New ursolic acid derivatives bearing 1,2,3-triazole moieties: design, synthesis and anti-inflammatory activity in vitro and in vivo

Article abstract of DOI:10.1007/s11030-021-10236-0

Abstract: In order to discover novel anti-inflammatory agents, three series of compounds obtained by appending 1,2,3-triazole moieties on ursolic acid were designed and synthesized. All compounds have been screened for their anti-inflammatory activity by using an ear edema model. The potent anti-inflammatory compound was subjected to in vitro cyclooxygenase COX-1/COX-2 inhibition assays. In general, the derivatives were found to be potent anti-inflammatory activity. Especially, the compound 11b exhibited the strongest activity of all of the compounds prepared, with 82.81% inhibition after intraperitoneal administration, which was better than celecoxib as a positive control. Molecular docking results unclose the rationale for the interaction of the compound 11b with COX-2 enzyme. Further studies revealed that compound 11b exhibited effective COX-2 inhibitory activity, with half-maximal inhibitor concentration (IC₅₀) value of 1.16?μM and selectivity index (SI = 64.66) value close to that of celecoxib (IC₅₀ = 0.93?μM, SI = 65.47). Taken together, these results could suggest a promising chemotype for development of new COX-2-targeting anti-inflammatory agent. Graphic abstract: [Figure not available: see fulltext.]

Full text of DOI:10.1007/s11030-021-10236-0

Molecular Diversity
https://doi.org/10.1007/s11030-021-10236-0
ORIGINAL ARTICLE
New ursolic acid derivatives bearing 1,2,3‑triazole moieties: design,
synthesis and anti‑infammatory activity in vitro and in vivo
Tian‑Yi Zhang1 · Chun‑Shi Li3 · Li‑Ting Cao1 · Xue‑Qian Bai1 · Dong‑Hai Zhao1 · Si‑Mei Sun2
Received: 19 March 2021 / Accepted: 22 May 2021
© The Author(s), under exclusive licence to Springer Nature Switzerland AG 2021
Abstract
In order to discover novel anti-infammatory agents, three series of compounds obtained by appending 1,2,3-triazole moieties
on ursolic acid were designed and synthesized. All compounds have been screened for their anti-infammatory activity by
using an ear edema model. The potent anti-infammatory compound was subjected to in vitro cyclooxygenase COX-1/COX-2
inhibition assays. In general, the derivatives were found to be potent anti-infammatory activity. Especially, the compound 11b
exhibited the strongest activity of all of the compounds prepared, with 82.81% inhibition after intraperitoneal administration,
which was better than celecoxib as a positive control. Molecular docking results unclose the rationale for the interaction of
the compound 11b with COX-2 enzyme. Further studies revealed that compound 11b exhibited efective COX-2 inhibitory
activity, with half-maximal inhibitor concentration (IC50) value of 1.16 µM and selectivity index (SI =64.66) value close
to that of celecoxib (IC50 =0.93 µM, SI=65.47). Taken together, these results could suggest a promising chemotype for
development of new COX-2-targeting anti-infammatory agent.
Graphic abstract
Keywords Ursolic acid · Anti-infammatory activity · Molecular docking · COX-1/COX-2
Tian-Yi Zhang and Chun-Shi Li have contributed equally to this
work.
2
* Dong-Hai Zhao
Zhoushan Hospital, Zhejiang University School
of Medicine, Zhoushan 316021, Zhejiang Province,
People’s Republic of China
donghaizhao2021@126.com
* Si-Mei Sun
3
sunsimei1024@163.com
The Third People’s Hospital of Dalian, Dalian 116000,
People’s Republic of China
1
Jilin Medical University, Jilin 132013, Jilin Province,
People’s Republic of China
Vol.:(0123456789)
1 3

Molecular Diversity
herbs. UA and their derivatives (compounds A-C) (Fig. 1)
have been investigated in detail by several groups, and many
interesting biological properties have been detected, among
them antitumor [8], antimicrobial [9] and anti-infammatory
[10]. Triazole-containing compounds (D-F) (Fig. 1) are able
to bind to a variety of enzymes and receptors by non-cova-
lent interactions, hence displaying a large array of biologi-
cal activities [11]. Among these triazole, 1,2,3-triazoles are
imperative class of heterocycles and play a major role in
medicinal chemistry with anticancer [12], anti-infammation
[13, 14] and antiobesity [15]. We hypothesized that several
natural sources derivatives (compounds G-I) (Fig. 1) have
benefcial efect on the anticancer [16, 17] and anti-infam-
mation efects [9, 18]. In continuity to our previous studies
and keeping in mind the medicinal potential of ursolic acid
and 1,2,3-triazoles fragments, it was planned to hybridize
these two fragments along with the incorporation of another
potent UA-based anti-infammation agents. Thence, three
novel series of UA derivatives (Fig. 2) were synthesized and
evaluated through biological experiments for their selective
target of COX-2 and for the anti-infammation activity.
Introduction
Infammation is a fundamental protective response of the
immune system to multiple injuries and diferent pathogens,
but the uncontrolled infammation is responsible for serious
and chronic problems [1, 2]. Nonsteroidal anti-infamma-
tory drugs demonstrated useful inhibition properties against
α-glycosidase, AR and SDH enzymes [3]. NSAIDs as indo-
methacin and ibuprofen work by inhibiting cyclooxygenase
(COX) enzymes, which catalyze prostaglandin biosynthesis
through arachidonic acid. Cyclooxygenase enzymes (COXs)
exist in two isoforms: constitutive form (COX-1) and induc-
ible form (COX-2) [4, 5]. Representing this category of
drugs celecoxib, rofecoxib and etoricoxib are/were some of
the highly efective COX-2 inhibitors [6, 7]. However, the
cardiovascular efects led to the withdrawal of rofecoxib and
etoricoxib from the market. Thus, new anti-infammatory
agent with high therapeutic efcacy and fewer side efects
are needed.
Ursolic acid (UA), a pentacyclic triterpene, exists abun-
dantly in the plant kingdom and as constituents of medicinal
(B)
(C)
(A)
(F)
(D)
(E)
(I)
(H)
(G)
Fig. 1 Chemical structures of reported triterpenes and 1,2,3-triazoles derivatives
1 3

Molecular Diversity
Fig. 2 Structures of target compounds 5a-d, 8a-i and 11a-h
was higher than that of celecoxib. The position or phys-
icochemical properties of the diferent substituents on the
phenyl ring had minimal impact on the anti-infammatory
properties of these compounds, indicating that the electronic
efect of the substituent on the benzene ring is not critical.
Furthermore, a comparison of the compounds in series 5 and
8 revealed that the introduction of a ketone moiety reduced
the anti-infammatory activity. To confrm the need for fur-
ther evaluation, we estimated the cytotoxicity of selected
compounds using the MTT assay on SGC-7901, BEL-7402
and LO2 cells. As shown in Table 2, compounds 8d, 8e, 11b
and 11e exhibited weak cytotoxicity.
Results and discussion
Chemistry
The synthetic route of all compounds is depicted in
Scheme 1. UA was treated with 1,3-dibromopropane in DMF
for 24 h to obtain intermediate 1, which was introduced with
sodium azide in DMF to obtain intermediate 2. The inter-
mediate 4 was synthesized by reacting propiolic acid with
substituted aniline in the presence of DCC and DMAP in
dry dichloromethane. Intermediate 7 was prepared by the
reaction of their respective aniline or phenol with propargyl
bromides in the presence of anhydrous potassium carbonate
in acetone for 12 h [19]. Intermediate 10 was synthesized
in the same way as intermediate 4 or intermediate 7. In the
fnal step, the compounds (4, 7 or 10) were further reacted
with intermediate 2 via copper-catalyzed azidealkyne (3+2)
cycloaddition reaction aforded desired products (5a-d, 8a-i
and 11a-h) [20]. All the synthesized compounds were thor-
oughly characterized by their spectral data.
Compound 11b was further assessment of its anti-infam-
matory activity. A dose of 100 mg/kg was administered via
the oral route at the time intervals of 1, 2, 3, 4, 5 and 24 h
after xylene application, and the inhibition rates were found
(Table 3). The activity of compound 11b (54.70%) was
found to the maximum after 2 h and was slightly higher to
that of celecoxib (51.01%) at this time point. Compound 11b
was also screened at concentrations of 100, 50 and 25 mg/
kg after 2-h oral administration. As shown in Table 4, com-
pound 11b exhibited a signifcant inhibitory efect toward
ear infammation inhibition (58.83%) at a dose of 100 mg/
kg, which was almost similar to that of celecoxib (58.77%).
Anti‑infammatory activity
Twenty-one of the UA derivatives and the reference drug
celecoxib were examined for their anti-infammatory activ-
ity using an in vivo xylene-induced ear edema model [21].
Most of the synthesized compounds exhibited potent anti-
infammatory activity at a dose of 100 mg/kg with inhibi-
tion rates ranging from 22.50% to 82.81% (Table 1). Six
compounds, namely 8d, 8e, 8 h, 8i, 11d and 11 g, showed
anti-infammatory activity with more than 65.00% inhibi-
tion, which was nearly equivalent to that of the inhibition
shown by celecoxib (65.00%). It is noteworthy that com-
pound 11b, with 82.81% inhibition, exhibited the strongest
anti-infammatory activity. Moreover, the activity of 11b
In vitro cyclooxygenase (COX‑1/2) inhibition assay
The ability of compound 11b to inhibit COX-1 and COX-2
enzymes was determined via measuring their peroxidase
activity using a colorimetric enzyme immune assay (EIA)
kit [22–24]. Furthermore, the COX-2 selectivity index
(COX-2 SI) value was calculated as [IC50 (COX-1)/IC50
(COX-2)] and compared to the reference drug (celecoxib as
a selective COX-2 inhibitor) [25]. Compound 11b displayed
a potency value (IC50 = 1.16 µM) close to the value for
1 3

Molecular Diversity
Scheme 1 Synthetic scheme for target compounds. Reagents and
conditions: a Br(CH2)3Br, K2CO3, KI, DMF, rt, 12 h; b NaN3, DMF,
rt, 2 h; c propiolic acid, DCC, DMAP, rt, 12 h; d propargyl bromide,
K2CO3, KI, acetone, refux, 6 h; e propargyl amine, DCC, DMAP, rt,
12 h; f sodium ascorbate, CuSO4·5H2O, DMF, H2O, rt, 12 h
celecoxib (IC50 =0.93 µM). Considering COX-2 selectivity
index values, compound 11b showed selectivity index value
(SI=64.66) close to that of celecoxib (SI=65.47) (Table 5).
Conclusions
Three structurally different series of compounds con-
sisting of ursolic acid and 1,2,3-triazole hybrids were
screened for the anti-infammatory activity. The results
clearly provided confdence for further development of
new analogues based on this chemical scafold. In pre-
liminary tests, compound 11b showed high edema inhibi-
tion percentage (82.81%) compared to celecoxib (65.00%).
Also, compound 11b was found to have potent COX-2
inhibitory activity (IC50 =1.16 µM, SI=64.66) close to the
COX-2 selective reference drug celecoxib (IC50 =0.93 µM,
SI = 65.47). Moreover, molecular modeling studies also
indicated toward the most probable binding site inter-
actions of compound 11b in the active site. In view of
the above, it may be concluded that compound 11b is an
efcient lead compound to development of better COX-2
inhibitors and anti-infammatory agents.
Molecular docking
To validate the data obtained by the in vitro COX-2 assay,
molecular docking studies of 11b were performed in com-
parison with celecoxib using COX-2 protein (PDB entry
4COX). AutoDock Vina was used to execute molecular
docking. Discovery Studio was used for interaction analy-
sis, binding conformation and making diagrams. The result
shows that compound 11b may bind well to the COX-2 pro-
tein and while interacting with Arg44 (5.44 Å and 2.57 Å),
Tyr122 (5.03 Å and 4.39 Å), and Gln372 (2.01 Å). It is
apparent that compound 11b (− 8.1 kcal/Mol) may bind
to 4COX in a way similar to the binding of celecoxib
(− 8.0 kcal/Mol) (Fig. 3). These molecular docking data
provide certain theoretical support for experimental results
and the subsequent structural optimization eforts.
1 3

Molecular Diversity
Table 1 Anti-infammatory activities of compounds 5a-d, 8a-i, 11a-h
and celecoxib following i.p. administration
Table 3 Anti-infammatory activity of compound 11b administered
orally at diferent times before xylene application
Dose (mg/kg) Number Edema mean S.D. Inhibition
Time (h)
Dose (mg/kg)
Number of Inhibition rate (%)
of mice (mg)
rate (%)
mice
11b
Celecoxib
DMSO
100
5
5
5
5
5
5
5
5
5
5
5
5
5
5
5
5
5
5
5
5
5
5
5
5
8.00 0.90
–
1
2
3
4
5
24
100
100
100
100
100
100
5
5
5
5
5
5
40.97**
54.70***
47.29***
31.27*
24.10*
3.08
30.62*
51.01***
43.39**
17.05
11.79
4.17
Celecoxib 100
2.80 0.65**
4.80 0.58**
6.20 0.60**
5.83 0.35
6.90 0.80*
4.95 1.15
65.00
55.34
22.50
27.19
13.75
38.13
61.25
33.75
56.56
65.00
68.75
52.50
57.50
65.63
66.56
55.31
82.81
49.69
70.63
60.00
25.63
65.63
64.06
UA
5a
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
5b
5c
5d
*0.01<p<0.05 compared with vehicle group, **p<0.01, ***
p<0.001 compared with a vehicle group
8a
3.10 1.00
5.30 1.65
8b
8c
3.48 1.15**
2.80 0.45**
2.50 0.70***
3.80 0.80*
3.40 0.45**
2.75 0.45**
2.68 0.65***
4.17 0.70**
1.38 0.65***
4.03 0.25*
2.35 0.85***
3.20 0.59*
5.95 0.45
8d
Table 4 Anti-infammatory activity of compound 11b administered
orally at diferent doses
8e
8f
Time (h)
Dose (mg/kg)
Number of Inhibition rate (%)
8 g
8 h
8i
mice
11b
Celecoxib
2
2
2
100
50
25
5
5
5
58.83***
30.88*
16.81*
58.77***
30.74*
17.11
11a
11b
11c
11d
11e
11f
11 g
11 h
*p<0.05, *** p<0.001 compared with a vehicle group
Table 5 In vitro COX-1/COX-2 enzyme inhibition assaysa
2.75 0.65**
2.88 1.00**
Cmpd
COX-1, IC50
(μM)a
COX-2, IC50
(μM)a
COX-2, SIb
*0.01<p<0.05 compared with vehicle group, **p<0.01, ***
p<0.001 compared with a vehicle group
–No anti-infammatory activity
11b
Celecoxib
75.01
60.89
1.16
0.93
64.66
65.47
aData are shown as IC50 (μM)
bThe in vitro COX-2 selectivity index (COX-1/COX-2)
Table 2 Cytotoxicity data (IC50, μmol/L) of compounds 8d, 8e, 11b
and 11e in SGC-7901, BEL-7402 and LO2 cells
Compound
IC50 (μmol/L)a
SGC-7901b
BEL7402c
LO2d
were uncorrected. The reactions were monitored by thin
1
layer chromatography (TLC). H and 13C-NMR spectra
8d
8e
11b
11e
>100
>100
>100
>100
>100
>100
>100
>100
>100
>100
>100
>100
were performed on an AV-300 or AV-400 spectrometer
(Bruker, Zurich, Switzerland) operating at 300 MHz for
1H and 101 MHz for 13C NMR while using TMS as the
internal standard. J values are given in Hertz. Mass spectra
were measured on an MALDI-TOF (Shimadzu, Japan).
High-resolution mass spectra were measured on a Thermo
Scientifc LTQ Orbitrap XL spectrometer.
aValues are the average of three independent experiments running in
b
triplicate. Variation was generally between 5 and 10%; human gas-
c
d
tric cancer cells; human liver cancer cells; human normal hepatic
cells
General procedure for the preparation of compound
1
Materials and methods
UA (1 mmol) was dissolved in DMF (15 mL), and K2CO3
(2 mmol), KI (0.5 mmol) and dibromoalkane (4 mmol)
were added. After being stirred for another 12 h, the reac-
tion mixture was poured onto 100 mL of distilled water
General procedures
Melting points were determined in open capillaries and
1 3

Molecular Diversity
Fig. 3 Binding modes of com-
pounds 11b and celecoxib with
4COX. a Compound 11b in
binding site of 4COX. b Dock-
ing study of compound 11b
with 4COX. c Docking study
of compound 11b with 4COX.
d Docked conformation of the
most active compound 11b in
COX-2. e Celecoxib in binding
site of 4COX. f Docking study
of celecoxib with 4COX. g
Docking study of celecoxib
with 4COX. h Docked con-
formation of the most active
celecoxib in COX-2
and partitioned with ethyl acetate. The organic layer was
washed with brine (3 × 10 mL), dried over anhydrous
Na2SO4 and purifed via silica gel column chromatogra-
phy with petroleum ether/ethyl acetate to yield the target
compound 1.
General procedure for the preparation of compound
2
Compound 1 (1 mmol) and sodium azide (2 mmol) were
weighed out and transferred to a vial to which DMF/H2O
1 3

Molecular Diversity
(1:1, 50 mL) was subsequently added. The reaction was
stirred at 80 °C for 24 h and then cooled to room tem-
perature. The reaction mixture was poured into water and
extracted with ethyl acetate. The combined organic layers
were dried with Na2SO4, fltered. After the removal of the
solvent under vacuum, compound 2 was obtained.
60.44, 55.24, 52.95, 48.42, 47.36, 42.26, 39.58, 39.07,
38.95, 37.07, 33.04, 31.90, 30.62, 29.50, 28.03, 27.24,
24.30, 23.52, 23.32, 22.70, 21.14, 20.92, 18.28, 17.15,
17.00, 15.54, 14.12. MS (MALDI-TOF) m/z 699 [M+H]+.
3-(4-((2-Fluorophenyl)carbamoyl)-1H-1,2,3-triazol-
1-yl)propyl(1S,2R,4aS,6aS,6bR,10S,12aR,12bR,14bS)-
10-hydroxy-1,2,6a,6b,9,9,12a-heptamethyl-1,3,4,5,6,6a,6
b,7,8,8a,9,10,11,12,12a,12b,13,14b-octadecahydropicene-
4a(2H)-carboxylate (5c)White solid. Yield: 68%; m.p.
General procedure for the preparation
of compounds 5a‑d, 8a‑i and 11a‑h
1
167–169 °C. H NMR (300 MHz, CDCl3) δ 9.20 (s, 1H,
Compound 2 (0.15 mmol) and the appropriate azide
NH), 8.45 (s, 1H, CH), 8.20 (s, 1H, Ar–H), 7.23–7.07 (m,
3H, Ar–H), 5.28 (s, 1H, CH), 4.54 (s, 2H, CH2), 4.10 (s, 2H,
CH2), 3.22 (s, 1H, CH), 2.32 (s, 2H, CH2), 2.21–0.73 (m,
44H, protons in UA skeleton). 13C NMR (101 MHz, CDCl3)
δ 125.97, 125.71, 124.85, 121.64, 115.26, 79.04, 60.44,
55.24, 53.00, 48.35, 42.09, 39.69, 39.18, 36.86, 32.97,
30.71, 29.66, 29.45, 28.09, 27.34, 24.27, 23.64, 21.08,
18.31, 17.29, 15.69. MS (MALDI-TOF) m/z 703 [M+H]+.
3-(4-(Phenylcarbamoyl)-1H-1,2,3-triazol-1-yl)propyl
(1S,2R,4aS,6aS,6bR,10S,12aR,12bR,14bS)-10-hydroxy-
1,2,6a,6b,9,9,12a-heptamethyl-1,3,4,5,6,6a,6b,7,8,8a,9,10
,11,12,12a,12b,13,14b-octadecahydropicene-4a(2H)-car-
.
(0.30 mmol) in CH2Cl2 (5 mL), H2O (5 mL), CuSO4 5H2O
(0.25 mmol) and Na-L-ascorbate (0.5 mmol) were added.
The reaction was stirred at room temperature overnight
where upon a saturated sodium chloride solution was added
and the mixture was extracted with dichloromethane (3×).
The combined organic extracts wsere washed with brine
(50 mL), dried over Na2SO4, fltered, and the solvent was
removed in a rotary evaporator under reduced pressure to
aford the crude product. The residue was purifed by fash
column chromatography to aford the target compounds 5a-
d, 8a-i and 11a-h each as a white solid.
1
3-(4-((3,4-Dimethylphenyl)carbamoyl)-1H-1,2,3-triazol-
1-yl)propyl(1S,2R,4aS,6aS,6bR,10S,12aR,12bR,14bS)-
10-hydroxy-1,2,6a,6b,9,9,12a-heptamethyl-1,3,4,5,6,6a,6
b,7,8,8a,9,10,11,12,12a,12b,13,14b-octadecahydropi-
cene-4a(2H)-carboxylate(5a) White solid. Yield: 75%;
boxylate (5d)White solid. Yield: 72%; m.p. 89–91 °C. H
NMR (300 MHz, CDCl3) δ 9.00 (s, 1H, NH), 8.21 (s, 1H,
CH), 7.72 (d, J = 8.1 Hz, 2H, Ar–H), 7.39 (d, J = 7.8 Hz,
2H, Ar–H), 7.17 (d, J = 7.4 Hz, 1H, Ar–H), 5.27 (s, 1H,
CH), 4.52 (d, J=5.7 Hz, 2H, CH2), 4.21–3.98 (m, 2H, CH2),
3.27–3.17 (m, 1H, CH), 2.36–2.26 (m, 2H, CH2), 2.20–0.73
(m, 44H, protons in UA skeleton). MS (MALDI-TOF) m/z
685 [M+H]+.
1
m.p. 179–181 °C. H NMR (300 MHz, CDCl3) δ 8.88 (s,
1H, NH), 8.17 (s, 1H, CH), 7.49 (s, 2H, Ar–H), 7.15 (d,
J=8.2 Hz, 1H, Ar–H), 5.28 (s, 1H, CH), 4.53 (s, 2H, CH2),
4.33 (t, J = 6.7 Hz, 1H, CH), 4.18–3.99 (m, 2H, CH2),
3.27–3.17 (m, 1H, CH), 2.29 (d, J=10.5 Hz, 8H, CH and
CH2), 2.19–0.74 (m, 43H, protons in UA skeleton and CH3).
13C NMR (101 MHz, CDCl3) δ 176.23, 137.28, 136.31,
131.93, 129.07, 124.68, 120.18, 116.38, 78.00, 59.39,
54.21, 51.95, 47.26, 46.85, 46.51, 41.11, 38.55, 38.07,
37.85, 37.73, 37.61, 35.97, 35.79, 32.01, 30.92, 29.58,
27.12, 26.98, 26.21, 23.28, 22.54, 22.30, 20.12, 18.91,
18.19, 17.29, 16.16, 15.97, 14.58, 14.43, 13.09. HRMS
(ESI) m/z calcd for C44H64N4NaO4 [M + Na]+: 735.4830,
found: 735.4830.
3-(4-((3-Methoxyphenoxy)methyl)-1H-1,2,3-triazol-
1-yl)propyl(1S,2R,4aS,6aS,6bR,10S,12aR,12bR,14bS)-
10-hydroxy-1,2,6a,6b,9,9,12a-heptamethyl-1,3,4,5,6,6a,6
b,7,8,8a,9,10,11,12,12a,12b,13,14b-octadecahydropicene-
4a(2H)-carboxylate (8a)White solid. Yield: 80%; m.p.
1
125–127 °C. H NMR (300 MHz, CDCl3) δ 7.76 (s, 1H,
CH), 7.21 (d, J = 8.4 Hz, 1H, Ar–H), 6.59 (d, J = 7.7 Hz,
3H, Ar–H), 5.26 (d, J= 13.6 Hz, 3H, CH and CH2), 4.47
(s, 2H, CH2), 4.20–3.96 (m, 2H, CH2), 3.81 (s, 3H, OCH3),
3.28–3.17 (m, 1H, CH), 2.26 (s, 2H, CH2), 1.92–0.75 (m,
44H, protons in UA skeleton). 13C NMR (101 MHz, CDCl3)
δ 130.14, 125.88, 123.03, 107.08, 101.45, 78.93, 60.73,
55.44, 53.07, 48.22, 47.64, 42.09, 39.52, 39.16, 37.07,
33.20, 31.01, 29.82, 28.25, 27.28, 24.40, 23.44, 21.25,
18.35, 17.35, 15.40. MS (MALDI-TOF) m/z 702 [M+H]+.
3-(4-((2,4-Dimethylphenoxy)methyl)-1H-1,2,3-triazol-
1-yl)propyl(1S,2R,4aS,6aS,6bR,10S,12aR,12bR,14bS)-
10-hydroxy-1,2,6a,6b,9,9,12a-heptamethyl-1,3,4,5,6,6a,6
b,7,8,8a,9,10,11,12,12a,12b,13,14b-octadecahydropicene-
4a(2H)-carboxylate (8b)White solid. Yield: 75%; m.p.
62–64 °C. 1H NMR (300 MHz, CDCl3) δ 7.62 (s, 1H, CH),
6.98 (s, 2H, Ar–H), 6.87 (s, 1H, Ar–H), 5.25 (d, J=15 Hz,
3H, CH and CH2), 4.46 (s, 2H, CH2), 4.08 (d, J = 5.7 Hz,
3-(4-(p-Tolylcarbamoyl)-1H-1,2,3-triazol-1-yl)propyl
(1S,2R,4aS,6aS,6bR,10S,12aR,12bR,14bS)-10-hydroxy-
1,2,6a,6b,9,9,12a-heptamethyl-1,3,4,5,6,6a,6b,7,8,8a,9,10
,11,12,12a,12b,13,14b-octadecahydropicene-4a(2H)-car-
1
boxylate (5b)White solid. Yield: 70%; m.p. 95–97 °C. H
NMR (300 MHz, CDCl3) δ 8.91 (s, 1H, NH), 8.18 (s, 1H,
CH), 7.60 (d, J = 8.3 Hz, 2H, Ar–H), 7.20 (d, J = 8.2 Hz,
2H, Ar–H), 5.28 (s, 1H, CH), 4.53 (s, 2H, CH2), 4.33 (t,
J=6.7 Hz, 1H, CH), 4.10 (s, 2H, CH2), 3.27–3.18 (m, 1H,
CH), 2.37 (s, 3H, CH3), 2.31 (d, J = 6.5 Hz, 2H, CH2),
2.22–0.74 (m, 43H, protons in UA skeleton). 13C NMR
(101 MHz, CDCl3) δ 134.11, 129.56, 125.90, 119.91, 78.99,
1 3

Molecular Diversity
2H, CH2), 3.28–3.16 (m, 1H, CH), 2.25 (m, 9H, CH, CH2
and CH3), 1.96–0.75 (m, 43H, protons in UA skeleton). 13
2H, Ar–H), 6.81 (s, 3H, Ar–H), 5.28 (s, 1H, CH), 4.51 (s,
2H, CH2), 4.41 (s, 2H, CH2), 4.33 (t, J=6.7 Hz, 1H, CH),
4.15–3.93 (m, 2H, CH2), 3.23 (d, J = 4.5 Hz, 1H, CH),
2.25–0.74 (m, 45H, protons in UA skeleton). MS (MALDI-
TOF) m/z 671 [M+H]+.
C
NMR (101 MHz, CDCl3) δ 177.31, 154.32, 138.30, 131.65,
130.24, 127.07, 126.69, 125.66, 111.63, 78.98, 62.63, 60.62,
55.21, 52.96, 48.26, 47.43, 42.13, 39.56, 39.09, 38.89,
38.75, 38.61, 36.90, 33.05, 30.63, 29.71, 29.49, 28.07,
27.23, 24.28, 23.56, 23.32, 21.17, 20.48, 18.29, 17.21,
17.02, 16.25, 15.63, 15.44. HRMS (ESI) m/z calcd for
C44H66N3O4 [M+H]+ 700.50478, found 700.50458.
3-(4-((3-(Trifuoromethyl)phenoxy)methyl)-1H-1,2,3-
triazol-1-yl)propyl(1S,2R,4aS,6aS,6bR,10S,12aR,12b
R,14bS)-10-hydroxy-1,2,6a,6b,9,9,12a-heptamethyl-
1,3,4,5,6,6a,6b,7,8,8a,9,10,11,12,12a,12b,13,14b-octadec-
ahydropicene-4a(2H)-carboxylate (8c)White solid. Yield:
75%; m.p. 151–153 °C. 1H NMR (300 MHz, CDCl3) δ 7.75
(s, 1H, CH), 7.43 (s, 1H, Ar–H), 7.25 (d, J= 8.6 Hz, 3H,
Ar–H), 5.30 (s, 3H, CH and CH2), 4.50 (s, 2H, CH2), 4.11
(s, 2H, CH2), 3.26 (s, 3H, CH and CH2), 2.25–0.75 (m,
44H, protons in UA skeleton). MS (MALDI-TOF) m/z 762
[M+Na]+.
3-(4-(((2-Fluorophenyl)amino)methyl)-1H-1,2,3-triazol-
1-yl)propyl(1S,2R,4aS,6aS,6bR,10S,12aR,12bR,14bS)-
10-hydroxy-1,2,6a,6b,9,9,12a-heptamethyl-1,3,4,5,6,6a,6
b,7,8,8a,9,10,11,12,12a,12b,13,14b-octadecahydropicene-
4a(2H)-carboxylate (8 g)White solid. Yield: 65%; m.p.
1
145–147 °C. H NMR (300 MHz, CDCl3) δ 7.78 (s, 1H,
CH), 7.37 (s, 2H, Ar–H), 7.18 (d, J = 9.4 Hz, 2H, Ar–H),
5.31 (s, 1H, CH), 4.53 (s, 2H, CH2), 4.07 (m, 2H, CH2), 3.73
(d, J=7.0 Hz, 1H, CH), 3.48 (s, 1H, CH), 3.25 (d, J=6.3 Hz,
1H, CH), 1.92–0.75 (m, 46H, protons in UA skeleton). 13
C
NMR (101 MHz, CDCl3) δ 177.31, 138.29, 125.63, 124.60,
121.72, 117.54, 114.66, 114.48, 112.60, 78.98, 60.58, 55.19,
52.94, 48.24, 47.49, 47.30, 42.11, 39.70, 39.54, 39.07,
38.86, 38.74, 38.59, 36.95, 36.80, 33.03, 30.60, 29.69,
29.48, 28.14, 27.96, 27.20, 24.25, 23.54, 23.30, 21.15,
18.28, 17.19, 17.00, 15.62, 15.42. HRMS (ESI) m/z calcd
for C42H62FN4O3 [M+H]+ 689.48005, found 689.48004.
3-(4-((p-Tolylamino)methyl)-1H-1,2,3-triazol-1-yl)prop
yl(1S,2R,4aS,6aS,6bR,10S,12aR,12bR,14bS)-10-hydroxy-
1,2,6a,6b,9,9,12a-heptamethyl-1,3,4,5,6,6a,6b,7,8,8a,9,10
,11,12,12a,12b,13,14b-octadecahydropicene-4a(2H)-car-
boxylate (8 h)White solid. Yield: 70%; m.p. 207–208 °C.
1H NMR (300 MHz, CDCl3) δ 7.97 (s, 1H, CH), 7.78 (s,
1H, NH), 7.15 (m, 4H, Ar–H), 5.28 (s, 1H, CH), 5.07 (s,
2H, CH2), 4.53 (s, 2H, CH2), 4.07 (m, 2H, CH2), 3.74 (d,
J = 7.0 Hz, 1H, CH), 3.49 (s, 1H, CH), 3.24 (s, 1H, CH),
2.38 (s, 3H, CH3), 1.93–0.75 (m, 44H, protons in UA skele-
ton). 13C NMR (101 MHz, CDCl3) δ 177.31, 146.01, 138.28,
129.83, 125.64, 121.89, 113.97, 78.98, 60.59, 55.19, 52.93,
48.24, 47.50, 47.26, 42.11, 40.66, 39.54, 39.07, 38.86,
38.74, 38.60, 36.96, 36.80, 33.03, 30.59, 29.68, 29.48,
28.14, 27.97, 27.21, 24.25, 23.54, 23.30, 21.15, 20.45,
18.29, 17.19, 17.00, 15.63, 15.45. MS (MALDI-TOF) m/z
684 [M+H]+.
3-(4-((2,4-Dichlorophenoxy)methyl)-1H-1,2,3-triazol-
1-yl)propyl(1S,2R,4aS,6aS,6bR,10S,12aR,12bR,14bS)-
10-hydroxy-1,2,6a,6b,9,9,12a-heptamethyl-1,3,4,5,6,6a,6
b,7,8,8a,9,10,11,12,12a,12b,13,14b-octadecahydropicene-
4a(2H)-carboxylate (8d)White solid. Yield: 70%; m.p.
59–61 °C. 1H NMR (300 MHz, CDCl3) δ 7.75 (d, J=6.1 Hz,
1H, CH), 7.39 (s, 1H, Ar–H), 7.21 (s, 1H, Ar–H), 7.10 (s,
1H, Ar–H), 5.31 (m, 3H, CH and CH2), 4.50 (s, 2H, CH2),
4.11 (s, 2H, CH2), 3.22 (s, 1H, CH), 2.81 (s, 2H, CH2),
2.25–0.74 (m, 44H, protons in UA skeleton). MS (MALDI-
TOF) m/z 762 [M+Na]+.
3-(4-(Phenoxymethyl)-1H-1,2,3-triazol-1-yl)propyl(
1S,2R,4aS,6aS,6bR,10S,12aR,12bR,14bS)-10-hydroxy-
1,2,6a,6b,9,9,12a-heptamethyl-1,3,4,5,6,6a,6b,7,8,8a,9,10
,11,12,12a,12b,13,14b-octadecahydropicene-4a(2H)-car-
boxylate (8e)White solid. Yield: 70%; m.p. 131–133 °C. 1H
NMR (300 MHz, CDCl3) δ 7.76 (s, 1H, CH), 7.37–7.29 (m,
2H, Ar–H), 7.06–6.96 (m, 3H, Ar–H), 5.30 (m, 3H, CH and
CH2), 4.50 (s, 2H, CH2), 4.08 (m, 2H, CH2), 3.53 (s, 2H,
CH2), 3.21 (s, 1H, CH), 2.37–0.76 (m, 44H, protons in UA
skeleton). 13C NMR (101 MHz, CDCl3) δ 177.33, 158.21,
138.32, 129.57, 125.65, 122.74, 121.30, 114.75, 78.99,
62.04, 60.61, 55.21, 52.96, 48.27, 47.44, 42.13, 39.56,
39.09, 38.88, 38.75, 38.61, 36.97, 36.83, 33.04, 30.62,
29.71, 29.50, 28.15, 27.99, 27.22, 24.27, 23.56, 23.32,
21.17, 18.30, 17.22, 17.02, 15.64, 15.45. MS (MALDI-TOF)
m/z 672 [M+H]+.
3-(4-(((3,4-Dimethylphenyl)amino)methyl)-1H-1,2,3-
triazol-1-yl)propyl(1S,2R,4aS,6aS,6bR,10S,12aR,12b
R,14bS)-10-hydroxy-1,2,6a,6b,9,9,12a-heptamethyl-
1,3,4,5,6,6a,6b,7,8,8a,9,10,11,12,12a,12b,13,14b-octadec-
ahydropicene-4a(2H)-carboxylate (8i).
1
White solid. Yield: 75%; m.p. 171–173 °C. H NMR
(300 MHz, CDCl3) δ 7.84–7.70 (m, 1H, CH), 7.56 (s, 1H,
Ar–H), 7.14 (s, 1H, Ar–H), 6.99 (s, 1H, Ar–H), 5.29 (s,
1H, CH), 5.00 (s, 1H, CH), 4.49 (s, 2H, CH2), 4.33 (s, 1H,
CH), 4.11 (m, 2H, CH2), 3.30–3.18 (m, 1H, CH), 2.28 (m,
9H, CH, CH2 and CH3), 1.93–0.78 (m, 43H, protons in UA
skeleton). 13C NMR (101 MHz, CDCl3) δ 177.31, 138.28,
137.46, 130.33, 125.64, 121.78, 115.46, 111.00, 79.00,
60.61, 55.20, 52.94, 48.24, 47.51, 47.25, 42.11, 40.55,
3-(4-((Phenylamino)methyl)-1H-1,2,3-triazol-1-yl)prop
yl(1S,2R,4aS,6aS,6bR,10S,12aR,12bR,14bS)-10-hydroxy-
1,2,6a,6b,9,9,12a-heptamethyl-1,3,4,5,6,6a,6b,7,8,8a,9,
10,11,12,12a,12b,13,14b-octadecahydropicene-4a(2H)-
carboxylate (8f)White solid. Yield: 68%; m.p. 102–104 °C.
1H NMR (300 MHz, CDCl3) δ 7.57 (s, 1H, CH), 7.23 (s,
1 3

Molecular Diversity
39.55, 39.07, 38.86, 38.74, 38.60, 36.97, 36.80, 33.03,
30.59, 29.68, 29.48, 28.14, 27.98, 27.21, 24.25, 23.54,
23.31, 21.15, 20.04, 18.75, 18.29, 17.20, 17.00, 15.62,
15.45. MS (MALDI-TOF) m/z 699 [M+H]+.
R,14bS)-10-hydroxy-1,2,6a,6b,9,9,12a-heptamethyl-
1,3,4,5,6,6a,6b,7,8,8a,9,10,11,12,12a,12b,13,14b-octa-
decahydropicene-4a(2H)-carboxylate (11e).
1
White solid. Yield: 68%; m.p. 61–63 °C. H NMR
3-(4-((Benzoyloxy)methyl)-1H-1,2,3-triazol-1-yl)propy
l(1S,2R,4aS,6aS,6bR,10S,12aR,12bR,14bS)-10-hydroxy-
1,2,6a,6b,9,9,12a-heptamethyl-1,3,4,5,6,6a,6b,7,8,8a,9,10
,11,12,12a,12b,13,14b-octadecahydropicene-4a(2H)-car-
boxylate (11a)White solid. Yield: 80%; m.p. 160–162 °C. 1H
NMR (300 MHz, CDCl3) δ 8.06 (d, J=7.1 Hz, 2H, Ar–H),
7.73 (s, 1H, CH), 7.56 (d, J=7.3 Hz, 1H, Ar–H), 7.44 (d,
J=7.5 Hz, 2H, Ar–H), 5.49 (s, 2H, CH2), 5.27 (s, 1H, CH),
4.46 (s, 2H, CH2), 4.07 (m, 2H, CH2), 3.21 (s, 1H, CH), 2.26
(d, J=6.7 Hz, 2H, CH2), 2.18–0.73 (m, 44H, protons in UA
skeleton). MS (MALDI-TOF) m/z 700 [M+H]+.
(300 MHz, CDCl3) δ 8.77 (d, J = 5.4 Hz, 1H, NH), 7.80
(s, 1H, CH), 7.67 (d, J = 1.6 Hz, 1H, Ar–H), 7.14 (t,
J = 8.0 Hz, 1H, Ar–H), 7.05 (d, J = 1.5 Hz, 1H, Ar–H),
5.25 (s, 1H, CH), 4.79 (d, J = 5.7 Hz, 2H, CH2), 4.44 (d,
J=4.6 Hz, 2H, CH2), 4.31 (s, 1H, CH), 4.10 (m, 2H, CH2),
3.90 (d, J = 9.0 Hz, 6H, OCH3), 3.21 (d, J = 4.6 Hz, 1H,
CH), 2.24 (m, 3H, CH and CH2), 2.34–2.25 (m, 2H, CH2),
2.04–0.72 (m, 42H, protons in UA skeleton). HRMS (ESI)
m/z calcd for C45H67N4O6 [M + H]+ 759.50551, found
759.50586.
3-(4-((4-Methylbenzamido)methyl)-1H-1,2,3-triazol-
1-yl)propyl(1S,2R,4aS,6aS,6bR,10S,12aR,12bR,14bS)-
10-hydroxy-1,2,6a,6b,9,9,12a-heptamethyl-1,3,4,5,6,6a,6
b,7,8,8a,9,10,11,12,12a,12b,13,14b-octadecahydropi-
cene-4a(2H)-carboxylate (11f)White solid. Yield: 76%;
3-(4-(((2,3-Dimethoxybenzoyl)oxy)methyl)-1H-1,2,3-
triazol-1-yl)propyl(1S,2R,4aS,6aS,6bR,10S,12aR,12b
R,14bS)-10-hydroxy-1,2,6a,6b,9,9,12a-heptamethyl-
1,3,4,5,6,6a,6b,7,8,8a,9,10,11,12,12a,12b,13,14b-octadec-
ahydropicene-4a(2H)-carboxylate (11b)White solid. Yield:
80%; m.p. 118–120 °C. 1H NMR (300 MHz, CDCl3) δ 7.82
(s, 1H, Ar–H), 7.38–7.32 (m, 1H, CH), 7.11–7.06 (m, 2H,
Ar–H), 5.52 (s, 2H, CH2), 5.27 (s, 1H, CH), 4.48 (s, 2H,
CH2), 4.07 (m, 2H, CH2), 3.88 (d, J=1.5 Hz, 6H, OCH3),
3.21 (s, 1H, CH), 3.00 (s, 1H, CH), 2.91 (s, 1H, CH), 2.24
(m, 3H, CH and CH2), 2.05–0.73 (m, 41H, protons in UA
skeleton). MS (MALDI-TOF) m/z 760 [M+H]+.
1
m.p. 112–114 °C. H NMR (300 MHz, CDCl3) δ 7.71
(d, J = 7.7 Hz, 2H, Ar–H), 7.66 (s, 1H, CH), 7.21 (d,
J=8.0 Hz, 2H, Ar–H), 5.28 (d, J=14.2 Hz, 1H, CH), 4.69
(d, J = 5.5 Hz, 2H, CH2), 4.41 (s, 2H, CH2), 4.05 (m, 2H,
CH2), 3.20 (s, 1H, CH), 2.38 (s, 3H, CH3), 2.17–0.72 (m,
46H, protons in UA skeleton). MS (MALDI-TOF) m/z 713
[M+H]+.
3-(4-((4-Methoxybenzamido)methyl)-1H-1,2,3-triazol-
1-yl)propyl(1S,2R,4aS,6aS,6bR,10S,12aR,12bR,14bS)-
10-hydroxy-1,2,6a,6b,9,9,12a-heptamethyl-1,3,4,5,6,6a,6
b,7,8,8a,9,10,11,12,12a,12b,13,14b-octadecahydropi-
cene-4a(2H)-carboxylate (11 g)White solid. Yield: 82%;
3-(4-(((4-Methoxybenzoyl)oxy)methyl)-1H-1,2,3-triazol-
1-yl)propyl(1S,2R,4aS,6aS,6bR,10S,12aR,12bR,14bS)-
10-hydroxy-1,2,6a,6b,9,9,12a-heptamethyl-1,3,4,5,6,6a,6
b,7,8,8a,9,10,11,12,12a,12b,13,14b-octadecahydropicene-
4a(2H)-carboxylate (11c)White solid. Yield: 65%; m.p.
63–65 °C. 1H NMR (300 MHz, CDCl3) δ 8.01 (d, J=8.9 Hz,
2H, Ar–H), 7.70 (s, 1H, CH), 6.91 (d, J=8.9 Hz, 2H, Ar–H),
5.45 (s, 2H, CH2), 5.27 (s, 1H, CH), 4.44 (d, J = 4.7 Hz,
2H, CH2), 4.07 (m, 2H, CH2), 3.86 (s, 3H, OCH3), 3.21
(s, 1H, CH), 2.25 (m, 3H, CH and CH2), 2.08–0.73 (m,
43H, protons in UA skeleton). HRMS (ESI) m/z calcd for
C44H64N3O6 [M+H]+ 730.47896, found 730.47919.
3-(4-(Benzamidomethyl)-1H-1,2,3-triazol-1-yl)propyl
(1S,2R,4aS,6aS,6bR,10S,12aR,12bR,14bS)-10-hydroxy-
1,2,6a,6b,9,9,12a-heptamethyl-1,3,4,5,6,6a,6b,7,8,8a,9,
10,11,12,12a,12b,13,14b-octadecahydropicene-4a(2H)-
carboxylate (11d)White solid. Yield: 72%; m.p. 95–97 °C.
1H NMR (300 MHz, CDCl3) δ 8.55 (s, 1H, NH), 8.16 (s,
1H, CH), 7.92 (d, J=7.6 Hz, 2H, Ar–H), 7.50–7.43 (m, 1H,
Ar–H), 7.39 (d, J = 7.3 Hz, 2H, Ar–H), 5.24 (s, 1H, CH),
4.84 (s, 2H, CH2), 4.51 (s, 2H, CH2), 4.17–3.98 (m, 2H,
CH2), 3.24 (d, J=5.1 Hz, 1H, CH), 2.34–2.25 (m, 2H, CH2),
2.17–0.71 (m, 44H, protons in UA skeleton). MS (MALDI-
TOF) m/z 699 [M+H]+.
1
m.p. 118–120 °C. H NMR (300 MHz, CDCl3) δ 7.80 (d,
J=8.8 Hz, 2H, Ar–H), 7.68 (s, 1H, CH), 6.86 (d, J=7.4 Hz,
2H, Ar–H), 5.23 (s, 1H, CH), 4.64 (d, J=5.3 Hz, 2H, CH2),
4.38 (s, 2H, CH2), 4.06 (m, 2H, CH2), 3.80 (s, 3H, OCH3),
3.18 (s, 1H, CH), 2.16 (m, 5H, CH and CH2), 2.02–0.70
(m, 41H, protons in UA skeleton). 13C NMR (101 MHz,
CDCl3) δ 177.34, 162.34, 138.27, 128.90, 125.67, 113.78,
78.99, 60.73, 55.44, 55.21, 52.94, 48.24, 47.51, 42.11,
39.55, 39.07, 38.86, 38.75, 38.62, 36.97, 36.83, 35.35,
33.04, 30.61, 29.70, 29.50, 28.17, 27.98, 27.22, 24.26,
23.55, 23.31, 21.16, 18.30, 17.20, 17.01, 15.67, 15.46. MS
(MALDI-TOF) m/z 751 [M+Na]+.
3-(4-((Furan-2-carboxamido)methyl)-1H-1,2,3-triazol-
1-yl)propyl(1S,2R,4aS,6aS,6bR,10S,12aR,12bR,14bS)-
10-hydroxy-1,2,6a,6b,9,9,12a-heptamethyl-1,3,4,5,6,6a,6
b,7,8,8a,9,10,11,12,12a,12b,13,14b-octadecahydropicene-
4a(2H)-carboxylate (11 h)White solid. Yield: 78%; m.p.
1
107–109 °C. H NMR (300 MHz, CDCl3) δ 7.63 (s, 1H,
NH), 7.45 (s, 1H, CH), 7.13 (d, J = 3.4 Hz, 2H, Fura-H),
6.50 (s, 1H, Fura-H), 5.27 (s, 1H, CH), 4.70 (d, J=5.9 Hz,
2H, CH2), 4.42 (s, 2H, CH2), 4.06 (m, 2H, CH2), 3.22 (s, 1H,
CH), 2.24 (s, 3H, CH and CH2), 2.06–0.74 (m, 44H, protons
3-(4-((2,3-Dimethoxybenzamido)methyl)-1H-1,2,3-
triazol-1-yl)propyl(1S,2R,4aS,6aS,6bR,10S,12aR,12b
1 3

Molecular Diversity
in UA skeleton). HRMS (ESI) m/z calcd for C41H61N4O5
[M+H]+ 689.46365, found 689.46356.
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Acknowledgements This research was supported by the Natural
Science Foundation of Jilin Province (No.20190201148JC); the
Department of Education of Jilin Province (No.JJKH20200461KJ
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