2009 Annual Report 1a.Objectives (from AD-416) 1) To synthesize N-coumaroyltyramine and its analogues and to identify their cellular effects and molecular mechanisms, particularly as related to cancer modulation,. 2)To assess anti-cancer and other health effects N-coumaroyltyramine and its analogues in an animal model. 1b.Approach (from AD-416) 1. Synthesis of phenylpropenic acid conjugates (N-coumaroyltyramine and its analogues). 2. Measurement of phenylpropenic acid conjugates in cell and biological samples. 3. Investigation of effects of phenylpropenic acid conjugates on cell cycle and growth of cancer cells. 4. Apoptosis study. 5. Hydrogen peroxide quenching study. 6. Investigation of effects of phenylpropenic acid conjugates on MAPK Kinases. 7. Investigation of effects of phenylpropenic acid conjugates on protein tyrosine kinase including EGFR. 8. Investigation of effects of phenylpropenic acid conjugates on xenograft tumor in nude mice. 3.Progress Report Several biological activities of synthesized and isolated phenolic conjugates found in coffee, cocoa, and tea (N-caffeoyltyramine, N-coumaroyltyramine, N-caffeoyldopamine, N-caffeoyltryptamine and their analogues) were evaluated using the Jurkat and U937 cell lines, and cells isolated from mouse blood. P-selectin is a membrane adhesion molecule involved in platelet-leukocyte interactions and platelet-endothelium interactions via binding to the P-selectin ligand (PSGL-1) on leukocytes and endothelium. Phenolic conjugates were able to suppress the expression of P-selectin by inhibiting the inflammatory pathway initiated by cyclooxygenase; however, the conjugates had no effect on the expression of PSGL-1. During the study, preliminary data about potential effects of the tested phenolic conjugates on other adhesion molecules, including CD61 (Integrin beta3), were also obtained. In addition, a study completed in mice showed bioavailability of caffedymine-type phenylpropenoic acid amides that inhibited P-selectin, but their presence had no long-term effects on body weight, food, water consumption, behavior, or toxicity. Over the lifespan of this project, phenolic conjugates were shown to activate cell signals via receptor binding. Phenolic conjugates found in coffee, cocoa, and tea bind to adrenoceptors and serotonin receptors, producing cAMP or suppressing the expression of interleukin-2, respectively. Caffedymine-type phenylpropenoic acid amides were able to increase cAMP production and suppress formation of P-selectin that regulates platelet aggregation, by the inhibition of cycloxygenase (COX) enzymes. Safflomide-type phenylpropenoic acid amides were found to bind to serotonin receptors resulting inhibiting formation of interleukin-2. These are both important factors in blood clot formation and cardiovascular disease. 4.Accomplishments 1. Phenolic conjugates activate cell signals via receptor binding. Phenolic conjugates found in coffee, cocoa, and tea bind to adrenoceptor, as well as serotonin receptor, thereby, producing cAMP or suppressing the expression of interleukin-2, respectively. Caffedymine-type phenylpropenoic acid amides were able to increase cAMP production and suppress the expression of P-selectin that regulates platelet aggregation, by the inhibition of cycloxygenase (COX) enzymes. Safflomide-type phenylpropenoic acid amides were found to bind to serotonin receptors, thereby, inhibiting the expression of interleukin-2. 6.Technology Transfer Number of the New/Active MTAs (providing only) 1 Number of Web Sites Managed 1 Review Publications Park, J.B. 2008. 5-Caffeoylquinic acid and caffeic acid orally administered suppresses P-selectin expression on mouse platelets. Journal of Nutritional Biochemistry. 20(10):800-805. Available http://dx.doi.org/10.1016/j.jnutbio.2008.07.009.