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Iridoids from Hymenodictyon floribundum

Abstracts

Two new iridoids floribundane A (1) and floribundane B (2) and the known compounds lupenone (3), scopoletin (4) and 4,5-dihydroblumenol A (5) were isolated from trunk bark and leaves of Hymenodictyon floribundum B.L. Rob. The structures of all compounds were established from their spectroscopic data.

Hymenodictyon floribundum; monoterpenes; secoiridoids; nitrogen-containing iridoid; C-9 iridoid


Das cascas do tronco e das folhas do Hymenodictyon floribundum B. L. Rob foram isolados dois novos iridoides, floribundano A (1) e floribundano B (2), e três compostos conhecidos, lupenona (3), escopoletina (4) e 4,5-di-hidroblumenol A (5). As estruturas de todos os compostos foram estabelecidas com base nos seus dados espectroscópicos.


ARTICLE

Iridoids from Hymenodictyon floribundum

Cristina M. P. BorgesI; Carlos DiakanawmaII; Dina I. M. D. de MendonçaIII,* * e-mail: disabel@ubi.pt

IDepartamento de Química, Universidade Agostinho Neto, Av. 4 de Fevereiro, 71, 2º andar, CP 3244, Luanda, Angola

IIDepartamento de Biologia, Universidade Agostinho Neto, Av. 4 de Fevereiro, 71, 2º andar, CP 3244, Luanda, Angola

IIIDepartamento de Química, Universidade da Beira Interior, Rua Marquês d'Ávila e Bolama, 6200-001 Covilhã, Portugal

ABSTRACT

Two new iridoids floribundane A (1) and floribundane B (2) and the known compounds lupenone (3), scopoletin (4) and 4,5-dihydroblumenol A (5) were isolated from trunk bark and leaves of Hymenodictyon floribundum B.L. Rob. The structures of all compounds were established from their spectroscopic data.

Keywords:Hymenodictyon floribundum, monoterpenes, secoiridoids, nitrogen-containing iridoid, C-9 iridoid

RESUMO

Das cascas do tronco e das folhas do Hymenodictyon floribundum B. L. Rob foram isolados dois novos iridoides, floribundano A (1) e floribundano B (2), e três compostos conhecidos, lupenona (3), escopoletina (4) e 4,5-di-hidroblumenol A (5). As estruturas de todos os compostos foram estabelecidas com base nos seus dados espectroscópicos.

Introduction

The Hymenodictyon genus comprises 22 species. Of these, 11 are endemic to Madagascar, 4 to Asia and 7 to Tropical Africa.1Hymenodictyon floribundum B.L. Rob. (Rubiaceae), endemic to Tropical Africa, is a small tree that grows in the mountains of the Huíla province and its traditional name is NDambi Yov'olwi, (omu)Lia-tyimeme. Its trunk bark is used in Angola folk medicine to treat fever.2

A previous study has shown that the trunk bark of this tree contains scopoletin, hymeselsin, scopolin and 3-O-β-D-glucopyranosyl-β-sitosterol.3 An exhaustive literature review revealed that, with the exception of an ongoing study examining the trunk bark and dried leaves of Hymendictyon floribundum (in the context of other medicinal plants of Angola),4 other parts of the tree have not yet to be studied.

In the aforementioned study, powdered trunk barks and dried leaves were extracted by maceration with methanol. The methanol extract was partitioned with hexane and chloroform. The hexane extract from trunk barks yielded lupenone (3)5 and the chloroform extract yielded scopoletin (4).6 The chloroform extract from dried leaves yielded floribundane A (1), 4,5-dihydroblumenol A (5)7,8 and floribundane B (2).

Results and Discussion

Floribundane A (1) was obtained as a colourless oil with = +25.31º. The HR-TOF- EI-MS showed a molecular ion peak at m/z 223.0843 [M]+(in agreement with the molecular formula C11H13NO4) implying 6 degrees of unsaturation. The UV maximum at 234 nm and the IR absorptions at 1724, 1633 and 1268 cm-1 suggested the presence of the chromophore -OOCC=CHO.9 The IR also showed the presence of a lactam ring: NH band (3403 cm-1), and the amide band I (1690 cm-1) with an absence of the amide II band.10 The 1H NMR spectrum of compound (1) (Table 1) displayed signals of two vinylic protons (H-3 δH 7.50, s and H-8 δH 6.85, q, J 7.2), a ABX system [H-6α, β δH 2.11, dd, J13.1 and 2.0; δH 2.06, dd, J13.1 and 2.6 (AB part) and H-5 δH 3.94, m (X part)] and a vinyl methyl group at δH 2.01 (d, J 7.2); typical signals of a secoiridoid nucleus.11 A few iridoid alkaloids have been isolated from natural sorces12-15 (gardenamide class), and some were obtained as metabolites produced by human16 and rat intestinal bacteria.17 Comparison with the published data of known iridoid alkaloids, such as (6) (Figure 1), and hemiacetal-secoiridoids11,18 like (7) (Figure 1) and compound (1), showed that the H-3 chemical shift (δH 7.50 ppm), despite the presence of a nitrogen atom in the molecule, was more compatible with the hemiacetal-secoiridoids (δHca.7.5 ppm) than with the gardenamide class and mor-117 (8) (Figure 1) (δHca. 7.3 ppm). Further, the upfield shift of H-1 (δH 5.46) relative to the corresponding signal of mor-1 (δH 5.11 ppm),17 and a downfield shift (δH 5.94 ppm) to the corresponding hemicetals,11,18 showed that (1) it was neither a hemiacetal nor a nitrogen-containing iridoid (N-2) but that it was compatible with a C-1(N)O-2 substitution. This was also supported by the presence of the 1H-1H COSY correlation between H-1/NH, rather than the H-1/NH, H-3/NH correlations expected in the iridoid alkaloids,17 and by UV and IR data indicating the presence of the chromophore -OOCC=CHO.9 The 13C NMR spectrum (Table 1) showed 11 carbons which were analysed as two methyls (one vinylic and a methyl ester), one methylene, four methines two of them vinylic, and four quaternary carbons (two vinylic and two carbonylic) from its DEPT spectrum. The 13C NMR chemical signals at δC 152.13 (C-3), 136.11 (C-8), 130.25 (C-9), the carbonyl group at δC 166.56, the methoxy signal at δC 51.31 and the vinyl methyl group at δC 13.96, were found to be consistent with a secoiridoid skeleton.11 The unusual downfield shift of C-1 to δC 76.97 was in agreement with the C-1(O)(N) substitution as observed on mor-1,17 but only the presence of O in the position 2 of the iridoid skeleton could explain the upfield shift of C-3 and C-4. The HMBC correlation between H-1/C-7 established the presence of the 1,7-d-lactam ring in the molecule. With the aid of 1H - 1H COSY, HSQC and HMBC data (Table 1, Figure 2) it was possible to establish the structure of (1) as a 7,8-secoiridoid, with C-11 as a methyl ester, a C-1(N)O-2 substitution and a 1,7-d-lactam ring. The NOESY correlations Ha-5/ Ha-6 and Hb-6/Hb-1 indicated that the 1,7-d-lactam ring is trans-fused. To our best knowledge, this is the first time that a secoiridoid has presented as a 1,7-trans-d-lactam ring. A similar structure, isosweroside, as a hemiacetal, was found in the roots of Sambucus ebulus L.19 Compound (1) was named floribundane A.



Floribundane B (2) was obtained as a colourless oil with = + 41.66º. The molecular formula C9H14O3 was established by HR-TOF-MS-EI which showed a molecular ion peak at m/z 170.0942 [M]+ (calculated for 170.2270), implying 3 degrees of unsaturation. The IR showed OH bands (3384 cm-1), an α,β-unsaturated lactone group (1692 cm-1, 1620 cm-1); the UV spectrum a α,β-unsaturated lactone system (230 nm). The 1H RMN spectrum (Table 1) showed signals of one diastereotopic oxymethylene (H-3) δH 4.23 (ddd, J111.3, J29.1, J34.4 Hz) and δH 4.42 (ddd, J111.3, J29.5, J33.6 Hz) that presented 1H - 1H COSY correlation with a diastereotopic methylene (H-4) δH 1.83 (m) and δH 2.12 (m); a oxymethylene multiplet δH 3.71 (H-7) that presented an 1H - 1H COSY correlation with another methylene δH 1.74 (m, H-6); a methine δH 3.18 (m, H-5) that showed cross peaks with H-4 and H-6 and a vinyl methine δH 7.02 (qd, J17.4, J21.2 Hz, H-8) coupled with a methyl δH 1.86 (d, J7.4 Hz, Me-10). This spectrum presents some similarities to that of compound (1): the presence of the vinyl double bond and the 1H - 1H COSY correlations between H-5, H-6 and H-7. Thus, it appears that the A ring, as in (1), is open as a 7,8-secoiridoid with a hydroxyl moiety at C-7. The 13C NMR spectrum (Table 1) showed 9 carbons which were analysed, from its DEPT spectrum, as one vinylic methyl (δC 14.18), four methylenes (two of them oxymethylenes, δC 27.51, 36.11, 59.97, 65.35), two methines (one vinylic, δC 28.93, 140.67), and two quaternary carbons (one vinilyc, δC 131.16, and the other carbonylic, δC 167.38). The lack of the two quaternary sp2 carbons belonging to the C-3 - C-4 double bond of (1) and the methine C-1, and the presence of signals characteristic of a methylene (δC 27.51), a oxymethylene (δC 65.35) and a sp2 carbon (δC 167.38) belonging to the α,β-unsaturated lactone group detected in the IR and UV spectra, is indicative of a major transformation in the B ring of (2). However, these new features of the B ring are in good agreement with those of the δ-lactone ring of C-9 iridoid derivatives viteoid II,20,21 7-hidroxyviteoid II22 and ovatolactone.23 The vinylic double bond can unambiguously be assigned to C-8 and C-9 and the lactone carbonyl to C-1, by means of 1H - 1H COSY, HSQC and HMBC data (Table 1, Figure 2). Several C-9 iridoid and secoiridoid (lactone ring opening) derivatives are known20-25 but to our best knowledge this is the first time that a C-9 7,8-secoiridoid derivative has been isolated. Thus, (2) was identified as a new natural product for which we propose the name floribundane B.

Conclusions

Although trunk bark components of H.floribundum have been well studied, there has as yet been no study which focuses solely on the leaves of this medicinal tree. The major compound trunk bark extract is scopoletin. The occurrence of iridoids has been observed for the first time in this genus. A new secoiridoid alkaloid, floribundane A, and a C-9 7,8-secoiridoid, floribundane B was isolated from the leaves of this tree.

Experimental

General experimental procedures

Optical rotations were obtained with a Bellingham+Stanley Ltd ADP 220 polarimeter. HREIMS measurements were carried out on a VG Autospec M and recorded at 70 eV. FTIR and UV spectra were measured in a Unicam Mattson 5000 FTIR and Unican Helios a respectively. NMR spectra were recorded in a Bruker Avance II, 600 MHz (1H NMR) and 150.9 MHz (13C NMR), in CDCl3. Chemical shifts are given in d ppm and are referenced to the residual CHCl3, 7.26 ppm for the 1H and 77.0 ppm for 13C. Two-dimensional experiments were performed with standard Bruker software. Column chromatography was carried out on silica gel (silica gel 60 (70-230 mesh), Merck, Darmstadt, Germany).

Plant material

Leaves and bark of Hymenodictyon floribundum were collected in the waterfall at the Comuna da Huíla, Huíla province, Angola in July 2001. A voucher specimen (3668) has been deposit at the Lubango Herbarium, Angola.

Extraction and isolation

Powdered trunk bark (3 kg) was extracted with methanol for a week at room temperature. The methanol extract was partitioned between MeOH-H2O (5:1) and hexane yielding 16.6 g of the hexane extract. The aq. methanolic fraction was concentrated, H2O added and extracted with chloroform to obtain the chloroform extract (36.8 g). A sample of the hexane extract (4.7 g) was fractionated in silica gel column with a hexane/EtOAc, EtOAc and EtOAc/MeOH gradients. The fraction eluted with hexane/EtOAc (95:5) was separated in silica gel column with a hexane/EtOAc gradient to yield lupenone (3) (20.3 mg). From the chloroform extract and by crystallization scopoletin (4) (30 g) was isolated.

Dried leaves (0.9 kg) were extracted with methanol for a week at room temperature. The methanol extract was partitioned between MeOH-H2O (5:1) and hexane yielding 34.02 g of the hexane extract. The aq. methanolic fraction was concentrated, H2O added and extracted with chloroform to obtain the chloroform extract (5.2 g). A sample of the chloroform extract (1.6 g) was fractionated in silica gel column with a hexane/EtOAc, EtOAc/CHCl3 and EtOAc/MeOH gradients. Two fractions were eluted with EtOAc/CHCl3 (7:3) (fraction I and II). Fraction I was separated in silica gel column with CHCl3 and a CHCl3/MeOH gradient to yield floribundane A (1) (10.2 mg) and 4,5-dihydroblumenol A (5) (5.6 mg). Fraction II was separated in silica gel column with CHCl3 and a CHCl3/MeOH gradient to yield floribundane B (2) (7.8 mg).

Floribundane A (1)

Oil; = +25.31º (CHCl3, c 0.079). FTIR (film) νmax/cm-1: 3403, 2959, 2930, 1724, 1690, 1633, 1379, 1268, 1196, 1092, 943, 756; UV (MeOH) λmax/nm (log e): 234 (4.00), 210.0 (4.15) 1H NMR (CDCl3, 600 MHz) and 13C NMR (CDCl3, 150.9 MHz): see Table 1; TOF-MS-EI pos: m/z 223 [M]+ (15), 206 (77), 190 (100), 164 (62), 163 (12), 162 (11), 149 (28), 146 (22), 134 (11), 122 (23), 121 (82), 120 (13), 93 (13), 83 (13), 79 (10); HREIMS: m/z 223.0843 [M]+ (calc. for C11H13NO4, 223.0845).

Floribundane B (2)

Oil; = +41.66º (CHCl3, c 0.048). FTIR (film) νmax/cm-1: 3384, 2962, 2926, 2866, 1692, 1620, 1262, 1154, 1094, 1060, 1026, 800, 760; UV (MeOH) λmax/nm (log e): 230 (3.16), 219 (3.34). 1H NMR (CDCl3, 600 MHz) and 13C NMR (CDCl3, 150.9 MHz) see Table 1; TOF-MS-EI pos: m/z 226 [M]+ (1), 168 (25), 137 (11), 135 (12), 128 (19), 126 (39), 125 (31), 124 (34), 123 (38), 122 (11), 121 (24), 111 (100), 110 (29), 109 (73), 108 (12), 107 (22), 99 (15), 98 (17), 97 (33), 96 (17), 95 (72), 93 (17), 91 (14), 85 (81), 84 (11), 83 (11), 83 (32), 82 (19), 81 (40), 79 (17), 77 (12), 71 (39), 69 (46), 67 (20); HREIMS: m/z 170.0942 [M]+ (calc. for C9H14O3, 170.2270).

Acknowledgments

This work was partially funded by the project POCTI/QUI/39380/2001 of Fundação para a Ciência e Tecnologia (FCT) with FEDER funding and Textile and Paper Materials Center. One of the authors (C.B.) gratefully acknowledges a GRICES PhD scholarship and INABE (Angola) for financial support.

Received: February 25, 2009

Web Release Date: March 22, 2010

Supplementary Information

1H NMR, 13C NMR, 1H-1H COSY, HSQC, HMBC, and NOESY NMR spectra of compounds 1 and 2 are available free of charge at http://jbcs.sbq.org.br, as PDF file.

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  • 1. Razafimandimbison, S. G.; Bremer, B.; Bot. J. Linn. Soc 2006, 152, 331.
  • 2. Bossard, E.; Medicine Traditionnelle au Centre et a l'Ouest de l'Angola, Vol. 1, Ministério da Ciência e Tecnologia, Instituto de Investigação Científica Tropical: Lisbon, 1996.
  • 3. Mitaine-Offer, A. C.; Tapondjou, L. A.; Djoukeng, J. D.; Bouda, H.; Lacaille-Dubois, M. A.; Biochem. Syst. Ecol. 2003, 31, 227.
  • 4. dos Santos, A. F.; Lopes, L. A.; Mata, R. C. S.; de Mendonça, D. I. M. D.; Sant'Ana, A. E. G.; Bioresour. Technol. 2007, 98, 135.
  • 5. Atta-Ur-Rahman; Pentacyclic Triterpenoids, Vol II, Elsevier Science B.V.: Amsterdam, 1994.
  • 6. Rao, P. S.; Asheervadam, Y; Khaleelullah, M.; Rao, N. S.; Murray, R. D. H.; J. Nat. Prod. 1988, 51, 959.
  • 7. de Marino, S.; Borbone, N.; Zollo, N.; Ianaro, A.; Di Meglio, P.; Iorizzi, M; J. Agric. Food Chem. 2004, 52, 7525.
  • 8. Gonzalez, A. G.; Guillermo, J. A.; Ravelo, A. G.; Jiménez, I. A.; Gupta, M. P.; J. Nat. Prod. 1994, 57, 400.
  • 9. Zuleta, L. M. C.; Cavalheiro, A. J.; Silva, D. H. S.; Furlan, M.; Young, M. C. M.; Albuquerque, S.; Castro-Gamboa, I.; Bolzani, V. S.; Phytochemistry 2003, 64, 549.
  • 10. Pretsch, E.; Seilb, J.; Simon, W.; Tablas para la Determinación Estrutural por Métodos Espectroscópicos, Springer-Verlag Ibérica:Madrid, 1998, p. I150.
  • 11. Shen, Y. C.; Lin, S. L.; Chein, C. C.; Phytochemistry 1996, 42, 1629.
  • 12. Machida, K.; Onodera, R.; Furuta, K.; Kikuchi, M.; Chem. Pharm. Bull. 1998, 46, 1295.
  • 13. Bringmann, G.; Hamm, A.; Kraus, J.; Ochse, M.; Noureldeen, A.; Jumbam, D. N.; Eur. J. Org. Chem 2001, 1983.
  • 14. Juma, B. F.; Majinda, R. R. T.; Nat. Prod. Res. 2007, 21, 121.
  • 15. Ono, M.; Ishimatsu, N.; Masouka, C.; Yoshimitsu, H.; Tsuchihashi, R.; Okawa, M.; Kinjo, J.; Ikeda, T.; Nohara, T.; Chem. Pharm. Bull. 2007, 55, 632.
  • 16. Kawata, Y.; Hattori, M.; Akao, T.; Kobashi, K.; Namba, T.; Planta Med. 1991, 57, 536.
  • 17. Li, X.; Huo, C.; Wang, Q.; Zhang, X.; Sheng, X.; Zhang, L.; J.Pharm. Biomed. Anal. 2007, 45, 268.
  • 18. Tanahashi, T.; Takenaka, Y.; Nagakura, N.; J. Nat. Prod. 1997, 60, 514.
  • 19. Gross, G. A.; Sticher, O.; Helv. Chim. Acta 1986, 69, 1113.
  • 20. Ono, M.; Ito, Y.; Kubo, S.; Nohara, T.; Chem. Pharm. Bull. 1997, 45, 1094.
  • 21. dos Santos, T. C.; Schripsema, J.; Monache, F. D.; Leitão, S. G.; J. Braz. Chem. Soc 2001, 12, 763.
  • 22. Gouda, Y. G.; Abdel-baky, A. M.; Darwish, F. M.; Mohamed, K. M.; Kasai, R.; Yamasaki, K.; Phytochemistry 2003, 63, 887.
  • 23. Machida, K.; Ando, M.; Yaoita, Y.; Kakuda, R.; Kikuchi, M.; Chem. Pharm. Bull. 2001, 49, 732.
  • 24. Marini-Bettolo, G. B.; Nicoletti, M.; Messana, I.; Patamia, M.; Galeffi, G.; Oguakwa, J. U.; Portalone, G.; Vaciago, A.; Tetrahedron 1983, 39, 323.
  • 25. Valladares, M. G.; Rios, M. Y.; J. Nat. Prod. 2007, 70, 100.
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  • Publication Dates

    • Publication in this collection
      19 July 2010
    • Date of issue
      2010

    History

    • Accepted
      22 Mar 2010
    • Received
      25 Feb 2009
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