1010 0009-3130/11/4606-1010  2011 Springer Science+Business Media, Inc. 1) Guangdong Key Laboratory of Marine Materia Medica, Research Center for Marine Microbes, South China Sea Institute of Oceanology, Chinese Academy of Sciences, Guangzhou 510-301, P. R. China, fax: +86 20 884451672, e-mail: yonghongliu@scscio.ac.cn; 2) Graduate University of the Chinese Academy of Sciences, Beijing 100-049, P. R. China; 3) College of Light Industry and Food Sciences, South China University of Technology, Guangzhou 510-641, P. R. China; 4) School of Chinese Materia Medica, Beijing University of Chinese Medicine, Beijing 100102, P. R. China. Published in Khimiya Prirodnykh Soedinenii, No. 6, pp. 858–859, November–December, 2010. Original article submitted May 28, 2009. Chemistry of Natural Compounds, Vol. 46, No. 6, 2011 NUCLEOSIDES FROM THE MARINE SPONGE Callyspongia SP. Riming Huang, 1,2 Xuefeng Zhou, 1 Yan Peng, 3 UDC 547.963.3 Xianwen Yang, 1 Tunhai Xu, 4 and Yonghong Liu 1* The genus Callyspongia (order Haplosclerida, family Callyspongiidae) is widely distributed and contains various bioactive constituents [1]. The wet marine sponge Callyspongia sp. (10 kg) was extracted with ethanol (90%). Ethanol was evaporated in vacuo to afford a syrupy residue that was suspended in distilled water and fractionated successively with petroleum ether, ethyl acetate, and n-butanol. Rechromatography of the ethyl acetate fraction over columns of silica gel, Sephadex LH-20, and ODS HPLC produced compounds 1–8. The isolated compounds were identified using spectral analysis (NMR and MS). Compounds 1–8 were identified as spongothymidine (1) [2], spongouridine (2) [3], 1-(2-deoxy--D-ribofuranosyl)thymine ( 3) [4], 2-deoxyuridine (4) [5], 2-deoxyinosine (5) [5], 2-deoxyadenosine (6) [6]. 1-(2-Deoxy--D-erythro-pentofuranosyl)-1H- 1,2,4-triazone (7) [7], and 1-(-D-ribofuranosyl)-1H-1,2,4-triazone (8) [8] by comparison of the results with the literature. Compounds 1–3, 5–8 were isolated for the first time from Callyspongia sp. Spongothymidine (1) . White solid. 1 H NMR (500 MHz, CD 3 OD, , J/Hz): 7.83 (1H, s, H-4), 6.30 (1H, t, J = 7.0, H-1), 4.42 (1H, m, H-3), 4.24 (1H, m, H-4), 3.93 (1H, m, H-2), 3.83 (1H, dd, J = 12.0, 3.0, H-5a), 3.80 (1H, dd, J = 12.0, 3.5, H-5b), 1.88 (3H, s, 5-CH 3 ). 13 C NMR (125 MHz, CD 3 OD): 163.7 (C-6), 150.4 (C-2), 140.4 (C-4), 109.7 (C-5), 95.3 (C-1), 87.0 (C-4), 73.3 (C-2), 70.2 (C-3), 60.1 (C-5), 13.1 (5-CH 3 ). Spongouridine (2). White solid. 1 H NMR (500 MHz, Py-d 5 , , J/Hz): 13.01 (1H, br.s, 1-NH), 8.53 (1H, d, J = 8.5, H-4), 7.73 (1H, br.s, 5-OH), 7.06 (2H, br.s, 2 and 3-OH), 6.82 (1H, d, J = 3.5, H-1), 5.80 (1H, d, J = 8.5, H-5), 4.91 (1H, m, H-3), 4.90 (1H, m, H-4), 4.65 (1H, m, H-2), 4.31 (1H, d, J = 12.0, H-5a), 4.20 (1H, d, J = 12.0, H-5b). 13 C NMR (125 MHz, Pyr-d 5 ): 164.3 (C-6), 152.1 (C-2), 141.0 (C-4), 102.4 (C-5), 90.4 (C-1), 86.2 (C-4), 75.9 (C-2), 71.1 (C-3), 61.7 (C-5). 1-(2-Deoxy--D-ribofuranosyl)thymine (3). White solid. 1 H NMR (500 MHz, DMSO-d 6 , , J/Hz): 11.27 (1H, br.s, 1-NH), 7.69 (1H, s, H-4), 6.16 (1H, t, J = 7.3, H-1), 5.22 (1H, d, J = 4.2, 3-OH), 5.01 (1H, t, J = 5.2, 5-OH), 4.22 (1H, m, H-4), 3.92 (1H, m, H-3), 3.56 (2H, m, H-5), 2.07 (2H, m, H-2), 1.76 (3H, s, 5-CH 3 ). 13 C NMR (125 MHz, DMSO-d 6 ): 163.5 (C-6), 150.6 (C-2), 139.4 (C-4), 110.7 (C-5), 94.3 (C-1), 87.1 (C-4), 70.2 (C-3), 61.1 (C-5), 40.3 (C-2), 13.2 (5-CH 3 ). 2-Deoxyuridine (4). White solid. 1 H NMR (500 MHz, CD 3 OD, , J/Hz): 8.00 (1H, d, J = 6.5, H-4), 6.82 (1H, t, J = 7.0, H-1), 5.72 (1H, d, J = 6.5, H-5), 4.41 (1H, m, H-4), 3.95 (1H, m, H-3), 3.79 (1H, dd, J = 12.0, 3.5, H-5a), 3.73 (1H, dd, J = 12.0, 4.0, H-5b), 2.31 (1H, m, H-2a), 2.21 (1H, m, H-2b). 13 C NMR (125 MHz, CD 3 OD): 163.3 (C-6), 151.1 (C-2), 141.0 (C-4), 103.4 (C-5), 92.4 (C-1), 87.2 (C-4), 71.2 (C-3), 61.3 (C-5), 40.3 (C-2). 2-Deoxyinosine (5). White solid. ESI-MS m/z 253 [M + H] + . 1 H NMR (500 MHz, DMSO-d 6 , , J/Hz): 12.11 (1H, br.s, 1-NH), 8.13 (1H, s, H-2), 8.07 (1H, s, H-8), 6.30 (1H, t, J = 6.9, H-1), 5.32 (1H, d, J = 4.0, 3-OH), 4.96 (1H, t, J = 5.5, 5-OH), 4.38 (1H, m, H-4), 3.94 (1H, m, H-3), 3.61 (1H, m, H-5a), 3.48 (1H, m, H-5b), 2.61 (2H, m, H-2 ). 13 C NMR (125 MHz, DMSO-d 6 ): 166.2 (C-6), 157.2 (C-4), 155.2 (C-2), 148.0 (C-8), 133.9 (C-5), 97.1 (C-1), 93.1 (C-4), 80.1 (C-3), 71.1 (C-5), 45.6 (C-2).