Aptamers Developing to Methamphetamine
Marit Nilsen-Hamilton, Biochemistry, Biophysics and Molecular Biology George Kraus, Chemistry
Project goals - Aptamers will be selected that specifically recognize methamphetamine for the development of a highly sensitive fluorescence-based assay Specific aims: 1) isolate aptamers specific for methamphetamine 2) characterize the isolated aptamers. Long-term goal - to produce aptamers that can be used as a screening tools for a variety of drugs to provide forensics investigators with a rapid accurate screening procedures for common drugs
The Methamphetamine Problem in Iowa
Iowa was number one in meth lab seizures per capita in 2004 Meth makers use ammonia, lithium from batteries and pseudoephedrine from cold remedies Over 12,000 people in Iowa over age 12 have reported meth use
HO Ph NHMe HCl Me
Li
Ph
NH3
NHMe Me
Meth reaction
Design of Inhibitors of Meth Production Objectives: The goal of this project is to identify inexpensive and nontoxic inhibitors of the chemical reaction that is used to produce meth. History: The chemical reaction that is most commonly used to make meth in Iowa and other states across the Midwest involves readily available chemicals. The lithium (in batteries) and the pseudoephedrine are available from stores. The ammonia is stolen from ammonia tanks at co-ops and in farmers' fields.
HO Ph NH2 HCl
Li
Ph
NH3
NH2
Model system
1. 2. 3. 4.
Laboratory test at ISU on model system Test at Division of Criminal Investigation (Nila) Tests at Drug Enforcement Administration testing DOT
After over 100 compounds, we have discovered five inorganic salts that inhibit the reduction of a model system. Calcium nitrate is the most effective. Their results have been confirmed by scientists at the Iowa Division of Criminal Investigation using ephedrine. One of the inhibitors (ammonium nitrate) caused corrosion of the storage tank.
Synthesis of Tethered meth molecule
NO2 HO OH H2 N O NO2 O N H O OR O HO NHMe Me
Deprotect
NO2 H2N O O N H
O NHMe Me
Attach amine to column
Aptamers are single stranded nucleic acids with characteristics of antibodies
Made of RNA or DNA
Bind target molecules with high specificity and high affinity
ATP-binding aptamer
SELEX
SELEX: Systematic evolution of ligands by exponential enrichment
The ATP aptamer attached to column matrix separates all chemical forms of adenosine
Deng etal 2001 Anal Chem 73 5415-21
A G-quartet-forming aptamer separates dipeptides Trp-Arg from Arg-Trp
Charles and McGown 2002 Electrophoresis 23 1599-604
Aptamer that recognizes cocaine
Stojanovic et al. 2001 J Am Chem Soc 123 4928-31
Homogeneous aptamer assay for cocaine
Stojanovic etal 2001 J Am Chem Soc 123 4928-31
Homogeneous aptamer assay for cocaine
Cocaine
Metabolites
Benzoyl-ecgonine
Cocaine
ecgonine methyl ester
Stojanovic etal 2001 J Am Chem Soc 123 4928-31
Colorimetric aptamer assay for cocaine
Stojanovic and Landry 2002 J Am Chem Soc 124 9678-9
Colorimetric aptamer assay for cocaine
Benzoyl-ecgonine
ecgonine methyl ester
Stojanovic and Landry 2002 J Am Chem Soc 124 9678-9