Discovery of New Iridoids as Farnesoid X Receptor Agonists from Morinda officinalis: Agonistic Potentials and Molecular Stimulation

Article abstract of DOI:10.1002/cjoc.202000654

The investigation of Morinda officinalis led to the isolation of twelve compounds (1—12), including three new iridoid glycosides morindalins A—C (1—3) and nine known compounds (4—12). Their structural identifications were conducted using HRMS, 1D and 2D NMR, and electronic circular dichroism (ECD) spectra as well as quantum chemical computations. Compound 6 displayed the most significantly agonistic activity against farnesoid X receptor (FXR) with an EC₅₀ value of 7.18 μM, and its agonistic effect was verified through the investigation of FXR downstream target genes including small heterodimer partner 1 (SHP1), bile salt export pump (BSEP), and organic solute transporter subunit alpha and beta (OSTα and OSTβ). The potential interaction of compound 6 with FXR was analyzed by molecular docking and molecular dynamics stimulation, revealing that amino acid residues Leu287, Thr288, and Ser332 played a crucial role in the activation of compound 6 towards FXR. These findings suggested that compound 6 could be regarded as a potential candidate for the development of FXR agonists.

Full text of DOI:10.1002/cjoc.202000654

Chinese Journal
of Chemistry
CJC 中 国 化 学 - An International Journal
Accepted Article
Title: Discovery of new iridoids as FXR agonists from Morinda officinalis: Agonistic
potentials and molecular stimulations
Authors: Zhi-Lin Luan, Fei Qiao, Wen-Yu Zhao, Wen-Hua Ming, Zhen-Long Yu, Jie
Liu, Sheng-Yun Dai, Shuang-Hui Jiang, Chao-Jie Lian, Cheng-Peng Sun,* Bao-Jing
Zhang, Jian Zheng,* Shuang-Cheng Ma,* and Xiao-Chi Ma
This manuscript has been accepted and appears as an Accepted Article online.
This work may now be cited as: Chin. J. Chem. 2020, 39, 10.1002/cjoc.202000654.
The final Version of Record (VoR) of it with formal page numbers will soon be
published online in Early View: http://dx.doi.org/10.1002/cjoc.202000654.
ISSN 1001-604X • CN 31-1547/O6
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Zhao Wenyu (Orcid ID: 0000-0002-3422-4913)
Cite this paper: Chin. J. Chem. 2021, 39, XXX—XXX. DOI: 10.1002/cjoc.202100XXX
Discovery of new iridoids as FXR agonists from Morinda offici-
nalis: Agonistic potentials and molecular stimulations
a,+
b,+
a,+
a
a
b
b
Zhi-Lin Luan, Fei Qiao, Wen-Yu Zhao, Wen-Hua Ming, Zhen-Long Yu, Jie Liu, Sheng-Yun Dai, Shuang-Hui
b
b
,a
a
,b
,a
a
Jiang, Chao-Jie Lian, Cheng-Peng Sun,* Bao-Jing Zhang, Jian Zheng,* Shuang-Cheng Ma,* and Xiao-Chi Ma
a
Dalian Key Laboratory of Metabolic Target Characterization and Traditional Chinese Medicine Intervention, College of Pharmacy, College
of Integrative Medicine, Dalian Medical University, Dalian, China.
b
National Institutes for Food and Drug Control, Beijing, China.
These authors contributed equally to this work.
+
Keywords
Morinda officinalis | Iridoid | FXR | Agonistic effect
Main observation and conclusion
The investigation of Morinda officinalis led to the isolation of twelve compounds (1-12), including three new iridoid glycosides, morinda-
lins A-C (1-3), and nine known compounds (4-12). Their structural identifications were conducted using HRMS, 1D and 2D NMR, and
electronic circular dichroism (ECD) spectra as well as quantum chemical computations. Compound 6 displayed most significantly ago-
nistic activity against farnesoid X receptor (FXR) with an EC50 value of 7.18 µM, and its agonistic effect was verified through the investi-
gation of FXR downstream target genes including small heterodimer partner 1 (SHP1), bile salt export pump (BSEP), and organic solute
transporter subunit alpha and beta (OSTα and OSTβ). The potential interaction of compound 6 with FXR was analyzed by molecular
docking and molecular dynamics stimulation, revealing that amino acid residues Leu287, Thr288, and Ser332 played a crucial role in
the activation of compound 6 towards FXR. These findings suggested that compound 6 could be regarded as a potential candidate for
the development of FXR agonists.
Comprehensive Graphic Content
*
E-mail: zhengjian@nifdc.org.cn (J. Zheng); masc@nifdc.org.cn (S.C. Ma);
View HTML Article
Supporting Information
suncp146@163.com (C.P. Sun).
Chin. J. Chem. 2021, 39, XXX－XXX©
2021
SIOC,
CAS,
Shanghai,
&
WILEY-VCH
GmbH
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differences between this version and the Version of Record. Please cite this article as doi:
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Zhi-Lin Luan et al.
Background and Originality Content
Results and Discussion
As a member of the nuclear receptor (NR) superfamily, far-
nesoid X receptor (FXR; NR1H4) is mainly expressed in the liver, il-
eum, and kidney, and plays critical roles in regulating the synthesis
and transport of bile acid, maintaining systemic energy homeosta-
Structural Characterization
Compound 1 was obtained as amorphous powders, and its mo-
lecular formula was defined as C16
H
22
O
10 according to a quasi-mo-
+
[1-3]
lecular ion peak at m/z 397.1111 [M + Na] (calcd. for C16
H
22NaO10
,
sis and integrating crosstalk between different organs.
FXR has
1
3
97.1111) in the HRMS spectrum. The H NMR data of 1 (Table 1)
been considered as a promising target for the treatment of various
metabolic disorders, such as cholestatic disorders, fatty liver dis-
eases, type 2 diabetes, dyslipidemia, obesity, atherosclerosis, and
H
displayed signals of three olefinic protons at δ 7.47 (1H, d, J = 1.5
Hz, H-3), 5.99 (1H, dt, J = 5.7, 2.1 Hz, H-6), and 5.66 (1H, dt, J = 5.7,
2.1 Hz, H-7), and one anomeric proton at δ 4.66 (1H, d, J = 7.9 Hz,
H-1′). The C NMR data of 1 (Table 1) showed 16 carbon signals,
including one carbonyl carbon at δ 170.9 (C-11), four olefinic car-
bons at δ 153.3 (C-3), 135.9 (C-6), 131.9 (C-7), and 111.8 (C-4), one
acetal carbon at δ 97.8 (C-1), and a group of glycose carbons at δ
00.6 (C-1′), 78.4 (C-5′), 78.1 (C-3′), 74.8 (C-2′), 71.6 (C-4′), and 62.7
[4-8]
H
renal diseases.
During the last decades, a lot of efforts have
13
been made for discovering and optimizing potential drug candi-
[
7, 9]
C
dates for FXR agonists.
C
The dried rhizomes of Morinda officinalis is Morindae Officinalis
Radix, known as “Ba Ji Tian” in Chinese, which is a traditional Chi-
nese medicine (TCM) initially recorded in “Shennong Ben Cao
C
C
1
1
[
10, 11]
(C-6′), which suggested that 1 was an iridoid glycoside. The H and
Jing”.
It possesses kidney-tonifying, strengthening tendons
1
3
C NMR data of 1 was closely resembling to those of 8-epiapo-
and bones, removing wind and dehumidification effects, therefore,
it has been used for the treatment of impotence, nocturnal emis-
sion, reproductive diseases, irregular menstruation, abdomen cry-
modynia, osteoarthritis, rheumatic arthralgia, and tendon atro-
[19]
dantheroside,
methoxy group [δ
the chemical shift value of C-11 was shielded from δ
epiapodantheroside to δ 170.9 in 1, revealing that 1 was the deriv-
with the following exception that signals of a
3.71 (3H, s) and δ 51.7] were absent in 1, and
169.1 in 8-
H
C
C
[10-12]
C
phy.
The phytochemical investigation of Morinda species led
ative of 8-epiapodantheroside. This deduction was confirmed
through an HMBC experiment showing long-range correlations of
H-3 with C-4/C-5/C-11 (Figure 2). In the NOESY spectrum of 1, cor-
relations of H-9 with H-5/H-10a and H-8 with H-1 (Figure 3) sug-
gested an β-orientation of H-1 and β-orientations of H-5, H-9, and
hydroxymethyl-8. The glycose linked at C-1 was assigned as β-D-
glucose through analysis of the hydrolyzed product of 1 using HPLC
to the isolation and identification of various secondary metabolites,
including mornaphthoates A-F, marinoids A-G, monotropein, as-
peruloside tetraacetate, asperuloside, asperulosidic acid, and poly-
[11-15]
saccharides, mono-, and oligosaccharides.
Some of them dis-
played a series of biological activities, such as anti-tumor, anti-in-
flammatory, hepatoprotective, anti-depression, and antibacterial
[12-17]
effects.
As our continuous work in the discovery of natural FXR agonists
[
20]
with an optical rotation detector and the coupling constant [δ
H
[9, 18]
4.66 (1H, d, J = 7.9 Hz, H-1′)]. The absolute configuration of 1 was
and revealing their potential interactions,
we found that M. of-
identified through a comparison of calculated and experimental
ficinalis exhibited a significantly agonistic effect towards FXR,
therefore, its phytochemical constituent was investigated through
the bioactivity-guided method to afford twelve compounds (Figure
[21, 22]
spectra of 1.
B3LYP/aug-cc-pVDZ level agreed with the experimental spectra of
, requiring (1R,5S,8S,9S)-configurations (Figure 4). Therefore, the
structure of 1 was shown in Figure 1, and named as morindalin A.
The calculated ECD of (1R,5S,8S,9S)-1 at the
1
1
), including three new iridoid glycosides morindalins A-C (1-3) and
nine known compounds (4-12). Their structural characterization
was performed by HRMS, 1D and 2D NMR, and electronic circular
dichroism (ECD) spectra as well as quantum chemical computations.
Some of them could activate FXR, and their agonistic effect was
confirmed by cell imaging technology. Molecular docking and mo-
lecular dynamics (MD) stimulation revealed their action mechanism
via the interaction with amino acid residues Leu287, Thr288, and
Ser332. This finding suggested that compound 6 could be served as
a potential candidate for the development of FXR agonists.
O
O
HO
O
O
H
H
H
OH
OH
O
HO
HO
HO
HO
H
H
H
O
O
O
O
O
O
O
O
O
O
O
HO
HO
HO
HO
O
HO
OH
OH
HO
OH
OH
OH
OH
1
1
H- H COSY
HMBC
1
2
3
1
1
Figure 2 Selected HMBC and H- H COSY correlations of compounds 1-3.
O
O
HO
O
O
HO
H
H
1
OH
H
OH
H
O
O
1
7
HO 10
9
5
3
HO
HO
1
H
HO
″
H
H
H
O
O
4
″
O
″ 1
″
O
O
H-6b
H-5
5
O
O
O
O
O
O
6
′
5′
O
1′
HO
HO
HO
2
O
O
HO
HO
O
HO
H-5
H-5
3
′
HO
HO
OH
OH
HO
OH
OH
H-8
H-9
OH
OH
OH
OH
1
H
5
2
3
4
H-1
H-9
H-8
H-9
OH
H
O
HO
O
O
O
OH
O
H
H-8
OH
H
O
H
O
OH
H-1
H-1
2
H-10a
H-5″a
HO
HO
HO
NOESY
H-2
″
H
H
H
O
O
O
O
O
O
O
O
1
3
O
O
O
O
HO
HO
HO
HO
HO
HO
HO
OH
HO
OH
OH
OH
OH
OH
O
OH
OH
6
7
8
HO
HO
HO
HO
O
Figure 3 Selected NOESY correlations of compounds 1-3.
H
OH
O
O
OH
NH
OH
NH
O
O
O
HO
H
O
O
O
O
2
O
HO
HO
N
N
OH
HO
HO
O
OH
H
H
OH
OH
OH 10
9
11
12
Figure 1 Chemical structures of compounds 1-12 isolated from M. offici-
nalis
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Chin. J. Chem. 2021, 39, XXX－XXX

New iridoids as FXR agonists from Morinda officinalis
Chin. J. Chem.
Figure 4 Experimental and calculated ECD spectra of compounds 1-3 (A-C)
at the B3LYP/aug-cc-pVDZ level.
loganin (6) found the downfield shift of C-6′ from δ
loganin (6) to δ 68.8 in 3 and the presence of a suit of an apiose (δ
10.3, 80.7, 78.1, 75.1, and 65.7), revealing that this apiose was
C
62.7 in secoxy-
C
C
[23]
1
1
OD) and 13C (150 MHz, CD
OD) NMR data of compounds 1-3
Table 1. H (600 MHz, CD
3
3
1
2
3
No.
δ
C
δ
H
(J in Hz)
δ
C
δ
H
(J in Hz)
δ
C
H
δ (J in Hz)
1
3
4
5
6
97.8
153.3
111.8
41.0
5.20 d (5.8)
7.47 d (1.5)
97.3
153.5
111.4
41.3
5.19 d (5.9)
7.49 d (1.6)
97.8
153.8
110.3
29.1
5.41 d (4.3)
7.47 d (1.8)
3.62 m
3.71 m
3.31 m
135.9
5.99 dt (5.7, 2.1)
129.1
5.76 dt (5.6, 2.4)
35.4
2.88 dd (16.4, 5.0)
2
.29 dd (16.4, 9.0)
7
8
9
131.9
51.5
44.6
65.3
5.66 dt (5.7, 2.1)
2.90 m
137.0
53.5
6.07 dt (5.6, 2.4)
3.60 m
176.4
134.6 5.64 ddd (17.3, 10.2, 9.6)
2.34 ddd (7.9, 5.7, 4.7)
3.60 m
44.8
2.83 ddd (7.7, 6.1, 4.8)
45.5
2.88 ddd (7.7, 6.1, 4.8)
5.28 dd (17.3, 1.7)
5.24 dd (10.2, 1.7)
1
0
176.4
120.8
3
.58 m
1
1
170.9
100.6
74.8
78.1
71.6
78.4
62.7
170.5
100.7
74.8
78.1
71.6
78.5
62.7
169.0
100.2
74.7
78.1
71.7
77.4
68.8
1
′
′
′
′
′
4.66 d (7.9)
3.22 m
4.66 d (7.9)
3.24 m
4.64 d (7.9)
3.20 dd (9.1, 7.9)
3.35 m
2
3
4
5
3.38 m
3.37 m
3.30 m
3.31 m
3.29 m
3.29 m
3.29 m
3.44 m
6
3.86 dd (11.8, 1.8)
3.86 dd (12.1, 2.2)
3.69 dd (12.1, 5.4)
4.00 d (11.3, 1.9)
3.63 dd (11.3, 6.3)
5.01 d (2.6)
3.91 d (2.6)
3
.68 dd (11.8, 5.4)
1
″
″
″
″
110.3
78.1
80.7
75.1
2
3
4
3.97 d (9.6)
3
.77 d (9.6)
3.57 s
5
″
65.7
5.18
OCH
3
-11
3.68 s
Compound 2 was isolated as amorphous powders, and has a
linked at C-6′ via a β-oriented glycosidic bond as the coupling con-
stant of the anomeric proton (δ 5.01, d, J = 2.6 Hz, H-1′′). This de-
molecular formula of C16
H
20
O
11 based on its HRMS data (m/z
11, 387.0927). A detailed com-
H
−
3
87.0938 [M − H] , calcd. for C16
H
19
O
duction was confirmed by an HMBC correlation of H-1″ with C-6′
(Figure 2). The configuration of this apiose was determined as D-
configuration according to a NOESY correlation of H-2″ with H-5a″
1
13
parison of H and C NMR data of 2 and 1 suggested that their dif-
ference was at C-10. The signals of the hydroxymethyl group at C-
[24-26]
1
0 [δ
H
3.60 (m) and 3.58 (m); δ
C
65.3] were absent in 2, and the
(Figure 3) as previously described.
The (1R,5S,8S,9S)-configu-
signal of a carbonyl group (δ
C
170.5) was present in 2, indicating
ration of 3 was assigned due to the agreement of the calculated ECD
spectrum of 1R,5S,8S,9S-3 and its experimental result (Figure 4).
Thus, the structure of 3 was defined as shown in Figure 1, and
named as morindalin C.
that 2 was a C-10 oxidate of 1. In the HMBC spectrum of 2, the cor-
relation of H-7 with C-10 (Figure 2) verified the above-mentioned
deduction. Furthermore, 2 had a D-glucose with a β-oriented glyco-
sidic bond according to the analysis of its hydrolyzed product by
Additionally, nine known compounds (4-12) were also iso-
[27]
HPLC-OR and the coupling constant [δ
H
4.66 (1H, d, J = 7.9 Hz, H-
lated from M. officinalis, namely geniposidic acid (4), deacetylas-
[
28]
[23]
[29]
1
′)]. The β-orientations of H-5, H-9, and COOH-10 and an α-orien-
perulosidic acid (5), secoxyloganin (6), apodantheroside (7),
[30]
[31]
[32]
tation of H-1 were determined based on NOESY cross-peaks of H-1
scandoside (8), ixoside (9), morinlongoside A (10), 1,2,3,4-
tetrahydro-β-carboline-3-carboxylic acid (11), and tryptophane
(12), respectively.
[33]
with H-8 and H-5 with H-9 (Figure 3). The absolute configuration of
[34]
2
1
was defined as 1S,5S,9R since the calculated ECD spectrum of
S,5S,9R-2 was similar to its experimental result (Figure 4). Accord-
The agonistic effects of all the isolated compounds towards
FXR
ingly, the structure of 2 was shown in Figure 1, and named as
morindalin B.
The molecular formula of 3 was assigned as C22
H
32
O
H
15 due to
32NaO15,
All the isolated compounds were assayed for their agonistic
effects of towards FXR examined by luciferase assay using FXR gene
promoter-driven element luciferase (FXRE) reporter, and the result
+
its HRMS data (m/z 559.1643 [M + Na] , calcd. for C22
1
13
5
59.1639). Comparison of the H and C NMR data of 3 and secoxy-
Chin. J. Chem. 2021, 39, XXX－XXX
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Report
Zhi-Lin Luan et al.
Figure 5 3D structures of compounds 3 (A) and 6 (B) with FXR.
Figure 6 2D interactions of compounds 3 (A) and 6 (B) with FXR.
was listed in Table 2. Compounds 3 and 6 displayed significant ago-
nistic activities against FXR with EC50 values of 36.48 μM and 7.18
μM, respectively. It should be noted that compounds 1-9 are iridoid
glycosides chemically, and while their activities show remarkable
differences. A preliminary conclusion regarding the structure-activ-
ity relationship (SAR) can be drawn that the seco-type iridoids (3
and 6) showed a better agonistic effect on FXR than their analogues.
These results reveal the reason at the molecular level why the ago-
nistic effect of compound 6 was more potent than that of com-
pound 3.
Table 2. The agonistic effects of compounds 1-12 against FXR.
Compound
EC50 (µM)
> 100
> 100
36.48
> 100
> 100
7.18
Compound
EC50 (µM)
> 100
> 100
> 100
> 100
> 100
> 100
1
2
3
4
5
6
7
8
Molecular docking
In order to reveal their potential interaction with FXR, the 3D
structure of FXR was downloaded from Protein Data Bank
9
10
11
12
(
http://www.rcsb.org/structure/3BEJ, ID 3BEJ), followed by the mo-
lecular docking analysis (Figures 5 and 6). Compound 3 bond in the
lignan-binding domain of FXR through the interaction of van der
Waals, attractive charge, hydrogen bond, alkyl, and π-alkyl. Amino
acid residue Arg331 interacted with compound 3 via two conven-
tional hydrogen bonds (Figure 6A). As shown in Figure 5B, com-
pound 6 also bond in the lignan-binding domain of FXR through sim-
ilar interactions described as compound 3. Both 3 and 6 possessed
a terminal olefinic bond that formed multiple interactions with FXR,
which partly explains why seco-type iridoids are more active than
common analogues. Furthermore, it was worth noting that three
conventional hydrogen bonds of amino acid residues Met290,
Ser332, and Tyr369 with compound 6 were observed in Figure 6B.
Molecular dynamics stimulation
The potential interaction of compound 6 with FXR was analyzed
by molecular docking, revealing that amino acid residues played a
crucial role in the activation of compound 6 towards FXR. To eluci-
date how they changed during 50 ns of MD stimulation, the final
docking result was applied for the investigation of the dynamical
interaction of compound 6 with FXR (Figure 7). During 50 ns of MD
stimulation, RMSD remained under 2 Å (Figure 7A) that was an im-
portant parameter for the evaluation of the stability of ligand with
protein, the radius of gyration of compound 6 with FXR remained
at the distance of 1.3-1.6 nm at three degrees of freedom (X, Y, and
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New iridoids as FXR agonists from Morinda officinalis
Chin. J. Chem.
Z axes, Figure 7C), and RMSF of residues was acquired to figure out
regions exhibiting higher flexibility with a distance of 1-5 Å (Figure
towards FXR, therefore, we investigated the mRNA and protein lev-
els of the well-known downstream targets of FXR in the liver (SHP1
and BSEP) and kidney (OSTα and OSTβ) after administration of com-
pound 6 (10 µM for 24 h) in HepG2 and HK2. CDCA, a defined FXR
agonist, was used as a positive control (40 µM for 24 h). After the
7
B). These results indicated the reliability of the MD stimulation be-
tween compound 6 and FXR. The potential energy of the complex
5
of compound 6 with FXR was about −5.85 × 10 kJ/mol (Figure 7D).
Figure 7 (A) RMSD of compound 6 with FXR for 50 ns of MD stimulation. (B) RMSF of compound 6 with FXR for 50 ns of MD stimulation. (C) Radius
of gyration of compound 6 with FXR for 50 ns of MD stimulation. (D) The potential energy of compound 6 with FXR for 50 ns of MD stimulation. (E)
The distance of compound 13 with amino acid residues Leu287, Thr288 (two hydrogen bonds), and Met290. (F) The distance of compound 6 with
amino acid residues Ala291, His294, Met328, and Ser332. (G) The distance of compound 6 with amino acid residues Leu348, Tyr369, His447, and
th
Trp469. (H) 3D structure of compound 6 with FXR at 50 ns of MD stimulation. (I) The interaction of compound 6 with amino acid residues Leu287,
th
Thr288, and Ser332 via four hydrogen bonds at 50 ns of MD stimulation.
Compared with the result of molecular docking, eleven amino acid
residues Leu287, Thr288, Met290, Ala291, His294, Met328, Ser332,
Leu348, Tyr369, His447, and Trp469 could form twelve hydrogen
bonds with compound 6 during 50 ns of MD stimulation (Figure 7E-
treatment with compound 6, the mRNA levels of SHP1, BSEP, OSTα,
and OSTβ were all significantly up-regulated (Figure 8A). In parallel,
the protein levels of these FXR targets were also increased by com-
pound 6 (Figure 8B-C). To further confirm whether the transcrip-
G), whereas some of the hydrogen bond interactions were unstable, tional regulation of FXR was modulated by compound 6, the cellular
including Met290, Ala291, His294, Met328, Leu348, Tyr369, His447, location of FXR was observed in HepG2 cells. After the administra-
and Trp469, because their distances with compound 6 were more
than 3.5 Å that was an acceptable distance to form a hydrogen
bond. As same as described in molecular docking, compound 6
could still bind to the Ligand binding domain of FXR through four
hydrogen bonds between the hydroxy group of Thr288 and COOH-
tion of compound 6, the cells showed clear translocation of FXR
from cytoplasm to nucleus (Figure 9). These results indicated that
compound 6 could activate FXR and transcriptionally regulated the
expression of FXR downstream targets, suggesting that compound
6 could serve as a candidate for the development of FXR agonists.
7
of compound 6, the carbonyl group of Leu287 and OH-2′ of com-
pound 6, and the hydroxy group of Ser332 and OH-4′ of compound
during this MD stimulation. These findings clarified that com-
Conclusions
6
pound 6 could activate FXR through the interactions with amino
acid residues Leu287, Thr288, and Ser332, indicating that com-
pound 6 could be regarded as an agent for the development of FXR
agonists.
In summary, the investigation of M. officinalis resulted in the
isolation and identification of three new iridoid glycosides morinda-
lins A-C (1-3) and nine known compounds (4-12). Compound 6 dis-
played significantly agonistic activities against FXR with an EC50
value of 7.18 µM. The result of cell imaging technology clarified
their agonistic effect against FXR at the cell level. Molecular docking
and MD stimulation revealed their action mechanism through the
Induction of FXR downstream targets by compound 6
Compound 6 displayed the most significantly agonistic effect
Chin. J. Chem. 2021, 39, XXX－XXX
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Zhi-Lin Luan et al.
Figure 8 Effect of compound 6 on FXR and its downstream target genes. (A) qPCR analysis showing the induction of FXR and its renal downstream
targets (SHP1, BSEP, OSTα, and OSTβ) in HepG2 cells treated with compound 6 (10 µM for 24 h). CDCA (40 µM for 24 h) was used as a positive control.
(B) Western blot analysis demonstrating a significant increase in the protein levels of FXR and its downstream targets in HepG2 cells treated with
compound 6. (C) Quantitative analysis of western blot results for FXR and its downstream targets in renal cells treated with compound 6. Data repre-
sent the mean ± SD, n = 3 for each group. *p<0.05, **p<0.01, ***p<0.001 compared to the DMSO group.
hydrogen bond interaction with amino acid residues Leu287,
Thr288, and Ser332. These findings suggested that compound 6
could be regarded as a potential candidate for the development of
Plant Material
FXR agonists.
Dried rhizomes of M. officinalis were purchased from Beijing
Tongrentang Co., Ltd. (Dalian, China) in June 2020, and identified
by Prof. Jing-Ming Jia (Shenyang Pharmaceutical University). A
voucher specimen (MO202006) has been deposited in the Depart-
ment of Medicinal Chemistry, Dalian Medical University.
Experimental
General Experimental Procedures
Extraction and Isolation
The optical rotations of isolated compounds were recorded on
a PerkinElmer 241 polarimeter. ECD spectra were recorded on a
Bio-Logic Science MOS-450 spectrometer. 1D and 2D NMR data
were recorded on a Bruker AV-600 spectrometer. Chemical shift
values are expressed in δ (ppm) using the peak signals of the sol-
M. officinalis (800 g) was extracted with 90% EtOH (8 L) for
three times to obtain a residue after removing the solvent. The res-
idue was suspended by 2 L water, and extracted with ethyl acetate
(
3 × 2 L), and n-BuOH (3 × 2 L), respectively.
vent CD
3
OD (δ
H
3.31 and δ
C
49.2) as references, and coupling con-
The n-BuOH extract (10 g) was separated by a silica gel (Qing-
stants (J in Hz) are given in parentheses. The antibody of FXR (PP-
A9033A-00) was purchased from R&D Systems. The antibodies of
small heterodimer partner 1 (SHP1, ab32559), and organic solute
transporter subunit beta (OSTβ, ab121285) were purchased from
Abcam. The antibody of bile salt export pump (BSEP, A8467) was
purchased from Abclonal. The antibody of organic solute trans-
porter subunit alpha (OSTα, bs-17529R) was purchased from BIOSS.
dao Marine Chemical Factory, Qingdao, China) column and eluted
with CH Cl -MeOH (from 50:1 to 1:1), resulting in the production of
2
2
eight fractions B1-B8. B5 (536 mg) was purified through preparative
HPLC with a YMC C18 column (250 mm × 10 mm, 5 µm) and eluted
with 10%-70% MeOH to afford three subfractions B51-B53. Separa-
tion of B53 (82 mg) through preparative HPLC (15% MeCN) led to
the isolation of compounds 3 (4.7 mg), 6 (4.9 mg), and 7 (2.7 mg).
Compounds 4 (1.7 mg), 5 (12.6 mg), and 8 (21 mg) were isolated
from B6 (120 mg) through preparative HPLC (10%-70% MeOH). B7
(
523 mg) was separated by preparative HPLC (10%-70% MeOH),
which was further purified by preparative HPLC (10%-20% MeOH)
to afford compounds 1 (3.7 mg), 2 (4.0 mg), 9 (4.8 mg), and 10 (5.0
mg). Finally, separation of B8 (135 mg) through preparative HPLC
(
10%-70% MeOH) afforded compounds 11 (3.4 mg) and 12 (4.3 mg).
20
Morindalin A (1): amorphous powder; [α] −16 (c 0.1, MeOH);
D
UV (MeOH) λmax (log ε) 240 (3.8) nm; ECD (MeOH) nm (Δε) 201
(
CD
1
13
+6.57), 233 (−3.88); H (600 MHz, CD
3
3
OD) and C NMR (150 MHz,
+
OD) data, see Table 1; HRMS m/z 397.1111 [M + Na] (calcd. for
C
16
H
22NaO10, 397.1111).
2
0
Morindalin B (2): amorphous powder; [α] −13 (c 0.1, MeOH);
D
Figure 9 Nuclear translocation of FXR stimulated by compound 6.
UV (MeOH) λmax (log ε) 237 (3.7) nm; ECD (MeOH) nm (Δε) 204
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New iridoids as FXR agonists from Morinda officinalis
Chin. J. Chem.
1
OD) and 13C NMR (150 MHz,
(
CD
C
+14.14), 237 (−8.81); H (600 MHz, CD
3
3
DMEM medium containing 10% fetal bovine serum (FBS). Human
proximal tubular epithelial cells (HK2) were maintained in
DMEM/F12 medium supplemented with 10% FBS. Cells were all
placed in an incubator at 37 °C and maintained in a humidified at-
−
OD) data, see Table 1; HRMS m/z 387.0938 [M − H] (calcd. for
16
H
19
O
11, 387.0927).
20
Morindalin C (3): amorphous powder; [α] −74 (c 0.1, MeOH);
D
UV (MeOH) λmax (log ε) 237 (3.6) nm; ECD (MeOH) nm (Δε) 223
2
mosphere containing 5% CO .
1
13
(
CD
−4.13), 319 (+1.10); H (600 MHz, CD
3
OD) and C NMR (150 MHz,
FXR activation bioassay
+
3
OD) data, see Table 1; HRMS m/z 559.1643 [M + Na] (calcd. for
22
C H
32NaO15, 559.1639).
The human FXR cDNA was cloned and inserted into pcDNA3.1
[9]
for the construction of FXR overexpression vector. FXRE reporter
was constructed by cloning a genomic DNA fragment upstream bile
salt export pump (BSEP) promoter into the luciferase vector
pGL3.0-basic. The human hepatoma HepG2 cell line was transiently
transfected with the expression plasmid of FXR, FXRE reporter, and
the null-Renilla luciferase plasmid as an internal control. After 24 h,
cells were treated with vehicle DMSO (0.1%) or corresponding com-
pounds at different concentrations. The cells were then harvested
for the determination of luciferase activity by Dual-Luciferase® Re-
porter Assay System (E1960, Promega, Madison, WI, USA).
ECD Calculations
To save consumption of calculations, model compounds with
the glycosyl units replaced with methyl groups were used for the
[35, 36]
ECD calculations.
The conformation search of compounds 1-3
was carried out at Merck Molecular Force Field (MMFF) by Discov-
ery Studio software, and the low energy conformers were kept with
an energy cutoff of < 5 kcal/mol.
[21, 22]
Geometries were optimized
using DFT at B3LYP/6-31G(d) level within a MeOH solvent simulated
using Gaussian 09 package (Gaussian 09. 2009). Harmonic vibra-
tional frequencies were also calculated at the same level of theory
to ensure that no imaginary values were present, confirming that
all the conformers were stable. The ECD calculations were run with
B3LYP functional and the aug-cc-pVDZ basis set of Gaussian 09
package with the same solvent as in the preceding DFT optimization
step. Depended on the Boltzmann distribution, the final ECD spec-
tra of compounds 1-3 were obtained by weighting spectra of all
conformers.
RNA isolation and real-time quantitative PCR
Total RNA was isolated from HepG2 and HK2 cells by the RNe-
asy Mini Kit (Qiagen, Germany), and quantified according to the
manufacturer's instructions. The quantity and purity of the ex-
tracted RNA were examined by using a Nanodrop spectrophotom-
eter (Thermo, USA). Reverse transcription was performed by re-
verse transcription kit (Tiangen, Beijing, China). The cDNA was used
as a template for real-time quantitative PCR on the Rrism 7500 real-
time PCR instrument with TransStart Tip Green qPCR SuperMix
(Transgen, China). Primer was designed by Software Oligo 6.0 (MBI
Inc., Colorado, USA) (Table S1).
The determination of the glycose of compounds 1-3
The absolute configurations of the glycose of compounds 1-3
[20]
were determined by HPLC-OR as previously described.
pounds 1-3 (1 mg) was hydrolyzed by 2M HCl (2.0 mL) at 90 C for
h to obtain the monosaccharide in the aqueous layer after the
CHCl extraction, and analyzed by HPLC with a Jasco LC-NetII/ADC
Com-
o
Western Blot
4
Cells were dissolved in RIPA Lysis buffer (Merck, Germany) with
protease inhibitor cocktail (Sigma-Aldrich, China), and then the su-
pernatants were collected. The concentrations of the protein sam-
ples were examined by bicinchoninic acid protein assay kit (Sigma-
Aldrich, China). Samples were heated for denaturation in the load-
ing buffer for 10 min at 95 °C. The denatured samples were then
separated by 10 % SDS-polyacrylamide gel electrophoresis (SDS-
PAGE), and transferred to polyvinylidene difluoride (PVDF) mem-
branes (Millipore, USA) which were then blocked with 10% (w/v)
skim milk and subsequently incubated with selected primary anti-
bodies overnight at 4 °C. After washes, the membranes were incu-
bated for 1 h at room temperature with HRP-conjugated secondary
antibodies. After washes, staining was developed using Tanon 5200
ECL detection system (Tanon, China).
3
system.
Molecular docking
The 3D structure of FXR was downloaded from Protein Data
Bank (http://www.rcsb.org/structure/3BEJ, ID 3BEJ), and then was
used to study the interactions between compounds 3 or 6 and FXR
[9]
by Discovery Studio 2016 (Accelrys, CA, USA). Firstly, the energy
optimization of 3D structures of compounds 3 and 6 was performed
at the CHARMm force field using the module of Prepare Ligand (Dis-
covery Studio 2016). The active pocket for substrate binding was
defined based on the site of the crystallographic ligand MFA-1. The
interaction of compound 6 with FXR was calculated by the module
of CDOCKER (Discovery Studio 2016), and the 3D interaction was
plotted by PyMOL software.
Immunohistochemistry
Molecular dynamics stimulation
GROMACS package was applied to study MD simulation at the
AMBER99SB force field.
Cells were firstly fixed with 4% PFA in PBS. After washed with
ice-cold PBS, cells were permeabilized with 0.3% Triton X-100 in
PBS for 10 min and blocked by 0.5% BSA in PBS for 30 mins. The
cells were then incubated with primary antibody overnight at 4 °C.
After washes, the cells were incubated with appropriate Dylight
488 (green) conjugated secondary antibody (Jackson Immuno Re-
search Laboratories) for 1 h at room temperature. Nuclei were
stained with DAPI. Images were obtained through a confocal micro-
scope.
[37, 38]
The restrained electrostatic potential
charges of ligand were performed at HF/6-31G* level using Gauss-
ian 09 program. The starting topology of the ligand was generated
by antechamber and ACPYPE programs. The best docking pose was
subjected to the steepest descent energy minimization to relax the
system. Then, NVT and NPT were used to equilibrate the minimized
system at the constant temperature of 300 K and the constant pres-
sure of 1 bar, respectively. In the last step, 50 ns of MD simulation
at 300 K and 1 bar pressure was performed, thus carrying out the
interaction analyses between the enzyme and ligand. The trajec-
tory file was analyzed by GROMACS to produce the root mean
square deviation (RSMD), root mean square fluctuations (RMSF),
and other parameters. The MD results were plotted by PyMOL and
GraphPad prism software.
Supporing Information
The supporting information for this article is available on the
WWW under https://doi.org/10.1002/cjoc.2021xxxxx.
Cell Culture
Acknowledgement
The human liver hepatocellular cells (HepG2) were cultured in
Chin. J. Chem. 2021, 39, XXX－XXX
© 2021 SIOC, CAS, Shanghai, & WILEY-VCH GmbH
www.cjc.wiley-vch.de

Report
Zhi-Lin Luan et al.
This work was supported by National Natural Science Founda-
reversing cholestasis effect by activation of farnesoid X receptor. Phy-
tomedicine. 2018, 42, 34-42.
Machida, K.; Takehara, E.; Kobayashi, H.; Kikuchi, M. Studies on the
constituents of Gardenia species. III. New iridoid glycosides from the
leaves of Gardenia jasminoides cv. fortuneana Hara. Chem. Pharm. Bull.
tion of China (No. 81703679), Liaoning Provincial Key Research and
Development Program (2019JH2/10300022), Natural Science Foun-
dation of Liaoning Province (2020-MS-256), and Dalian Young Star
of Science and Technology.
(Tokyo). 2003, 51, 1417-1419.
Sun, C.P.; Yuan, T.; Wang, L.; Kang, N.; Zhao, F.; Chen, L.X.; Qiu, F. Anti-
inflammatory labdane-type diterpenoids from Physalis angulata. Rsc
Adv. 2016, 6, 76838-76847.
References
Forman, B.M.; Goode, E.; Chen, J.; Oro, A.E.; Bradley, D.J.; Perlmann,
T.; Noonan, D.J.; Burka, L.T.; Mcmorris, T.; Lamph, W.W.; Evans, R.M.;
Weinberger, C. Identification of a nuclear receptor that is activated by
farnesol metabolites. Cell. 1995, 81, 687-693.
Lee, F.Y.; Lee, H.; Hubbert, M.L.; Edwards, P.A.; Zhang, Y. FXR, a multi-
purpose nuclear receptor. Trends Biochem Sci. 2006, 31, 572-580.
Sinal, C.J.; Tohkin, M.; Miyata, M.; Ward, J.M.; Lambert, G.; Gonzalez,
F.J. Targeted disruption of the nuclear receptor FXR/BAR impairs bile
acid and lipid homeostasis. Cell. 2000, 102, 731-744.
Liang, J.H.; Luan, Z.L.; Tian, X.G.; Zhao, W.Y.; Wang, Y.L.; Sun, C.P.; Huo,
X.K.; Deng, S.; Zhang, B.J.; Zhang, Z.J.; Ma, X.C. Uncarialins A-I, Mono-
terpenoid Indole Alkaloids from Uncaria rhynchophylla as Natural Ag-
onists of the 5-HT1A Receptor. J. Nat. Prod. 2019, 82, 3302-3310.
Wang, C.; Huo, X.K.; Luan, Z.L.; Cao, F.; Tian, X.G.; Zhao, X.Y.; Sun, C.P.;
Feng, L.; Ning, J.; Zhang, B.J.; Ma, X.C. Alismanin A, a Triterpenoid with
a C34 Skeleton from Alisma orientale as a Natural Agonist of Human
Pregnane X Receptor. Org. Lett. 2017, 19, 5645-5648.
Hiroshi, K.; Masami, M.; Kiyokazu, T.; Kenichiro, I.; Tetsuro, F.; He, Z.D.;
Yang, C.R. Secoiridoid, coumarin and secoiridoid-coumarin glucosides
from Fraxinus chinensis. Phytochemistry. 1992, 31, 1277-1280
Peng, K.; Yang, L.; Zhao, S.; Chen, L.; Zhao, F.; Qiu, F. Chemical constit-
uents from the fruit of Gardenia jasminoides and their inhibitory ef-
fects on nitric oxide production. Bioorg. Med. Chem. Lett. 2013, 23,
Chavez-Talavera, O.; Tailleux, A.; Lefebvre, P.; Staels, B. Bile Acid Con-
trol of Metabolism and Inflammation in Obesity, Type 2 Diabetes,
Dyslipidemia, and Nonalcoholic Fatty Liver Disease. Gastroenterology.
2
017, 152, 1679-1694 e1673.
Levi, M. Nuclear receptors in renal disease. Biochim. Biophys. Acta.
011, 1812, 1061-1067.
2
1
127-1131.
Moris, D.; Giaginis, C.; Tsourouflis, G.; Theocharis, S. Farnesoid-X Re-
ceptor (FXR) as a Promising Pharmaceutical Target in Atherosclerosis.
Curr. Med. Chem. 2017, 24, 1147-1157.
Sepe, V.; Distrutti, E.; Fiorucci, S.; Zampella, A. Farnesoid X receptor
modulators 2014-present: a patent review. Expert Opin. Ther. Pat.
Takayanagi, T.; Ishikawa, T.; Kitajima, J. Sesquiterpene lactone gluco-
sides and alkyl glycosides from the fruit of cumin. Phytochemistry.
2
003, 63, 479-484.
Zhou, K.; Zhao, F.; Liu, Z.; Zhuang, Y.; Chen, L.; Qiu, F. Triterpenoids and
flavonoids from celery (Apium graveolens). J. Nat. Prod. 2009, 72,
2
018, 28, 351-364.
1
563-1567.
Teodoro, J.S.; Rolo, A.P.; Palmeira, C.M. Hepatic FXR: key regulator of
whole-body energy metabolism. Trends Endocrinol. Metab. 2011, 22,
Tao, S.H.; Wu, J.; Qi, S.H.; Zhang, S.; Li, M.Y.; Li, Q.X. Scyphiphorins A
and B, two new iridoid glycosides from the stem bark of a Chinese
mangrove Scyphiphora hydrophyllacea. Helv. Chim. Acta. 2007, 90,
4
58-466.
Luan, Z.L.; Huo, X.K.; Dong, P.P.; Tian, X.G.; Sun, C.P.; Lv, X.; Feng, L.;
Ning, J.; Wang, C.; Zhang, B.J.; Ma, X.C. Highly potent non-steroidal FXR
agonists protostane-type triterpenoids: Structure-activity relationship
and mechanism. Eur. J. Med. Chem. 2019, 182, 111652.
Singh, B.; Sharma, R.A. Indian Morinda species: A review. Phytother.
Research. 2020, 34, 924-1007.
Zhang, J.H.; Xin, H.L.; Xu, Y.M.; Shen, Y.; He, Y.Q.; Hsien-Yeh; Lin, B.;
Song, H.T.; Juan-Liu; Yang, H.Y.; Qin, L.P.; Zhang, Q.Y.; Du, J. Morinda
officinalis How. - A comprehensive review of traditional uses, phyto-
chemistry and pharmacology. J. Ethnopharmacol. 2018, 213, 230-255.
Yu, J.H.; Zhai, H.J.; Yu, Z.P.; Zhang, Q.Q.; Ge, Y.X.; Zhang, Y.Y.; Jiang, C.S.;
Zhang, H. Methyl 2-naphthoates from a traditional Chinese herb
Morinda officinalis var. officinalis. Tetrahedron. 2019, 75, 3793-3801.
Xu, H.L.; Liu, L.L.; Chen, Y.X.; Ma, H.; Li, M.; Qu, W.S.; Yin, J.; Zhang, X.;
Gao, Y.T.; Shan, J.J.; Gao, Y.Q. The chemical character of polysaccha-
rides from processed Morindae officinalis and their effects on anti-
liver damage. Int. J. Biol. Macromol. 2019, 141, 410-421.
Zhai, H.J.; Ye, G.H.; Xue, J.J.; Yu, J.H.; Zhang, Y.Y.; Zhang, H. Two new
naphthoate derivatives from Morinda officinalis. J. Asian Nat. Prod.
Res. 2019.
Zhai, H.J.; Yu, J.H.; Zhang, Q.Q.; Liu, H.S.; Zhang, J.S.; Song, X.Q.; Zhang,
Y.Y.; Zhang, H. Cytotoxic and antibacterial triterpenoids from the roots
of Morinda officinalis var. officinalis. Fitoterapia. 2019, 133, 56-61.
Jiang, K.M.; Huang, D.; Zhang, D.W.; Wang, X.L.; Cao, H.J.; Zhang, Q.;
Yan, C.Y. Investigation of inulins from the roots of Morinda officinalis
for potential therapeutic application as anti-osteoporosis agent. Int. J.
Biol. Macromol. 2018, 120, 170-179.
1
718-1722.
Esakkimuthu, S.; Nagulkumar, S.; Darvin, S.S.; Buvanesvaragurunathan,
K.; Sathya, T.N.; Navaneethakrishnan, K.R.; Kumaravel, T.S.; Murugan,
S.S.; Shirota, O.; Balakrishna, K.; Pandikumar, P.; Ignacimuthu, S. Anti-
hyperlipidemic effect of iridoid glycoside deacetylasperulosidic acid
isolated from the seeds of Spermacoce hispida L. - A traditional anti-
obesity herb. J. Ethnopharmacol. 2019, 245.
Francois, B.; Pierre, D.; Michel, K. Apodantheroside, an iridoid gluco-
side from Feretia apodanthera. Phytochemistry. 1980, 19, 2763-2764.
Kohei, K.; Yasuhiro, F.; Yasuhisa, S.; Endang, H.; Umar, M.; Toshiko, S.
Studies on the constituents of Indonesian Borreria latifolia. Heterocy-
cles. 2002, 56, 537-544.
Takeda, Y.; Nishimura, H.; Inouye, H. Two new iridoid glucosides from
Ixora chinensis. Phytochemistry. 1975, 14, 2647-2650.
Cuong, N.M.; Huong, T.T.; Son, N.T.; Cuong, T.D.; Van, D.T.; Khanh, P.N.;
Ha, V.T.; Tram, N.C.; Long, P.Q.; Kim, Y.H. Morinlongosides A-C, Two
New Naphthalene Glycoside and a New Iridoid Glycoside from the
Roots of Morinda longissima. Chem. Pharm. Bull. (Tokyo). 2016, 64,
1
230-1234.
Xie, G.Y.; Qin, X.Y.; Chen, Y.J.; Wen, R.; Wu, S.S.; Qin, M.J. Alkaloids
from the Rhizomes of Iris germanica. Chem. Nat. Compd. 2017, 53,
1
96-198.
Lu, H.L.; Ouyang, F.; Su, Y.L.; Ji, T.F.; Liu, H. Study on chemical constit-
uents of the fruits of Nitraria tangutorum Bobr. Northwest Pharm. J.
2
013, 28 221-223.
Tatsis, E.C.; Schaumlöffel, A.; Warskulat, A.C.; Massiot, G.; Schneider,
B.; Bringmann, G. Nudicaulins, Yellow Flower Pigments of Papaver nu-
dicaule: Revised Constitution and Assignment of Absolute Configura-
tion. Org. Lett. 2013, 15, 156-159.
Zhao, W.; Song, X.; Zhao, L.; Zou, C.; Zhou, W.; Lin, B.; Yao, G.; Huang,
X.; Song, S. Quassinoids from Picrasma quassioides and their neuro-
protective effects. J. Nat. Prod. 2019, 82, 714-723.
Yan, C.Y.; Huang, D.; Shen, X.; Qin, N.B.; Jiang, K.M.; Zhang, D.W.;
Zhang, Q. Identification and characterization of a polysaccharide from
the roots of Morinda officinalis, as an inducer of bone formation by
up-regulation of target gene expression. Int. J. Biol. Macromol. 2019,
1
33, 446-456.
Huo, X.K.; Liu, J.; Yu, Z.L.; Wang, Y.F.; Wang, C.; Tian, X.G.; Ning, J.; Feng,
L.; Sun, C.P.; Zhang, B.J.; Ma, X.C. Alisma orientale extract exerts the
Sun, C.P.; Yan, J.K.; Yi, J.; Zhang, X.Y.; Yu, Z.L.; Huo, X.K.; Liang, J.H.; Ning,
J.; Feng, L.; Wang, C.; Zhang, B.J.; Tian, X.G.; Zhang, L.; Ma, X. The study
www.cjc.wiley-vch.de
© 2021 SIOC, CAS, Shanghai, & WILEY-VCH GmbH
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New iridoids as FXR agonists from Morinda officinalis
Chin. J. Chem.
of inhibitory effect of natural flavonoids toward beta-glucuronidase
and interaction of flavonoids with beta-glucuronidase. Int. J. Biol. Mac-
romol. 2020, 143, 349-358.
Yun, W.J.; Zhang, X.Y.; Liu, T.T.; Liang, J.H.; Sun, C.P.; Yan, J.K.; Huo, X.K.;
Tian, X.G.; Zhang, B.J.; Huang, H.L.; Ma, X.C. The inhibition effect of
uncarialin A on voltage-dependent L-type calcium channel subunit al-
pha-1C: Inhibition potential and molecular stimulation. Int. J. Biol.
Macromol. 2020, 159, 1022-1030.
(The following will be filled in by the editorial staff)
Manuscript received: XXXX, 2021
Manuscript revised: XXXX, 2021
Manuscript accepted: XXXX, 2021
Accepted manuscript online: XXXX, 2021
Version of record online: XXXX, 2021
Chin. J. Chem. 2021, 39, XXX－XXX
© 2021 SIOC, CAS, Shanghai, & WILEY-VCH GmbH
www.cjc.wiley-vch.de

Running title
Chin. J. Chem.
Entry for the Table of Contents
Discovery of new iridoids as FXR agonists from Morinda officinalis: Agonistic potentials and molecular stimulations
+
+
+
Zhi-Lin Luan, Fei Qiao, Wen-Yu Zhao, Wen-Hua Ming, Zhen-Long Yu, Jie Liu, Sheng-Yun Dai, Shuang-Hui Jiang, Chao-Jie Lian, Cheng-Peng Sun,* Bao-
Jing Zhang, Jian Zheng,* Shuang-Cheng Ma,* and Xiao-Chi Ma
Chin. J. Chem. 2021, 39, XXX—XXX. DOI: 10.1002/cjoc.202100XXX
New iridoids from Traditional Chinese Medicine M. officinalis showed significant agonistic activity against FXR at both gene and protein levels.
a
Dalian Key Laboratory of Metabolic Target Characterization and
Traditional Chinese Medicine Intervention, College of Pharmacy,
College of Integrative Medicine, Dalian Medical University, Dalian,
China.
b
National Institutes for Food and Drug Control, Beijing, China
Chin. J. Chem. 2018, template© 2018 SIOC, CAS, Shanghai,
&
WILEY-VCH Verlag GmbH
&
Co. KGaA,
Weinheim