Analiz Konpozisyon Cistanche Tubulosa: Separasyon ak Estrikti Phenylethanoid Glycoside ak Neolignan Glycoside

Kontakte:joanna.jia@wecistanche.com

Konstitiyan yo nan Hook Cistanche Tubulosa (Schrenk). F. II.1) Izolasyon ak estrikti yon nouvo glikozid feniletanoid ak yon nouvo glikozid Neolignan

Fumio Yoshizawa,* yon Takeshi Deyama/ Nobuo Takizawa/ Khan UsmanghaniJ ak Mansoor Ahmad6

Laboratwa Rechèch Santral yo, Yomeishu Seizo Co., Ltd., yon 2132-37 Nakaminowa, Minowa-machi, Kamiina-gun, Nagano 399-46, Japon ak Depatman Pharmacognosy, Fakilte Famasi, University of Karachi? Karachi-32, Pakistan. Li te resevwa 9 novanm 1989

Yon nouvophenylethanoid glikozidyo te rele tubuloside E (I), ak yon nouvoneolignan glikozid, alkòl dehydrodiconiferyl yO-0-D-glucopyranoside (II), yo te izole nan tout plant yo nanCistanche tubulosa (Schrenk) Hook. f. (Orobanchaceae), together with dehydrodiconiferyl alcohol 4-O-^-D-glucopyranoside (III), syringalide A 3'-a-L- rhamnopyranoside (IV), isosyringalide S'-a-L-rhamnopyranoside (V), (+ )-syringaresinol O-^-D-glucopyranoside (VI), ( + )-pinoresinol O-^-D-glucopyranoside (VII), liriodendron (VIII), 6-deoxycatalpol (IX), 8-epiIoganic acid (X), 20-hydroxyecdysone (XI), 8-hydroxygeraniol l-j?-D-glucopyranoside (XII) and syringin (XIII). The structure of tubuloside E (I) was established as 2-(3,4-dihydroxy phenyl)ethyl O-a-L-rhamnopyranosyl-(l ->3)-2-O-acetyl-4-Op-coumaroyl-^-D-glucopyranoside sou baz prèv chimik ak done espèk.

Mo kle Cistanche tubulosa-, Orobanchaceae;phenylethanoid glikozid;iridoid; tubuloside E; dehydrodiconiferyl alkòl glikozid; siringalid rhamnoside;neolignan glikozid; 13C-NMRMOE

phenylethanoid glikozid nan cistanche

Nan yon papye anvan, nou te rapòte izolasyon ak detèminasyon estriktirèl nouvoglikozid feniletanoid, tubulosides A一D soti nan plant yo antye nanCistanche tubulosa(Schrenk) Hook. f. (Orobachaceae) nan Pakistan.

Papye sa a tranzaksyon ak izòlman ak elisid estriktirèl nan yon nouvophenylethanoid glikozid, yo te rele tubuloside E (I), ak yon nouvoneolignan glikozid, dehydrodiconiferyl alkòl ■/-QgD-glucopyranoside (II), osi byen ke izolasyon an nan 11 konpoze li te ye, dehydrodiconiferyl alkòl 4-OgD-glucopyranoside (III), syringalide A 3'-aL-rhamnopyranoside (IV), isosyringalide 3'-aL-rhamnopyranoside (V), (plis)-syringaresinol OgD-glucopyranoside(VI), (plis)-pinoresinol OgD-glucopyranoside (VII), liriodendron (VIII), 6-deoxycatalpol (IX) , 8- asid epiloganik (X), 20-hydroxyecdysone (XI), 8-hydroxygeraniol 1-gD-glucopyranoside (XII) ak siringin (XIII). Konpoze VI, VIII一X, XII, ak XIII te deja izole nan salsa Cistanche?'

Tubuloside E (I) te izole kòm yon poud amorphe, C31H38O15 -3/2H2O, [a】D -134.0 degre (MeOH). Enfrawouj (IR) spectre sijere prezans nan gwoup idroksil (3430cm{-1), ester konjige (1734cm-x), yon lyezon doub (1634cm-1) ak bag aromat ( 1608 ak 1518cm-1). Pwoton nikleyè rezonans mayetik 0H-NMR) spectre I te montre siyal akòz yon gwoup methyl nan ramnose 0 1.07 (3H, d, J= 6 Hz)], yon gwoup asetoksil {{22} }.98 (3H, s)], benzylic methylene protons 0 2.72 (2H, t, J—1 Hz)], yon pwoton glikoseanomeric 0 4.54 (1H, d, J{{ 32}} Hz)], yon proton ramnoseanomeric 0 5.00 (1H, br s)], de pwoton olefinik [<5 6.34,="" 7.66="" (1h="" each,="" d,="" j="16" hz)]="" and="" aromatic="" protons="" 0="" 6.50—7.46="">

Sou asetilasyon, konpoze I bay octaacetate (la), ki gen 】H-NMR spectre te montre senk gwoup asetoksil alifatik 0 1.87, 1.95, 2.02 (3H chak, s), 2.10x2 (6H, s)] ak twa gwoup asetoksil aromat [<5 2.27,="" 2.28,="" 2.31="" (3h="" each,="" s)].="" as="" shown="" in="" table="" i,="" the="" carbon="" nuclear="" magnetic="" resonance="" (13c-nmr)="" spectrum="" of="" i="" was="" almost="" identical="" with="" that="" of="" 2'-acetylacteoside="" (ib)「)except="" for="" the="" signals="" due="" to="" the="" ^-coumaric="" acid="">

Sou methanolysis I ak klori asetil nan metanol, methyl p-coumarate, ak 3,4-dihydroxyphenethyl alkòl yo te detekte sou chromatografi kouch mens (TLC) ak segondè-pèfòmans likid chromatografi (HPLC).

Idwoliz asid I ak 10 pousan asid silfirik bay glikoz ak ramnoz nan yon rapò 1 a 1.

From the above results, the structure of tubuloside E (I) was established as 2-(3,4-dihydroxy phenyl)ethy 1 O-a-L- rhamnopyranosyl-( 1 -^3)-2-(?-acetyl-4-O-/>-coumaroyl-^- D-glucopyranoside.

Konpoze III te izole kòm yon poud amorphe, [a] D - 56.4 degre (MeOH). Idroliz anzimatik III bay yon aglikon (Illb) kòm yon poud amorphe, [0]270 —11300, [a] D —31.6 degre (MeOH), ak glikoz. Konpoze Illb ak III yo te idantifye kòm alkòl dehydrodiconiferyl3,4) ak 4-O-#-D-glucopyranoside,3,4) li yo respektivman, pa konparezon ak echantiyon natif natal (TR,】H- ak 13C-NMR spectre) .

Konpoze II te izole kòm yon poud amorphe, [a]D 一 7.4 degre (MeOH), ki gen spectre IR sijere prezans nan yon gwoup idroksil (3430 cm-1) ak yon bag aromat (1614,1522,1502 cm). T). 〔H-NMR nan II a te montre siyal akòz de gwoup methoxyl aromat 0 3.74, 3.80 (3H chak, s)], yon pwoton benzylik 0 5.45 (1H, d, J{{ 16}}Hz)], de pwoton olefinik trans 0 6.17 (1H, dt, J= 16, 7 Hz), d 6.58 (1H, d, J{= 16 Hz)] ak senk pwoton aromat 0 6.7一7.1 (5H, m)]. Sou asetilasyon, konpoze II bay hexaacetate (Ila), ki gen 'H-NMR spectre te montre senk siyal asetil alifatik 0 2.01, 2.03, 2.09, 2.06 (3H, 3H, 3H, 6H)]. Idroliz anzimatik II bay aglycone (lib) ak glikoz. Lib konpoze te izole kòm yon poud amorphe, [a] D plis 34.8 degre (MeOH), ki gen IR, iH- ak 13C-NMR spectre yo te prèske idantik ak sa yo ki nan Illb, eksepte pou [a] D valè ak dichroism sikilè (CD). ) spectre.

Konfigirasyon absoli lib la te elucide pa konparezon nan spectre CD li yo ak sa yo ki nan Illb ak III, ki gen stereochimi konfime nan done pibliye. Yon lòt bò, maksimòm pozitif Illb te montre A& -11300 (270) ak -10990 (283) kòm maksimòm negatif. Reyalite sa yo sijere ke lib ak Illb se enantiomers youn ak lòt. Estereochimi lib la dwe montre nan Tablo 1.

Kidonk, konpoze II te etabli yo dwe dehydrodico niferyl alkòl monoglukozid. Pozisyon glikoz la

respektivman. Chanjman nan kabòn ki koresponn yo nan ale nan II soti nan lib yo te -4.4 ak plis 7.3 ppm, ki endike ke yon moso glikosil nan II te lye ak gwoup la idroksil nan atòm Cy'.5)

Soti nan rezilta ki anwo yo, konpoze II te etabli kòm dehydrodiconiferyl alkòl y'-O-^-D-glucopyranoside.

Konpoze IV ak V yo te izole kòm yon poud amorphe (IV, C29H36O14 • 5/2H2O; V, C29H36O14- 1/2H2O). Spectres IR yo sijere prezans gwoup idwoksil (3400cm7), yon lyezon doub (1634cm-1), ak bag aromat (1606 ak 1518cm-1). Sou asetilasyon, konpoze IV ak V bay octaacetates yo ( IVa, Va), ki gen spectre 1H-NMR te montre senk asetoksil alifatik ak twa siyal asetoksil aromat, respektivman. Konpoze IVa ak Va yo te idantifye kòm syringalide A 3'-aL-rhamnopyranoside octaacetate ak isosyringalide 3'-aL-rhamnopyranoside octaacetate, respektivman pa konparezon nan (以 IR ak 】H-NMR) ak echantiyon natif natal.6) 'H- ak Spectre 13C-NMR nan IV ak V sipòte estrikti sa yo (gade Eksperimantal). Konpoze VI-XIII yo te idantifye kòm (plis)-sereng-esinol O-少-D-glucopyranoside (VI),2c) (plis)-pinoresinol O- jS-D-glucopyranoside (VII),7) liriodendrin (VIII),2Z,) 6-de-oxycatalpol (IX),2c) 8-asid epiloganik (X),2fl) {{50} }hydroxyecdysone (XI),8) 8-hydroxygeranyol 1-0-D-glucopyrano-side (XII) ak siringin (XIII),2d) respektivman, pa konparezon ak echantiyon natif natal

Eksperimantal

Pwen k ap fonn yo te detèmine sou yon aparèy mikwo-pwen k ap fonn Mitamura epi yo pa korije. Wotasyon optik yo te mezire ak yon polarimèt dijital JASCO DIP-140. Yo te anrejistre espèk IR ak yon espektrofotomèt IR Hitachi 279-30 ak espektrofotomèt iltravyolèt (UV) ak yon espektrofotomèt Hitachi 200-20. Spectre CD yo te anrejistre ak yon JASCO J-20A CD spectrophotometer ak mas spectra (MS) ak yon JEOL JMS-100. 'H-NMR ak 13C-NMR spectre yo te anrejistre ak yon machin JEOL FX-90Q (89.55 ak 22.5 MHz, respektivman).

Chanjman chimik yo bay sou echèl d (ppm) ak tetramethylsilane (TMS) kòm yon estanda entèn. Abreviyasyon yo itilize pou done rezonans mayetik nikleyè (NMR) yo jan sa a; s, singlet; d, doubt; t, triple; q, quartet; br, laj. Kwomatografi gaz (GC) te kouri sou yon aparèy Shimadzu GC-4CM ak yon detektè iyonizasyon flanm dife. HPLC te fèt sou yon machin Hitachi 655A-11. Yo te itilize jèl silica (Wako jèl C-300) pou kwomatografi kolòn. Silica jèl 60 F254 (Merck) plak prekouch yo te itilize pou TLC ak deteksyon te pote soti nan flite 10 pousan H2SO4 ki te swiv pa chofaj.

An karantèn

Plant antye nancistanche tubulosa(7 kg), yo te kolekte nan mwa desanm 1986, nan Karachi, Pakistan, yo te ekstrè ak MeOH (20 1 x 2) anba rflu. Ekstrè a te konsantre anba presyon redwi epi rezidi a te sispann nan dlo. Sispansyon sa a te ekstrè ak EtOAc satire ak dlo. Kouch dlo a te sibi yon kolòn Diaion HP -20 (Nippon Rensui Co.) epi lave ak H2O, ak Lè sa a, eluye ak MeOH. MeOH eluate (140 g) te chromatographed sou yon kolòn polyamid C -200 (Wako Pure Chemical) lè l sèvi avèk H2O ak Lè sa a, MeOH bay de fraksyon (fraksyon 1, H2O eluate; fraksyon 2, MeOH eluate). Fraksyon 1 te konsantre pou bay yon rezidi. Yo te chromatographed rezidi a sou yon kolòn jèl silica lè l sèvi avèk CHCl3-MeOH-H2O repete. Apre kwomatografi HPLC repete (kolòn, Develosil ODS-10, 20 x 250 mm; sòlvan; H2O-CH3CN oswa H2O-MeOH), 10 konpoze yo te izole [II (45mg), III (40mg), VI (35mg). ), VII (43mg), VIII (12mg), IX (8mg), X (10mg), XI (20mg), XII (10mg) ak XIII (3.6mg)]. Fraksyon 2 te trete menm jan ak fraksyon 1 epi li te bay konpoze I (150 mg), IV (10 mg), ak V (lOmg).

Tubuloside E (I)

Poud amorphe, [a]p3 —134.0 degre (c= 1.5, MeOH). Anal. Kalki pou C31H38O15 -3/2H2O: C, 54,98; H, 6,10. Yo te jwenn: C, 55,01; H, 5,98. IR cm-1: 3430,1732, 1634, 1608,1518. ]H・NMR (MeOH-4) 3: 1.07 (3H, d, J=6Hz, CH3 nan ramnose), 1.98 (3H, s, OAc), 2.72 (2H, t, J{ {37}}Hz, Ar-CH2~), 4.54 (1H, d, J=8Hz, Hl glikoz), 5.00 (1H, brs, Hl nan ramnoz), 6.34 ( 1H, d, J= 16Hz, Ar-CH=CH-), 6.5一6.8 (3H, aromat H), 6.80 (2H, d, J-9Hz, H{ {65}}; H-5'), 7.46 (2H, d, J=9Hz, H{-2; H{-6Z), 7.66 (1H, d, J= 16 Hz, Ar—CH {= CH—). 13C-NMR: Tablo L

Dehydrodiconiferyl alkòl y^-O-^-D-GIucopyranoside (II)

Poud amorphe, [a]^5 —7.4 degre (c= 1.6, MeOH). IR cm"1: 3430, 1614, 1522, 1502. UV h nm: 227 (sh), 280. H・NMR (DMSO-rf6) J: 3.74, 3.80 (3H chak, s, OCH3), 5.45 (1H, d,丿=6Hz, Ha), 6.17 (1H, dt,丿=16, 7Hz, H-#), 6.58 (1H, d〃= 16Hz; H-az) , 6.7-7.1 (5H, m, aromat H) 13C-NMR: Tablo I.

Dehydrodiconiferyl alkòl 4-Oj8- D-Glucopyranoside (III)

Poud amorphe, [a]^5 -56.4.(c= 1.1, MeOH). IR cm -1: 3400, 1604, 1566, 1502. UV 11 nm; 222, 277. XH-NMR (DMSO-(76) 6: 3.76, 3.82 (3H chak, s, OCH3), 5.52 (1H, d, J-7 Hz, Ha), 6.18 (1H, dt, J=16, 7 Hz, H-^), 6.49 (1H, d,<7=16 hz,="" h-"),="" 6.7—7.2="" (5h,="" m,="" aromatic="" h).="" 13c-nmr:="" table="">

Syringalide A 3'-(xL-Rhamnopyranoside (IV)

Amorphe poud, Anal. Kalki pou C29H36O14 • 5/2H2O: C, 56,11; H, 5,94. Jwenn: C, 56.4{{107}}; H, 6,04. IR cm— pousan 3400, 1606, 1518. 】H・NMR (MeOH-J4) d: 1.15 (3H, d, J-6 Hz, CH3 nan rhamnose), 2.90 (2H, t, J{{27} }} Hz, Ar-CH2-), 4.46 (1H, d, J=8Hz, Hl nan glikoz), 5.27 (1H, brs, Hl nan ramnoz), 6.35 (1H, d, J-16Hz, Ar—CH {= CH—), 6.6—7.3 (3H, aromat H), 7.68 (1H, d, J{=16Hz, Ar-CH {{54} }} CH-). 13C-NMR (CD3OD) 8: 130.6 (Cl), 116.1 (C-2), 130.8 (C{-3, 156.6 (C{-4), 116.1 (C{-6 ), 72.1 (Ca, rham-2, 3), 36.2 (C{-8), 127.6 (Cr), 114.6 (C2), 149.6 (C・3'), 146.6 (C{{94). }}'),116.4 (C-5'), 123.1 (C-6'),168.2 (C・a'),115.2 (C书),147.0 104.0 (glu-1), 76.0

(glu-2), 81.5 (glu-3), 70.3 (glu{-4), 75.9 (glu-5), 62.3 (glu{-6), 102.8 (rham-1), 73.7 (rham{-4), 70.5 (rham{-5), 18.3 (rham{-6).

Isosyringalide S^aL-Rhamnopyranoside (V)

Amorphe poud, Anal. Kalki pou C29H36O14-1/2H2O: C, 56,31; H, 5,96. Jwenn; C, 56,4{{70}}; H, 6,04. IR 唸cm-1: 3420, 1608, 1518. XH-NMR (MeOH-J4) d: 1.09 (3H, d,丿=6 Hz, CH3 nan ramnose), 2.80 (2H, t, J{28}} Hz, Ar-CH2-), 4.40 (1H, d, J=8Hz, Hl nan glikoz), 5.23 (1H, br s, Hl nan ramnose), 6.37 (1H, d, J-18Hz, AsCH=C旦'一),6.6—7.2 (3H, aromat H), 7.71 (1H, d, 丿{=18Hz, Ar -CH=CH-y 13C-NMR (CD3OD) S: 131.4 (Cl), 116.3 (C{-2), 144.5 (C{-3 X 146.0 (C{-4) ), 117.0 (C-5), 121.2 (C{-6), 72.2 (Ca), 36.5 (C/), 127.1 (Cr), 116.8 (C{-2\ 69, 131.2 (C-3\ 5'), 161.2 (C-4'), 168.2 (CH), 114.7 (C-^), 147.5 (Cy), 104.1 (glu-1), 76.1 (glu-2), 81.5 (glu{-3),

70.2 (glu-4), 75.9 (glu{-5), 62.3 (glu{-6), 102.8 (rham{-1), 72.1 (rham{-2, 3 ), 73.7 (rham-4), 70.6 (rham-5), 18.4 (rham{-6).

(plis)-Syringaresinol Oj?-D-Glucopyranoside (VI)

Poud amorphe, [a]g5 —14.4.(c=0.1, MeOH). Anal. Kalki pou C28H36O13: C, 57,93; H, 6,25. Yo te jwenn: C, 58,01; H, 6,26. IRv^cm-1: 3430, 1600, 1518. 】H・NMR (DMSO"6).: 3.78 (12H, s?OCH3 x 4), 6.60,6.66 (2H chak, s, aromat H).

(plis)-Pinoresinol O-^-D-Glucopyranoside (VII)

Poud amorphe, [a]^5 plis 38.9 degre (c=0.6, MeOH). Anal. Kalki pou C26H32O11 T/2H2O: C, 58,97; H, 6,28. Yo te jwenn: 58,94; H, 6,29. IR cm -1: 3425, 1606, 1516. 】H-NMR (C5D5N) W 3.77, 3.80 (3H chak, s, OCH3), 6.9-7.7 (6H, m, aromat H).

Liriodendron (VIII)

Zegwi san koulè (MeOH), mp 255一257 degre, [a]p5 -9.2 degre (c=L3, piridin). IR cm-1: 3400, 1598, 1510. XH-NMR (DMSO/6)<5: 3.78="" (12h,="" s,="" och3x4),="" 6.67="" (4h,="" s,="" aromatic="">

6-Deoxycatalpol (IX)

Zegwi san koulè (MeOH), mp 204—206 degre, [a]p5 -50.1 degre (c=0.7, MeOH). Anal, Kalki pou C15H22O9: C, 52,02; H, 6,40. Yo te jwenn: C, 52,12; H, 6,41. IR 唸cm-1: 3400, 1658. 】H・NMR (D2O) 3:

l. 3—1.9 (1H, m, H-6), 2.1—2.7 (3H, m, H-5, H-6, H-9), 3.65 (1H, s-tankou, H-7), 3.90, 4.35 (2H, ABsystem, J=13Hz, H-10), 4.92 (1H, d, J=7 Hz, aromerik H), 5.10 (1H, dd, J=6, 4Hz, H{-4), 5.12 (1H, d, J{-10Hz, Hl), 6.38 (1H, dd, 1Hz, H-3).

{{0}}Asid Epiloganik (X) Zegwi san koulè (MeOH), mp 147一149 degre, [a]p5 — 135.0 degre (c= 1.0, piridin ). Anal. Kalki pou C16H24O10 H2O: C, 48,73; H, 6,65. Yo te jwenn: C, 48,60; H, 6,70. IR v;cm-1: 3400, 1680, 1640, 1430. 】H・NMR (C5D5N)<5: 1.18="" (3h,="" d,="" j="7Hz," ch3),="" 2.2—2.5="">

m, H-6), 2.60 (1H, m, H-8), 3.08 (1H, dt, J=3, 8.5Hz, H-9), 3.58 ( 1H, m, H-5), 5.40 (1H, d, J=7.5Hz, anomerik H), 5.91 (1H, d〃{=3Hz, Hl), 7.92 (1H , s, H-3).

20-Hydroxyecdysone (XI)

Solid kristalin, mp 241一242 degre, [a]p5 plis 70.0 degre (c= LI, MeOH). Anal. Kalki pou C27H44O7: C, 67,47; H, 9,23. Yo te jwenn: C, 67,28; H, 9,18. IR cm -1: 3440, 1646. XH-NMR (C5D5N) 1.06 (3H, s, H{-19), 1.20 (3H, s, H{-18), 1.36 (6H, s, H-26, H-27)? 1.58 (3H, s, H-21), 6.24 (lH,d, J{=2Hz, H{-7). 13C-NMR (C5D5N): 38.0 (Cl),

68.2 (C-2, C-3), 32.5 (C・4), 51.5 (C-5), 203.7 (C-6), 121.8 (C-7), 166.3 (C-8), 34.6 (C{-9), 38.8 (C{-10), 21.3 (C-ll), 31.9 (C {{30}}), 48.2 (C-13), 84.4 (C-14), 32.1 (C{-15), 21.6 (C{-16 ), 50.2 (C-17), 18.0 (C{-18), 24.6 (C{-19), 77.0 (C{-20), 21.8 (C{-21 ), 77.7 (C{-22), 27.6 (C{-23), 42.7 (C{-24), 69.8 (C{-25), 30.1 (C{-26 , 27).

8-Idroksijeraniol 1-^-D-GIucopyranoside (XII)

Zegwi san koulè (MeOH), mp 58—60 degre , [a]^5 —40.1.(c=L2, MeOH). Anal. Kalki pou C16H28O71/2H2O: C, 56,29; H, 8,56. Yo te jwenn: C, 56,40; H, 8,50. IR cm"1: 3350, 1670, 1446. XH-NMR (C5D5N)<5: 1.72,="" 1.59="" (3h="" each,="" s,="" ch3),="" 2.04="" (4h,="" brm,="" h-4,="" h-5),="" 4.80="" (1h,="" d,="" j="7.5Hz," anomeric="" h),="" 5.56="" (2h,="" brt,="" j="7Hz,">

Seringin (XIII)

Zegwi san koulè (MeOH), mp 188一189 degre, [a]^5 — 15.3 degre (c=0.4, MeOH). Anal. Kalki pou C17H24O9: C, 55,13; H, 6,53. Yo te jwenn: C, 54,83; H, 6,50. IR cm -1: 3400, 1592, 1514. 'H-NMR (DMSO/6)W 3.78 (6H, s, OCH3 x 2), 6.3—6.5 (2H, m, —CH=CH—), 6.72 (2H, s, aromat H).

Acetylation nan I Compound I (50 mg) te fonn nan piridin (l ml) ak anhydride acetic (1 ml) epi kite nan tanperati chanm lannwit lan. Te melanj reyaksyon an vide nan dlo glas, ak Lè sa a, ekstrè ak EtOAc. Ekstrè EtOAc la te konsantre nan vacuo epi yo te chromatographed rezidi a sou yon kolòn jèl silica lè l sèvi avèk benzèn-asetòn (5:1) bay octaacetate (la) (35mg). IR cm"1: 1760, 1638, 1604, 1508. iH-NMR(CDC13)(5: 1.02 (3H, d, J=6Hz, CH3 nan rhamnose), 1.87, 1.95, 2.02 (3H chak, s, OAc), 2.10 (6H, s, OAc x 2), 2.27, 2.28, 2.31 (3H chak, s, Ar-OAc), 2.90 (2H, t, J=7Hz, Ar-CH{ {43}}), 6.38 (1H, d, J=16Hz, AR ~ CH=CH-), 6.80—7.15 (3H, m, aromat H), 7.24 (2H, d, J-9 Hz, H{-3\ H・5'), 7.55 (2H, d,丿=9Hz, H-2, H-6), 7.72 (1H, d, J{70}}Hz, Ar-CH=CH-).

Asetilasyon II ak III

Konpoze II ak III (20 mg chak) yo te acetylated nan menm fason ki dekri pou konpoze I epi yo te bay hexaacetates yo [Ila (12mg), Illa (15mg)]. Ila: IR 唸顋 cm-1: 1754, 1608, 1506. 】H・NMR (CDC13) 5: 2.01, 2.03, 2.09 (3H chak, s, OAc), 2.06 (6H, s, OAc x 2.09) ), 2.30 (3H, s, OAc), 3.84 (3.95 (3H chak, s, OCH3), 5.54 (1H, d,丿=6Hz, Ha), 6.05 (1H5 dt, J{{38} }, 7Hz, H-^), 6.53 (1H, d, «/= 16Hz, H-a'), 6.8一7.1 (5H, m, aromat H). Illa: IR v^cm 1: 1760, 1604, 1516. 】H-NMR (CDC13)<5: 2.05="" (9h,="" s,="" oac="" x="" 3),="" 2.08="" (6h,="" s,="" oac="" x="" 2),="" 2.11="" (3h,="" s,="" oac),="" 3.80,="" 3.92="" (3h="" each,="" s,="" och3),="" 5.51="" (1h,="" d,="" j="7Hz," h-a),="" 6.14="" (1h,="" dt,="" j="16," 7hz,="" h-#),="" 6.62="" (1h,="" d,丿="16Hz,">

(5H, m, aromat H).

Asetilasyon nan IV ak V

Konpoze IV ak V yo te acetylated nan menm fason ki dekri pou konpoze I epi yo te bay octaacetates yo [IVa, Va]. IVa: IR 唸児 cmT: 1754, 1636, 1514. 】H・NMR (CDC,) W 1.03 (3H? d, J=6Hz, CH3 nan rhamnose), 1.87, 1.94, 1.99 (3H) chak s, OAc), 2.09 (6H, s, OAcx2), 2.28 (6H, s, OAcx2), 2.30 (3H, s, OAc), 2.86 (2H, t, J=6 .6Hz, Ar■-CH?—), 3.67 (2H, t, J=7.2Hz Ar-CH2~CH2-), 6.35 (1H, d, J{=16Hz , AsCH=CU—), 6.87—7.35 (3H, m, Ar—7.09 (4H, q, J{=8.6Hz, A2/B2/tip), 7.65 (1H, d, J =16Hz, Ar CH=CH). Va: IR cm-1: 1750, 1636, 1608, 1514. ^-NMR (CDC13) 1.03 (3H, d,<7= 6="" hz,="" ch3="" for="" rhamnose),="" 1.87,="" 1.94,="" 2.02="" (3h="" each,="" s,="" oac),="" 2.09="" (6h,="" s,="" oac="" x="" 2),="" 2.28="" (6h,="" s,="" oac="" x="" 2),="" 2.31="" (3h,="" s,="" oac),="" 2.86="" (2h,="" t,="" j="6.6" hz,="" ar-ch2-),="" 3.73="" (2h,="" t,=""><7=6.6 hz,="" ar="" ch2-ch2-),="" 6.34="" (lh,d,="" j="16" hz,="" ar—ch="" —="" ch—),="" 6.78—7.20="" (3h,="" m,="" ar—h),="" 7.31="" (4h,="" q,=""><7=8.5>

A2zB/tip), 7.71 (1H, d, «/=16Hz, Ar-CH=CH ).

Metanoliz nan I

Konpoze I (apeprè 1 mg) te refluxed ak metanolik 5 pousan CH3COC1 (2 ml) pou 30 min, ak Lè sa a, reyaktif la te evapore nan vacuo. TLC [CHCl3-MeOH (10:1)] ak HPLC [kolòn, Develosil ODS] te detekte prezans methyl p-koumarat ak 3,4-dihydroxyphenethyl alkòl nan rezidi a. -7 (4.6 x 250 mm); sòlvan, 55 pousan MeOH; detektè (UV), 220 nm; to koule, 1.0ml/min]. Methyl p-koumarat; Rf 0.82, ;R (min) 8.8, 3,4-dihydroxyphenethyl alkòl;母0.31, tR (min) 3.2.

Idwoliz asid nan IA

solisyon konpoze I (2 mg) nan 10 pousan H2SO4 (1 ml) te chofe nan yon beny dlo bouyi pou 30 min. Solisyon an te pase nan yon kolòn Amberlite IR-45 epi konsantre pou bay yon rezidi, ki te redwi ak borohydride sodyòm (apeprè 3 mg) pou 1 èdtan. Yo te pase melanj reyaksyon an nan yon kolòn Amberlite IR-120 epi konsantre nan sechrès. Asid borik te retire pa distilasyon ak MeOH epi rezidi a te acetylated ak anidrid acetic (1 gout) ak piridin (1 gout) nan 100 degre pou 1 èdtan. Reyaktif yo te evapore nan vacuo. Glucitol acetate ak rhamnitol acetate yo te detekte nan yon rapò de 1 a 1 soti nan konpoze I pa GC. Kondisyon: kolòn, 1.5 pousan OV-17, 3 mm x 1.5 m; tanperati kolòn, 180 degre; gaz konpayi asirans, N2 (30ml/min). tR (min): 2.0 (rhamnitol acetate), 5.5 (glucitol acetate).

Idroliz anzimatik nan II ak III Konpoze II

(13 mg) oswa III (40 mg) te idrolize ak selulaz (40 mg) nan dlo (1 ml) pou 2h nan 37 degre. Te melanj reyaksyon an ekstrè ak EtOAc. Ekstrè EtOAc la te konsantre nan vacuo pou bay rezidi a, ki te chromatographed sou HPLC [kolòn, Develosil ODS-10 (20 x 250 mm); sòlvan. II: H2O-CH3CN (77.5:22.5); III: H2O-CH3CN (80:20); detektè (UV), 205 nm; to koule, 6.0ml/min] bay lib (3.3 mg), Illb (5.7 mg). lib: [a]^5 plis 34.8 degre (c=0.3, MeOH). CD (c=3.3mg/10ml, MeOH) [0]25 (nm): 1740 (330), 15200 (283), 14100 (270), 0 (247), - 16700 (232). ). IR 唸E; cm"1: 3380, 1606, 1520, 1500. UV nm: 276. MS m/z (pousan): 358 (M plis, 53), 340 (M plis -H2O, 100), 338 (58), 325 ( 47), 310 (23), 162 (41), 151 (36), 137 (59) XH-NMR (asetòn-J6)<5: 3.55="" (1h,="" brt,=""><7=6hz, h-g),="" 3.84,="" 3.87="" (3h="" each,="" s,="" och3),="" 4.19="" (2h,="" brd,="" j-5hz,="" h-yz),="" 5.56="" (1h,="" d,="" j="6," 5="" hz,="" h-a),="" 6.23="" (1h,="" dt,=""><7=16, 5="" hz,="" h-#),="" 6.56="" (1h,="" brd,="" j="16Hz," hh),="" 6.7—7.1="" (5h,="" m,="" aromatich).="" 13c-nmr:="" tablel.iiib:="" [a]-31.6°="" (c="0.6," meoh).="" cd="" (c="2.28mg/lOml," meoh)="" [0]25="" (nm):="" 2200="" (330),="" 0="" (310),="" -10990="" (283),="" -11300="" (270),="" 0="" (242),="" 5650="" (232).="" 0-nmr="" (acetone-j6)=""><5: 3.84,="" 3.87="" (3h="" each,="" s,="" och3="" x="" 2),="" 4.19="" (2h;="" brd,="" j="5Hz," h"'),5.56="" (1h,="" d,丿="6.5" hz,="" h-a),="" 6.23="" (1h,="" dt,丿="16," 5="" hz,="" h・#),="" 6.52="" (1h,="" d,="" j="16" hz,="" h-az),="" 6.7一7.1="" (5h,="" m,="" aromatic="" h).="" 13c-nmr:="" table="" i.="" the="" ir="" and="" uv="" spectra="" were="" similar="" with="" that="" of="">

RekonesansNou rekonesan anvè Prof. A. Ueno ak Doktè T. Miyase, University of Shizuoka, pou mezi RMN ak spectre CD yo, ak pou sijesyon ki gen anpil valè yo, ak Dr. M. Kikuchi, Tohoku College of Pharmacy, pou bay echantiyon natif natal.

Referans ak Nòt

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8) a) H. Hikino, Y. Hikino, Fortschritted. Chem. òg. Naturst., 28, 256 (1970); b) RW Miller, J. Clardy, J. Kozlowski, KL Mikolajczak, RD Plattner, RG Powell, CR Smith, D. Weisleder, ak Q.-T. Zheng, Planta Medica, 1985, 40.