Advanced
Two New Diterpenoids from Thuja orientalis and Their Cytotoxicity
Two New Diterpenoids from Thuja orientalis and Their Cytotoxicity
Bulletin of the Korean Chemical Society. 2014. Sep, 35(9): 2855-2858
Copyright © 2014, Korea Chemical Society
  • Received : March 28, 2014
  • Accepted : May 19, 2014
  • Published : September 20, 2014
Download
PDF
e-PUB
PubReader
PPT
Export by style
Article
Author
Metrics
Cited by
TagCloud
About the Authors
Chung Sub Kim
Won Se Suh
Sang Un Choi
Korea Research Institute of Chemical Technology, Daejeon 305-343, Korea
Ki Hyun Kim
Kang Ro Lee

Abstract
Keywords
PPT Slide
Lager Image
PPT Slide
Lager Image
PPT Slide
Lager Image
PPT Slide
Lager Image
PPT Slide
Lager Image
Experimental Section
General . Optical rotations were obtained on a JASCO P-1020 Polarimeter. IR spectra were recorded on a Bruker Vector 22 IR spectrophotometer. NMR spectra, including 1 H- 1 H COSY, HMQC, HMBC and NOESY experiments, were recorded on a Varian UNITY INOVA 500 NMR spectrometer operating at 500 MHz ( 1 H) and 125 MHz ( 13 C). HRFAB and HREI mass spectra were obtained on a JEOL JMS700 mass spectrometer. Preparative HPLC was performed using a Gilson 306 pump with a Shodex refractive index detector. Silica gel 60 (230-400 mesh) and RP-C 18 silica gel (230-400 mesh) were used for the column chromatography. TLC was performed using the precoated Silica gel F 254 plates and RP-18 F 254s plates (Merck). Spots were detected on TLC by heating after spraying with 10% H 2 SO 4 in EtOH (v/v).
Plant Material . The leaves of T. orientalis were collected in Yeongcheon City, Korea during May 2009. The plant was identified by one of the authors (K. R. Lee). A voucher specimen (SKKU-NPL 0819) of the plant was deposited at the herbarium of the School of Pharmacy at the Sungkyunkwan University in Suwon, Korea.
Extraction and Isolation . An amount of 4 kg leaves of T . orientalis was extracted at room temperature with 80% MeOH and evaporated under reduced pressure with 405 g residue. The residue was dissolved in water (800 mL × 2) and solvent-partitioned to n -hexane (73 g), CHCl 3 (41 g), EtOAc (42 g) and n -BuOH (104 g) layers. The n -hexane-soluble layer (36 g) was chromatographed on a silica gel column (230-400 mesh, 600 g, 5 × 60 cm) eluted with n -hexane:EtOAc (5:1 ~ 1:1, gradient system) to yield four fractions (H1-H4). Fraction H1 (3.6 g) was separated over a RP-C 18 silica gel column (230-400 mesh, 150 g, 3 × 30 cm) with 90% MeOH to five subfractions (H11-H15). Fraction H13 (620 mg) was separated over a silica gel column (230-400 mesh, 50 g, 2 × 25 cm) with CHCl 3 :MeOH (150:1) and purified with a RP-C 18 prep. HPLC with 75% CH 3 CN at a flow rate of 2.0 mL/min (Econosil RP-18 10 μm column; 250 × 10 mm; 10 μ particle size; Shodex refractive index detector) to yield 1 (4 mg, t R = 17.1 min) and 2 (3 mg, t R = 18.1 min). Fraction H13 (620 mg) was separated over a silica gel column (230-400 mesh, 50 g, 2 × 25 cm) with CHCl 3 :MeOH (150:1) and purified with a RP-C 18 prep. HPLC with 75% CH 3 CN to yield 3 (4 mg, t R = 19.8 min). Fraction H14 (440 mg) was further separated on a silica gel (230-440 mesh, 50 g, 2 × 25 cm) eluted with n -hexane:EtOAc (20:1) to yield four subfractions (H141-H144). Fraction H141 (50 mg) was purified with a RP-C 18 prep. HPLC with 85% CH 3 CN to yield 5 (4 mg, t R = 20.2 min), 6 (8 mg, t R = 23.1 min) and 7 (4 mg t R = 29.8 min). Fraction H142 (80 mg) was purified with a RP-C 18 prep. HPLC with 85 % CH 3 CN to yield 9 (32 mg, t R = 27.2 min). Fraction H144 (90 mg) was purified with a RP-C 18 prep. HPLC with 90% CH 3 CN to yield 8 (27 mg, t R =20.2 min). All fractions were purified as described above. Fraction H2 (3.6 g) was separated over a RP-C 18 silica gel column (230-400 mesh, 150 g, 3 × 30 cm) with 90% MeOH to give two subfractions (H21-H22). Fraction H21 (150 mg) was purified with a silica gel prep. HPLC with n -hexane: EtOAc (4:1) at a flow rate of 2.0 mL/min (Apollo Silica 5 μm column; 250 × 10 mm; 5 μ particle size; Shodex refractive index detector) to yield compound 4 (15 mg, t R = 15.8 min).
Thujuoric Acid A (1) : Colorless gum;
PPT Slide
Lager Image
+11.0° ( c 0.4, MeOH); IR (KBr) ν max 3078, 2932, 2845, 1691, 1641, 1263, 1164, 1099, 1029 cm −1 ; 1 H NMR see Table 1 ; 13 C NMR see Table 2 ; HRFABMS m/z 401.2300 [M + Na] + (calcd for C 22 H 34 NaO 5 , 401.2304).
Thujuoric Acid B (2) : Colorless gum;
PPT Slide
Lager Image
+5.0° ( c 0.4, MeOH); IR (KBr) ν max : 3079, 2928, 2844, 1693, 1645, 1266, 1163, 1100, 1027 cm −1 ; 1 H NMR see Table 1 ; 13 C NMR see Table 2 ; HRFABMS m/z 401.2302 [M + Na] + (calcd for C 22 H 34 NaO 5 , 401.2304).
Thujuoric Acid C (3) : Colorless gum;
PPT Slide
Lager Image
+36.6° ( c 0.2, CHCl 3 ); IR (KBr) ν max : 3078, 2932, 2845, 1691, 1641, 1263, 1164, 1099, 1029 cm −1 ; 1 H NMR see Table 1 ; 13 C NMR see Table 2 ; HRFABMS m/z 401.2304 [M + Na] + (calcd for C 22 H 34 NaO 5 , 401.2304).
8β,18-Dihydroxysandaracopimar-15-ene (4) : Colorless oil;
PPT Slide
Lager Image
−2.0° ( c 0.35, CHCl 3 ); IR (KBr) ν max 3424, 2922, 1635, 1444, 1385, 1034, 909, 757 cm −1 ; 1 H NMR see Table 1 ; 13 C NMR see Table 2 ; HREIMS m/z 306.2557 [M] + (calcd for C 20 H 34 O 2 , 306.2559).
Cytotoxicity Assay . A SRB bioassay was used to determine the cytotoxicity of each compound isolated against four cultured human tumor cell lines. 16 The assays were performed at the Korea Research Institute of Chemical Technology. The cell lines which were used were A549, SK-OV-3, A498 and HCT-15. Doxorubicin was used as a positive control. The cytotoxicities of doxorubicin against the A549, SK-OV-3, A498, and HCT-15 cell lines were IC 50 0.0007, 0.1274, 0.0094, and 0.2149 μM, respectively.
Supporting Information. The spectral data of compounds 1, 2 and 4 are available on request from the corresponding author.
Acknowledgements
This research was supported by the Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Education, Science and Technology (2012R1A5A2A28671860). We are thankful to the Korea Basic Science Institute (KBSI) for the measurements of NMR and MS spectra.
References
Sung S. H. , Koo K. A. , Lim H. K. , Lee H. S. , Cho J. H. , Kim H. S. , Kim Y. C. 1998 Korean J. Pharmacognosy 29 347 -
Xu G. H. , Ryoo I. J. , Kim Y. H. , Choo S. J. , Yoo I. D. 2009 Arch. Pharm. Res. 32 275 -    DOI : 10.1007/s12272-009-1233-y
Yoon J. S. , Koo K. A. , Ma C. J. , Sung S. H. , Kim Y. C. 2008 Nat. Pro. Sci. 14 167 -
Fan S.-Y. , Zeng H.-W. , Pei Y.-H. , Li L. , Ye J. , Pan Y.-X. , Zhang J.-G. , Yuan X. , Zhang W.-D. 2012 J. Ethnopharmacol. 141 647 -    DOI : 10.1016/j.jep.2011.05.019
Kosuge T. , Yokota M. , Sugiyama K. , Saito M. , Iwata Y. , Nakura M. , Yamamoto T. 1985 Chem. Pharm. Bull. 33 5565 -    DOI : 10.1248/cpb.33.5565
Kim C. S. , Choi S. U. , Lee K. R. 2012 Planta Med. 78 485 -    DOI : 10.1055/s-0031-1298215
Yoshida K. , Fueno T. 1987 Bull. Chem. Soc. Jpn. 60 229 -    DOI : 10.1246/bcsj.60.229
Bae J. , Jeon J. , Lee Y.-J. , Lee H.-S. , Sim C. J. , Oh K.-B. , Shin J. 2011 J. Nat. Prod. 74 1805 -    DOI : 10.1021/np200492k
Demarco P. V. , Farkas E. , Doddrell D. , Mylari B. L. , Wenkert E. 1968 J. Am. Chem. Soc. 90 5480 -    DOI : 10.1021/ja01022a027
Matsuo A. , Uto S. , Nakayama M. , Hayashi S. 1976 Tetrahedron Lett. 17 2451 -    DOI : 10.1016/0040-4039(76)90017-4
Itokawa H. , Yoshimoto S. , Morata H. 1988 Phytochemistry 27 435 -    DOI : 10.1016/0031-9422(88)83115-7
Heymann H. , Tezuka Y. , Kikuchi T. , Supriyatna S. 1994 Chem. Pharm. Bull. 42 138 -    DOI : 10.1248/cpb.42.138
Barrero A. F. , Alvarez-Manzaneda E. , Lara A. 1996 Tetrahedron Lett. 37 3757 -    DOI : 10.1016/0040-4039(96)00640-5
Joseph-Nathan P. , Santillan R. L. , Gutièrrez A. 1984 J. Nat. Prod. 47 924 -    DOI : 10.1021/np50036a003
Fang J.-M. , Chen Y.-C. , Wang B.-W. , Cheng Y.-S. 1996 Phytochemistry 41 1361 -    DOI : 10.1016/0031-9422(95)00795-4
Skehan P. , Storeng R. , Scudiero D. , Monks A. , Mcmahon J. , Vistica D. , Warren J. T. , Bokesch H. , Kenney S. , Boyd M. R. 1990 J. Natl. Cancer Inst. 82 1107 -    DOI : 10.1093/jnci/82.13.1107