DSpace at VNU: Two new flavonol glycosides from the leaves of Cleome viscosa L.

Phytochemistry Letters 18 (2016) 10–13 Contents lists available at ScienceDirect Phytochemistry Letters journal homepage: www.elsevier.com/locate/phytol Short communication Two new flavonol glycosides from the leaves of Cleome viscosa L Nhat Minh Phana,* , Tan Phat Nguyena , Tien Dung Lea , Thanh Chi Maia , Mai Thanh Phongb , Dinh Tri Maia a b Institute of Chemical Technology, Vietnam Academy of Science and Technology, Ho Chi Minh city, Viet Nam University of Technology, Vietnam National University, Ho Chi Minh city, Viet Nam A R T I C L E I N F O Article history: Received 25 May 2016 Received in revised form 18 August 2016 Accepted 26 August 2016 Available online xxx Keywords: Cleome viscosa Capparaceae Visconoside A Visconoside B A B S T R A C T From the leaves of Cleome viscosa L., two new flavonol glycosides, named visconoside A (1) and visconoside B (2), together with six known flavonol glycosides, vincetoxicoside A (3), vincetoxicoside B (4), kaempferitrin (5), kaempferide 3-O-b-D-glucopyranoside 7-O-a-L-rhamnopyranoside (6), kaempferol 3-O-b-D-glucopyranoside 7-O-a-L-rhamnopyranoside (7), and isorhamnetin 3-O-b-D-glucopyranoside (8) were isolated by various chromatography methods Its chemical structure was elucidated by IR, UV, HR-ESI-MS, NMR 1D and 2D experiments and compared with literatures ã 2016 Phytochemical Society of Europe Published by Elsevier Ltd All rights reserved Introduction Results and discussion Cleome viscosa L commonly known as “wild or dog mustard”, is an annual, sticky herb belonging to family Capparaceae found as a common weed throughout the tropics of the world The whole plant is used as drugs by the traditional medical practitioners with beneficial action for the treatment of diarrhoea, fever, inflammation, liver diseases, bronchitis, skin diseases, and malarial fever (Mali, 2010) This plant has been screened scientifically for various pharmacological activities such as anthelmintic, antimicrobial, anti-inflammatory, hepatoprotective (Mali, 2010), antifibrotic (Kumar et al., 2009), anticonvulsant (Mishra et al., 2010), antioxidant (Gupta et al., 2011), antinociceptive, antibacterial (Bose et al., 2011), antitumor (Gopal et al., 2012) and antibacterial activities (Donkor et al., 2014) Previous phytochemical investigations of C viscosa have yielded a number of flavanones (Srivastava et al., 1979), flavonol (Chauhan et al., 1979; Senthamilselvi et al., 2012), sterol (Srivastava, 1980), coumarinolignoid (Ray et al., 1985; Kumar et al., 1988), diterpene (Jente et al., 1990), lactam derivative (Jana and Biswas, 2011), and dipyridodiazepinone derivative (Chatterjee et al., 2013) In this paper, we describe the isolation and structure elucidation of two new flavonol glycosides (1, 2) and six known flavonol glycosides (3–8) from Cleome viscosa L collected in Ben Cat, Binh Duong province, Viet Nam The MeOH extract from the dried leaves of Cleome viscosa L was subjected to column chromatography over silica gel normal-phase and reversed-phase RP-18 to give two new flavonol glycosides, named visconosides A (1), visconosides B (2), and six known flavonol glycosides, vincetoxicoside A (3) (Gaind et al., 1981), vincetoxicoside B (4) (Ishiguro et al., 1991), kaempferitrin (5) (Rao et al., 2009), kaempferide 3-O-b-D-glucopyranoside 7-O-a-Lrhamnopyranoside (6) (Imperato, 1984), kaempferol 3-O-b-Dglucopyranoside 7-O-a-L-rhamnopyranoside (7) (Song et al., 2007), and isorhamnetin 3-O-b-D-glucopyranoside (8) (Wang et al., 2012) Compound (1) was isolated as a yellow amorphous powder, and the HR-ESI–MS showed the quasimolecular ion at m/z 821.2116 [M + Na]+ consistent with the molecular formula C35H42O21 The aglycone of was identified as quercetin, according to observation of fifteen carbon signals in 13C-NMR and DEPT spectrum (Table 1) including one carbonyl carbon at dC 178.0 (C-4), seven oxygenated aromatic carbons, two quaternary aromatic carbons and five nonoxygenated aromatic carbons together with two AX-type aromatic protons at dH 6.45 (d, 2.0, H-6) and 6.76 ; three ABX-type aromatic protons at dH 6.93 (d, 8.5, H-50 ), 7.30 (d, 2.0, H-60 ) and 7.31 (dd, 2.0, 8.5, H-20 ) in 1H-NMR data Moreover, three anomeric carbons at dH 5.36 (br s, H-1”), 4.27 (d, 8.0, H-000 ), 5.54 (d, 1.0, H-10000 ) corresponded to three anomeric carbons at dC 101.1 (C-100 ), 104.8 (C-1000 ), 96.8 (C-10000 ) were assigned to a-L-rhamnose (Rha I), b-D-glucose (Glc) and a-L-rhamnose (Rha II) units, respectively The COSY and HSQC spectrum allowed analysis of their spin systems and assignment of * Corresponding author E-mail address: pnhatminh@gmail.com (N.M Phan) http://dx.doi.org/10.1016/j.phytol.2016.08.020 1874-3900/ã 2016 Phytochemical Society of Europe Published by Elsevier Ltd All rights reserved N.M Phan et al / Phytochemistry Letters 18 (2016) 10–13 Table 1 H (500 Hz) and Aglycone 13 C (125 Hz) NMR spectral data for compounds and in DMSO-d6 Sugar dH dC dH dC – – – – 6.45 d (2.0) – 6.76 d (2.0) 158.2 133.9 178.0 161.1 99.8 161.9 94.8 – – – – 6.27 d (1.5) – 6.50 br s 156.8 133.2 177.4 160.5 99.5 161.7 93.6 Glc 1000 2000 3000 4000 5000 6000 10 1' 2' 3' 4' 5' 6' – – – 7.31 dd (2.0, 8.5) – – 6.93 d (8.5) 7.30 d (2.0) 156.3 105.9 120.4 115.8 145.5 149.1 115.8 121.6 – 7.26 d (1.5) – – 6.93 d (7.5) 7.33 dd (2.0, 8.0) 155.6 105.3 120.5 115.6 145.4 148.8 115.2 121.5 Rha II 1”” 2”” 3”” 4”” 5”” 6”” Sinapinoyl 5.36 br s 4.20 s 3.87 d (2.5) 4.82 t (10.0) 3.37 dd (6.5, 10.0) 0.74 d (6.5) – 1.96 s 101.2 69.7 76.9 71.5 68.5 17.3 170.2 21.0 5.54 br s 4.08 br s 3.86 d (2.5) 4.79 t (10.0) 3.07 m 0.66 d (6.5) – 1.94 s 99.9 69.7 76.4 71.3 68.4 17.1 169.8 20.8 Rha I 1” 2” 3” 4” 5” 6” 4”-OAc 11 their proton resonances to determine clearly every sugar unit Beside, the sugar moiety was identified as L-rhamnose and Dglucose by the acidic hydrolysis and using TLC to compare the hydrolysate with the authentic sugars (see Experimental) Further, revealed an acetyl group [dC 170.2–21.0 with methyl proton at dH 1.96 (s, CH3CO)] The HMBC spectrum (Fig 1), showed correlations between anomeric proton at dH 5.36 (br s, H-100 ) of Rha I and carbons at dC 133.9 (C-3) of aglycone; between anomeric proton at dH 4.27 (d, 8.0, H-1000 ) of Glc and carbons at dC 76.9 (C-300 ) of Rha I; between oxygenated methine proton at dH 3.87 (d, 2.5, H-300 ) and dH dC dH dC 4.27 d (8.0) 2.99 t (8.5) 3.17 dd (8.5, 9.0) 3.10 t (9.0) 3.22 m 3.72 dd (1.5, 11.5) 3.54 dd (9.0, 11.5) 104.8 73.4 77.0 70.1 76.6 61.2 4.32 d (7.5) 3.07 m 3.24 t (9.0) 3.07 m 3.63 m 4.52 dd (8.5, 11.0) 4.25 dd (2.0, 11.0) 105.0 73.0 76.7 70.6 74.1 63.6 5.54 d (1.0) 3.84 br s 3.63 dd (3.5, 9.5) 3.31 t (9.5) 3.44 dd (3.5, 6.0) 1.11 d(6.0) 98.6 70.0 70.4 71.8 70.3 18.1 5.54 br s 3.89 d (2.5) 3.63 m 3.31 t (9.0) 3.40 dd (6.0, 9.0) 1.15 d (6.0) 98.6 69.8 70.3 71.7 69.9 18.0 6.30 d (15.5) 7.41 d (15.5) – 6.50 s – – – 6.50 s – 3.54 d (6.5) 114.3 145.1 123.8 104.9 147.3 137.8 147.3 104.9 166.4 55.3 a b 1000 00 2000 00 3000 00 4000 00 5000 00 6000 00 ÀÀCOO ÀÀOCH3 carbon acetal at dC 104.8 (C-1000 ) On the other hands, anomeric proton at dH 5.54 (d, 1.0, H-10000 ) correlated with oxygenated aromatic carbon at dC 161.9 (C-7) Based on data of HR-ESI-MS, 1D, 2D-NMR and compared with previous published data (Minh et al., 2015), the structure of was determined as quercetin 3-O-[b-Dglucopyranosyl-(1 ! 3)]-a-L-(4-O-acetyl)-rhamnopyranoside 7-Oa-L-rhamnopyranoside, and named visconoside A Compound (2) was obtained as a yellow amorphous powder The molecular formula was established as C46H52O25 by HR-ESI-MS data ([M + Na]+ m/z 1027.2680) The 1H and 13C-NMR data (Table 1) Fig Chemical structures and selected HMBC and COSY correlations of compounds and 12 N.M Phan et al / Phytochemistry Letters 18 (2016) 10–13 demonstrated that has the same aglycone and sugar chains as 1, except for presence of an sinapoyl unit, including two oxymethyl at dC 55.3 (–OCH3), one carbonyl carbon at dC 166.0 (–COO), three oxygenated aromatic carbons at dC 147.3 (C-300000 , C-50000 '), 137.8 (C400000 ), one quaternary aromatic carbon at dC 123.8 (C-100000 ), four olefinic carbons at dC 104.9 (C-200000 , C-600000 ), 114.3 (C-a), 145.1 (C-b) correlated with two aromatic protons at dH 6.50 (s, H-200000 , H-600000 ), two trans protons at dH 6.30 (d, 15.5, H-a) and 7.41 (d, 15.5, H-b), respectively Moreover, the sinapoyl moiety was also confirmed by correlations observed in the HMBC spectrum (Fig 1), between protons at dH 6.30 (H-a), 7.41 (H-b) and carbons at dC 166.0 (-COO), 123.8 (C-100000 ), between protons at dH 6.50 (H-200000 , H-600000 ) and carbons at dC 145.1 (C-b), 123.8 (C-100000 ), 137.8 (C-400000 ) On the other hands, two methylene protons at dH 4.52 (dd, 8.5, 11.0, H-6a000 ), 4.25 (dd, 2.0, 11.0, H-6b000 ) correlated with carbonyl at dC 166.0 So, the sinapoyl moiety attached to C-6 of the glucose unit Based on data of HR-ESI-MS, 1D, 2D-NMR and compared with previous published data (Minh et al., 2015; Han et al., 2015); the structure of was identified as quercetin 3-O-[sinapoyl-(1 ! 6)-b-D-glucopyranosyl(1 ! 3)]-a-L-(4-O-acetyl)-rhamnopyranoside 7-O-a-L-rhamnopyranoside, and named visconoside B (25 mg) Fraction M3 (5 g) was separated by silica gel chromatographic column using CHCl3-MeOH (5:1), and further separated by RP-18 using gradient mixtures of MeOH–H2O (5:1, v/v) to affrord compound (268 mg) The same manner was applied to fraction M4 (75 g), eluted CHCl3-MeOH (6:1 ! 3:1) to give four subfractions (M4.1–M4.4) Subfraction M4.1 (12 g) was further purifed by RP-18 with MeOH–H2O (4:1, v/v) to give compounds (25 mg) and (56 mg) Subfraction M4.2 (18 g) was done as the same manner, further separated by RP-18 with MeOH– H2O (4:1, v/v) to affrord (250 mg) and (7 g) Fraction M5 (5 g) was applied on a silica gel chromatographic column and eluted with CHCl3-MeOH (2:1) in silica gel column chromatography to yield compound (40 mg) Experimental 3.3.2 Visconoside B (2) 3.1 General experimental procedures The optical rotations were measured on a ADP220 polarimeter (Bellingham + Stanley Ltd., RG224BA, UK) The IR data were recorded on a Bruker Tensor 27 FT-IR spectrometer (Bremen, Germany) The UV spectra were performed with a Jasco V-630 spectrophotometer (Tokyo, Japan) The high resolution electrospray ionisation mass spectroscopy (HR-ESI-MS) was recorded on a Bruker MicroTOF-QII spectrometer (Bruker Daltonik GmbH, Bremen, Germany) The 1H-NMR (500 MHz), 13C-NMR (125 MHz), DEPT, COSY, HSQC and HMBC spectra were recorded on a Bruker AM500 FT-NMR spectrometer using tetramethylsilane (TMS) as internal standard Column chromatography was carried out using silica gel normal-phase (230–400 mesh) and reversed-phase (Merck KGaA, 64271 Darmstadt, Germany) Analytical TLC was carried out in silica gel plates (Kieselgel 60 F254, Merck) Compounds were visualized by spraying with aqueous 10% H2SO4 and heating for 3–5 3.2 Plant material The leaves of Cleome viscosa L were collected in Ben Cat, Binh Duong province, Viet Nam, in May 2015; and identified by Prof Vo Van Chi A voucher specimen (No VH/MINH-0515) was deposited in the Institute of Chemical Technology, Vietnam Academy of Science and Technology 3.3 Extraction and isolation Powdered leaves of Cleome viscosa L (8 kg) were extracted with 95% EtOH for three times (3  30 L, total amount 90 L) at room temperature, filtered residue, removed solvents under low pressure, obtained crude extract (980 g) Then, crude extract was applied to solid-phase extraction procedures and successively partitioned into n-hexane (70 g), CHCl3 (150 g), EtOAc (260 g) and MeOH (450 g) The MeOH extract was subjected to silica gel column chromatography and eluted with gradient solvent system of chloroform – methanol (95:5–5:95) to afford seven fractions: M1 (25 g), M2 (30 g), M3 (86 g), M4 (75 g), M5 (60 g), and M6 (72 g) Fraction M2 (5 g) was chromatographed on silica gel and eluted with CHCl3-MeOH (10:1) to obtain compounds (72 mg) and 3.3.1 Visconoside A (1) Yellow amorphous powder; ½aŠ25 D À 0.94 (c 0.01, MeOH); IR nmax (MeOH): 3317, 2943, 2831, 1449, 1417, 1114, 1022 cmÀ1; UV (MeOH) lmax: 257 and 348 nm; HR-ESI–MS: m/z 821.2116 [M+Na]+ (calcd for C35H42O21Na, 821.2116); 1H and 13C NMR data (DMSO-d6), see Table Yellow amorphous powder; ½aŠ25 D À 1.23 (c 0.01, MeOH); IR nmax (MeOH): 3317, 2943, 2831, 1449, 1416, 1115, 1022 cmÀ1; UV (MeOH) lmax: 249 and 336 nm; HR-ESI–MS: m/z 1027.2680 [M+Na]+ (calcd for C46H52O25Na, 1027.2695); 1H and 13C NMR data (DMSO-d6), see Table 3.4 Acid hydrolysis Each new compounds (2 mg) was refluxed with 2N aq CH3COOH (5 mL) for h at 100  C After extraction with CH3Cl (3  mL), the aqueous layer was repeatedly evaporated to dryness with MeOH until neutral, and then analyzed by TLC over silica gel (MeCOEt–isoPrOH–Me2CO–H2O 20:10:7:6) by comparison with authentic samples (L-rhamnose Rf 0.65; D-glucose Rf 0.40) (Nguyen et al., 2015) Acknowledgments This research work has been financially supported by the Vietnam Academy of Science and Technology, Project No VAST DLT.09/16-17 References Bose, U., Bala, V., Ghosh, T.N., Gunasekaran, K., Rahman, A.A., 2011 Antinociceptive, cytotoxic and antibacterial activities of Cleome viscosa leaves Braz J Pharmacog 21, 165–169 Chatterjee, A., Chattopadhyay, S.K., Tandon, S., Kaur, R., Gupta, A.K., Malik, R., Kant, R., 2013 Isolation of a unique dipyridodiazepinone metabolite nevirapine during large scale extraction of cliv-92 from the seeds of Cleome viscosa Ind Crops Prod 45, 395–400 Chauhan, J.S., Srivastava, S.K., Srivastava, S.D., 1979 Kaempferide 3-glucuronide from the roots of Cleome viscosa Phytochemistry 18, 69 Donkor, A.M., Bugri, K.G., Atindaana, E.A., 2014 Evaluation of antibacterial potentiation of crude extracts of Phyllanthus amarus, Tamarindus indica and Cleome viscosa and their formulation Int J Plant Res 4, 23–28 Gaind, K.N., Singla, A.K., Wallace, J.W., 1981 Flavonoid glycoside of Kalanchoe spathulata Phytochemistry 20, 530–531 Gopal, V.Y., Ravindernath, A., Kalpana, G., Reddy, P.V., 2012 Antitumor activity of Cleome viscosa against ehrlich ascites carcinoma (EAC) in swiss albino mice Int J Phytopharm 2, 51–55 Gupta, P.C., Nisha Sharma, N., Rao Ch, V., 2011 Comparison of the antioxidant activity and total phenolic, flavonoid content of aerial part of Cleome viscosa L Int J Phytomed 3, 386–391 Han, L., Shi, P., Dong, Y., Wang, T., Li, X., Hao, J., Zhang, Y., Wang, T., 2015 New rare sinapoyl acylated flavonoid glycosides obtained from the seeds of Lepidium apetalum Willd Molecules 20, 13982–13996 N.M Phan et al / Phytochemistry Letters 18 (2016) 10–13 Imperato, F., 1984 A new kaempferide 3,7-diglycoside from the fern Asplenium bulbiferum Chem Ind 18, 667–668 Ishiguro, K., Nagata, S., Fukumoto, H., Yamaki, M., Takagi, S., Isoi, K., 1991 A flavanonol rhamnoside from Hypericum japonicum Phytochemistry 30, 3152–3153 Jana, A., Biswas, S.M., 2011 Lactam nonanic acid, a new substance from Cleome viscosa with allelopathic and antimicrobial properties J Biosci 36, 27–35 Jente, R., Jakupovic, J., Olatunji, G.A., 1990 A cembranoid diterpene from Cleome viscosa Phytochemistry 29, 666–667 Kumar, S., Ray, A.B., Konno, C., Oshima, Y., Hikino, H., 1988 Cleomiscosin D, a coumarino-lignan from seeds of Cleome viscosa Phytochemistry 27, 636–638 Kumar, S.V., Christina, A.J.M., GeethaRani, P.V., Nalini, G., Chidambaranathan, N., 2009 Antifibrotic effect of Cleome viscosa Linn on Carbon tetra chloride (CCl4) induced liver fibrosis Der Pharma Chem 1, 92–96 Mali, R.G., 2010 Cleome viscosa (wild mustard): a review on ethnobotany, phytochemistry, and pharmacology Pharm Biol 48, 105–112 Minh, P.N., Tri, M.D., Phat, N.T., Dat, B.T., Hanh, N.N., Luan, N.Q., Thanh, M.T., Huynh, C.H., 2015 Two new flavonol glycosides from the leaves of Cleome chelidonii L.f J Asian Nat Prod Res 17, 338–342 Mishra, A., Mishra, A.K., Jain, S.K., 2010 Anticonvulsant activity of Cleome viscosa seed extracts in swiss albino mice Int J Pharm Pharm Sci 2, 177–181 13 Nguyen, T.P., Le, T.V.H., Mai, D.T., Le, T.D., Phan, N.M., Bui, T.D., 2015 Two new oleanane-type triterpene saponins from the leaves of Schefflera sessiliflora De P.V Phytochem Lett 11, 102–105 Rao, Y.K., Geethangili, M., Chan, H.S., Wu, W.S., Tzeng, Y.M., 2009 High-performance liquid chromatographic determination of kaempferol glycosides in Cinnamomum osmophloeum leaves Int J Appl Sci Eng 7, 1–9 Ray, A.B., Chattopadhyay, S.K., Kumar, S., 1985 Structures of cleomiscosins, coumarinolignoids of Cleome viscosa seeds Tetrahedron 41, 209–214 Senthamilselvi, M.M., Kesavan, D., Sulochana, N., 2012 An anti-inflammatory and anti-microbial flavone glycoside from flowers of Cleome viscosa Org Med Chem Lett 2, 1–5 Song, N., Xu, W., Guan, H., Liu, X., Wang, Y., Nie, X., 2007 Several flavonoids from Capsella bursa-pastoris (L.) medic Asian J Tradit Med 2, 218–222 Srivastava, S.K., Chauhan, J.S., Srivastava, S.D., 1979 A new naringenin glycoside from Cleome viscosa Phytochemistry 18, 2057–2058 Srivastava, S.K., 1980 Stigmasta-5,24(28)-diene 3b-O-a-L-rhamnoside from Cleome viscosa Phytochemistry 19, 2510–2511 Wang, D.M., Pu, W.J., Wang, Y.H., Zhang, Y.J., Wang, S.S., 2012 A new isorhamnetin glycoside and other phenolic compounds from Callianthemum taipaicum Molecules 17, 4595–4603  ... C.H., 2015 Two new flavonol glycosides from the leaves of Cleome chelidonii L.f J Asian Nat Prod Res 17, 338–342 Mishra, A., Mishra, A.K., Jain, S.K., 2010 Anticonvulsant activity of Cleome viscosa. .. extraction of cliv-92 from the seeds of Cleome viscosa Ind Crops Prod 45, 395–400 Chauhan, J.S., Srivastava, S.K., Srivastava, S.D., 1979 Kaempferide 3-glucuronide from the roots of Cleome viscosa. .. A.M., Bugri, K.G., Atindaana, E.A., 2014 Evaluation of antibacterial potentiation of crude extracts of Phyllanthus amarus, Tamarindus indica and Cleome viscosa and their formulation Int J Plant