Pmlet0129 91.93

Casearia velutina Blume (Flacourtiaceae/Salicaceae sensu lato) [1]are shrubs or small arbors that are distributed mainly in the Yunnan and Hainan provinces of China [2]. It is used in folk med- icines for the treatment of profluvium, gastrelcosis, and inflam-matory infections [3]. However, no phytochemical investigations on this plant were reported before our research on it [4].
Aiming to discover new or bioactive compounds from Flacourtia-ceae/Salicaceae s. l. and to provide useful chemotaxonomic refer- Xing-Yun Chai1,2, Fei-Fei Li1, Chang-Cai Bai1, Zheng-Ren Xu1, ence for them, the stems of Casearia velutina were chemically in- vestigated. Herein, we describe the isolation and structural eluci- 1 State Key Laboratory of Natural and Biomimetic Drugs, School of dation of three new acylated glucosides (1–3), together with 13 Pharmaceutical Sciences, Peking University Health Science Center, known compounds, and the protective effect against H2O2-in- duced impairment in PC12 cells and the inhibition against snake 2 Key Laboratory of Marine Bio-resources Sustainable Utilization, venom phosphodiesterase I (PDE I) of the isolated compounds. In South China Sea Institute of Oceanology, Chinese Academy of addition, the chemotaxonomy of the genus Casearia is briefly discussed.
Compound 1 was obtained as a colorless gelatinous solid. Its mo- lecular formula was determined as C18H26O11 by the positive HR‑ESI‑MS m/z = 441.1362 [M + Na]+. GC analysis showed the Chemical investigation of the stems of Casearia velutina led to the presence of D-glucose after acid hydrolysis of 1. The 1H‑NMR isolation and structural elucidation of three new acylated glyco- " Table 1) displayed a benzoyl moiety [δH = 7.98 (2H, sides, casearicosides A–C (1–3), together with 13 known com- d, J = 7.5 Hz), 7.55 (1H, t, J = 7.5 Hz), and 7.42 (2H, t, J = 7.5 Hz)], a pounds. The structures of the new compounds were established β-glucose moiety (δH = 4.32, 1H, d, J = 8.0 Hz, H-1′), and the frag- by spectroscopic and chemical methods. These isolates were ment of 2-hydroxymethylbutane-1,2,3,4-tetrol [5], including evaluated for protective effects against H2O2-induced impair- two hydroxymethylene singlets [δH = 3.57 (2H, s, H-1), 3.54 (2H, ment in PC12 cells and inhibitory activity against snake venom s, H-5)], a hydroxymethene multiplet [δH = 4.11 (1H, dd, J = 10.5, phosphodiesterase I. A brief chemotaxonomy of the genus Casea- 3.0 Hz, H-4a), δH = 3.48 − 3.60 (1H, m, H-4b)], and a hydromethine [δH = 3.85 (1H, dd, J = 8.5, 3.0 Hz, H-3)], which were supported bythe 13C‑NMR data and confirmed by the HMBCs from H-4 to C-3 (δC = 73.4) and C-2 (δC = 76.0) and from H-1, H-5 to C-3. Further Casearia velutina Blume · Salicaceae · casearicosides A – C · Fla- HMBCs from H-1′ to C-4, H-6′ to carbonyl (δC = 167.9) of the ben- zoyl moiety suggested that the benzoyl is attached at C-6′ and thesugar is linked with the aglycone at C-4 (l Supporting information available online at Compound 1 was hydrolyzed and purified by preparative TLC http://www.thieme-connect.de/ejournals/toc/plantamedica (CHCl3 − MeOH − H2O 6 : 4 : 1, Rf = 0.28) to afford the aglycone.
NMR data of compounds 1–3 (500/125 MHz; δ in ppm, J in Hz) (a Measured in CD3OD; b Measured in DMSO‑d6; c Partly overlapped).
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Chai X-Y et al. Three New Acylated … Planta Med 2010; 76: 91–93 (9) [13], isotachioside (10) [13], 3-methoxyl-4-hydroxyl-1-O-β- D-furanapiose-(1′′→6′)-β-D-glyucopyranose phenglycoside (11) [14], (+)-aviculinol 3α-O-α-L-rhamnoside (12) [15], (+)-lyoniresi-nol 3α-O-α-L-rhamnoside (13) [16], (−)-lyoniresinol 3α-O-α-L-rhamnoside (14) [17], (−)-lyoniresinol 3α-O-β-D-xylopyranoside(15) [18], and 5-O-trans-caffeoyl quinic acid (16) [19] were iden-tified by comparing their NMR data with references. Copies of theoriginal spectra can be obtained from the corresponding author.
Previous studies on the phylogeny of the Flacourtiaceae, based onone- or three-gene sequences [20, 21], have shown that the fam-ily is polyphyletic and recognized by two major clades, one withclose affinities to the Salicaceae (including Poliothyrsis, Xylosma,Scolopia, Homalium, Idesia, and Casearia), and the other genera(i.e., Gynocardia) with cyclopentenoid cyanogenic glycosideswere separated into the Achariaceae. However, the genus Casea- The ESI‑MS m/z = 175.0 [M + Na]+ data and the specific rotation of ria was weakly supported and disputed, as the existing research the aglycone [+ 4.5 (c 0.002, MeOH)] indicated that the absolute either provides no direct biosynthetic link to Salicaceae, which configuration of C-3 was R (lit. value [R] + 4.3) rather than S (lit.
richly produces the characteristic glucosides of 2,5-dihydroxy- value [R] −4.0) [5]. Thus, the aglycone of 1 is (3R)-2-(hydroxy- benzyl alcohol, or cannot demonstrate that this genus contains methyl)butane-1,2,3,4-tetrol, and 1 was elucidated as (3R)-2-(hy- cyanohydrin [22]. Thus, the isolation of 4 and 5, together with droxymethyl) butane-1,2,3,4-tetrol-4-β-D-(6-O-benzyl)glucopyr- their possible precursor 6, from C. velutina as the first two phenyl cyanogenic glycosides isolated from Casearia will provide power- Compounds 2 and 3 were obtained as amorphous white pow- ful proof that this genus is obviously different from the other genera of Salicaceae, although more evidence is necessary to sup- C23H28O11 by the positive HR‑ESI‑MS, indicating that they are port Casearia being united in the Achariaceae. Apart from the cy- isomers. The D-glucose was detected by TLC and GC analysis after anohydrins, other isolates such as the phenolic glycosides 9–11 acid hydrolysis of both compounds. The 1H‑NMR and 13C‑NMR (benzyl alcohol type) cannot support this genus showing close af- " Table 1) of 2 and 3 show high similarity, exhibiting a ben- zoyl moiety, a 1,2,4-trisubstituted benzene, a 6-O-acylated gluco- Compounds 1–4, 6–11, and 16 were evaluated for protective ef- syl moiety, and a methoxy, a hydroxymethylene, and two hy- fects against H2O2-induced impairment in PC12 cells. Com- droxymethines in each compound. HMBCs from H-9 (δH = 3.71, pounds 1, 4, 6, and 16 exhibited significant protective effects 3.10) to C-7 (δC = 72.6), from H-8 (δH = 3.63) to C-1 (δC = 133.5), " Table 2), with cell viability of 77.0–97.1%, compared with the from H-5 (δH = 6.62) to C-3 (δC = 147.0), from H-6 (δH = 6.68) to C- H2O group (control, 100%) and the H2O2 group (model, 54.4%).
4 (δC = 145.5), and from -OCH3 (δH = 3.69) to C-3 established the Compounds 4, 6, and 8–11 were tested for their inhibition effects aglycone as 3-methoxy-4-hydroxy-phenylpropanetriol (arylgly- against snake venom phosphodiesterase I (PDE I). Compound 8 cerol) for both 2 and 3. In addition, HMBCs from H-1′ to C-9 and showed the highest inhibitory rate, 5.7 % at 5 µM, which is equal from H-6′ to C-7′′ (carbonyl) established the structure (without to that of the weak control L-cysteine (5.6 %) at 5 µM.
stereochemistry) of 2 and 3 as 3-methoxy-4-hydroxy-phenylpro-panetriol-9-O-β-D-(6-O-benzoyl)glucopyranoside.
A comparison of the NMR data of 2 and 3 indicated the differ- " Table 1), suggesting them to be ste- The dried stems (7.0 kg) of C. velutina were extracted three times reoisomers at C-7 and C-8. According to the literature [6], erythro with 80 % EtOH (3 × 42 L). After evaporation of the solvent, the arylglycerols measured in DMSO-d6 exhibit a 1.3 ppm 13C‑NMR soluble fraction was suspended in H2O and extracted successively Downloaded by: Peking University Health Science Center. Copyrighted material.
difference for C-8 and C-7 (ΔδC8−C7), but for threo arylglycerols, with petroleum ether, EtOAc, and n-BuOH. The n-BuOH extract the ΔδC8−C7 difference value is 2.9 ppm. Another proposed meth- (120 g) was subjected to a silica gel chromatography column od to determine the relative stereochemistry of this type of com- eluted with a gradient of CHCl3−MeOH to afford 9 fractions (Frs.
pound is by comparing the coupling constant of H-7 and H-8 I−IX). Fr. IV afforded compounds 2 (10 mg), 3 (7.0 mg), 4 (90 mg), 5 (J = 6.3 Hz as erythro and J = 5.0 Hz as threo) [7, 8]. Therefore, the (5.0 mg), 8 (25 mg), 12 (16 mg), 13 (80 mg), 14 (5.0 mg), and 15 ΔδC8−C7 values of 1.4 ppm for 2 and of 0.3 ppm for 3 determine (28 mg). Fr. VI afforded compounds 1 (13 mg) and 6 (203 mg).
that they are both in the same erythro form, which is also in good Compounds 7 (25 mg), 9 (5.0 mg), 10 (6.0 mg), 11 (11 mg), and accordance with their coupling constants of H-7 and H-8 16 (10 mg) were obtained from Fr. VII.
(J = 5.0 Hz). Based on the above NMR interpretation combined Determination of the configuration of D-glucose was carried out by with their opposite optical rotation values ([α]25 TLC and GC analysis according to the procedure described in a pre- − 2.5 of 3), 2 and 3 were finally elucidated as (+)(erythro)-3-meth- vious paper [23]. The procedure reported by Denizot [24] was used 4-hydroxy-phenylpropanetriol-9-O-β-D-(6-O-benzoyl)glu- for the measurement of protective effects against H2O2-induced copyranoside, named casearicoside B (2), and as (−)(erythro)-3- impairment in PC12 cells, and the method described in a previous 4-hydroxy-phenylpropanetriol-9-O-β-D-(6-O-benzo- paper [25] was used for the measurement of inhibition against yl)glucopyranoside, named casearicoside C (3).
PDE I. For a detailed protocol, see the Supporting Information.
Additionally, the 13 known compounds (2R)-prunasin (4) [9], Casearicoside A (1): Colorless gelatinous solid; [α]25 (2S)-sambunigrin (5) [10], 2-β-D-glucopyranosyl-2-phenylacetic MeOH); UV (MeOH): λmax (log ε) = 285 (3.48), 325 (sh) (2.56) nm; acid amide (6) [11], 2-O-β-D-glucopyranosyl-2-hydroxy-phenyl- IR (KBr): νmax = 3383, 2891, 1715, 1602, 1453, 1319, 1282, 1177, acetic acid (7) [12], benzylalcohol glucoside (8) [9], tachioside Chai X-Y et al. Three New Acylated … Planta Med 2010; 76: 91–93 9 Seigler DS, Pauli GF, Nahrstedt A, Leen R. Cyanogenic allosides and gluco- Protective effects of compounds 1, 4, 6, and 16 against H2O2-in- sides from Passiflora edulis and Carica papaya. Phytochemistry 2002; 10 Towers GHN, Mcinnes AG, Neish AC. The absolute configuration of the phenolic cyanogenic glucosides taxiphyllin and dhurrin. Tetrahedron 11 Nahrstedt A, Rockenbach J. Occurrence of the cyanogenic glucoside pru- nasin and its corresponding mandelic acid amide glucoside in Olinia species (Oliniaceae). Phytochemistry 1993; 34: 433 12 Abrosca BD, DellaGreca M, Fiorentino A, Monaco P, Previtera L, Simonet AM, Zarreli A. Potential allelochemicals from Sambucus nigra. Phyto- a Measured in 150 µM; b measured in 10 µM with purity > 95% (x ± s); * p < 0.01 com- 13 Zhong XN, Otsuka H, Ide T, Hirata E, Takeda Y. Hydroquinone diglycoside acyl esters from the leaves of Myrsine seguinii. Phytochemistry 1999; pared with 150 µM H2O2 group (model); ** p < 0.01 compared with uncheated group 14 Kanchanapoom T, Kasai R, Yamasaki K. Iridoid and phenolic diglycosides from Canthium berberidifolium. Phytochemistry 2002; 61: 461–464 15 Kim HJ, Woo ER, Park H. A novel lignan and flavonoids from Polygonum 1071, 911, 807, 715, 628 cm−1; HR‑ESI‑MS: m/z = 441.1362 [M + aviculare. J Nat Prod 1994; 57: 581–586 16 Smite E, Pan H, Lundgren LN. Lignan glycosides from inner bark of Betu- Na]+ (calcd. for C18H26O11Na: 441.1367); 1H‑ and 13C‑NMR data: la pendula. Phytochemistry 1995; 40: 341–343 17 Kaneda N, Kinghorn AD, Farnsworth NR, Tuchinda P, Udchachon J, Santi- Casearicoside B (2): Amorphous white powder; [α]25 suk T, Reutrakul V. Two diarylheptanoids and a lignan from casuarina junghuhniana. Phytochemistry 1990; 29: 3366-3368 18 Hiroyuki F, Tetsuya S, Nobutoshi T. Chemical evaluation of Betula spe- max = 3422, 2252, 2127, 1647, 1282, 1049, 1026, 1002, 827, 766, cies in Japan. I. Constituents of Betula ermanii. Chem Pharm Bull 629 cm−1; HR‑ESI‑MS: m/z = 503.1519 [M + Na]+ (calcd. for C23H28O11Na: 503.1524); 1H‑ and 13C‑NMR data: see l 19 Lu Y, Sun Y, Foo LY, McNabb WC, Molan AL. Phenolic glycosides of forage Casearicoside C (3): Amorphous white powder; [α]25 legume Onobrychis viciifolia. Phytochemistry 2000; 55: 67–75 20 Soltis DE, Soltis PS, Chase MW, Mort ME, Albach DC, Zanis M, Savolainen V, Hahn WH, Hoot SB, Fay MF, Axtell M, Swensen SM, Prince LM, Kress WJ, max = 3418, 2251, 2127, 1601, 1283, 1050, 1027, 1003, 827, 76, Nixon KC, Farris JA. Angiosperm phylogeny inferred from 18S rDNA, 524 cm−1; HR‑ESI‑MS: m/z = 503.1519 [M + Na]+ (calcd. for rbcL, and atpB sequences. Bot J Linn Soc 2000; 133: 381–461 C23H28O11Na: 503.1524); 1H‑ and 13C‑NMR data: see l 21 Chase MW, Zmartzty S, Lledó MD, Wurdack KJ, Swensen SM, Fay MF.
When in doubt, put it in Flacourtiaceae: a molecular phylogenetic anal- ysis based on plastid rbcL DNA sequences. Kew Bull 2002; 57: 141–181 22 Mosaddik MA, Forster PI, Booth R, Waterman PG. Phenolic glycosides The 1H- and 13C‑NMR spectra for compounds 1–3, detailed isola- from some Australian species of Flacourtiaceae (Salicaceae sensu lato).
tion protocols, acid hydrolysis and sugar analysis, bioactivity as- say procedures, and structures of compounds 4–16 are available 23 Chai XY, Song YL, Xu ZR, Shi HM, Bai CC, Bi D, Wen J, Li FF, Tu P. Itosides J– N from Itoa orientalis and the structure-anti-COX‑2 activity relation-ship of phenolic glycosides. J Nat Prod 2008; 71: 814–819 24 Denizot FLR. Rapid colorimetric assay for cell growth and survival mod- ifications to the tetrazolium dye procedure giving improved sensitivity and reliability. J Immunol Methods 1986; 89: 271–277 The authors wish to thank Prof. Xiao-Ping Pu (Peking University 25 Chai XY, Ren HY, Xu ZR, Bai CC, Zhou FR, Ling SK, Pu XP, Li FF, Tu PF. Inves- Health Sciences Center) for her kind help in the inhibition assay tigation of two Flacourtiaceae plants: Bennettiodendroon leprosipesand Flacourtia ramonchi. Planta Med advance online publication against PDE I. This work was supported by the program Chang- jiang Scholar and Innovative Team in University (grant number985-2-063-112).
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1 The Angiosperm Phylogeny Group. An update of the angiosperm phylog- eny group classification for the orders and families of flowering plants:APG II. Bot J Linn Soc 2003; 141: 399–436 2 Editorial Board of Flora Reipublicae Popularis Sinicae of Chinese Science Academy. Flora Reipublicae Popularis Sinicae, Vol. 52. Beijing: Beijing 3 Li FF, Chai XY, Xu ZR, Ren HY, Tu PF. Chemical constituents from stems of Georg Thieme Verlag KG Stuttgart · New York · Casearia velutina. Chin J Chin Mater Med 2008; 39: 984–986 4 Chai XY, Lu YN, Ren HY, Tu PF. Advances in studies on chemical constit- uents and bioactivities of plants from Flacourtiaceae. Chin J Chin Mater 5 Kitajima J, Suzuki N, Ishikawa T, Tanaka Y. New hemiterpenoid pentol State Key Laboratory of Natural and Biomimetic Drugs and monoterpenoid glycoside of Torillis japonica fruit and considera- tion of the origin of apiose. Chem Pharm Bull 1998; 46: 1583–1586 6 Otsuka H, Takeuchi M, Inoshiri S, Sato T, Yamasaki K. Phenolic com- pounds from Coix lachrymal-jobi var. ma-yuen. Phytochemistry 1989; 7 Seidel V, Bailleul F, Waterman PG. Novel oligorhamnosides from the Fax: + 86 10 82 80 27 [email protected] stem bark of Cleistopholis glauca. J Nat Prod 2000; 63: 6–11 8 Masataka S, Masao K. Studies on the constituents of Osmanthus spe- cies. X. Structures of phenolic glucosides from the leaves of Osmanthusasiaticus. Chem Pharm Bull 1992; 40: 325–326 Chai X-Y et al. Three New Acylated … Planta Med 2010; 76: 91–93

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