2.  Bignoniaceae family › 120 genera, 800 species  Found in Central and South America › Neotropical liana  Versatile plant › Horticulture, food, crafts, timber, dyes, rituals, and medicine Arevalo, et al. 2011, Gentry and Cook 1984, Gentry 1992, Witherup, et al. 1995

3.  M. Obovata and M. iquitosensis › 2 most known species › 9 species total for Martinella  M. iquitosensis known to produce martinelline and martinellic acid

4.  Chemically unique › Natural pyrroloquinoline ring structures › Synthesis is sought after  Biologically unique › Natural product (not peptide) that potently antagonize bradykinin receptors (GPCRs) Pyrroloquinoline quinone Davies, et al. 2013, Lovely and Mahmud 1999,, Ma, et al. 2001Ma, et al. 2003 Powell and Batey 2002, Shirai, et al. 2008, Yee, et al. 2006, Zhang, et al. 2007

5.  Bark removal and extraction  TLC and FCC › Four fractions  IR spectra  1H and 13 C NMR › 2D more helpful… Solvents Used Extraction: CH 2 Cl 2 /MeOH FCC: Hex/EtOAC gradient

7.  This proved to be more of a learning experience than productive  HPLC difficulties › Long method time for little purified product › Inconsistent results for first duplicate

8.  Solubility tests › H 2 O – no, EtOH (95%) – somewhat, DMSO (99.5%) – yes  Disk diffusion antibiotic sensitivity testing and MIC microtiter assays  Bacteria used › Escherichia coli, Pseudomonas aeruginosa, Bacillus cereus, and Staphylococcus aureus

9.  Disk diffusion showed that only NB1-6-1, NB1-6-6, and NB1-6-7 merited further investigation  Clear results: › EtOH (10.5%) + 1 mg/mL NB1-6-7 - S. aureus › EtOH (5.25%) +.5 mg/mL NB1-6-7 - B. cereus › DMSO (24.9%) + 1.5 mg/mL NB1-6-7 E. coli › DMSO (24.9%) + 1.5 mg/mL NB1-6-7 S. aureus

10.  Will run 2D NMRs overnight this week 2/3/14 › Hope to ID NB1-6-7 › Pyrroloquinoline ring in any sample?  If I had another semester › Bradykinin receptor antagonism investigation › LC-MS analysis to determine mass (no preexisting method)

11.  (1) Arevalo, C.; Ruiz, I.; Piccinelli, A.; Campone, L.; Rastrelli, L. Phenolic derivatives from the leaves of Martinella obovata (Bignoniaceae). Natural Product Communications, 2011, 6:7, 957-960.  (2) Gentry, A. H. A synopsis of Bignoniaceae ethnobotany and economic botany. Annals of the Missouri Botanical Garden, 1992, 79, 53-64.  (3) Gentry, A.H.; Cook, K. Martinella (Bignoniaceae): A widely used eye medicine of South America. Journal of Ethnopharmacology, 1984, 11, 337-343.  (4) Witherup, K.; Ransom, R.; Graham, A.; Bernard, A.; Salvatore, M.; Lumma, W.; Anderson, P.; Pitzenberger, S.; Varga, S. Martinelline and martinellic acid, novel G-protein linked receptor antagonists from the tropical plant Martinella iquitosensis (Bignoniaceae). Journal of the American Chemical Society, 1995, 117, 6682-6685.  (5) Ma, D.; Xia, C.; Jiang, J.; Zhang, J. First Total Synthesis of Martinellic Acid, a Naturally Occurring Bradykinin Receptor Antagonist. Organic Letters, 2001, 3:14, 2189-2191.  (6) Zhang, Z.; Zhang, Q.; Yan, Z.; Liu, Q. One-Step Synthesis of the Tricyclic Core of Martinellic Acid from 2-(Cyanomethyl)-3-oxo- N -arylbutanamides. Journal of Organic Chemistry, 2007, 72, 9808-9810.  (7) Ma, D.; Xia, C.; Jiang, J.; Zhang, J.; Tang, W. Aromatic Nucleophilic Substitution or CuI-Catalyzed Coupling Route to Martinellic Acid. Journal of Organic Chemistry, 2003, 68, 442-451.  (8) Davies, S.; Fletcher, Ai.; Lee, J.; Lorkin, T.; Roberts, P.; Thomson, J. Asymmetric Synthesis of (-)- Martinellic Acid. Organic Letters, 2013, 15:8, 2050-2053.  (9) Powell, D.; Batey, R. Total Synthesis of the Alkaloids Martinelline and Martinellic Acid via a Hetero Diels-Alder Multicomponent Coupling Reaction. Organic Letters, 2002, 4:17, 2913-2916.  (10) Yee Ng, Pui.; Masse, C.; Shaw, J. Cycloaddition Reactions of Imines with 3-Thiosuccinic Anhydrides: Synthesis of the Tricyclic Core of Martinellic Acid. Organic Letters, 2006, 8:18, 3999-4002.  (11) Shirai, A.; Miyata, O.; Tohnai, N.; Miyata, M.; Procter, D.; Sucunza, D.; Naito, T. Total Synthesis of (-)- Martinellic Acid via Radical Addition-Cyclization-Elimination Reaction. Journal of Organic Chemistry, 2008, 73, 4464-4475.  (12) Lovely, C.; Mahmud, H. An approach to the pyrroloquinoline core of martinelline and martinellic acid. Tetrahedron Letters, 1999, 40, 2079-2082.