Studies On The Chemical Constituents Of Cistanche Tubulosa
Mar 19, 2022
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Song Zhihong, Mo Shaohong, Chen Yan, Tu Pengfei, Zhao Yuying, Zheng Junhua
Summary
Objective: To study the chemical constituents of domestic Cistanche tubulosa. Methods: Various chromatographic techniques including HPLC were used to separate the chemical components, and the structures were identified based on physicochemical properties and spectral data. RESULTS: Four iridoid glycosides, one lignan glycoside and one monoterpene were isolated, which were pentagonal acid ( Ⅰ ) , 8-epilogic acid ( Ⅱ ) and geoponic acid ( Ⅲ ) , auro cyanidin ( Ⅳ ) , eugenol glucoside ( Ⅴ ) and 8-hydroxy geraniol ( Ⅵ ) . Conclusion: Compounds I and VI were isolated from this genus for the first time, and compounds III, IV, and V were isolated from this plant for the first time.
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Introduction
The traditional Chinese medicine cistanche is the dry fleshy stem with scaly leaves of the genus Cistanche in the family Lidangaceae. It is one of the commonly used traditional Chinese medicines. Cold knee pain, blood dry constipation embolism. There are 4 species and 1 variant of Cistanche in China. The 2000 edition of the Chinese Pharmacopoeia contains Cistanche deserticola Y.C.Ma. Due to the shortage of drug sources, other species of this genus are also used in various places. Cistanche tubulosa (Schenk) R. Wight is mainly produced in Xinjiang, my country. It is rich in resources and is the main source plant of Cistanche deserticola. In Pakistan, India and other countries, there is also a kind of Cistanche tubulosa C.tububx (Schrenk) Hook.f. used as a medicinal material, but it is quite different from the domestic tube flower in terms of morphology, chemical composition and host plants, and is considered to be is different species 4. Japanese scholars have carried out in-depth research on Cistanche tubulosa produced in Pakistan. Phenyl ethanol glycosides, iridoid glycosides, lignan glycosides and other components have been isolated [3.4]. Domestic scholars have conducted preliminary research on the chemical constituents of the domestic Cistanche tubulosa, and isolated and identified 9 compounds54. In order to elucidate the chemical composition of domestic Cistanche tubulosa and provide the scientific basis for the development and utilization of new sources of traditional Chinese medicine Cistanche, we studied its chemical composition. Using various chromatographic techniques including HPLC, 4 iridoid glycosides, 1 lignan glycoside and 1 monoterpenoid were isolated from the normal alcohol extraction fraction of 95% ethanol extract, based on physicochemical properties and spectral data The identified structures are adoxosidic acid (I), 8-eploganic acid (8-eploganic acid, II), genposidic acid.D, mussamosidic acid (mussamosidic aid) IIV, eugenol glucoside [(+)-sringaresinl-Oβ-D-glucopyranoside. V] and 8-hydroxy geraniol (8-hydroxy-geraniol, VI. Compounds I and VI were isolated from this genus for the first time Obtained. Compounds III IIV and V were isolated from this plant for the first time.
1 Instrument and materials
XT-4A micro melting point apparatus (uncorrected). Infrared spectrometer: Perkin-Elmer Model 983 (KBr tablet). Nuclear Magnetic Resonance Spectrometer: Bruker ARX-400, Varian UNITY-500 (TMS or solvent as internal standard). Mass spectrometer: KYKY-ZHP-5* type, Zab-SpecE type. High-performance liquid chromatography: Waters 600 model (600 pumps 486 UV-vis detector); Gilson 712 preparative (306 pumps, 118 UV-vis detector). HPLC column: a. Phenomenex Prodigy, 5'm, ODS(3), 4.6mm × 250 mm; b. Alltech Econosl Cis. 10 Pm. 22 mm × 250 mm, chromatographically pure methanol, after re-distilling deionized water Filter use. TLC silica gel (silica gel H.60 type) is a product of Qingdao Ocean Chemical Group; reversed-phase silica gel (RP-18, 100-120 mesh) is a product of Beijing Eurasian New Technology Company; Sephadex LH-20 is a Pharmacia product; Do1 macroporous adsorption The resin is the product of Tianjin Nankai University Chemical Plant; the Diaion HP20 macroporous adsorption resin is the product of Japan's Mitsubishi Chemical; the high-efficiency thin-layer board and the RP-18F254 thin-layer board are Merck products; the polyamide film is the product of Zhejiang Siqing Biochemical Materials Factory.
The dried fleshy stems of Cistanche tubulosa C.tubulos (Schenk) R.Wight were purchased from Xinjiang Yutian County Pharmaceutical Company, identified by Professor Tu Pengfei, and the samples were kept in our laboratory.
2 Extraction and separation
Cistanche tubulosa crude drug 36.0 kg, crushed and extracted with 95% EtOH 4 times. The combined filtrate was concentrated under reduced pressure to obtain an ethanol extract, suspended with an appropriate amount of H20, and extracted with P.ether, EtOAc and n-BuOH in turn. The n-BuOH extract (500 g) was subjected to Dpou macroporous adsorption resin column chromatography, eluting with H2O and 10%, 30%, 50%, and 70% EtOH successively. The H2O fraction (140.0g) was subjected to column chromatography on Diaion HP20 macroporous adsorption resin and eluted with H2O-MeOH gradient to obtain 7 fractions (FrI ~ V, FrIV (3.2g), which were subjected to Sephadex LH-20 column chromatography (H2O-MeOH gradient elution). Fr4 (0.2 g) was separated by HPLC [Cs column b H2O-MeOH-HCOOH (88:12:0.5), 10 ml'min~1, 260 ml, to obtain compound I (24 mg), II (24 mg), III 70 mg) and IIV (15 mg).
The 50% EtOH fraction (10.0 g) was subjected to reverse-phase silica gel column chromatography (gradient elution of 30%~85% MeOH) to obtain 11 fractions (FrI~X, FrVVIII 1.5g), which were subjected to Sephadex LH-20 column chromatography (40%~85% MeOH gradient elution). 70% McOH gradient elution), preparative thin-layer chromatography on silica gel CHCl-MeOH (3÷1) to obtain compounds V (70 mg) and VI (10 mg), 3 Structure identification
Compound I off-white powder, mp 132～134 ℃. IR (KBr) cm~1:3361 (hydroxyl), 2924, 1674 and 1627 (αβ-unsaturated acid). 'H NMR (400 MHz, DMSO-dk) δ4.48 (IH, d.7.8Hz, GH-1), 5.08 (IH, d6.4Hz, H-1), 7.33 (1H, s, H-3) . See Table 1 for 13c NMR data. FAB-MS (posit) m/z: 399(M+Na+.377(M+H)+, 215(M+H-Gle). The molecular formula is C6H24O10. After analyzing the above data, I is consistent with the Wufuhua reported in the literature [7] Glycosides.
Table 1 CNMR data of compound L.III.V
Compound Ⅱ White needle crystal (McOH), mp 147～149 ℃ IR (KBr) cm~1; 3430 (hydroxyl). 1677 and 1646 (α, β-unsaturated acid), 'H NMR (500 M Hz, DMSO- d) 0.90(3H,d7.1 Hz,CH3),4.44(1H,d,7.8Hz,GH-1), 5.18(1H,d,3.6Hz,H-1),6.92(1H,s,H- 3). FAB-MS (neg) m/z: 375 (M-H) 7,212 (M-H-Gle). The molecular formula is CsH2xO10. Analysis of the above data. IIV is in line with the 8-epoxy strychnic acid reported in the literature [4. The TLC identification is consistent with the standard product.
Compound III Self-coloring powder, mp 139～141 ℃. IR (KBr) cm-1: 3389 (hydroxyl), 2918.1675 and 1627 (α,β-unsaturated acid).'H NMR (500 MHz. DMSO-de) à 3.98 (1H,d,14.5Hz,H- 10),4.14(1H,d14.5Hz,H-10),4.53(1H,d,7.3 Hz,GH-1),5.05(IH,d7.1 Hz,H-1),5.67(H,s, H-7), 7.34 (IH, sH-3)."c NMR data are shown in Table 1. FAB-MS (posit) m/z: 375 (M+H)+, 213 (M+H-Gle)+. The molecular formula is CidH22O0. Analysis of the above data, III is consistent with the geniposide reported in the literature [8].
Compound IV White powder, mp 140～142 ℃. IR (KBr) m-1: 3383 (hydroxyl), 29221675 and 1635 (α,β-unsaturated acid). 'H NMR (500 MHz, DMSO-da) δ1.19 (3H,s,CH3), 4.49 (1H,d,8.0Hz,GH-1), 5.31 (1H,d,4.0Hz,H-1), 7.28(IH,s,H-3.'c NMR data are shown in Table 1. FAB-MS (posit) m/z: 415(M+K)+, 399(M+Na)+, 377(M+H+, 215( M+H-Gk)+. The molecular formula is CuH2aO10. Analysis of the above data shows that IV is in line with the jasmine anthocyanin reported in the literature [8].
Compound V is light yellow powder, mp 173～174 ℃, Mol ish reaction is positive. IR (KBr) cm~1: 3428 (hydroxyl), 2919, 2850, 1589 and 1513 (benzene ring), H NMR (500 MHz, CD:OD) 63.88(6H,s,3',5'-OCH3), 3.90(6H,s.3, 5-oCH3),4.76(1H.d4.5Hz,H-2),4.81(1H,d, 4.0Hz,H-6),4.90(1H,d, 7.5Hz,GH -1),6.70(2H,s,H-2.6).6.76(2H,s,H-2,6.3c NMR (125 MHz, CD; OD) many signals appear in pairs): 55.8/56.0(C- 1,5), 57.1/57.4(4×OCH3), 62.8(GC-6, 71.6(GC-4), 73.15/73.21(C-4,8), 76.0(GC-2), 78.1(GC-3 ), 78.6(GC-5), 87.5/87.9(C-2,6), 104.8(C-2'.6), 105.1(C-2,6), 105.6(GC-1), 133.3(C- 1), 135.8(C-1", 136.5(C-4), 139.8(C-4), 149.6(C-3',5), 154.7(C-3,5), FAB-MS (posit) m /z: 603 (M+Na), 418 (M+H-Gle), the molecular formula is C2H36O13. Analysis of the above data. V is consistent with the eugenol glucoside reported in the literature [9].
Compound VI Colorless needle crystal (MeOH), mp 85～Compound VI Colorless needle crystal (MeOH), mp 85～87 ℃, 'H NMR (500 MHz, CD: OD) a1.65, 1.67 (2 × 3H. s.2×CH3),2.07(2H,m,H-4),2.17(2H,m,H-5),3.91(2H,s,H-8),4.08(2H,d,6.0Hz,H -1), 5.38(2Hbrt.6.0Hz, H-2.6, CNMR(125 MHz, CD:OD) 613.7(C-10, 16.2(C-9), 27.0(C-5), 40.3 (C-4) , 59.4 (C-1), 68.9 (C-8), 125.1 (C-6). 126.2 (C-2), 135.2 (C-7), 138.9 (C-3). The above data and literature [10] The reported 8-hydroxygeraniol is consistent.
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