This is a medicinal chemistry proposal designed to develop new methods for inhibiting an important class of protease, those that are allosterically activated by substrate. Inhibitors of the botulinum toxin metalloproteases will be designed and synthesized based on two approaches: 1. design and synthesis of peptidyl transition-state analog inhibitors; and 2. Design based on the geometry of the zinc binding domain in the X-ray crystal structure of Botulinum toxin A. Botulinum toxin (BoNT) and tetanus toxin (TeNT) are converted in vivo to zinc metalloproteases which cleave single amide bonds in VAMP/synaptobrevin, SNAP25 and syntaxin. These metallo-proteases are the most selective proteases yet identified. Structure-activity data obtained with peptide substrates have established that substrate converts the BoNT metalloproteases from an inactive form to an active form. Examination of the 3.1 A X-ray crystal structure of native BoNT/A toxin provides insights into this unique substrate selectivity and also suggests how novel structures can be designed that will bind tightly to the INACTIVE protease, thereby preventing proteolysis. We postulate that it should be possible to stabilize the inactive form of the various BoNT metalloproteases; these inhibitors would be expected to have unique specificities and should not inhibit endogenous mammalian proteases. This new approach to the design of novel protease inhibitors will be applicable to other substrate activated proteases, many of which function in human biology and are implicated in disease.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
1R01GM059956-01
Application #
2898365
Study Section
Bio-Organic and Natural Products Chemistry Study Section (BNP)
Project Start
1999-09-01
Project End
2003-08-31
Budget Start
1999-09-01
Budget End
2000-08-31
Support Year
1
Fiscal Year
1999
Total Cost
Indirect Cost
Name
University of Wisconsin Madison
Department
Type
Schools of Pharmacy
DUNS #
161202122
City
Madison
State
WI
Country
United States
Zip Code
53715
Brewer, Matthias; James, Clint A; Rich, Daniel H (2004) Synthesis of a tripeptide derivative containing the gln-arg hydroxyethylene dipeptide isostere. Org Lett 6:4779-82
Sukonpan, C; Oost, T; Goodnough, M et al. (2004) Synthesis of substrates and inhibitors of botulinum neurotoxin type A metalloprotease. J Pept Res 63:181-93
Oost, Thorsten; Sukonpan, Chanokporn; Brewer, Matthias et al. (2003) Design and synthesis of substrate-based inhibitors of botulinum neurotoxin type B metalloprotease. Biopolymers 71:602-19
Brewer, Matthias; Oost, Thorsten; Sukonpan, Chanokporn et al. (2002) Sequencing hydroxyethylamine-containing peptides via Edman degradation. Org Lett 4:3469-72
Brewer, M; Rich, D H (2001) Synthesis of a tripeptide derivative containing the Phe-Arg hydroxyethylene dipeptide isostere. Org Lett 3:945-8