Phenolic glycoside constituents from the rhizomes of atractylodes macrocephala koidz

lournal o f MedicinalMaterials, 2022, VoL 27, No (pp - 90) PHENOLIC GLYCOSIDE CONSTITUENTS FROM THE RHIZOMES OF A TRA CTYLODES MA CROCEPHALA KOIDZ Cao Thanh Hai1’2, Nguyên Thi Luyen3, Pham The Chinh2, Ngo Thi Thuy Ngan4, Nguyên Tuong Vy5, Nguyên Tien Dat1’3, Dang VietHau1,3’* 'Graduate University o f Science and Technology, Vỉetnam Academy o f Science and Technology (VAST), Hanoi, Vỉetnam; 2Thai Nguyên Universiíy o f Sciences, Tan Thinh Ward, Thai Nguyên City, Thai Nguyên, Vỉetnam; 3Centerfo r Research and Technology Trans/er, VAST, Hanoi, Vỉetnam; 4Thai Nguyên University o f Medicine and Pharmacy, Thai Nguyên, Vietnam; sHanoi University o f Science, Vietnam *Corresponding author: hauhoahock20@gmail.com (Received March 25th, 2022) Summary Phenolic Glycoside Constituents from the Rhizomes of Atractylodes macrocephala Koidz Four compounds were isolated from Atractylodes macrocephaỉa rhizomes as syringin (1), syringaresinol-4-O-^-Dglucoside (2), ewf-syringylglycerol-8-0-4'-sinapyl ether 4-0-/?-D-glucopyranoside (3), vy«-syrìngylglycerol-8-0-4’-sinapyl ether 4-O-yĩ-D-glucopyranoside (4) Thr structures were elucidated by ESI-MS, NMR spectroscopic evidence and in comparison with published data Compounds 2, 3, and have been isolated írom this plant for the first time Keywords: Atractylodes macrocephala; Asteraceae; Syringin, Syringaresinol-4-O-P-D-glucoside, Picraquassiosìde c Introduction Atractylodes macrocephala rhizomes is one of the most highly used medicinal plants in East Asia and it has been used treatment diarrhea, peptic ulcer disease [1] Up to now, more than 200 secondary 86 compounds have been isolated from the plant {2] A lot of research indicated that A macrocephala rhizomes had various bioactivities, including antibacterial, anti-inílammatoiy, anti-tumor, anticancer, immunomodulatory, and other activities Journal ofM edicinal Materials, 2022, Vol 27, No Previous studies were also demonstrated that sesquiterpenes, polyacetylenes together witìi their glycosides were ứie main Chemical components and were responsible for creating the biological activities of this plant [1],[2],[3],[4],[5],[6],[7] In this study, we found three lignan glycosides and one phenylpropanoid glycoside from A macrocephala rhizomes Fig Structure of compounds 1-4 Experimental 2.1 General procedures ESI-MS: Agilent LC-MSD-Trap SL 'H-NMR (500 MHZ) and I3C-NMR (125 MHZ) spectral data were measured on a Bruker Avance 500 Ultrashield NMR Spectrometer at 25°c Chemical shiữs were reported as values with reference to TMS as the intemal Standard for 'H (8 = ppm) TLC: Silicơ gel 60 F254 (0.25mm, Merck); reversed phase RP18F254S (0.25mm, Merck) CC: Silica gel 60 (230-400 mesh, Merck) for the íirst column, silica gel 60, 40-63 pm (Merck) for the following columns Optical rotations were recorded in a JASCO P-2000 polarimeter 2.2 Pỉaní materials The Atractyỉodes macrocephala Koidz rhizomes were collected in May 2021 in Quan Ba district - Ha Giang province and identiíìed by Dr Ngun The Cuong, Institute of Ecology and Biological Resources, Vietnam Academy of Science and Technology This plant was grown ừi Quan Ba district - Ha Giang province The voucher specimens (code: Ha Giang 06) were deposited at the Agro-Pharmaceutical Department, Center for Research and Technology Transfer, Vietnam Academy of Science and Technology 2.3 Extractìon and isoỉation The powder of A macrocephala Koidz rhizomes (2.5 kg) were extracted with methanol at room temperature (3 times, each ovemight) Aíter concentrating under reduced pressure to give 200.1 gram crude extract was suspended in 1.000 mL and then sequentially partitioned with dichloromethane The organic solvent was evaporated in vacuum to afford CH2CI2 (45.02 g) Water layer was loaded on the diaion column with a solvent System as MeOH:H20 (0:100 to 100:0, v:v) to yield ửactions as AMW1, AMW2 and AMW3 Fraction AMW3 was separated on a silica gel column, eluting with a gradient of CH2Cl2:MeOH (100:1 to 0:100, v:v) to give ữactions as AMWB1.10 and AMWB1.11 Fraction AMWB1.11 was separated on a silica gel column, eluting with solvent Systems CH2Cl2:Me0H:H20 (10:1:0.01, v:v:v) to aốòrd ữactions as AMWB2.4, AMWB2.7, AMWB2.9 Fraction AMWB2.4 was subjecteđ to silica gel column chromatography with a mobile phase as CH2Cl2:CH3COCH3 (1:1, v:v) to give a ửaction AMWB 3.4 Fraction AMWB3.4 was separated on a sephadex LH20 column solvent System as CH30H:H20 (1:2, v:v) to a obtain (9.7 mg) Fraction AMWB2.7 was íiirther puriííed on an RP18 column using the solvent System CH3C0CH3:H20 (1:6, v:v) to yield (9.3 mg) and (4.2 mg) Fraction AMWB2.9 was separated on an RP-18 column with a mobile phase as CH3C0CH3:H20 (1:6, v:v) to aíĩord (8.0 mg) Syringin (1) Colorless semisolid Melting point: 191-192°c [ a ]D = -18.0° (H20 ) ESI-MS (positive); m/z 373.4 [M+H]+ 'H- and l3C-NMR: Table Journal o f MedicinalMaterials, 2022, Vol 27, No 87 SyringaresinpI-4-ơ-yặ-D-glucopyranoside (2) Amorphous powdcr Melting point: 170- 173°c [a]20D= —21.5 (c Ọ.20, MeOH) ESI-MS (positive): m/z 581.1 [M+H]+ ,‘H- and 3*13C-NMR: Table e/ư-syringylglycerol-8-0-4'-sinapyl ether 40-p-D-g\ucoọyrsLnoside (3) Colorless senũsọỊid ESlvMS (positive): m/z 599.6 [M+H]+ ‘H-and 13C-NMR: Table syn-syringylglycerol-8-0-4 '-siitapyl eíber4ỡ-/f-D-gĩucopyranoside (4) Colọrless semisqlid ẸSI-MS (positive): m/z 599.4 [M+H]+ ‘H- and 13C-NMR: Table Table [H and 13C-NMR data of compounds 1-4 in CD3 OD Positi on Comooundl ỖH õc 136.3 - 105.9 154.7 135.7 154.7 105.9 131.7 1305 6.77(1H,s) 6.58(1H,t) 6.35(1Hi dt) 135.9 105.5 154.4 135.3 154.4 105.5 131.3 130.0 424(2H,dd) 63.6 6.77(1H,S) 33.88(3H,s) OCH] 53.88(3H,s) OCHi 1’ Conwound2 õc ÕH 0L74Í1HS)' 6.74(2H,S) 4.79(1H,d) 3.16(2H,m) 4.30(1H,m) 64.0 3.94(1H,dd) 729 682(1H, s) 5.03(1H,d) :4.25(1H,m) 3.85(1H,m) 349 (1H, m) õc~ ,ÕH õc õc~ 139.3 105.8 153.8 135.5 153.8 105.8 74.0 88.0 1392 105.7 153.7 135.4 153.7 105.7 74.0 87.9’ 6.77(2H.S) 139.5 105.7 153.8 135.6 153.8 105.7 74.0 87.0 139.7 106.0 1534 1352 1534 106.0 73.6 87.6 61.9 61.9 618 612 6.77(2H,s) 4.95(1H,d) 4.30(1H,dd) 3.95(1H,m) 3.62(1H,m) 57.1 56.8 3.86(3H,s) 57.0 56.9 3.83(31,s) 578 565 57.0 57.4 387(6H,s) 57.1 56.8 3.86(6B,s) 57.0 56.9 3.83PijS) 57.0 565 133.0 104.6 149.3 136.3 149.3 133.1 104.5 149.5 136.3 149.5 1348 104.8 1542 136.9 1542 134.7 104.7 1542 136.8 1542 134.7 104.9 1545 136.4 154.5 133.8 1044 153.9 1362 153.9 104.6 104.5 6.67(1H,s) ị 6.67(2H,s) T 4.74(1Hid) 3.16(1H;m) 4.30(1H,m) 3.94(2Hdd) 3.86(3H,s] 13.86(3Hs) 72.9 6.82(1H,s) õc 57.4 3.87(3H,S) V 8' 139.5 104.9 154.4 1356 154.4 104.9 87.2 55.7 ÕH 57.0 37 4' I rCenạix>tind4 Compoumt3 &139.6 104.9 154.5 135.7 154.5 104.9 872 557 õc 6.73(1H,s) 6.75(1H,S) 104.8 104.7 í 6.75(1H,s) 104.9 1044 87.6 55.5 87.6 55.5 6.73(1H,S,H36] 657(1H,brd) 6.35(1H,dt) 131.3 130.0 1312 129.9 6S5(1H,brd) 6.35(1H,dt) 131.3 129.9 1312 129.3 72.9 72.9 425 (2H, m) 63.5 63.5 425(2H, t) ,63.5 627 56.7 562 3.86(3H,s) 56.7 3,86(3H,S) 562 4.80 105.6 105.3 Gtc-r 4.89(1H,d) 105.4 105.4 4.80(1H,d) 10S.7 105.6 (1H,overlap] 1058 4.88(1H,d) 105.4 75.7 76.1 75.7 350(1H,ni) T 3.50(1H,m) 75.7 ’75.7 3.49(1Hi«n) 75.7 76.0 3.50(1H,m) 75.7 77.8 78.4 3.42(1H,m) 3" 3.44(1H,m) 77.8 78.1: 3.44(114,mi 77.8 77.8 778 78.1 3,44(1H,m) 71.4 71.6 71.3 71.3 3.44(1H,m) 71.4 71.3 3.43(ỊH,mj 4" 3.41 1H,m 71.6 3.42(1H, mi 71.3 78.4 78.6 sr 78.4 323(1H,m) 78.4 ,788 322(1H,m) 322 (1H,m) i 784 78.6 322 (1H, m) 78.3 3,87(1H,m) 3.79(1H,m) 3.81(1H,dd) 62.6 3.80(1H,dd) 62.6 626 62.5 3.70(1H,m) 62.6 62.6 6■ 63.0 62.6 3.69(1H,íld) 3.69(1H,dd) 3.69(1H,dd) *Sc of syringin (125 MHz, CD3 OD) [8], **& ofSyringaresinol-4-0-/ỉ-D-glucopyranoside (125 MHz, CD3 OD) [10 ***8c of Picraquassioside c (150 MHz, CD3OD) [12], '"'ỗ c of Pìeraquassioside c (150 MHz, C5D5N) [13] OCH3 y-cot 56.9 568 57.0 57.0 Results and dỉscussion Compound was obtained as a colorless semisolid from the water layer The molecule formula was determinedtobe C 17H24O9 based on the molecular ion positive peak ES1-MS at m/z 373.4 [M+H]+ The 'H-NMR spectrum showed characteristic signals of two íraM.s-olefìnic protons at &6.58 (1H, t, J = 15.5 Hz, H-7) and 6.35 (1H, dt, J = 15.5,6.0 Hz, H-8)and two aromtiẹ protons at ƠH6.77 (2H, s, H-2, H-6) The signàls of two methoxy groups at ỗH 3.88 (6H, s, -OCH3, 5OOH3), and a down-fíeld shitted methylene at ỗH 88 3.87(3H,s) 3.87(3H,s) 568 56.6 56.6 56.6 4.24 (2H, dd, J = 6.0, 1.5 Hz, H-9) were recognized Addiliooally, an anomeric-pmton signàl was observed at ÔH4.89 (1H, d , J = 7.5 Hz, H -l') and 13C-NMR were contirmed the presence of a ý?-glucopyranosyl moiety Analysis of the 13c NMR spectnim o f revealẽd the presence of an aromatic ring at ỏc 154.4 (C-3), ĩ'54.4 (C-5),135.9 (C-D, -135.3 (C 4), 105.5 (0 ), 105.5 (€-6), an àllýl at ỗc 131.3 (C-7), 130.0 (G-8), 63í6 (C-9), a glucopyranosyl moiety at ỏc 105.4 (C-l'), 78.4 (C5'), 77.8 (C-3'), 75.7 (0 '), 71.3 (0 '), 62.6 (C6') and two methoxyl groups at Ỗc51.0 (03/C -5, JournaLófMedicinal Materials, 2022, VoL 27, No CH3) Anạlyzing tìie NMR spectra of methoxy groups were rẹcognized at (5//3.87 (6H, corabined witìí knịwledge ofi its; chemotaxonomy s, 3',5'-OCH3),3.86 (6H, s , 3,5- OCHj) iThe 13Cand comparison with the literature [8], was NMR and 'HSQC spectra were ớbserved the suggested as syringin This compound was presence of twenty-eight carbon signâls including isolated hom this plant [9] eight quatemary carbons, thirteen methines, three Compound was obtained as an amorphous methylenes and four methoxys The HMBC powder from the water layer The ESI-MS of coưelations observed between H -l" [ÔH4.80 (ilH, showed a molecular ion at mJz 581.1 [M+H]+ and íd, J = 7.5 Hz, H-l")], H-2, H-6 [ổ//6.82 (2H, s, H1D, 2D-NMR spectra, consistent with the, íormula of :2, H-6)] and C-4 (ôc 135.5) indicated that the C28H 36O 13 Its 'H-NMR spectrum appeared signals linkage position of yể-Đ-glycopyranosyl located at of fọụr aromatic protons at ỗH6.74 (2H, s, H-2/H-6), C-4 In addiíion, the correlations from H-7', H-8' 6.67 (2H, H-2YH-6') and characteristic signals of [ỖH 6,57 (1H, br d, J = 16.0 Hz, H-7'), 6.35 (1H, two tetrahydroíuran rings at Ôh 4.19 (1H, d, J= 4.5 dt, J= 16.0,5.5 Hz, H-8')]to C -í' (ỏc 134.8), from Hz, H-7), 4.74 (1H, J = 4.5 Hz, H-7’), 4.30 (2H, m, H-2', H6' [ÔH6.73 (2H, s, H-2', H-6')].to C-4' (ổc H-9b, H-9h), 3.94'(2H,»dd, J= 9.0,3.0 Hz, H-9a, H- 136.9) were coníirmed allyl alcohol moiety at C9'a), 3.16 (2H, H-8,,H-8') together with signals for 1' The ent- conííguration of at C-7 and C-8 was foúr methòxy groups at Á/3.87 (6H, s, 3-ỎCH3, 5determined by the coupling constant J 7,8-value OCH3), 3.86 (6H, s, 3'-OGH3, 5'-OCH3) In additìon, (5.0 Hz) in the ‘H-NMR spectrum [11] an anomeric-ựLOtott signal was recognized at Ổh 4.88 Comparison of their NMR data with those (1H, d, J = 7.5 Hz, H-l") Analysis combined 13Creported [12] in the literatures was identified entNMR and HSQC spectra displayed the signals of syringylglycerol-8-ỡ-4'-sinapyl ether 4-Ơ-/1-Dtwenty-eight carbon including tvvo benzene rings.at glucopyran/úte This is the íìrst time this ỏc 154.4 (C-3), 154.4 (C-5), 149.3 (C-5'), 149.3Ỉ(Ccompound has been isolated from Atractylodes ỹ), 139.5 (C-í), 136.3 (c4Ĩ, 135.6(04), 133.0(0 macrocephala rhizomes It belongs to the cỉass of 1% 104.9 (0 ), 104.9 (0 ), 104.6 (O2'),104.6 (O organic as phenolic glycosides 6'), two teừahydroíuran rings at ỏc 87.6 (C-7'), 87.2 Compound was isolated as a colorless (0 7), 72.9 (0 ), 72.9 (0 , 55.7 (0 ), 55.5 (Csemisolid from the water layer of É macrocephala 80, íour.methoxys at ỏc 57.1 (3 -OCH3), 57.1 (5rhizomes The ‘H and 13C-NMR spectra of OGH3), 56.9 (3,-ÓCH3), 56.9 (S^OCHs) and caibon were similar to those of with some compound signals for a /?-glycopyranosyl moiety at ỏc 105.4 (Cdiffeĩienees being that the Chemical shift of H-7 and 1'0,78.3 (C-5"), 77.8 (C-3'O, 75.7 (C-2"),71.3 (CC-8 in changed from downfíeld at Ỗfí5.03 (1H, d, 47)1 62.6 (C-6") TheHMBC correlations ữom H -l" Ổ//4.88 (1H, d,J = 75 Hz, H-l") to C ỏc 135.6 (C- J = 5.0 Hz, H-7),