tensive analysis of spectroscopic data, and comparison with data in May 1997. A voucher specimen (No. CUZ-026527) is deposited
in the literature. The leishmanicidal, antimicrobial, cytotoxic, in the Vargas Herbarium, Facultad of Biology, Universidad Nacio-
and aldose reductase inhibitory activities of some of the com- nal de San Antonio Abad del Cusco, Per.
pounds are described.
Copaifera paupera oil (29.4 g) was chromatographed by dry flash
chromatography on silica gel, using n-hexane-EtOAc mixtures of
increasing polarity to afford 67 100 ml frs., which were reduced
to 5 frs. by TLC: A (0±5%, n-hexane-EtOAc), B (5±15%), C (15±
25%), D (25±45%), and E (45±100%). A 40 mg sample of fraction
A (16.2 g) was rechromatographed on semi-preparative HPLC
using n-hexane as eluent to give 14 (tR 22.6 min), and 15 (tR 25.0
min). Fraction B (6.9 g) was rechromatographed on Sephadex LH-
20, using n-hexane-CHCl3-MeOH (2:1:1) as eluent to afford frs.
1±3. Fr. 1 (1.5 g) was then chromatographed on silica gel using
Cl2CH2-acetone (from 10:0 to 5:5) as eluent to give 6 (Rf 0.61,
n-hexane-EtOAc, 6:2). Fr. 2 (2.0 g) was subjected to a silica gel-
column with n-hexane-EtOAc (from 10:0 to 5:5) as eluent to af-
ford 13 and 1 (Rf 0.72, and 0.65, respectively, n-hexane-EtOAc,
6:4). Chromatography of fr. 3 (3.2 g) under the same conditions
yielded 2, 4, and 10 (Rf 0.60, 0.55, and 0.42, respectively, n-hex-
ane-EtOAc, 9:1). Fraction C (2.8 g), after chromatography on Se-
phadex LH-20 (n-hexane-CHCl3-MeOH, 2:1:1), gave rise to
compounds 5 and 11 (Rf 0.53, and 0.40, respectively, n-hexane-
EtOAc, 7:3). Fraction D (2.1 g) was purified by silica gel column,
using Cl2CH2-acetone (from 10:0 to 8:2) as eluent to give 3, 8, 9
and 17 (Rf 0.34, 0.60, 0.51, and 0.58, respectively, Cl2CH2-acetone,
9:1). Chromatography of fraction E (1.4 g) on Sephadex LH-20
(n-hexane-CHCl3-MeOH, 2:1:1) afforded 7 (Rf 0.53, n-hexane-
EtOAc, 6:4), and four additional fractions. Fr. 2 (50 mg) of these
were further purified by silica gel column using n-hexane-EtOAc
(from 10:0 to 0:10) yielded 16 (Rf 0.38, n-hexane-EtOAc, 7:3).
(±)-Copalic acid 1 (856.0 mg), [a]D25: ±15.38 (c 1.5, CHCl3); (±)-me-
thyl copalate 2 (15.4 mg), [a]D25: ±18.58 (c 1.8 , CHCl3); agathic acid
15-methyl ester 3 (17.7 mg), [a]D25: ±18.58 (c 0.4, CHCl3); agathic
acid 15,19-dimethyl ester 4 (2.3 mg), [a]D25: ±15.68 (c 0.2, CHCl3);
(±)-polyalthic acid 5 (145.8 mg), [a]D25: ±34.38 (c 1.8, CHCl3); (±)-
methyl polyalthate 6 (8.8 mg), [a]D25: ±38.98 (c 2.4, CHCl3); (±)-pi-
nifolic acid 7 (51.7 mg), [a]D25: ±27.98 (c 2.1, CHCl3); methyl 3b-hy-
droxy-labda-8(17),13-dien-15-ate 8 (4.3 mg), [a]D25: + 24.88 (c
809
Materials and Methods
General
0.31, CHCl3); (±)-methyl-18-hydroxy-copaiferolate 9 (12.4 mg),
[a]D25: ±21.58 (c 0.2, CHCl3); (+)-14,15-bisnorlabd-8(17)-en-13-
one 10 (3.4 mg), [a]D25: + 31.98 (c 0.4, CHCl3); (±)-kaurenic acid 11
Optical rotations were measured on a Perkin-Elmer 241 auto- (25.5 mg), [a]D25: ±71.78 (c 6.7, CHCl3); 16b-kauran-19-oic acid 13
matic polarimeter and the [a]D are given in 10±1 deg cm2 g±1
.
(65.2 mg), [a]D25: ±51.78 (c 1.2, CHCl3); b-bisabolene 14 (11.5 mg),
IR spectra were recorded in CHCl3 on a Bruker IFS 55 spectro- [a]D25: + 68.18 (c 0.5, CHCl3); caryophyllene oxide 15 (5.5 mg),
photometer and UV spectra were collected in absolute EtOH [a]D25: ±5.98 (c 0.2, CHCl3); 3-methyl-5-(2,2,6-trimethyl-6-hydro-
on a JASCO V-560. 1H- and 13C-NMR spectra were recorded on xy-cyclohexyl)-pentanoic acid 16 (10.0 mg), [a]D25: ±1.98 (c 0.9,
a Bruker Avance 400 NMR spectrometer at 400 MHz and 100 CHCl3); with all other spectroscopic and chemical data identical
MHz, respectively. EIMS and HREIMS were recorded an a Mi- with previously reported data [1], [5], [6], [7], [8], [9], [10], [11],
cromass Autospec spectrometer. TLC 1500/LS 25 Schleicher [12], [13], [14], [15].
and Schüell foils were used for thin-layer chromatography. Si-
lica gel (particle size 40±63 mM, Merk) and Sephadex LH-20 Pauperol (17). Colorless lacquer. [a]D25: ±7.48 (c 0.1, CHCl3). UV
(Pharmacia), were used for column chromatography, and semi- (EtOH): lmax = 220, 211, 202 nm. IR (film): lmax = 3500, 2900,
preparative m-porasil column (Waters, 10 mM, 19 mm 25 cm) 2850, 1750, 1680, 1450, 1430, 1370, 1215, 1140, 1080, 880,
for HPLC.
750 cm±1. 1H-NMR (400 MHz): d, see Table 1. 13C-NMR (100
MHz): d, see Table 1. EI-MS: m/z (%) = 572 (M+, 1), 556 (11), 554
(1), 524 (5), 317 (83), 285 (40), 257 (26), 203 (29), 147 (42), 95
Plant material and isolation
Copaifera paupera crude oil obtained from the trunk of the tree (65), 69 (100); HR-EI-MS: calcd. for C36H60O5 572.44408, found
was collected in Madre de Dios, Provincia de Tambopata, Per, 572.44621.
Tincussi B M et al. Antimicrobial Terpenoids from¼ Planta Med 2002; 68: 808±812