201600030 FULL PAPER (+)-N-Formylnorglaucine Rotamers from Unonopsis stipitata DIELS by Felipe M. A. da Silva* a ), Francinaldo A. da Silva Filho a ), Bruna R. de Lima a ), Richardson A. de Almeida a ), Dacio M. Mendonc ßa b ), Raimundo C. Pereira Junior a ), L ıvia M. Dutra c ), Andersson Barison c ), Hector H. F. Koolen d ), Afonso D. L. de Souza a ), and Maria Lucia B. Pinheiro a ) a ) Department of Chemistry, Federal University of Amazonas, Manaus-AM, 69077-000, Brazil (phone: +55 92 982517341; e-mail: felipemas@ufam.edu.br) b ) Amazon Biotechnology Center, Manaus-AM, 69075-351, Brazil c ) NMR Center, Federal Universisty of Parana, Curitiba-PR, 81531-990, Brazil d ) DeMpSter Mass Spectrometry Group, Amazonas State University, Manaus-AM, 69050-010, Brazil (+)-N-formylnorglaucine (1), an aporphine alkaloid containing a formyl group linked to the heterocyclic nitrogen, was isolated from the leaves of Unonopsis stipitata, an Amazon medicinal plant. The chemical structure was characterized based on 1D- and 2D-NMR spectroscopy and HR-ESI-MS. NMR spectra revealed that 1 is composed of two rotamers (1a and 1b) in a ratio of approximately 2:1. In addition, the fragmentation behavior of 1 displayed an unusual fragmentation pattern compared to regular aporphine alkaloids. Thus, this compound is reported for the first time as a natural product in this study. Keywords: Unonopsis stipitata, Aporphine alkaloid, Rotamers, (+)-N-Formylnorglaucine, Annonaceae. Introduction Unonopsis (Annonaceae) is a neotropical genus with broad distribution through the Amazon region [1]. Some species of Unonopsis have been described due to its medicinal applications [2][3], including Unonopsis stipitata whose pulverized leaves are employed in the treatment of brain disorders [3]. A recent study indicated that U. stipi- tata is a promising source of aporphine alkaloids [4], which are extensively explored in synthetic studies due to promising biological activities observed for several apor- phinoid structures [5 8]. Besides its attractive biological properties, the fast recognition and dereplication of this class in complex matrices by mass spectrometry (MS) has accelerated the research for new compounds [9 11]. This favorable sit- uation is possible due to the advances on the gas-phase chemistry of aporphines through key fragmentation recognition via collision-induced dissociation (CID) experiments [12]. Aiming the discovery of new alkaloids from Amazon medicinal plants, we carried out a chemical investigation of the leaves of U. stipitata, which resulted in the isolation of a new natural aporphine compound. In the present work, we chemically characterized this substance through 1D- and 2D-NMR techniques and MS. In addition, the fragmentation behavior by CID was investigated through high-resolution MS. Results and Discussion Structure Elucidation Compound 1 was obtained as a yellow amorphous solid with molecular formula C 21 H 24 NO 5 (m/z 370.1627), as determined by HR-ESI-MS. The 1 H-NMR spectrum indi- cated duplicate aromatic, CH, methoxyl, and CH 2 signals in CDCl 3 (Tables 1 and 2) at a ratio of approximately 2:1, suggesting the presence of two rotameric isomers (1a and 1b) [13 15]. This ratio was estimated based on the inte- gration of aromatic signals. Therefore, each signal of the two rotamers was individually assigned. In the 1 H-NMR spectrum, signals of eight deshielded H-atoms were observed at d(H) 6.62 (s,HC(3)), 6.80 (s, HC(8)), 8.14 (s,HC(11)), and 8.26 (s,NCHO), which are relative to the main rotamer 1a, and at d(H) 6.65 (s, HC(3 0 )), 6.77 (s,HC(8 0 )), 8.15 (s,HC(11 0 )), and 8.39 (s,NCHO’) for the minor rotamer 1b. Besides, four MeO groups were observed at d(H) 3.68 (s, MeO C(1)), 3.91 (s, MeOC(2)), 3.91 (s, MeOC(10)), and 3.92 (s, MeOC(9)) for 1a, whereas MeO group signals at d (H) 3.67 (s, MeOC(1 0 )), 3.91 (s, MeOC(2 0 )), 3.92 (s, MeOC(10 0 )), and 3.94 (s, MeOC(9 0 )) were attributed to 1b. A typical CH signal was observed at d(H) 4.92 (dd, J = 14.2, HC(6a)) for 1a and d(H) 4.49 (dd, J = 14.4, 4.5, HC(6a 0 )) for 1b, suggesting that 1 has an aporphine skeleton [10][13 15]. Through HSQC experiments, it DOI: 10.1002/hlca.201600030 © 2016 Verlag Helvetica Chimica Acta AG, Zurich Helv. Chim. Acta 2016, 99, 1–5 1