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SUPPLEMENTARY MATERIAL
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Three new seco-ursadiene triterpenoids from Salvia syriaca
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Amal MF. Al-Aboudi*a, Musa H. Abu Zarga*a, Barakat E. Abu-Irmailehb,
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Firas F. Awwadia and Monther A. Khanfara
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8
a
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11942, Jordan.
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Chemistry Department, Faculty of Science, The University of Jordan, Amman
b
Department of Plant Protection, Faculty of Agriculture, The
University of Jordan, Amman 11942, Jordan.
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* Corresponding authors:
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Musa H. Abu Zarga Tel: +962 6 5355000 ext 22131; email: musaaz@ju.edu.jo
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Amal MF. Al-Aboudi Tel: +962 6 5355000 ext 22156; email: amal001@hotmail.com
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1
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Three new seco-ursadiene triterpenoids from Salvia syriaca
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Abstract
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Three new seco-ursadiene triterpenoids 1 – 3 together with 11 known compounds were
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isolated from Salvia syriaca of Jordanian origin. The compounds were identified by
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NMR spectroscopy including extensive 2D NMR experiments and mass spectrometry.
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The structure of compound 3 was confirmed by X-ray, and the information thus
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obtained was used to confirm the stereochemistry of compounds 1 and 2. This is the
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second report of 17,22-seco-17(28),12-ursadien-22-oic acids.
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Keywords: Salvia syriaca; Labiateae; triterpenoids; seco-ursadiene; 17,22-seco-
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17(28),12-ursadien-22-oic acid, X-ray.
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S3. Experimental
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S3.1. Instrumentation and general experimental procedures
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Melting points were determined using an Electrothermal IA 9300 Digital Melting
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Point apparatus and were uncorrected. EIMS were obtained at 70 eV on a Finnigan
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MAT TSQ 70-triple quadruple instrument. HREIMS were obtained (at 70 eV) on a
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Finnigan Mat 711 A sector field instrument modified by AMD Intecta. HR-ESI-MS
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for compound 3 was measured in positive ion mode using electro-spray ionization
37
technique on a Bruker APEX-2 instrument (Bremen,Germany). Specific rotations
38
were measured on MCP 200 (Anton paar, Seelz, Germany). NMR spectra were
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carried out on a Bruker DPX300 MHz (at 300 MHz for 1H and 75 MHz for
40
CDCl3 or DMSO-d6 using TMS as internal standard. Column chromatography was
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carried out on Silica gel 60 (0.063-0.200 mm, Fluka) or on neutral alumina
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(Brockmann Activity No. 1, Macherey Nagel, Germany). TLC was performed on
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silica gel GU254 pre-coated glass plates (0.25 or 0.50 mm in thickness, Machery
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Nagel, USA). Compounds were visualized under UV light or spraying with sulfuric
45
acid - anisaldehyde spraying reagent followed by heating at 120 0C. Methylation of
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the acids was achieved by treatment of 15 mg of the acid with excess ethereal
47
diazomethane. Acetylation of the methyl ester derivatives was carried out using
48
excess Ac2O / pyridine at room temperature for 24h. Final purification of the
49
derivatives was achieved by TLC.
2
13
C) in
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S3.2. Isolation of compounds 1-14
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Fraction I (120 g) was chromatographed on fine silica column (240 g) using benzene
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– ethyl acetate mixtures of increasing polarity for elution. The resulting fractions were
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collected into five sub-fractions (I-1 – I-5). Treatment of sub-fractions I-2 and I-3
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with methanol afforded 7,4'-dimethoxy-5-hydroxyflavone 4 (8.5 g) (Hanawa &
55
Towers 2003) and salvigenin 5 (6.2 g) (Hatam & Yousif 1992), respectively. Fraction
56
II (150 g) was purified on a fine silica column (300 g) eluting with benzene – ethyl
57
acetate mixtures of increasing polarity to give six collective sub-fractions (II-1 – II-6).
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Treatment of sub-fractions II-2, II-3 and II-5 with methanol gave genkwanin 6 (3.5 g)
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(Wagner & Chari 1976), cirsimaritin 7 (1.2 gm) (Abdalla & Abu Zarga 1987) and
60
oleanolic acid 8 (22 g) (Seebacher et al. 2003), respectively. Fractions III and IV
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were combined (110 g) and treated with methanol to give a solid (98 g) and a mother
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liquor (12 g). The mother liquor was chromatographed on a neutral alumina column
63
(200 g) which was eluted with chloroform-methanol mixtures of increasing polarity.
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The resulting fractions were followed by TLC and similar fractions were combined to
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give a total of six collective sub-fractions. A sample of the solid (0.60 g), obtained
66
from the combined fractions III and IV, was purified by TLC using methanol-
67
chloroform (5:95 v/v) as solvent to give salvigenin 5 (120 mg), cirsimaritin 7 (80 mg),
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cirsilineol 10 (70 mg) (Barberan et al. 1985), and a mixture (280 mg) of oleanolic acid
69
8 and ursolic acid 11 (Seebacher et al. 2003). Fraction V (50 g) was treated with
70
methanol to give a solid which was washed several times with the same solvent to
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give apigenin 12 (600 mg) (Miski et al. 1983). Fraction VI (60 g) was
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chromatographed on a column of fine silica (120 g) eluting with a gradient of benzene
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– ethyl acetate to give five sub-fractions. Treatment of subtraction VI-4 with methanol
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precipitated a solid which was purified further by recrystallization from methanol to
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give 7-O-methylluteolin 13 (120 mg) (Ahmad et al. 2001). Fraction VII (80 g) was
76
treated with methanol to give an impure solid which was purified by recrystallization
77
from methanol to give -sitosteryl glucoside 14 (2.3 g) (Jiang et al. 2012).
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S3.3.1. Methylation of compound 1
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Compound 1 (15 mg) in 3 mL of methanol, was treated with excess diazomethane in
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ether. After evaporation of solvent and excess reagent, the residue was purified by
81
TLC to give the methyl ester 1a: 1H NMR (CDCl3): δ 3.65 (3H, s, COOCH3), 3.27
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(3H, s, 11-OCH3), 3.22 (1H, dd, J = 6.5, 9.9 Hz, H-3α), 3.74 (1H, dd, J = 3.3, 8.3 Hz,
3
83
H-11β). 13C NMR (CDCl3): δ 173.8 (COOCH3), 51.3 (COOCH3); EIMS: m/z 500 (6)
84
[M]+.,, 468 (3), 371 (13), 357 (21), 339 (10), 129 (100).
85
S3.3.2. Acetylation of compound 1a
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Acetylation of 1a was achieved by treatment with Ac2O-pyridine at room temperature
87
for 24 hours. The solvent and excess reagent were evaporated and the residue was
88
purified by TLC to afford the monoacetyl derivative 1b: 1H NMR (CDCl3):  2.05
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(3H, s, 3-OCOCH3), 3.79 (1H, dd, J = 3.5, 8.5 Hz, H-11β), 4.55 (1H, dd, J = 3.3, 8.2
90
Hz, H-3); EIMS: m/z 542 (4) [M]+., 413 (10), 399 (10), 381 (7), 129 (100).
91
92
S3.4.1. Methylation of compound 2
93
Treatment of 2 (15 mg) in methanol with excess diazomethane in ether afforded its
94
methyl ester 2a: 1H NMR (CDCl3):  3.66 (3H, s, COOCH3); 3.28 (1H, dd, J = 4.4,
95
12.2 Hz, H-3), 3.41 (1H, ddd, J = 2.8, 3.6, 11.0 Hz, H-1), 4.18(1H, dd, J = 3.8, 8.3
96
Hz, H-11β), 5.91 (1H, d, J =2.8 Hz, 1-OH). 13C NMR (CDCl3):  173.7 (COOCH3),
97
51.4 (COOCH3); EIMS: m/z [M]+., 516 (4), 484 (100), 387 (22), 355 (72), 337 (20),
98
319 (13), 129 (22).
99
S3.4.2. Acetylation of compound 2a
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Acetylation of 2a with Ac2O-pyridine afforded the monoacetyl derivative 2b: 1H
101
NMR (CDCl3):  2.03 (3H, s, 3- OCOCH3), 3.45 (1H, ddd, J = 3.0, 4.3, 11.0 Hz, H-
102
1), 4.18 (1H, dd, J = 3.8, 8.4 Hz, H-11β), 4.54 (1H, dd, J = 4.6, 12.3 Hz, H-3),
103
5.87( 1H, d, J = 3.0 Hz, 1-OH ) ; EIMS: m/z 558 (3) [M]+., 526 (10), 429 (7), 337
104
(13), 319 (8), 239 (21) 185 (10), 129 (100).
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S3.5.1. Methylation of compound 3
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Treatment of 3 (15 mg) with ethereal diazomethane provided the methyl ester 3a. 1H
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NMR (CDCl3):  3.66 (3H, s, COOCH3), 3.28 (1H, dd, J = 4.3, 12.0 Hz, H-3), 3.55
108
(1H, dd, J = 4.5, 11.4 Hz, H-1), 4.31 (1H, dd, J = 4.0, 7.7 Hz, H-11β).
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(CDCl3):  173.7 (COOCH3), 51.4 (COOCH3); EIMS: m/z 502 (2) [M]+., 484 (36),
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355 (37), 337 (18), 319 (10), 225 (13), 217 (22), 185 (31), 129 (100). HREIMS: m/z
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502.3641 (calcd. for [M]+., C31H50O5, 562.3658).
112
S3.5.2. Acetylation of compound 3a
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Acetylation of 3a with Ac2O - pyridine afforded the monoacetyl derivative 3b: 1H
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NMR (CDCl3):  2.04 (3H, s, OCOCH3), 3.60 (1H, m, H-1), 4.33 (1H, dd, J = 3.9,
4
13
C NMR
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7.7 Hz, H-11β), 4.54 (1H, dd, J = 4.4, 12.1 Hz, H-3). EIMS: m/z 526 (46) [M–
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H2O]+., 433 (7), 397 (36), 337 (88), 319 (22), 248 (41), 129 (100).
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S3.5.3. Crystal structure determination of compound 3
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The diffraction data set of compound 3 was collected at room temperature using an
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Oxford Xcalibur diffractometer (Mo Kα radiation, λ = 0.7107 Å). Data were acquired
120
and processed to give hkl files using CrysAlisPro software*. The structure was solved
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by direct methods and refined by least-squares method on F2 using the SHELXTL
122
program package**. Carbon bound
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positions and refined isotropically using a riding model. Oxygen bound hydrogen
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atoms were found using Fourier difference map and refined isotropically using
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riding model except for water hydrogen atoms, the hydrogen atoms of the water
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molecule were refined istropically without restrains and constraints.
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hydrogen atoms were refined anisotropically. Details of the data collection and
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refinement are given in Table S1. CCDC 1007856 contains the supplementary
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crystallographic data for 3. Compound 3 crystallizes as a hydrate; the molecular
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structure is shown in figure 2. The water molecule is bonded to the carboxyl group of
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3 via O-HO hydrogen bonding interactions.
hydrogen atoms were placed in calculated
All non-
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* CrysAlisPro, Oxford Diffraction Ltd., Version 1.171.35.19 (release 27-10-2011
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CrysAlis171.NET)
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**SHELXTL (XPREP, X., XL, XP, XCIF), version 6.10; Bruker AXS Inc.: Madison,
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WI. 2002.
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S3.6 7,4'-Dimethoxy-5-hydroxyflavone 4
138
1
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= 2.1 Hz, H-6), 6.44 (1H, d, J = 2.1 Hz, H-8 ), 6.55 (1H, s, H-3), 6.98 (2H, d, J = 8.9
140
Hz, H-3`& H-5`), 7.80 (2H, d, J = 8.9 Hz, H-2`& H-6`), 12.78 (1H, s, 5-OH).
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NMR (CDCl3)  ppm: 55.6 (4`-OCH3), 55.9 (7- OCH3), 92.6 (C-8), 98.1 (C-6), 104.1
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(C-3), 105.6 (C-10), 114.5 (C-3`& C-5`), 123.6 (C-1`), 128.0 (C-2' & C-6'), 157.7 (C-
143
5), 162.2 (C-9),162.6 ( C-4'), 164.0 (C-2), 165.5 (C-7), 182.5 (C-4)
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S3.7 Salvigenin 5
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1
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6-OCH3), 6.51 (1H, s, H-8), 6.54 (1H, s, H-3), 6.98 (2H, d, J = 8.9 Hz, H-3`& H-5`),
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7.80 (2H, d, J = 8.9 Hz, H-2`& H-6`), 12.75 (1H, s, 5-OH). 13C-NMR (CDCl3)  ppm:
H-NMR (CDCl3)  ppm: 3.85 (3H, s, 4'-OCH3), 3.86 (3H, s, 7- OCH3), 6.32 (1H, d, J
13
C-
H-NMR (CDCl3)  ppm: 3.86 (3H, s, 4`-OCH3), 3.89 (3H, s, 7-OCH3), 3.94 (3H, s,
5
148
55.6 (4`-OCH3), 56.4 (7-OCH3), 60.9 (6-OCH3), 90.6 (C-8), 104.1 (C-3), 106.1 (C-
149
10), 114.5 (C-3`& C-5`), 123.5 (C-1`), 128.0 (C-2`& C-6`), 132.6 (C-6), 153.1 (C-5),
150
153.2 (C-9), 158.7 (C-7), 162.6 (C-4`), 164.0 (C-2), 182.7 (C-4).
151
S3.8 Genkwanin 6
152
1
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6.74 (1H, d, J = 2.2 Hz, H-8), 6.82 (1H, s, H-3), 6.90 (2H, d, J = 8.8 Hz, H-3'& H-5'),
154
7.93 (2H, d, J = 8.8 Hz, H-2' & H-6'), 10.37 (1H, bs, 4'-OH), 12.94 (1H, s, 5-OH).
155
13
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105.2 (C-10), 116.2 (C-3'& C-5'), 121.6 (C-1'), 129.1 (C-2`& C-6`), 157.7 (C-5),
157
161.7 (C-9), 161.8 (C-4'), 164.6 (C-2), 182.4 (C-4).
158
S3.9 Cirsimaritin 7
159
1
160
3), 6.90 (1H, s, H-8), 6.93 (2H, d, J = 8.6 Hz, H-3`& H-5`), 7.92 (2H, d, J = 8.6 Hz,
161
H-2`& H-6`), 12.65 (1H, brs, 5-OH).
162
60.5 (6-CH3), 92.1 (C-8), 103.1 (C-3), 105.5 (C-10), 116.5 (C-3`& C-5`), 121.4 (C-
163
1`), 128.9 (C-2`& C-6`), 132.3 (C-6), 152.6 (C-7), 153.1 (C-5), 159.1 (C-9), 162.0 (C-
164
4`), 164.6 (C-2), 182.7 (C-4).
165
S3.10 Oleanolic acid 8
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1
167
Me’s), 2.80 (1H, dd, J = 3.8, 13.6 Hz, H-18), 3.20 (1H, dd, J = 5.0 , 10.7 Hz, H-3),
168
5.25 (1H, brs, H-12). 13C-NMR (DMSO-d6)  ppm: 15.3 (25-CH3), 15.5 (24-CH3),
169
17.1 (26-CH3), 18.3 (C-6), 22.9 (11-CH3), 23.4 (C-16), 23.6 (29-CH3), 25.9 (27-
170
CH3), 27.1 (C-15), 27.7 (C-2), 28.1 (C-23), 30.7 (C-20), 32.4 (C-22), 32.6 (C-7), 33.1
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(30-CH3), 33.8 (C-21), 37.1 (C-10), 38.4 (C-1), 38.8 (C-4), 39.3 (C-8), 41.0 (C-18),
172
41.6 (C-14), 45.8 (C-19), 46.6 (C-17), 47.6 (C-9), 55.2 (C-5), 79.0 (C-3), 122.0 (C-
173
12), 143.6 (C-13), 183.6 (C-28).
174
S3.11 3β,11α-Dihydroxy-17,22-seco-17(28),12-ursadien-22-oic acid 9
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1
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CH3), 0.69 (3H, d, J = 6.8 Hz, 29-CH3), 0.73 (3H,s, 23-CH3), 0.85 (3H, s, 24-CH3),
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0.90 (3H, s, 26-CH3), 1.12 (3H, s, 27-CH3), 1.44 (1H, d, J = 8.5 Hz, H-9), 1.77 (1H,
178
m, H-19), 2.35 (1H, d, J = 11.2 Hz, H-18), 2.94 (1H, m, H-3), 3.32 (1H, brs, 3-OH),
179
4.20 (1H, d, J = 3.0 Hz, 11-OH), 4.55 (1H, d, J = 2.4 Hz, H-28a), 4.66 (1H, brs, H-
180
28b), 5.05 (1H, d, J = 3.1 Hz, H-12). 13C-NMR (DMSO-d6)  ppm: 11.2 (C-29), 12.5
H-NMR (DMSO-d6)  ppm: 3.83 (3H, s, 7-OCH3), 6.34 (1H, d, J = 2.2 Hz, H-6),
C-NMR (DMSO-d6)  ppm: 56.5 (7-OCH3), 93.2 (C-8), 98.5 (C-6), 103.5 (C-3),
H-NMR (DMSO-d6)  ppm: 3.69 (3H, s, 7-CH3), 3.88 (3H, s, 6-CH3), 6.81 (1H, s, H13
C-NMR (DMSO-d6)  ppm: 57.0 (7-CH3),
H-NMR (DMSO-d6)  ppm: 0.73, 0.75, 0.88, 0.94, 0.98, 1.11, 1.23 (each 3H, s, 7
H-NMR (DMSO-d6)  ppm: 0.62 (3H, d, J = 5.94 Hz, 30-CH3), 0.63 (3H, s, 25-
6
181
(C-30), 16.4 (C-25), 16.9 (C-26), 18.4 (C-6), 19.2 (C-24), 23.0 (C-27), 27.8 (C-15),
182
28.1 (C-16), 29.0 (C-23), 30.0 (C-20), 33.6 (C-2), 34.2 (C-1), 36.8 (C-19), 38.7 (C-
183
10), 41.0 (C-7), 41.2 (C-21), 42.4 (C-8), 42.8 (C-4), 43.2 (C-14), 55.0 (C-9), 55.7 (C-
184
5), 56.2 (C-18), 66.0 (C-11), 77.5 (C-3), 108.9 (C-28), 129.6 (C-12), 141.5 (C-13),
185
151.1 (C-17), 174.7 (C-22).
186
S3.11 Cirsilineol 10
187
1
188
s, 6-OCH3), 6.79 (1H, s, H-8), 6.87 (1H, s, H-3), 7.06 (1H, d, J = 8.6 Hz, H-5`), 7.43
189
(1H, d, J = 2.1 Hz, H-2`), 7.54 (1H, dd, J = 2.1, 8.6 Hz, H-6`), 9.41 (1H, s, 4`-OH),
190
12.87 (1H, s, 5-OH). 13C-NMR (DMSO-d6)  ppm: 56.3 (3`-OCH3), 57.0 (7-OCH3),
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60.5 (6-OCH3), 92.0 (C-8), 102.2 (C-3), 105.6 (C-10), 112.6 (C-2`), 113.6 (C-3`),
192
119.3 (C-6`), 123.4 (C-1`), 132.4 (C-6), 147.8 (C-4`), 151.7 (C-3`), 152.6 (C-9), 153.1
193
(C-5), 159.2 (C-7), 164.4 (C-2), 182.7 (C-4).
194
S3.12 Ursolic acid 11
195
1
196
= 6.2Hz, 29-CH3), 0.88(3H, d, J = 5.9 Hz, 28- CH3), 0.90 (6H, overlapping s & d, 24-
197
CH3, 30-CH3, respectively) 1.00 (3H, s, 27-CH3), 2.11 (1H, d, J = 11.4 Hz 18-CH3),
198
3.03(1H, m, H-3), 4.30 (1H, d, J = 4.8 Hz, 3-OH), 5.13 (1H, brs, H-12), 11.90 (1H,
199
brs, COOH). 13C–NMR: (CDCl3): δ15.7(C-25), 16.6(C-23), 17.4(C-26), 17.4(C-29),
200
21.6(C-30), 23.4(C-27), 28.8 (C-24), 18.4(C-11), 23.3(C-6), 24.3(C-16), 27.9(C-2),
201
27.4(C-15), 30.6(C-21), 33.1(C-7), 37.0(C-22), 38.7(C-1), 38.9(C-20), 39.0(C-19),
202
47.3(C-9), 52.8(C-18), 55.3(C-5), 77.3(C-3), 125.1(C-12), 36.8(C-10), 38.7(C-4),
203
39.1(C-8), 41.8(C-14), 47.5(C-17), 138.7(C-13), 178.8(C-28).
204
S3.13 Apigenin 12
205
1
206
H-8), 6.70 (1H, s, H-3), 6.75 (2H, d, J = 8.7 Hz, H-3' & H-5'), 7.85 (2H, d, J = 8.7 Hz,
207
H-2' & H-6'), 10.55 (2H, bds, 7-OH & 4'-OH), 12.90 (1H, s, 5-OH).13C–NMR
208
(DMSO – d6) δ ppm: 94.3 (C-8), 99.3 (C-6), 103.3 (C-3), 104.2 (C-10), 116.4 (C-3'
209
& C-5'), 121.7 (C-1'), 129.0 (C-2' & C-6'), 157.8 (C-9), 161.7 (C - 4'), 161.9 (C-5),
210
164.2 (C-2), 164.6 (C-7), 182.2 (C-4).
211
S3.14 7-O-Methylluteolin 13
212
1
213
6.72 (1H, d, J = 2.2 Hz, H-8), 6.79 (1H, s, H-3), 7.06 (1H, d, J = 8.6 Hz, H-5`), 7.43
H-NMR (DMSO-d6)  ppm: 3.69 (3H, s, 3`-OCH3), 3.84 (3H, s,7-OCH3), 3.89 (3H,
H–NMR (CDCl3): δ ppm: 0.69(3H, s, 23-CH3), 0.76 (3H, s, 26-CH3), 0.78 (3H, d, J
H–NMR (DMSO – d6) δ ppm = 6.15 (1H, d, J= 1.7 Hz, H-6), 6.43 (1H, d, J= 1.7 Hz,
H-NMR (DMSO-d6)  ppm: 3.83 (3H, s, 7-OCH3), 6.35 (1H, d, J = 2.2 Hz, H-6),
7
214
(1H, d, J = 2.2 Hz, H-2`), 7.54 (1H, dd, J = 2.2, 8.6 Hz, H-6`), 9.43 (1H, s, 4`-OH),
215
12.91 (1H, s, 5-OH). 13C-NMR (DMSO-d6)  ppm: 56.6 (7-OCH3), 93.1 (C-8), 98.5
216
(C-6), 104.2 (C-3), 105.3 (C-10), 112.6 (C-2`), 113.6 (C-6`), 119.3 (C-5`), 123.4 (C-
217
1`), 147.3 (C-3`), 151.7 (C-4`), 157.7 (C-9), 161.7 (C-2), 164.4 (C-5), 165.7 (C-7),
218
182.4 (C-4).
219
S3.15 β-sitosterol-D-glycoside 14
220
1
221
(3H, d, J = 6.0 Hz, 21-CH3), 0.96 (3H, s, 19-CH3) 1.00 (6H, d, J = 6.5 Hz, 26-CH3,
222
27-CH3), 0.90 – 2.50 (complex CH2 and CH), 3.63 (1H, m, H-3), 5.33 (1H, m, H-6),
223
4.21 (1H, d, J = 8.0 Hz, H-1′), 3.30 – 4.40 (overlapping signals of the sugar
224
hydrogns).
225
19), 19.6(C-27), 20.2(C-26), 21.1 (C-11), 23.1(C-28), 24.3 (C-15), 25.9 (C-23), 28.3
226
(C-16), 29.2 (C-25), 29.7 (C-2), 31.9 (C-8), 33.8 (C-22), 36.0 (C-20), 36.7 (C-10),
227
38.8(C-4), 39.7 (C-12), 42.4(C-13) 45.7 (C-24), 50.1(C-9), 55.9(C-17), 56.7 (C-14),
228
61.6 (C-6′), 70.6(C-3′), 74.0 (C-2′), 77.2 (C-4′), 77.3 (C-5′), 77.5 (C-3), 101.3 (C-1′),
229
121.6 (C-6), 141.0 (C-5).
H–NMR (DMSO-d6): δ 0.65 (3H, s, 18-CH3), 0.80 (3H, t, J = 5.2 Hz, 29-CH3), 0.90
13
C–NMR (DMSO-d6): δ 11.9 (C-18), 12.3 (C-29), 19.1 (C-21), 19.4(C-
230
8
CH3
H3C
H
H3CO
COOH
H
H
CH3
CH2
CH3
H
CH3
HO
H
H3C
CH3
231
232
233
Figure S1. Important HMBC correlations for compound 1
234
9
235
CH3
236
237
H3C
238
239
H
H3CO
240
241
H
O
242
COOH
H
H
CH3
243
244
CH3
H
CH3
245
246
247
248
HO
H
H3C
CH3
249
250
251
Figure S2. KeyHMBC correlations for compound 2
252
10
CH2
253
Table S1. Crystal data and structure refinement for compound 3.
254
Empirical formula
C30H48O5
255
Formula weight
506.70
256
Temperature
293(2) K
257
Wavelength
0.71073 Å
258
Crystal system
Orthorhombic
259
Space group
P212121
260
Unit cell dimensions
a = 7.9443(9) Å
261
b = 13.1414(11) Å
262
c = 26.895(3) Å.
263
Volume
2807.9(5) Å3
264
Z
4
265
Density (calculated)
1.199 Mg/m3
266
Absorption coefficient
0.081 mm-1
267
F(000)
1112
268
Crystal size
0.3 x 0.2 x 0.1 mm3
269
Theta range for data collection
2.98 to 25.00°.
270
Reflections collected
9313
271
Independent reflections
4688 [R(int) = 0.0634]
272
Completeness to theta = 25.00°
99.8 %
273
Absorption correction
Semi-empirical from equivalents
274
Max. and min. transmission
1.00000 and 0.96829
275
Refinement method
Full-matrix least-squares on F2
276
Data / restraints / parameters
4688 / 0 / 334
277
Goodness-of-fit on F2
0.984
278
Final R indices [I>2sigma(I)]
a
R1 = 0.0829, bwR2 = 0.1468
279
R indices (all data)
a
R1 = 0.1641, bwR2 = 0.1751
280
Largest diff. peak and hole
0.251 and -0.211 e. Å -3
281
--------------------------------------------------------------------------------------------------
282
-------------------
283
284
a
R1 = Fo-Fc / Fo.
b
wR2 ={[w(Fo2- Fc2 )2] / [w(Fo2)2]}1/2
285
11
HO
a
HO
OH
OH b
HO
HO
OH
O
HO
c
-amyrin
O
22
OH
HO
20
HO
OH
19
OH
d
28
O
O
H
H H
17
HO
HO
Urmiensolide
Compound 3
286
287
288
Scheme S1: A proposed biogenetic pathway of compound 3.
289
a: Cytochrom P-450 dependent monooxygenase, O2, NADPH; b: Dehydrogenase
290
NAD+; c: Flavin-peroxide ( Baeyer-Villiger) oxidation; d: Deprotonation and ring
291
cleavage.
292
293
12
[C6H10O2R4] +
+
m/z 115: R4 = H
m/z 129: R4 = CH3
COOR4
R3O
Ion D
R2
R2
– C6H10O2R4
R1O
R1O
Ion C
– R3OH
– R3OH
+
R2
COOR4
R2
– C6H10O2R4
R1O
R1O
Ion B
Ion A
294
295
296
SchemeS2. Mass fragmentation pattern for seco-ursadienes 1 – 3 and their methyl
297
esters.
298
13
299
300
301
302
Figure S3: 1H-NMR spectrum of 3-hydroxy-11-methoxy-17,22-seco-17(28),12ursadien-22-oic acid (1).
303
14
304
305
1
306
Figure S4:
H-NMR spectrum of 1,3-dihydroxy-11-methoxy-17,22-seco-
307
17(28),12-ursadien-22-oic acid (2).
308
309
310
311
15
312
313
1
314
Figure S5:
H-NMR spectrum of 1,3,11-trihydroxy-17,22-seco-17(28),12-
315
ursadien-22-oic acid (3).
316
317
16
318
319
13
320
Figure S6:
C-NMR spectrum of 3-hydroxy-11-methoxy-17,22-seco-17(28),12-
321
ursadien-22-oic acid (1).
322
323
17
324
325
13
326
Figure S7:
C -NMR spectrum of 1,3-dihydroxy-11-methoxy-17,22-seco-
327
17(28),12-ursadien-22-oic acid (2).
328
329
330
331
332
18
333
334
13
335
Figure S8:
C -NMR spectrum of 1,3,11-trihydroxy-17,22-seco-17(28),12-
336
ursadien-22-oic acid (3).
337
338
339
19
340
341
342
343
Figure S9: EIMS of 3-hydroxy-11-methoxy-17,22-seco-17(28),12-ursadien-22-oic
344
acid (1).
345
346
347
20
348
349
350
Figure S10: EIMS of 1,3-dihydroxy-11-methoxy-17,22-seco-17(28),12-ursadien-
351
22-oic acid (2).
352
353
354
21
355
356
357
358
359
Figure S11: EIMS of 11,3,11-trihydroxy-17,22-seco-17(28),12-ursadien-22-oic
360
acid (3).
361
22
362
363
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364
365
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23