New Lanostanoids from the Fungus Ganoderma concinna Antonio G. Gonza ´ lez, † Francisco Leo ´n, † Augusto Rivera, ‡ Juan I. Padro ´n, † Javier Gonza ´ lez-Plata, § Juan C. Zuluaga, ⊥ Jose ´ Quintana, | Francisco Este ´vez, | and Jaime Bermejo* ,† Instituto Universitario de Bio-Orga ´ nica “Antonio Gonza ´ lez”, Instituto de Productos Naturales y Agrobiologı ´a.-CSIC, Avenida Astrofı ´sico F. Sa ´ nchez 3, 38206 La Laguna, Tenerife, Spain, Departamento de Quı ´mica, Facultad de Ciencias, Universidad Nacional de Colombia, Apartado Ae ´ reo 14490, Bogota ´ , D.C., Colombia, Departamento de Fı ´sica Fundamental, Universidad de La Laguna, Avenida Astrofı ´sico F. Sa ´ nchez 4, 38206 La Laguna, Tenerife, Spain, Departamento de Quı ´mica, Universidad de Co ´ rdoba, Monterı ´a, Colombia, and Departamento de Bioquı ´mica, Facultad de Medicina, Universidad de Las Palmas de Gran Canaria, Avenida S. Cristo ´ bal, 35016 Las Palmas de Gran Canaria, Spain Received March 16, 2001 Three new compounds, 5R-lanosta-7,9(11),24-triene-3-hydroxy-26-al (1), 5R-lanosta-7,9(11),24-triene- 15R-26-dihydroxy-3-one (2), and 8R,9R-epoxy-4,4,14R-trimethyl-3,7,11,15,20-pentaoxo-5R-pregnane (3), were isolated from Ganoderma concinna along with 12 known compounds. The structures of compounds 1 and 2 were determined on the basis of MS and NMR studies. The structure of 3 was determined by MS, NMR, and single-crystal X-ray diffraction. Compounds 1, 2, and 3 induce apoptosis in human promyelocytic leukemia HL-60 cells, as indicated by examining the morphological features of cells and detection of DNA fragmentation by gel electrophoresis. In connection with our earlier studies 1,2 of Polyporaceae (Basidiomycetes) metabolites, we have investigated the constituents of the EtOAc extract of the fruit bodies of Ganoderma concinna Ryv. Nov. sp. (Ganodermataceae). After extraction by the usual methods, the triterpenoids and the sterols in the EtOAc extract were separated by column chromatography, Sephadex LH-20, and prepara- tive TLC methods. Twelve compounds of the 15 iso- lates were identified as known compounds. These include ganoderal A 3 (4), whose 13 C NMR spectral data (Table 2) have not previously been reported in the literature, gano- dermenonol, 4 ganodermadiol, 5 ganoderic acid Y, 6 ganoderiol F, 7 ganodermatriol, 8 ganodermanontriol, 8 ganoderiol A, 8 ganoderiol B, 8 ergosta-7,22-dien-3-one, 9 fungisterol, 10 and ergosterol peroxide. 11 The structures of the known com- pounds were confirmed by comparison of their spectroscopic data (MS, 1 H and 13 C NMR) with literature references. Two of the remaining three compounds were new lano- stanoid triterpenes, which we named 5R-lanosta-7,9(11),24- triene-3-hydroxy-26-al (1), 5R-lanosta-7,9(11),24-triene- 15R-26-dihydroxy-3-one (2), and 8R,9R-epoxy-4,4,14R- trimethyl-3,7,11,15,20-pentaoxo-5R-pregnane (3). Compound 1 showed a positive Lieberman-Burchard (LB) reaction, and a hydroxyl (3406 cm -1 ) and an R-- unsaturated aldehyde (1688 cm -1 ) absorption were ob- served in its IR spectrum. The HREIMS spectrum of 1 showed a molecular ion at m/z 438.3426 corresponding to the molecular formula C 30 H 46 O 2 (calcd 438.3497). The 1 H NMR spectrum of 1 (Table 1) showed signals for tertiary methyl groups at δ 0.55, 2 × 0.86, 0.96, and 0.98 and a secondary methyl group at δ 0.93 (d, J ) 6.6 Hz), as required by the lanostane skeleton. A vinyl methyl and olefinic signals were observed at δ 1.73, 5.30, 5.46, and 6.47, respectively. The 13 C NMR spectrum of 1 (Table 2) showed signals due a conjugated diene group at δ 120.3, 142.4, 145.9, 116.0, which suggested a 7,9(11)-diene lanostane skeleton. Comparison of the 13 C NMR spectral data with those reported for ganodermadiol 5 allowed the location of the aldehyde group. 2 These data established the structure of 1 as 5R-lanosta-7,9(11),24-triene-3-hydroxy-26-al. HREIMS and 13 C NMR data of 2 indicated the molecular formula C 30 H 46 O 3 . The IR spectrum of this compound showed the presence of a hydroxyl group (3444 cm -1 ), a carbonyl group (1703 cm -1 ), and an unsaturated carbon (2968 cm -1 ). The UV, 1 H NMR, and 13 C NMR spectra of 2 were similar to those of 1, suggesting that 2 may have a 5R-lanosta-7,9(11),24-triene structure. However in the 13 C NMR spectrum the carbonyl carbon signal was observed at low field at δ 216.6. Also, the 1 H NMR spectrum did not exhibit any signal around δ 3.23 (H-3 of 1) for the presence of a hydroxyl group. These observations suggested the presence of a carbonyl group at C-3. A singlet at δ 3.98 integrated for two hydrogens and the signal at δ 4.28 (dd, J ) 9.2, 4.9 Hz) integrated for one hydrogen, in accord with those reported in the literature. 2,6 The shifts correspond to the protons at C-26 and C-15, respectively, on the basic of NOESY and HMBC spectra (Figure 1). Thus, compound * To whom correspondence should be addressed. Tel: (34) 922-250766. Fax: (34) 922-318571. E-mail: jbermejo@ull.es. † Instituto Universitario, Instituto Productos Naturales. ‡ Departamento de Quı ´mica, Universidad Nacional de Colombia. § Departamento de Fı ´sica, Universidad de La Laguna. ⊥ Departamento de Quı ´mica, Universidad de Co ´rdoba. | Departamento de Bioquı ´mica, Universidad de las Palmas de Gran Canaria. 417 J. Nat. Prod. 2002, 65, 417-421 10.1021/np010143e CCC: $22.00 © 2002 American Chemical Society and American Society of Pharmacognosy Published on Web 03/03/2002