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Natural Product Research

Formerly Natural Product Letters
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Four New Diarylheptanoid Glycosides From Alnus Rubra Bark

Ruben F. Gonzalez-laredo a; Jie Chen b; Yvette M. Karchesy b; Joseph J. Karchesy b a institoto Tecnolgico de Durango. Durango. Mxico b Department of Forest Products, Oregon State University. Corvallis, OR

Online Publication Date: 01 March 1999 To cite this Article: Gonzalez-laredo, Ruben F., Chen, Jie, Karchesy, Yvette M. and Karchesy, Joseph J. , (1999) 'Four New Diarylheptanoid Glycosides From Alnus Rubra Bark', Natural Product Research, 13:2, 75 - 80 To link to this article: DOI: 10.1080/10575639908048826 URL: http://dx.doi.org/10.1080/10575639908048826

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R i n d i Malaysia n

FOUR NEW DIARYLHEPTANOJDGLYCOSIDES FROMALMISRUBRA BARK

RUBEN F. GONZALEZ-LAREDO, JIE CHEW, YWTIE M. KARCHFSY AND JOSEPH J. KARCHESY 'Institute Tecnol6giw & h a n g o , 34080 Durango, -0.. Mikico; Depmhnent of Forest Products, Oregon State University, Corvdlh, OR 97331
(Received 10th June 1998)

Abstrrct: Four nm diarykptanoid glymides, rubranosidesA3,C and D (1-4). w r isolated fiom d aldcr (Alms tubm)bark. All contain the COmmoIL ee aglyconc moiety, rubran01 (5). Keywordr:diqaqbmid, Alnus rubra, Betulaceae,rubranosides, mbmol, red aldcr.
INTRODUCTION
A l m s d r a , Bong (red alder) is the major h r w o species p w k g in the adod pacific ~ 0 r t h ~ when . I ~ ~ t harvested for timber,its bark is forests used for fucl or as a plywood adhesive cxttnder. Indigenous historyofus~tbebarkformedicalandcoloringpurposts. Previousresearch spcs have a has shownthe diarylkptanoid xyloside orcgonin t be a precmorto the redo orange mining phenomenon which gives this tree its cornmollname. I frnther n investigationof the bark, four n w diarylheptanoidgly&dcs have becn isolattd e and their strwhms elucidated by spectrometric techniques.
RESULTS AND DISCUSSION

An acetone-watex extract of freshly collected red alder bark was chromatographed over Sephadcx LH-20 and silica gel to give four diarybptanoid glymsides, the rubranosides 1-4. H I3CNMR spectra (Tables 1 and 2) indicated that these and glycosidcs had a common aglyconc whose NMR signals wcre very similar t o
75

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76

R.F. GONZALEZ-LAREDO et al.

thoseofcentrolotml and its Slycosidts ( s c c r ~ s i d t s ) . ~e centrolotal the Lk i

diarykphnoid chains i 1-4 CoIlSisttd of six C H z groups and one hydmxy n methine group at C-3. contrastto ccntrolobol, the two aromatic ring systems of In the Qarylhepcanoidunits were wtshol group as evidenced by the characteristic d i h y h x y bmmg carboa s i & at 144.0-144.2 145.9-146.1 and ppm 3,4and aromatic proton signals i an ABX pa#ern with ortho and meta coupling n
constants.

Incompoundl,theCNMRseriesofsignalsat103.2,78.2,77.8,75.4, 71.8and 62.9pprn supporttd the assignment of a f3-wglucOsylm i t to the oey structun.The &glucosyi Idage was collfifmed by the doublet ( J = 7.6Hz)from the aaomcric p t o n signal a 6 4.30. position of the glywsidic lrnkage was t Ttae contimed by an HMBC spectnnn, which s h o d Connectivity of the anomeric proton signal to the hcptanoid C-3signal at 79.5 ppm. Also, the anomeric glycosyl signal at 103.2 ppm s h o d Connectivity to the hcptanoid H-3 signal at 6 3.70. Hydrolysis of 1 using fj-glucosidasegave t agiyconc 5, rubranol, and glucosc. & FAB-MS of 1 gave an intcnse ion peak at m/z 493 confirming its m l c l r weight. The assignment of an 8 dguration at C-3was done b oeua application of the I3CNMR glycosylation shift rule for secondary alcohols.7-9 Compsrison of shift values for C-2 (-2.6 pjm) and C-4 (-3.4 ppm) of 1 and S showedtk pr0-S carbon to be C-4and C-3thus & The structurt of rubranoside A (1) was thus assigned as (3~1,7-bis-(314-dihydroxyphcnyl)-hept glucopyranoside. The N M R spectra of compound 2 d i f f m d kom 1 only i the signals for n the glycosyl m i t (Tables 1 and 2). The 13CNMR signals at 104.3,75.3,78.1, oey 71.5and 67.0ppm indicated a &D-xylopyranoside.The ammcric proton signal appcarad a 6 4.22as a doublet ( J = 74 Hz)confimung the ~nfonnation t . Acidic hydrolysis of 2 gave 5 and xylost. An [M-HJ- ion peak at d z 463 confirmed the molecular weight The conformation at G was assigned on the 3 basis of the glycosylationrule since thc shift at C-4 (-3.1ppm, pro-S)was larger than the shift at C-2(2.3p) compatiscm of the 13CNMR spectrum of2 and on 5. Tht structun of rubranoside B (2), was thus assigned as (3E)-1,7-bis-(3,4~droxyphenyl)-hcptan-3-O-B-Dxylo~ide.

w-q-

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FOUR NEW DIARYLHEFTANOIDGLYCOSIDES

71

Compound 3 showed t o sets of 1 3 NMR sics for the g~ycosyl w ~ portion ofthespectnrmindicatingittobediglycosidc. Signalsat 103.8,75.5,78.2,71.7, 77.8 and 62.7 ppm wcrc consistent with a f3-D-glwpyranoside p u p . Signals at 105.1,74.4,87.5,70.1 and 66.4 ppm were suggestive of a ~-~-xylopyranosyl hf t group with a glycosylation induced s i t a the xylose C-3 (87.5 ppm). Since the glucose signals showed no such shift, this was a strong indicator that the glucosyl group was attached ((2-14-3) to the xylosyl group. An m-H]-ion peak at d z 625 confinnedthe molecular weight of 3. The assignment of the c o d g u t i o n of the C-3 position of the aglycone as was again done by comparing the gly~o~ylati~n Of C-3 (-2.3 p ~ m and C-4 (-3.2 p ~ m )The structure of shift ) . rubranoside C (3), was thus assigned as (3B)-1,7-bis (3,4dihy&oxyphenyl> heptan-3-O-f3-~glmpyranosyl(l-+3) j3-mylopymmidc. Compound 4 also exhibited I3CNMR signals for a diglycoside moiety. Signals at 110.9,78.1,80.7,75.1 and 65.8 ppm were attributed to a p-P apiofuranosyl m i t while those at 103.5,75.5,78.2,71.7,76.6 and 68.5 ppm oey were attriiuted to a j3-Dglucowranoside unit. Glycosylation at the glucose C-6 position was based on the down field shift of 5.6 ppn when compared to the signal for the unsubstituted glucose C-6 in compound 1. An HMBC spcctnrm confined attachment of the glucose unit to the heptaaoid C-3 position by showing correlation of the C-3 13CNMR signal (80.3 ppm) with the glucose anomeric proton (6 4.27). In addition, correlation was observed between the glucost C-6 carbon signal (68.5 ppm) and the apiose anomcric proton signal at S 5.03 (4 J = 2.2 Hz).Hydrolysis of 4 gave 5, apiose and glucosc. FAB-M!3 gave an w-H]-ion peak at m/z 625 confirming the molecular weight. The assignment of an configurationat C-3 was again based on the larger glycosylation s i t at C-4 (-3.1 hf ppm) cumpared to C-2 (-2.3 ppm). Compound 4, rubrawside D, was assigned the
structure (3E) 1,7-bis (3,4-dihydroxyphenyl>ptan-3-O-(~~apio~yl

( 1 4 ) P-D-glucopyranoside.

EXPERIMENTAL
was done oe using a Kratos m d l MS-5OTC m a s spectrometer with samples dissolved in a mixture of dithiothreitol and dithioerythntol. Optical rotations w e measured using a JASCO model DIP-370 polarimeter. Freshly collected Alnus nrbro bark (63 kg, voucher specimen deposited at Oregon State University Herbarium) from McDonald State Forest was extracted wt acetone-water (7:3) @ RT to give a reddish brown powder (6.4 kg) on ih evaporation. The crudc extract (100 g) was chromatographed over Sephadex LH20 (1 1 x 45 cm ) using an aq.MeOH @en& (O+ 100%MCOH). The fraction

NMR spectra were recorded on a B&er AM300 s e t o e e FAB-M!3 pcrmtr

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78

R.F. GONZALEZ-LAREDO et al.

eluted wt 20% MeOH (26 g) was rechromatographed(0.5 g portion) over ih Scphadex LH-20 using H2GMeOH (1: 1) to give compound 4 (70 mg) [ah,-5 1.7" (MeOR c 2.96). and o q o n i n (341 m ) Thc fraction eluted wt 50% MeOH in g. ih the first column (8.4 g) was e rc (248 mg) over S using C&,i EtOAc-MeOH (1:1:0.5) to give compounds 1(78 mg) [ab-28.3" (MeOH; c 0.3), 2 (18 mg) [ab-22.5 O (MeO% c 0.4). and 3 (50 mg) [=ID-30.3O(MeoH; c 0.75). R u b m l [ab-2.5O (MeOH, c 0.65), w-H]-m/z 331, RtO.60 (Si, =-Me20MeOH 6:3: 1) was obtained as an oil (13 mg) upon enqmatic hydrolysis of compound 1(22 mg) using p-glucosidase (35'C. 48 hrs). The hydrolysate was frcaedried and extracted wt ethyl acetate. The chirality of sugars was ih determined by GC of their trirncthylsbted (-)-2-butyl glywside derivatives.'o The sugars were obtained by acidic hydrolysis and their identification confirmed by TLC in cellulose (EtOAc-Pyr-Water 8:2: 1, and aniline phtalate spray reagent).

ACKNOWLEDGEMENT
The first author acknowledges permission leave from lTD and fin:ancial support from SEP-SElT-COSNET (Grants No.299.96-E 218.97-E). and

REFERENCES
1.
2.

3.
4.
5.

6.
7. 8.

9.

10.

E. Gmthcr. (1973). Ethnobotany of Western Washington.p. 33, R e v i d Edition, University of Washington Press, W e . D.R. Forlines, T. Tavenner, J.C.S. a and J.J. Karchesy. (1992). In: Wn Plant P o l y p k d s , (R. W. Htming~ay P.E. and Laks,Eds.) p. 767-782, Plenum Press, New York. J. J. Kmhesy, ML.Laver, D.F. Barofsky and E. Barofsky. (1974). J.C S.Chem Commun. 649-650. . A.A. Craveiro, A.C. Prado, O.R. Gottlieb, and P.C. Welerson (1970). Phytockmishy 9,1869-1875. M.N, N. Kenmochi, M.Fujita, N. Furukawa and T. Inoue. (1986). @ Chem P h a m Bull. 34,1056-1060. E. Smite, L. Lundgren and R. Andersson.(193). Phytochem 32,365-369. P.K.Agrawal. (1992). Phytochemisrry 31,3307-3330. S Seo,Y. Tomita, K. Ton and Y.Yoshimura (1978). J. Am Chem Soc, . 100,3331-3339. G. Speranza, P. Manitto, D. Monti and D. Pezzuto. (1992). J. Nut. Prod 55,723-729. GJGri, . . e w gJ.P.Kamerling and J F G V i g t a t . . . l e n h r(1978). Carbohydrare . Reseurch, 62,349-357.

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FOUR NEW DIARYLHEPTANOID GLYCOSIDES

79

n T b e 1. "C NMR data for rubranosides A-D(1-4) and rubran01(5) i CD30D. al

7-

1 2 3 4 5 6 7

31.8 37.9 79.5 34.8 2. 56 33.0 36.2

3. 21 38.2 80.2 35.1 25.8 33.1 3. 63

32.0 38.2 80.2 35.0 25.6 32.9 36.2

32.1 38.2 80.3 35.1 25.8 33.1 36.2

32.4 40.5 7. 18 38.2 2. 63 32.9 36.2

, 1" ' l 2," '2 3,3" ' 4,4" 5',5" 6',6"

135.8,135.8 117.0,116.7 146.0,1 5 9 4. 144.0,144.0 116.4,116.4 121.0,120.8

135.8,135.8 116.8,116.7 146.1,146.1 144.2,144.2 116.4,116.4 120.8,120.8

135.7,135.8 116.7,116.7 146.0,146.0 144.0,lM.O 1 6 4 116.4 1., 1 0 9 120.9 2.,

1 5 8 1 5 9 135.4,135.7 3., 3. 1 6 7 116.7 1 6 3 116.2 1., 1., 1460,146.0 146.1,la.O 1 4 1 l . 144.1,lM.O 4.,Ml 116.4, 116.4 116.6, 116.6 120.8,121.0 120.7,120.7

103.2 75.4 78.2 71.8 7. 78 62.9

143 0. 75.3 7. 81 7. 15 67.0

103.8 75.5 78.2 71.7 77.8 62.7

135 0. 75.4 78.1 71.7 76.6 68.5

105.1 74.4 8. 75 7. 01 66.4

110.9 7. 81 8. 07 75.1 65.8

Table 2.
2
2.52 m 1.72m 3.61 m 1.52 m 1.37m 1.U)m 2.43 t (7.4) 2.54 m 1.72 m 3.56 - 3.65 m ' 1.53 m 1.38m 1.53 m 2.44 m 2.55 t 0 L76m 3.66 m 1.55m 1.38 m 1.53 m 2.44 m 2.56 m 1.63 m 350 m 1.35- 1.Mm 1.53m 2.44 t (7.5)

'H NMR data for rubranmides A-D (1-4) and rubranol(5) in C D 3 O D .

3
4 5

2.53 t (7.5) 1.73m 3.70 m 1.55 m 1.35 m 1.50 m 2.41 t (7.5)

s,5"
~~ ~

2'. 2"
6.60 d (2). 6.62 d (2) 6.65 d (8), 6.66 d (8) 6.47 dd (2, a), 6.48 dd (2.8)

6'. 6"

6.59d(1.8).6.69d(1.8) 6.64 d (8)' 6.48dd(1.8.8)'

6.59 d (2), 6.61 d (2) 6.61 d (2). 6.65 d (2) 6.59 d (1.8).6.61 d (2) 6.64 d (8),6.65 d (8) 6.66 d (8.1). 6.68 d (8.1) 6.64 d (8),6.65 d (8) 6.47 dd (2,8)' 6.47 dd (2.8.I), 6.54 & (2.8.1) 6.47 dd (I .8.8). 6.49 dd (2,s) I Downloaded By: [Educacin Superior Tecnolgica] At: 07:46 28 June 2007
~~

9
~.

I 2-5 64 6b

4.30 d (7.6) ' 3.20 3.41 m 3.89 dd (2, 11.7) 3.72 dd (5.3, 11.7)

4.55 d (7.8) 3.17 -3.41 m' 3.88 dd (2, 11.8) 3.56 3.65 m '

4.27 d (7.l) 3.17-3.38m 3.99 dd (1.5. 10.8) 3.64 m

~~ ~

rJIu

1 2 3 4 Sa 5b

4.22 d (7.4) 3.l8m 3.32 m 3.50 m 3.86 dd (5.4, 11.4) 3.l8m

4.26 d (7.6) 3. I7 - 3.41 me 3.48 t (8.8) 3.56 3.65 m' 3.91 dd (5.4, 11.4) 3.17 3.41 m '

4-potmskml
5.03 d (2.2) 3.92 (2.2) 3.94 d (9.6). 3.75 d (9.6) 3.60 d (1 I). 3.53 d (I 1) b)f-pm(onsignal c) six p + m a i p l
a)

2 4d4b W5b