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Evaluation of Pacific Northwest Native Plants For the Presence of ACE Inhibitors Andrea M Garcia Working with Dr. Joe Karchesy
What is ACE • ACE, Angiotensin Converting Enzyme • An exopeptidase that catalyses the conversion of Angiotensin I to Angiotensin II, in the Renin- Angiotensin pathway
What is the Renin- Angiotensin pathway Angiotensinogen Renin Angiotensin I ACE Angiotensin II Hypertension
High blood pressure is considered to be a silent killer. • Nearly one in three U.S. adults have high blood pressure. • Uncontrolled high blood pressure can lead to stroke, heart attack, heart failure or kidney failure. • Once high blood pressure develops, it usually lasts a lifetime and it can be prevented and controlled by taking action.
Synthetic drugs Exercise and good eating habits. Use of natural products, teas or supplements. Plant products that exhibit ACE inhibition. hawthornberries Preventive measures
What are ACE inhibitors? • A group of pharmaceuticals or compounds that slow or inhibit the activity of ACE. • Based on research from the venom of a Brazilian pit viper that led to the development of the first ACE inhibitor drug to treat hypertension.
Ace inhibitors in plants • There are various secondary metabolites with inhibitory activity towards ACE. • Found in flavonoids, proanthocyanidins and small peptides. • There are well known plant extracts that have been used by indigenous peoples.
Reason for investigating ACE inhibition in plants • Herb drug interactions. • Low cost compared to synthetic drugs. • They have a wide range of therapeutic use and are suitable for chronic treatments • Find better and more selective compounds. • Possibility of indicating new molecular structure with ACE activity.
In Vitro Screening of Inhibitors • Collect specimens of plants to be studied. • Isolate compounds. • React plant ACE inhibitors with rabbit lung enzyme assay. • Test for ACE activity, using the HPLC method (Hansen et al. 1995). • Further testing for the plants that exhibit ACE compounds will be done for us through the college of pharmacy.
Reaction • Use a Titer plate • Rabbit lung ACE enzyme • Plant solution • Dansyltriglycine substrate • Incubate • Stop the reaction and analyze with HPLC.
ACE – Catalyzed Cleavage of Dansylytriglycine. • Dansyltriglycine (dansyl-Gly-Gly-Gly), UV labeled tripeptide. • Presence of ACE • Dansylglycine (dansyl-Gly) • Diglycine (Gly-Gly) From Elbl and Wagner, 1991
High Performance Liquid Chromatography • Dansyl-L-glutamine: the internal standard • Dansylglycine, product of enzyme reaction • Dansyltriglycine, substrate of enzyme reaction. • Ratio of the two is the measurement of inhibition. From Hansen etal, 1995
Status • Analysis currently underway for ACE inhibition • Promising compounds will be forwarded to Pharmacy for further characterization.
Acknowledgements • Jaworski Scholarship, HHMI, Garden Club of America Joe Karchesy, Kevin Ahern, Debbie Maynard