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Evaluation of Pacific Northwest Native Plants For the Presence of ACE Inhibitors Andrea M Garcia Working with Dr. Joe Karchesy.

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Presentation on theme: "Evaluation of Pacific Northwest Native Plants For the Presence of ACE Inhibitors Andrea M Garcia Working with Dr. Joe Karchesy."— Presentation transcript:

1 Evaluation of Pacific Northwest Native Plants For the Presence of ACE Inhibitors Andrea M Garcia Working with Dr. Joe Karchesy

2 What is ACE ACE, Angiotensin Converting Enzyme ACE, Angiotensin Converting Enzyme An exopeptidase An exopeptidase that catalyses the conversion of Angiotensin I to Angiotensin II, in the Renin- Angiotensin pathway

3 Renin- Angiotensin pathway

4 What is the Renin- Angiotensin pathway Angiotensinogen Renin Renin Angiotensin I ACE ACE Angiotensin II Hypertension

5 High blood pressure is considered to be a silent killer. Nearly one in three U.S. adults have high blood pressure. 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. 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. Once high blood pressure develops, it usually lasts a lifetime and it can be prevented and controlled by taking action.

6 Preventive measures Synthetic drugs Synthetic drugs Exercise and good eating habits. Exercise and good eating habits. Use of natural products, teas or supplements. Use of natural products, teas or supplements. Plant products that exhibit ACE inhibition. Plant products that exhibit ACE inhibition. hawthornberries hawthornberries

7 What are ACE inhibitors? A group of pharmaceuticals or compounds that slow or inhibit the activity of ACE. 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. 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.

8 Ace inhibitors in plants There are various secondary metabolites with inhibitory activity towards ACE. There are various secondary metabolites with inhibitory activity towards ACE. Found in flavonoids, proanthocyanidins and small peptides. Found in flavonoids, proanthocyanidins and small peptides. There are well known plant extracts that have been used by indigenous peoples. There are well known plant extracts that have been used by indigenous peoples.

9 Reason for investigating ACE inhibition in plants Herb drug interactions. Herb drug interactions. Low cost compared to synthetic drugs. Low cost compared to synthetic drugs. They have a wide range of therapeutic use and are suitable for chronic treatments They have a wide range of therapeutic use and are suitable for chronic treatments Find better and more selective compounds. Find better and more selective compounds. Possibility of indicating new molecular structure with ACE activity. Possibility of indicating new molecular structure with ACE activity.

10 List of plants under study that we think may contain ACE inhibition Black Cohosh Cimicifuga elata Baneberry Actea Rubra Prince Pine Pipsisewa Ocean Spray Holodiscis discolor Red Alder Alnus Rubra Devil’s Club Oplopanax Horridum Soapberry Shepherdia canadenis Buckbrush Amelanchier Alnifolia

11 In Vitro Screening of Inhibitors Collect specimens of plants to be studied. Collect specimens of plants to be studied. Isolate compounds. Isolate compounds. React plant ACE inhibitors with rabbit lung enzyme assay. React plant ACE inhibitors with rabbit lung enzyme assay. Test for ACE activity, using the HPLC method (Hansen et al. 1995). 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. Further testing for the plants that exhibit ACE compounds will be done for us through the college of pharmacy.

12 Reaction Use a Titer plate Use a Titer plate –Rabbit lung ACE enzyme –Plant solution –Dansyltriglycine substrate Incubate Incubate Stop the reaction and analyze with HPLC. Stop the reaction and analyze with HPLC.

13 ACE – Catalyzed Cleavage of Dansylytriglycine. Dansyltriglycine (dansyl- Gly-Gly-Gly), UV labeled tripeptide. Dansyltriglycine (dansyl- Gly-Gly-Gly), UV labeled tripeptide. Presence of ACE Presence of ACE Dansylglycine (dansyl- Gly) Dansylglycine (dansyl- Gly) Diglycine (Gly-Gly) Diglycine (Gly-Gly) From Elbl and Wagner, 1991

14 High Performance Liquid Chromatography Dansyl-L-glutamine: the internal standard Dansyl-L-glutamine: the internal standard Dansylglycine, product of enzyme reaction Dansylglycine, product of enzyme reaction Dansyltriglycine, substrate of enzyme reaction. Dansyltriglycine, substrate of enzyme reaction. Ratio of the two is the measurement of inhibition. Ratio of the two is the measurement of inhibition. From Hansen etal, 1995

15 Status Analysis currently underway for ACE inhibition Analysis currently underway for ACE inhibition Promising compounds will be forwarded to Pharmacy for further characterization. Promising compounds will be forwarded to Pharmacy for further characterization.

16 Acknowledgements Acknowledgements Jaworski Scholarship, HHMI, Garden Club of America Jaworski Scholarship, HHMI, Garden Club of America Joe Karchesy, Kevin Ahern, Debbie Maynard


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