A New Cadinane Sesquiterpene from the Marine Brown Alga Dictyopteris divaricata

A sample of the marine brown alga D. divaricata collected off the coast of Yantai (P.R. China) was dried, powdered, and extracted with the mixture of CHCl3 and MeOH (1:1, v/v). By a combination of silica gel and Sephadex LH-20 column chromatography and preparative TLC, a new cadinane sesquiterpene 1,4-epoxymuurolan-5β-ol (1) was isolated from this species. Its structure was established by detailed MS and NMR spectroscopic analysis, as well as comparison with literature data.


Introduction
Marine brown algae of the genus Dictyopteris are prolific sources of sesquiterpenes, and cadinane is a main carbon skeleton type [1][2][3][4][5][6][7][8]. In our investigations on the structurally interesting and biologically active terpenes from Chinese marine algae, we examined the chemical constituents of D.

OPEN ACCESS
divaricata collected off the coast of Yantai and as a result, a new cadinane sesquiterpene, 1,4epoxymuurolan-5β-ol (1) has been isolated and characterized for the first time. This paper reports the isolation and structural elucidation of compound 1 (Figure 1).

Results and Discussion
The dried and powdered alga D. divaricata was extracted with a mixture of CHCl 3 and MeOH (1:1, v/v). The concentrated extracts were partitioned between H 2 O and EtOAc. The EtOAc-soluble fraction was further purified by a combination of silica gel and Sephadex LH-20 column chromatography, as well as preparative TLC, to yield compound 1.  (Table 1) displayed one methyl singlet, three methyl doublets, and one broad singlet, attributed to an oxygenated methine. The 13 C-NMR spectrum (Table  1), along with the DEPT and HSQC experiments revealed the presence of four methyl, four methylene, five methine, and two quaternary carbon atoms. A detailed comparison of the above spectra data with those reported for 1,4-epoxymuurolan-5α-ol revealed that 1 differed from this last compound mainly at C-5 (δ C 85.5 d) [9], suggesting that compound 1 may be a C-5 isomer of 1,4-epoxymuurolan-5α-ol. The 1 H-1 H COSY correlations as shown in Table 1 and the observed HMBC correlations from H-12 to C-7, C-11, and C-13, from H-13 to C-7, C-11, and C-12, from H-14 to C-1, C-9, and C-10, from H-15 to C-3, C-4, and C-5, and from H-5 to C-3, C-4, C-6, C-7, and C-15 confirmed the planar structure of 1. The relative configuration of 1 was determined by analysis of NOESY spectrum and coupling constants. The NOESY correlations between H-5 and H-7, H-2a indicated H-5, H-7 and C-2 to be located on the same face of the molecule. The same orientation of C-14 and C-2 was suggested on the basis of the NOESY correlation between H-14 and H-2b. H-6 and C-15 were assigned on the same face according to the observed NOESY correlation between H-6 and H-15. H-6 was located on the opposite face of H-7 based on the large coupling constant (11.6 Hz) between them. The above evidence established the structure of 1 to be 1,4-epoxymuurolan-5β-ol (Figure 1), the C-5 epimer of 1,4-epoxymuurolan-5α-ol [9]. Compound 1 was tested for the toxicity against brine shrimp (Artemia salina) [10]. However, it exhibited no toxicity against brine shrimp at 100 μg/mL.

Experimental
General NMR spectra were recorded in CDCl 3 with TMS as internal standard on a Bruker Avance 500 MHz NMR spectrometer operating at 500 and 125 MHz for 1 H and 13 C, respectively. Low and high resolution mass spectra were determined on a VG Autospec 3000 mass spectrometer. The IR spectrum was obtained on a JASCO FT/IR-4100 Fourier Transform infrared spectrometer. Optical rotation was measured on a JASCO P-1020 polarimeter. Column chromatography was performed with silica gel (200-300 mesh, Qingdao Haiyang Chemical Co., Qingdao, P.R. China), RP-18 reversed-phase silica gel (YMC), and Sephadex LH-20 (Pharmacia). TLC was carried out with precoated silica gel plates (GF-254, Qingdao Haiyang Chemical Co., Qingdao, P.R. China). All solvents were of analytical grade.

Algal Material
The brown alga Dictyopteris divaricata was collected off the coast of Yantai (lat. 37°31'15"N, long. 121°26'59"E), Shandong Province, P. R. China, in July 2008, and a voucher specimen (MBA0807) has been deposited at the Bio-Resource Laboratory of Yantai Institute of Coastal Zone Research for Sustainable Development, Chinese Academy of Sciences.

Brine Shrimp Assays
Brine shrimp assays were performed as previously described [10].