennig, A.; Roth, D.; Enderle, T.; Nau, W. M., ChemBioChem 2006, 7, 733-737.
[5] Hennig, A.; Florea, M.; Roth, D.; Enderle, T.; Nau, W. M., Anal. Biochem. 2007, 360, 255-
265.
PSA 74
Synthesis of large polyazamacrocycles by the [3+3] cyclocondensation
Jana Hodaová a,b , Petr Hadrava a , Jií Hlinka a
a Institute of Organic Chemistry and Biochemistry, Academy of Sciences of the Czech Republic,
Flemingovo nám. 2, 166 10 Prague 6, Czech Republic
b Department of Organic Chemistry, Institute of Chemical Technology, 166 28 Prague 6, Czech
Republic
Condensation reaction of diamines with dicarbonyl compounds has been frequently used
for the preparation of macrocyclic Schiff bases. Usually, the [2+2] cyclocondensation prevails
leading to the cyclic tetra Schiff bases. The macrocycle formation is often supported by use of a
metal cation template. [1-3] Only recently, the first example of the [3+3] cyclocondensation
product obtained in the absence of a template has been reported. [4] Finding that the reaction of
trans-1,2-diaminocyclohexane with terephthaldehyde affords the triangular [3+3] hexa Schiff
base in practically quantitative yield in the absence of a template [5,6] attracted our attention to
the [3+3] cyclocondensation reaction studies. Due to reversible character of the condensation
reaction, the equilibrium composition in solution can differ from that in the solid, which is
obtained upon evaporation of the solvent. Our studies have revealed that the rigid rod-like
dialdehydes give on the reaction with trans-1,2-diaminocyclohexane exclusively the [3+3] hexa
Schiff bases both in solution and in the solid phase. The cyclocondensations have been carried
out in the absence of a template, and without use of the high-dilution method. The results
indicate superior thermodynamic stability of the [3+3] cyclocondensation products.
The [3+3] cyclocondensation of the rigid rod-like aromatic dialdehydes with trans-1,2diaminocyclohexane
is a highly efficient atom-economy approach to the preparations of large
polyazamacrocycles. Various rigid dialdehydes have been used in the cyclisation reaction in
order to extend systematically the macrocycle size as well as to introduce the additional binding
sites into the macrocycle structure. Novel macrocyclic [3+3] hexa Schiff bases have been
reduced to the corresponding hexaamines. These macrocyclic polyamines can be utilised in
anion binding, [7] chiral molecular recognition, and catalysis.
[1] S. M. Nelson, Pure Appl. Chem. 1980, 52, 2461-2476.
[2] P. Guerriero, P. A. Vigato, D. E. Fenton, P. C. Hellier, Acta Chem. Scand. 1992, 46, 1025-
1046.
[3] W. Radecka-Paryzek, V. Patroniak, J. Lisowski, Coord. Chem. Rev. 2005, 249, 2156-2175.
[4] S. R. Korupoju, P. S. Zacharias, Chem. Commun. 1998, 1267-1268.
[5] J. Gawronski, H. Kolbon, M. Kwit, A. Katrusiak, J. Org. Chem. 2000, 65, 5768-5773.
[6] M. Chadim, M. Budšínský, J. Hodaová, J. Závada, P. C. Junk, Tetrahedron Asymmetry
2001, 12, 127-133.
[7] J. Hodaová, M. Chadim, J. Závada, J. Aquilar, E. García-España, S. V. Luis, J. F. Miravet,
J. Org. Chem. 2005, 70, 2042-2047.
The authors thank the Czech Science Foundation (Reg. No. 203/05/0702) for financial support
of this research project.