Kev Ntsuam Xyuas Kev Ntsuas Ntawm Cistanche Tubulosa: Kev Sib Cais Thiab Cov Qauv Ntawm Phenylethanoid Glycoside Thiab Neolignan Glycoside

Hu rau:joanna.jia@wecistanche.com

Cov khoom xyaw ntawm Cistanche Tubulosa (Schrenk) Hook. F. II.1) Kev rho tawm thiab cov qauv ntawm Ib Tug Tshiab Phenylethanoid Glycoside Thiab Ib Tug Tshiab Neolignan Glycoside

Fumio Yoshizawa, * a Takeshi Deyama / Nobuo Takizawa / Khan UsmanghaniJ thiab Mansoor Ahmad6

Central Research Laboratories, Yomeishu Seizo Co., Ltd., a 2132-37 Nakaminowa, Minowa-machi, Kamiina-gun, Nagano 399-46, Nyiv thiab Department of Pharmacognosy, Kws Qhia Ntawv ntawm Pharmacy, University of Karachi? Karachi -32, Pakistan. Tau txais lub Kaum Ib Hlis 9, 1989

Ib tug tshiabphenylethanoid glycosidenpe tubuloside E (I), thiab ib qho tshiabneolignan glycosides, dehydrodiconiferyl cawv yO-0-D-glucopyranoside (II), raug cais tawm ntawm tag nrho cov nroj tsuag ntawmCistanche 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 los ntawm cov pov thawj tshuaj thiab cov ntaub ntawv spectral.

Ntsiab lus Cistanche tubulosa- Orobanchaceae;phenylethanoid glycoside;iridoid; tubuloside E; dehydrodiconiferyl cawv glycoside; syringalide rhamnoside;neolignan glycosides; 13C-NMRMOE

phenylethanoid glycoside hauv cistanche

Hauv ib daim ntawv dhau los, peb tau tshaj tawm txog kev cais tawm thiab kev txiav txim siab ntawm cov qauv tshiabphenylethanoid glycosides, tubulosides A一D los ntawm tag nrho cov nroj tsuag ntawmCistanche tubulosa(Schrenk) Hoob. f. (Orobanchaceae) hauv Pakistan.

Daim ntawv no hais txog kev sib cais thiab kev tsim qauv ntawm ib qho tshiabphenylethanoid glycoside, hu ua tubuloside E (I), thiab ib qho tshiabneolignan glycosides, dehydrodiconiferyl cawv ■/-QgD-glucopyranoside (II), nrog rau kev sib cais ntawm 11 cov tshuaj paub, dehydrodiconiferyl cawv 4-OgD-glucopyranoside (III), syringalide A 3'-aL-rhamnopyranoside (IV), isosyringalide 3'-aL-rhamnopyranoside (V), (ntxiv rau)-syringaresinol OgD- glucopyranoside (VI), (plus)-pinoresinol OgD-glucopyrano- side (VII), liriodendron (VIII), 6-deoxycatalpol (IX) , 8- epiloganic acid (X), 20-hydroxyecdysone (XI), 8-hydroxygeraniol 1-gD-glucopyranoside (XII) thiab syringin (XIII). Cov tebchaw VI, VIII一X, XII, thiab XIII yav dhau los tau raug cais tawm ntawm Cistanche salsa?'

Tubuloside E (I) raug cais raws li amorphous hmoov, C31H38O15 -3/2H2O, [a】D -134.0 degree (MeOH). Lub infrared (IR) spectrum qhia tias muaj cov pab pawg hydroxyl (3430cm-1), conjugated ester (1734cm-x), ob daim ntawv cog lus (1634cm-1) thiab aromatic rings ( 1608 thiab 1518cm-1). Lub proton nuclear magnetic resonance 0H-NMR) spectrum ntawm kuv pom cov teeb liab vim yog ib pawg methyl ntawm rhamnose 0 1.07 (3H, d, J= 6 Hz)], ib pawg acetoxyl {{22} }.98 (3H, s)], benzylic methylene protons 0 2.72 (2H, t, J—1 Hz)], glucoseanomeric proton 0 4.54 (1H, d, J{{ 32}} Hz)], ib rhamnoseanomeric proton 0 5.00 (1H, br s)], ob olefinic protons [<5 6.34,="" 7.66="" (1h="" each,="" d,="" j="16" hz)]="" and="" aromatic="" protons="" 0="" 6.50—7.46="">

Ntawm acetylation, compound kuv them rau octaacetate (la), uas nws 】 H-NMR spectrum pom tsib aliphatic acetoxyl pawg 0 1.87, 1.95, 2.02 (3H txhua, s), 2.10x2 (6H, s)] thiab peb pawg acetoxyl aromatic [<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="">

Ntawm methanolysis ntawm I nrog acetyl chloride hauv methanol, methyl p-coumarate, thiab 3, 4-dihydroxyphenethyl cawv tau kuaj pom ntawm nyias txheej chromatography (TLC) thiab cov kua dej chromatography (HPLC).

Acid hydrolysis ntawm kuv nrog 10 feem pua ​​​​sulfuric acid them nyiaj qabzib thiab rhamnose hauv qhov piv ntawm 1 txog 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.

Compound III raug cais raws li cov hmoov amorphous, [a]D - 56.4 degree (MeOH). Enzymatic hydrolysis ntawm III tau them nyiaj rau aglycone (Illb) ua cov hmoov amorphous, [0] 270 -11300, [a]D -31.6 degree (MeOH), thiab qabzib. Cov tshuaj Illb thiab III tau txheeb xyuas tias yog dehydrodiconiferyl cawv 3,4) thiab nws 4-O-#-D-glucopyranoside, 3,4) feem, los ntawm kev sib piv nrog cov qauv tseeb (TR, 】H- thiab 13C-NMR spectra) .

Compound II raug cais raws li cov hmoov amorphous, [a]D 一 7.4 degree (MeOH), uas nws IR spectrum qhia tias muaj ib pawg hydroxyl (3430 cm-1) thiab ib lub nplhaib aromatic (1614,1522,1502 cm. T). Lub 〔H-NMR ntawm II tau pom cov cim vim yog ob pawg methoxyl aromatic 0 3.74, 3.80 (3H txhua, s)], benzylic proton 0 5.45 (1H, d, J{{ 16}}Hz)], ob trans olefinic protons 0 6.17 (1H, dt, J= 16, 7 Hz), d 6.58 (1H, d, J= 16 Hz)] thiab tsib tus ntxhiab protons 0 6.7一7.1 (5H, m)]. Ntawm acetylation, compound II tau them nyiaj rau hexaacetate (Ila), uas nws 'H-NMR spectrum pom tsib aliphatic acetyl signals 0 2.01, 2.03, 2.09, 2.06 (3H, 3H, 3H, 6H)]. Enzymatic hydrolysis ntawm II them rau aglycone (lib) thiab qabzib. Compound lib raug cais raws li cov hmoov amorphous, [a]D ntxiv rau 34.8 degree (MeOH), uas nws IR, iH- thiab 13C-NMR spectra yuav luag zoo ib yam nrog cov Illb, tshwj tsis yog rau [a]D qhov tseem ceeb thiab ncig dichroism (CD ) spectrum.

Qhov tseeb configuration ntawm lib yog elucidated los ntawm kev sib piv ntawm nws CD spectrum nrog cov Illb thiab III, uas nws stereochemistry tau lees paub nyob rau hauv cov ntaub ntawv luam tawm.4) Lub CD spectrum ntawm lib qhia A & ntxiv 14100 (270) thiab ntxiv rau 15200 (283) raws li zoo maxima, ntawm qhov tod tes, uas ntawm Illb pom A& -11300 (270) thiab -10990 (283) as negative maxima. Cov lus tseeb no qhia tias lib thiab Illb yog enantiomers ntawm ib leeg. Lub stereochemistry ntawm lib yuav tsum yog qhov uas qhia hauv Daim Ntawv 1.

Yog li, compound II tau tsim los ua dehydrodico niferyl cawv monoglucoside. Txoj haujlwm ntawm cov piam thaj

raws. Kev hloov pauv ntawm cov pa roj carbon zoo sib xws hauv kev mus rau II los ntawm lib yog -4.4 thiab ntxiv rau 7.3 ppm, uas tau qhia tias glucosyl moiety hauv II tau txuas rau pawg hydroxyl ntawm Cy' atom.5)

Los ntawm cov txiaj ntsig saum toj no, compound II tau tsim los ua dehydrodiconiferyl cawv y'-O-^-D-glucopyranoside.

Cov tshuaj IV thiab V raug cais raws li cov hmoov amorphous (IV, C29H36O14 • 5/2H2O; V, C29H36O14- 1/2H2O). IR spectra qhia tias muaj cov pab pawg hydroxyl (3400cm7), ob daim ntawv cog lus (1634cm{14}}), thiab cov nplhaib uas muaj ntxhiab (1606 thiab 1518cm-1). Ntawm acetylation, cov tebchaw IV thiab V tau them nyiaj rau octaacetates ( IVa, Va), uas nws 1H-NMR spectra pom tsib aliphatic acetoxyl thiab peb aromatic acetoxyl signals, feem. Cov tshuaj IVa thiab Va raug txheeb xyuas tias yog syringalide A 3'-aL-rhamnopyranoside octaacetate thiab isosyringalide 3'-aL-rhamnopyranoside octaacetate, raws li kev sib piv ntawm (以 IR thiab 】H-NMR) nrog cov qauv tseeb. 13C-NMR spectrum ntawm IV thiab V txhawb nqa cov qauv no (saib Kev sim). Cov tshuaj VI-XIII tau txheeb xyuas tias (ntxiv rau )-syringes- esinol O-少-D-glucopyranoside (VI), 2c) (ntxiv )-pinoresinol O- jS-D-glucopyranoside (VII), 7) liriodendrin (VIII), 2Z,) 6-de- oxycatalpol (IX), 2c) 8-epiloganic acid (X), 2fl) {{50} }hydro- xyecdysone (XI), 8) 8-hydroxygeraniol 1-0-D-glucopyrano- side (XII) thiab syringin (XIII), 2d) raws li kev sib piv nrog cov qauv tseeb

Kev sim

Cov ntsiab lus melting tau txiav txim siab ntawm Mitamura micro-melting point apparatus thiab tsis raug kho. Kev hloov kho qhov muag tau ntsuas nrog JASCO DIP-140 digital polarmeter. IR spectra raug kaw nrog Hitachi 279-30 IR spectrophotometer thiab ultraviolet (UV) spectra nrog Hitachi 200-20 spectrophotometer. CD spectra tau kaw nrog JASCO J-20A CD spectrophotometer thiab mass spectra (MS) nrog JEOL JMS-100. 'H-NMR thiab 13C-NMR spectra tau kaw nrog JEOL FX-90Q tshuab (89.55 thiab 22.5 MHz, feem).

Cov tshuaj hloov pauv tau muab rau ntawm qhov ntsuas d (ppm) nrog tetramethylsilane (TMS) raws li tus qauv sab hauv. Cov ntawv luv uas siv rau cov ntaub ntawv nuclear magnetic resonance (NMR) yog raws li hauv qab no; s, ib leeg; d, ob;. t, peb; q ,qw;. br, loj. Gas chromatography (GC) tau khiav ntawm Shimadzu GC-4CM apparatus nrog lub nplaim ionization ntes. HPLC tau ua tiav ntawm Hitachi 655A-11 tshuab. Silica gel (Wako gel C-300) tau siv rau kab chromatography. Silica gel 60 F254 (Merck) precoated daim hlau tau siv rau TLC thiab kev kuaj pom tau ua los ntawm kev txau 10 feem pua ​​​​H2SO4 tom qab cua sov.

Muab cais nyob ib leeg

Tag nrho cov nroj tsuag ntawmcistanche tubulosa(7 kg), sau thaum lub Kaum Ob Hlis 1986, hauv Karachi, Pakistan, tau muab rho tawm nrog MeOH (20 1 x 2) nyob rau hauv reflux. Cov extract tau concentrated nyob rau hauv lub siab txo thiab cov residue raug tshem tawm hauv dej. Qhov kev ncua no tau muab rho tawm nrog EtOAc saturated nrog dej. Cov txheej dej tau raug rau Diaion HP-20 (Nippon Rensui Co.) kem thiab ntxuav nrog H2O, thiab tom qab ntawd eluted nrog MeOH. Cov MeOH eluate (140 g) tau chromatographed ntawm polyamide C-200 (Wako Pure Chemical) kem siv H2O thiab tom qab ntawd MeOH muab ob feem (feem 1, H2O eluate; feem 2, MeOH eluate). Tshooj 1 tau concentrated los muab ib qho residue. Cov residue tau chromatographed ntawm silica gel kem siv CHCl3-MeOH-H2O ntau zaus. Tom qab rov chromatography ntawm HPLC (kem, Develosil ODS-10, 20 x 250 mm; hnyav; H2O-CH3CN lossis H2O-MeOH), 10 cov tebchaw raug cais [II (45mg), III (40mg), VI (35mg. ), VII (43mg), VIII (12mg), IX (8mg), X (10mg), XI (20mg), XII (10mg) thiab XIII (3.6mg)]. Tshooj 2 tau kho tib yam li feem 1 thiab muab cov tshuaj sib xyaw ua ke I (150 mg), IV (10 mg), thiab V (lOmg).

Tubuloside E (I)

Amorphous hmoov, [a]p3 —134.0 degree (c= 1.5, MeOH). Qhov quav. Calcd rau C31H38O15 -3/2H2O: C, 54.98; H, 6.10 Nws. Pom tau: C, 55.01; ib 5,98. IR cm-1: 3430,1732,1634,1608,1518. ]H・NMR (MeOH-4) 3: 1.07 (3H, d, J=6Hz, CH3 of rhamnose), 1.98 (3H, s, OAc), 2.72 (2H, t, J{ {37}}Hz, Ar-CH2~), 4.54 (1H, d, J=8Hz, Hl ntawm qabzib), 5.00 (1H, brs, Hl ntawm rhamnose), 6.34 ( 1H, d, J= 16Hz, Ar-CH=CH-), 6.5一6.8 (3H, aromatic 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: Tab L

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

Amorphous hmoov, [a]^5 - 7.4 degree (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 txhua, 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, aromatic H) 13C-NMR: Table I.

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

Amorphous hmoov, [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 ib, 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))

Amorphous hmoov, Anal. Calcd rau C29H36O14 • 5/2H2O: C, 56.11; ib 5,94. Pom: C, 56.4{{107}}; H, 6.04 Nws. IR cm — feem pua ​​3400, 1606, 1518. 】H・NMR (MeOH-J4) d: 1.15 (3H, d, J-6 Hz, CH3 ntawm rhamnose), 2.90 (2H, t, J{27 }} Hz, Ar-CH2-), 4.46 (1H, d, J=8Hz, Hl ntawm qabzib), 5.27 (1H, brs, Hl ntawm rhamnose), 6.35 (1H, d, J-16Hz, Ar—CH=CH—), 6.6–7.3 (3H, aromatic 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{108}}), 76.0

(glu-2), 81.5 (glu-3), 70.3 (glu-4), 75.9 (glu-5), 62.3 (glu-6), 102.8 (rham{15}}), 73.7 (rham-4), 70.5 (rham-5), 18.3 (rham-6).

Isosyringalide S^aL-Rhamnopyranoside (V)

Amorphous hmoov, Anal. Calcd rau C29H36O14-1/2H2O: C, 56.31; ib 5,96. Pom tau; C, 56.4{{70}}; H, 6.04 Nws. IR 唸cm-1: 3420, 1608, 1518. XH-NMR (MeOH-J4) d: 1.09 (3H, d, 丿=6Hz, CH3 ntawm rhamnose), 2.80 (2H, t, J-6 Hz, Ar-CH2-), 4.40 (1H, d, J=8Hz, Hl ntawm qabzib), 5.23 (1H, br s, Hl ntawm rhamnose), 6.37 (1H, d, J-18Hz, AsCH=C旦'一), 6.6-7.2 (3H, aromatic 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-3X 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{107}}), 76.1 (glu{110}}), 81.5 (glu{113}}),

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{24}}).

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

Amorphous hmoov, [a]g5 — 14.4.(c=0.1, MeOH). Qhov quav. Calcd rau C28H36O13: C, 57.93; H, 6.25 Nws. Pom tau: 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 txhua, s, aromatic H).

(ntxiv rau)-Pinoresinol O-^-D-Glucopyranoside (VII)

Amorphous hmoov, [a]^5 ntxiv rau 38.9 degree (c=0.6, MeOH). Qhov quav. Calcd rau C26H32O11 T/2H2O: C, 58.97; H., 6.28. pom: 58,94; H, 6.29 Nws. IR cm-1: 3425, 1606, 1516. 】 H-NMR (C5D5N) W 3.77, 3.80 (3H txhua, s, OCH3), 6.9-7.7 (6H, m, aromatic H).

Liriodendron (VIII)

Cov koob tsis muaj xim (MeOH), mp 255一257 degree, [a]p5 -9.2 degree (c=L3, pyridine). IR cm-1: 3400, 1598, 1510. XH-NMR (DMSO/6)<5: 3.78="" (12h,="" s,="" och3x4),="" 6.67="" (4h,="" s,="" aromatic="">

6-Deoxycatalpol (IX)

Cov koob tsis muaj xim (MeOH), mp 204-206 degree, [a]p5 -50.1 degree (c=0.7, MeOH). Anal, Calcd rau C15H22O9: C, 52.02; H, 6.40 Nws. Pom: C, 52.12; ib 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{10}}, H-6, H-9), 3.65 (1H, s-like, H-7), 3.90, 4.35 (2H, ABsystem, J=13Hz, H-10), 4.92 (1H, d, J=7 Hz, aromeric H), 5.10 (1H, dd, J=6, 4Hz, H-4), 5.12 (1H, d, J-10}Hz, Hl), 6.38 (1H, dd, 1Hz, H-3).

{{0}}Epiloganic Acid (X) Cov koob tsis muaj xim (MeOH), mp 147一149 degree , [a]p5 — 135.0 degree (c= 1.0, pyridine ). Qhov quav. Calcd rau C16H24O10 H2O: C, 48.73; H 6, 65. Pom: C, 48.60; ib 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{15}}), 5.40 (1H, d, J=7.5Hz, anomeric H), 5.91 (1H, d〃=3}Hz, Hl), 7.92 (1H , s, H-3 ib.

20-Hydroxyecdysone (XI)

Crystalline solid, mp 241一242 degree , [a]p5 plus 70.0 degree (c= LI, MeOH). Qhov quav. Calcd rau C27H44O7: C, 67.47; H, 9.23 Nws. Pom: C, 67.28; H, 9.18 Nws. 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{57}}) 77.7 (C-22), 27.6 (C-23), 42.7 (C-24), 69.8 (C-25), 30.1 (C-26) , 27).

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

Koob tsis muaj xim (MeOH), mp 58–60 degree , [a]^5 —40.1.(c=L2, MeOH). Qhov quav. Calcd rau C16H28O71/2H2O: C, 56.29; ib 8,56. Pom tau: C, 56.40; H, 8.50 Nws. 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,">

Syringin (XIII)

Cov koob tsis muaj xim (MeOH), mp 188一189 degree , [a]^5 — 15.3 degree (c=0.4, MeOH). Qhov quav. Calcd rau C17H24O9: C, 55.13; ib 6,53. Pom: C, 54.83; H, 6.50 Nws. 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, aromatic H).

Acetylation ntawm I Compound I (50 mg) tau yaj hauv pyridine (l ml) thiab acetic anhydride (1 ml) thiab sab laug hauv chav sov thaum hmo ntuj. Cov tshuaj tiv thaiv sib xyaw yog nchuav rau hauv dej khov, thiab tom qab ntawd muab rho tawm nrog EtOAc. Lub EtOAc extract tau concentrated nyob rau hauv vacuo thiab cov residue tau chromatographed ntawm silica gel kem siv benzene-acetone (5:1) muab cov octaacetate (la) (35mg). IR cm"1: 1760, 1638, 1604, 1508. iH-NMR(CDC13)(5: 1.02 (3H, d, J=6}Hz, CH3 ntawm rhamnose), 1.87, 1.95, 2.02 (3H txhua, s, OAc), 2.10 (6H, s, OAc x 2), 2.27, 2.28, 2.31 (3H txhua, 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, aromatic 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=16Hz, Ar-CH=CH-).

Acetylation ntawm II thiab III

Cov tshuaj II thiab III (20 mg txhua) tau acetylated tib yam li tau piav qhia rau cov tshuaj sib xyaw I thiab muab cov hexaacetates [Ila (12mg), Illa (15mg)]. Ila: IR 唸顋 cm-1: 1754, 1608, 1506. 】H・NMR (CDC13) 5: 2.01, 2.03, 2.09 (3H txhua, s, OAc), 2.06 (6H, s, OAc x 2 ), 2.30 (3H, s, OAc), 3.84 (3.95 (3H txhua, s, OCH3), 5.54 (1H, d, 丿=6}Hz, Ha), 6.05 (1H5 dt, J=16 }, 7Hz, H-^), 6.53 (1H, d, «/= 16Hz, H-a'), 6.8一7.1 (5H, m, aromatic 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, aromatic H).

Acetylation ntawm IV thiab V

Cov tshuaj IV thiab V tau acetylated tib yam li tau piav qhia rau compound I thiab muab cov octaacetates [IVa, Va]. IVa: IR 唸児 cmT: 1754, 1636, 1514. 】H・NMR (CDC,) W 1.03 (3H? d, J=6Hz, CH3 ntawm rhamnose), 1.87, 1.94, 1.99 (3H txhua, 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/type), 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/type), 7.71 (1H, d, «/=16Hz, Ar-CH=CH ).

Methanolysis ntawm I

Compound I (ca. 1 mg) tau refluxed nrog methanolic 5 feem pua ​​​​CH3COC1 (2 ml) rau 30 min, thiab tom qab ntawd cov reagent tau evaporated tawm hauv vacuo. Lub xub ntiag ntawm methyl p-coumarate thiab 3, 4-dihydroxyphenethyl cawv nyob rau hauv cov seem tau kuaj pom los ntawm TLC [CHCl3-MeOH (10: 1)] thiab HPLC [kem, Develosil ODS -7 (4.6 x 25{27}} mm); hnyav, 55% MeOH; ntes (UV), 220 nm; Flow tus nqi, 1.0ml / min]. Methyl p-coumarate; Rf 0.82, ;R (min) 8.8, 3, 4-dihydroxyphenethyl cawv; 母0.31, tR (min) 3.2.

Acid Hydrolysis ntawm IA

tov ntawm compound I (2 mg) nyob rau hauv 10 feem pua ​​H2SO4 (1 ml) tau rhuab nyob rau hauv boiling dej da dej rau 30 feeb. Cov tshuaj tau dhau los ntawm Amberlite IR-45 kem thiab muaj zog kom muab cov khoom seem, uas tau txo nrog sodium borohydride (ca. 3 mg) rau 1 h. Cov tshuaj tiv thaiv sib xyaw tau dhau los ntawm Amberlite IR-120 kem thiab ua rau kom qhuav. Boric acid raug tshem tawm los ntawm distillation nrog MeOH thiab cov seem yog acetylated nrog acetic anhydride (1 poob) thiab pyridine (1 poob) ntawm 100 degree rau 1 h. Cov reagents tau evaporated tawm hauv vacuo. Glucitol acetate thiab rhamnitol acetate tau kuaj pom hauv qhov piv ntawm 1 txog 1 los ntawm compound I los ntawm GC. Qhov xwm txheej: kem, 1.5 feem pua ​​OV-17, 3 hli x 1.5 m; sab temp., 180 degree; cov pa roj, N2 (30ml / min). tR (min): 2.0 (rhamnitol acetate), 5.5 (glucitol acetate).

Enzymatic Hydrolysis ntawm II thiab III Compound II

(13 mg) lossis III (4{23}} mg) tau hydrolyzed nrog cellulase (40 mg) hauv dej (1 ml) rau 2h ntawm 37 degree . Cov tshuaj tiv thaiv sib xyaw tau muab rho tawm nrog EtOAc. Lub EtOAc extract tau concentrated nyob rau hauv vacuo muab cov residue, uas yog chromatographed ntawm HPLC [kem, Develosil ODS-10 (20 x 250 mm); kuab tshuaj. II: H2O-CH3CN (77.5:22.5); III: H2O-CH3CN (80:20); ntes (UV), 205 nm; flow rate, 6.0ml/min] muab lib (3.3 mg), Illb (5.7 mg). lib: [a]^5 plus 34.8 degree (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 (%) : 358 (M plus , 53), 340 (M plus -H2O, 100), 338 (58), 325 ( 47), 310 (23), 162 (41), 151 (36), 137 (59). XH-NMR (acetone-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="">

Kev lees paubPeb ua tsaug rau Prof. A. Ueno thiab Dr. T. Miyase, University of Shizuoka, rau kev ntsuas ntawm NMR thiab CD spectra, thiab rau lawv cov lus qhia tseem ceeb, thiab rau Dr. M. Kikuchi, Tohoku College of Pharmacy, rau kev muab cov qauv tseeb.

Cov ntaub ntawv thiab Cov Lus Qhia

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