COMMUNICATIONS
Table 1. Inhibition of p38 MAP kinase, cytokine release, and cytochrome P450 [4] a) G. Mloston, T. Gendek, H. Heimgartner, Helv. Chim. Acta 1998, 81,
isoforms byselected compounds.
1585 1595; b) R. Bartnik, W. Hahn, G. Mloston, Rocz. Chem. 1977, 51,
1747.
[5] S. Laufer, H.-G. Striegel, K. Neher (Merckle GmbH),
DE 19842833 A1, 1998 (Chem. Abstr. 2000, 132, 210154).
[6] H. Lettau, Z. Chem. 1970, 10, 462.
IC50 Æ SEM [mm][a]
Inhibition [%] of
P450 isoforms[b]
Compound p38
TNF-a
IL-1b
2D6
3A4
[7] a) T. Ravindranathan, R. D. Wakharkar, A. B. Landge, Org. Prep.
Proced. Int. 1986, 18, 95 98; b) L. Testaferri, M. Tiecco, M. Tingoli, D.
Chianelli, M. Montanucci, Synthesis 1983, 751 755.
[8] a) G. Mloston, M. Celeda, G. K. S. Prakash, G. A. Olah, H. Heimgart-
ner, Helv. Chim. Acta 2000, 83, 728 738; b) I. J. Ferguson, K. Schofield,
J. Chem. Soc. Perkin Trans. 1 1975, 275 277; c) D. Wang, J. Haseltine,
J. Heterocycl. Chem. 1994, 31, 1637 1639.
SB 203580 0.29 Æ 0.03 (7) 0.59 Æ 0.09 (21) 0.037 Æ 0.006 (20) 73.1
76.6
87.1
28.3
16.5
28.8
ML 3163
4b
4.0 Æ 1.0
2.2 (1)
1.1 Æ 0.4 (4)
2.2 Æ 0.9
0.38 Æ 0.13 (4)
0.45 Æ 0.03
71.8
7.8
4c
4d
0.50 (1)
2.2 (1)
0.51 Æ 0.24 (4) 0.11 Æ 0.03 (4)
13.4
0.7
1.1 Æ 0.3
0.38 Æ 0.04
[a] Tests were carried out in duplicate, except where the number in brackets denotes
otherwise. SEM Standard error of measurement. [b] Results are from one
experiment each, carried out at a test-compound concentration of 10 mm in
phosphate buffer (pH 7.4) with DMSO (0.1%).
[9] M. Spatzenegger, W. Jaeger, Drug Metab. Rev. 1995, 27, 397 417.
favorable cell-penetration properties. In the whole-blood
assay, IC50 values (mm) for the most active derivatives 4b
(TNF-a: 5.6 Æ 0.95, IL-1b: 1.5 Æ 0.7), 4c (TNF-a: 0.51 Æ 0.24,
IL-1b: 0.11 Æ 0.03), and 4d (TNF-a: 5.1 Æ 0.4, IL-1b: 1.1 Æ 0.7)
were lower than those of lead compound ML 3163 (TNF-a:
20.3 Æ 4.8, IL-1b: 2.78 Æ 0.13), and close to the nanomolar
range. Finally, the most promising results came from the
toxicityscreen, in which 4b d (Table 1) onlymoderately
interacted with those P450 isoforms most important for drug
metabolism.[9] This profile gives 4b d a clear advantage over
both SB 203580 and ML 3163, and makes them strong
candidates for further development as anti-inflammatory
drugs.
Tetrafunctional Photoaffinity Labels Based on
Nakanishi×s m-Nitroalkoxy-Substituted
Phenyltrifluoromethyldiazirine**
Mohammed Daghish, Lothar Hennig,
Matthias Findeisen, Sabine Giesa, Frank Schumer,
Horst Hennig, Annette G. Beck-Sickinger, and
Peter Welzel*
Photoaffinitylabeling has been demonstrated to be a
remarkablyefficient method for studying the interactions of
biologicallysignificant compounds (ligands) with their target
macromolecules.[1] The method allows the identification of the
targets (for example, binding proteins) and, also the binding
domain within the target protein. An appropriate photo-
affinity-labeling compound should contain three structural
elements:
a) a ligand which directs the label to the binding site on the
protein,
b) a photolabile group for attaching to the protein,
c) an indicator that allows the identification of the labeled
peptides after enzymatic digestion of the labeled protein.
Received: November 6, 2001 [Z18173]
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[*] Prof. Dr. P. Welzel, M. Daghish, Dr. L. Hennig
Institut f¸r Organische Chemie
Universit‰t Leipzig
Johannisallee 29, 04103 Leipzig (Germany)
Fax : (49)341-9736-559
Dr. M. Findeisen, Dr. S. Giesa
Institut f¸r Analytische Chemie
Universit‰t Leipzig
Johannisallee 29, 04103 Leipzig (Germany)
F. Schumer, Prof. Dr. H. Hennig
Institut f¸r Anorganische Chemie
Universit‰t Leipzig
Johannisallee 29, 04103 Leipzig (Germany)
Prof. Dr. A. G. Beck-Sickinger
Institut f¸r Biochemie
Universit‰t Leipzig
Talstrasse 33, 04103 Leipzig (Germany)
[**] We are grateful to Mrs. R. Herold and Mrs. R. Oehme for technical
assistance. Financial support bythe Deutsche Forschungsgemein-
schaft, BC Biochemie GmbH, and the Fonds der Chemischen
Industrie is gratefullyacknowledged.
Supporting information for this article is available on the WWW under
Angew. Chem. Int. Ed. 2002, 41, No. 13
¹ WILEY-VCH Verlag GmbH, 69451 Weinheim, Germany, 2002
1433-7851/02/4113-2293 $ 20.00+.50/0
2293