Chemistry of Natural Compounds, Vol. 43, No.

6, 2007

DISTRIBUTION OF COUMARINS IN THE TRIBE

PLUCHEEAE, GENUS Pterocaulon

A. C. Stein,1 D. Fritz,1 L. F. Paiva Lima,2 N. I. Matzenbacher,2 UDC 547.972

J. Schripsema,3 V. Pires,4 P. Sonnet,4 and G. von Poser 1

The genus Pterocaulon (Asteraceae) is included in the tribe Plucheeae [1]. About half of the Pterocaulon species have
been analyzed previously and many compounds have been isolated: caffeoylquinic acids from P. virgatum [2]; thiophene
acetylenes and flavanols from P. virgatum [3]; polyacetylenes from P. alopecuroides, P. balansae, P. lanatum, and P. rugosum
[4]; hydroxy-α-caryophyllene from P. serrulatum [5]; flavonoids from P. sphacelatum [6], P. purpurascens [7], and
P. alopecuroides [8]; and coumarins from P. balansae, P. lanatum [4], P. purpurascens [9–11], P. redolens [12], P. virgatum
[13–16], P. polystachyum [17], and P. alopecuroides [8]. Several species of the genus are used in folk medicine as insecticides
and agents against snake bites [15].
In this work we investigate the occurrence of coumarins in exudates of the species of Pterocaulon native to Brazil and
isolate the most abundant compounds. The taxonomic significance of these compounds is discussed.
In Plucheeae the only genus that contains coumarins is Pterocaulon in which 40 different coumarins were previously
found in ten species. In this work we analyzed five other species native to South Brazil, and coumarins were found in all of
them. These data indicate that these compounds could be considered chemotaxonomic markers for the genus Pterocaulon.
Although the species of Pterocaulon exhibit other classes of compounds, coumarins are characteristic of the genus. In
Pterocaulon, coumarins can be di-, tri-, or tetraoxygenated, and all of them are 6,7 oxygenated.
Pterocaulon alopecuroides and P. polystachyum were studied previously, but in this work we investigated the exudate
instead of the extract of the powdered plant. The exudate of P. alopecuroides afforded 5-methoxy-6,7-methylenedioxycoumarin
(1) and 7-(2,3-epoxy-3-methyl-3-butyloxy)-6-methoxycoumarin (2), previously found in P. virgatum [13, 18]. To our knowledge
this is the first report on the occurrence of coumarins 1 and 2 in Pterocaulon alopecuroides. From this plant, the compounds
7-(2,3-dihydroxy-3-methylbutyloxy)-6-methoxycoumarin and 7-(2,3-dihydroxy-3-methylbutyloxy)-5-hydroxy-6-coumarin were
previously obtained [8]. This difference could be due to the extraction method employed and to the fact that the plants were
collected in different places. Nevertheless, it is worthwhile to state that the plant exudate contained several coumarins of which
only the most abundant were isolated.
The exudate of P. polystachyum, besides 5-methoxy-6,7-methylenedioxycoumarin (1), afforded three other coumarins,
namely ayapin (3), prenyletin (4), and prenyletin methyl-ether (5). These compounds were previously isolated from the
chloroform extract of the powdered plant material [17, 19].
R
O MeO RO

O O O O O O O O O
O
1, 3 2 4, 5

1: R = OMe; 3: R = H; 4: R = H; 5: R = Me

1) PPG-Ciencias Farmaceuticas, UFRGS, Porto Alegre, RS, Brazil, fax (051) 33165610, e-mail:
[email protected]; 2) PPG-Botanica, ICBS, 91509-900 Porto Alegre, RS, Brazil, fax (051) 33167670; 3) Grupo
Metabolomica, LCQUI/CCT, Universidade Estadual do Norte Fluminense, 28015-620 Campos dos Goytacazes, RJ, Brazil;
4) Faculte de Pharmacie, Universite de Picardie Jules Verne, 80037 Amiens, France. Published in Khimiya Prirodnykh
Soedinenii, No. 6, pp. 572-574, November-December, 2007. Original article submitted August 1, 2006.

0009-3130/07/4306-0691 ©2007 Springer Science+Business Media, Inc. 691

TABLE 1. 1H NMR Data of Compounds 1-5 (coupling constants in Hz)

Atom 1* 2* 3 4 5
H-3 6.22 (d, J = 9.7) 6.27 (d, J = 9.4) 6.27 (d, J = 9.5) 6.20 (d, J = 9.4) 6.22 (d, J = 9.4)
H-4 7.95 (d, J = 9.7) 7.62 (d, J = 9.4) 7.58 (d, J = 9.5) 7.83 (d, J = 9.4) 7.77 (d, J = 9.4)
H-5 - 6.90 (s) 6.82 (s) 7.07 (s) 6.93 (s)
H-8 6.54 (s) 6.87 (s) 6.83 (s) 6.89 (s) 6.88 (s)
5-OMe 4.15 (s) - - - -
6-7-OCH2O- 6.00 (s) - 6.07 (s) - -
6-OH - - - 5.50 (t) -
6-OMe - 3.90 (s) - - 3.82 (s)
H-1′a - 4.12 (dd, J = 11.4; 6.0) - 4.62 (d, J = 6.7) 4.67 (d, J = 6.4)
H-1′b - 4.34 (dd, J = 11.4; 4.0) - 4.62 (d, J = 6.7) 4.67 (d, J = 6.4)
H-2′ - 3.19 (dd, J = 6.0; 4.0) - 5.46 (t) 5.51 (t)
H-4′ - 1.39 (s) - 1.76 (s) 1.78 (s)
H-5′ - 1.38 (s) - 1.78 (s) 1.78 (s)
______
Experimental data (400 MHz; CDCl3). *Experimental data (500 MHz; CDCl3).

TABLE 2. 13C NMR Data of Compounds 1-5

Atom 1* 2* 3 4 5
C-2 161.27 161.22 161.13 163.82 164.00
C-3 111.72 113.79 113.36 112.89 113.40
C-4 138.76 143.17 143.40 145.87 145.80
C-5 138.04 108.65 105.05 110.05 108.15
C-6 131.74 146.67 144.89 139.97 145.50
C-7 152.65 151.82 151.25 150.15 152.47
C-8 92.36 101.89 98.35 102.11 101.75
C-9 151.51 149.77 151.25 148.36 148.36
C-10 106.60 111.97 112.67 112.98 112.98
5-OCH3 59.92 - - - -
6-7-OCH2O- 101.80 102.31 - - -
6-OCH3 - 56.44 - - 56.83
C-1′ - 68.51 - 67.20 67.20
C-2′ - 60.83 - 120.11 120.27
C-3′ - 58.25 - 139.76 139.76
C-4′ - 24.53 - 25.84 25.84
C-5′ - 19.07 - 18.28 18.28
______
Experimental data (100 MHz; CDCl3). *Experimental data (125 MHz; CDCl3).

The presence of high amounts of coumarins in the exudate of these plants suggests that the compounds are accumulated
in the epicuticular wax or in the trichomes or in both. In order to determine the exact localization of the compounds, a morpho-
anatomical study is being carried out.
The presence of some secondary metabolites can be a useful character in the revision of taxonomic systems.
Nevertheless sometimes the available data are not sufficient. As until now the only chemical character used in cladistic analysis
of the tribe Plucheeae was the presence or absence of the sesquiterpene cuauhtematl (cuauhtemon) and its derivatives, the data
presented here could be useful.
Some coumarins such as ayapin isolated from several species are described as phytoalexins, protecting the plant against
microorganisms attach to it, especially fungus. In the case of Pterocaulon the method employed for the extraction of the
compounds could indicate that the substances, if not all, the majority, are located in the surface of the leaves and branches where
they could act as phytoalexins. Interestingly enough, native species of Pterocaulon are used in some rural areas in South Brazil

692
to treat animal skin diseases popularly diagnosed as mycoses. In a recent study our group investigated the antifungal activity
of crude extracts obtained from P. alopecuroides, P. balansae, and P. polysthachyum, finding significant results [20].
Plant Material. Aerial parts of the plants were collected in the Rio Grande do Sul state, in March, 2003. The voucher
specimens are deposited in the Herbarium of the Federal University of Rio Grande do Sul (ICN) (Pterocaulon alopecuroides
(Lam.) DC. (136583); P. angustifolium DC. (136587); P. balansae Chodat (136589); P. cordobense O.K. (136588); P. lorentzii
Malme (136586); P. polypterum (DC.) Cabrera (136585); P. polystachyum DC. (136584) and P. rugosum (Vahl) Malme
(136590).
Chemical Methods. Whole leaves and branches of the plants (ca. 50 g) were carefully dried and extracted twice by
immersion in hexane:dichloromethane (1:1) for 15 s. This short extraction period proved sufficient for obtaining the compounds
that coated the twigs and leaves without extracting other components such as chlorophyll from those parts of the plant. The
extracts were evaporated to dryness under reduced pressure and analyzed by TLC on silicagel using elution with CH2Cl2 100%
up to CH2Cl2–MeOH (95:5). From P. alopecuroides and P. polystachyum, the main products in each case were isolated by
column chromatography on silica gel using a chloroform-methanol gradient system followed by preparative TLC on silica gel
with chloroform–methanol (98:2), to afford 5-methoxy-6,7-methylenedioxycoumarin (1) and 7-(2,3-epoxy-3-methyl-3-butyloxy)-
6-methoxycoumarin (2) from P. alopecuroides. From P. polystachyum, besides compound 1, ayapin (3), prenyletin (4), and
prenyletin methyl-ether (5) were isolated. The products were identified by 1H NMR and 13C NMR spectroscopy (Tables 1, 2)
and comparison with literature data [14, 18, 21, 22].

ACKNOWLEDGMENT

The authors thank CNPq and CAPES-COFECUB for financial support.

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