Controllable thioester-based hydrogen sulfide slow-releasing donors as cardioprotective agents

Article abstract of DOI:10.1039/c9cc02829c

Hydrogen sulfide (H₂S) is an important signaling molecule with promising protective effects in many physiological and pathological processes. However, the study of H₂S has been impeded by the lack of appropriate H₂S donors that could mimic its slow-releasing process in vivo. Herein, we report the rational design, synthesis, and biological evaluation of a series of thioester-based H₂S donors. These cysteine-activated H₂S donors release H₂S in a slow and controllable manner. Most of the donors comprising an allyl moiety showed significant cytoprotective effects in H9c2 cellular models of oxidative damage. The most potent donor 5e decreased the mitochondrial membrane potential (MMP) loss and lactate dehydrogenase (LDH) release in H₂O₂-stimulated H9c2 cells. More importantly, donor 5e exhibited a potent cardioprotective effect in an in vivo myocardial infarction (MI) mouse model by reducing myocardial infarct size and cardiomyocyte apoptosis. Taken together, our studies demonstrated that these new allyl thioesters are potential cardioprotective agents by releasing H₂S.

Full text of DOI:10.1039/c9cc02829c

View Article Online
View Journal
ChemComm
Accepted Manuscript
This article can be cited before page numbers have been issued, to do this please use: H. Yao, S. Luo, J.
Liu, Y. Liu, S. Xie, J. Xu, Z. Zhu and S. Xu, Chem. Commun., 2019, DOI: 10.1039/C9CC02829C.
This is an Accepted Manuscript, which has been through the
Volume 52 Number
1 4 January 2016 Pages 1–216
Royal Society of Chemistry peer review process and has been
accepted for publication.
ChemComm
Accepted Manuscripts are published online shortly after
Chemical Communications
www.rsc.org/chemcomm
acceptance, before technical editing, formatting and proof reading.
Using this free service, authors can make their results available
to the community, in citable form, before we publish the edited
article. We will replace this Accepted Manuscript with the edited
and formatted Advance Article as soon as it is available.
You can find more information about Accepted Manuscripts in the
author guidelines.
Please note that technical editing may introduce minor changes
to the text and/or graphics, which may alter content. The journal’s
standard Terms & Conditions and the ethical guidelines, outlined
in our author and reviewer resource centre, still apply. In no
event shall the Royal Society of Chemistry be held responsible
for any errors or omissions in this Accepted Manuscript or any
consequences arising from the use of any information it contains.
ISSN 1359-7345
COMMUNICATION
Marilyn M. Olmstead, Alan L. Balch, Josep M. Poblet, Luis Echegoyenet al.
Reactivity differences of Sc3N@C2n (2n 68 and 80). Synthesis of the
first methanofullerene derivatives of Sc N@D5h-C80
=
3
rsc.li/chemcomm

Page 1 of 5
ChemComm
Please do not adjust margins
View Article Online
DOI: 10.1039/C9CC02829C
Journal Name
ARTICLE
Controllable Thioester-Based Hydrogen Sulfide Slow-releasing
Donors as Cardioprotective Agents
Hong Yao,‡a Shanshan Luo,‡b Junkai Liu,a Shaowen Xie,a Yanpeng Liu,a Jinyi Xu,a Zheying Zhu,c and
Received 00th January 20xx,
Accepted 00th January 20xx
Shengtao Xua
DOI: 10.1039/x0xx00000x
www.rsc.org/
diallyl trisulfide (DATS), derivative of Lawesson’s reagent (GYY4137),
dithiolethione (ADT-OH), geminal-dithiol species (gem-dithiol) and
so on (Figure 1).[5] More recently, donors based on esterase-
activation, pH-modulation, ROS-activation and cysteine-activation as
well as their H2S-related biological effects have been reported.[6]
S
Hydrogen sulfide (H2S) is an important signaling molecule with promising
protective effects in many physiological and pathological processes.
However, the study on H2S has been impeded by the lack of appropriate H2S
donors that could mimic its slow-releasing process in vivo. Herein, we report
the rational design, synthesis, and biological evaluation of a series of
thioester-based H2S donors. These cysteine-activated H2S donors release
H2S in a slow and controllable manner. Most of the donors comprising allyl
moiety showed significant cytoprotective effects in H9c2 cellular models of
oxidative damage. The most potent donor 5e decreased the mitochondrial
membrane potential (MMP) loss and lactate dehydrogenase (LDH) release
in H2O2-stimulated H9c2 cells. More importantly, donor 5e exhibited potent
cardioprotective effect in an in vivo myocardial infarction (MI) mice model
by reducing myocardial infarct size and cardiomyocytes apoptosis. Taken
together, our studies demonstrated that these new allyl thioesters are
potential cardioprotective agents by releasing H2S.
R1 R2
S
S
MeO
P
N
S
S
S
S
S
H2N
O
NO2
O2N
S
S
HO
O
DATS
GYY4137
ADT-OH
Gem-dithiol
Figure 1. Representative H2S donors.
It should be noted that although a number of H2S donors have
been reported, ideal donors with controllable and slow H2S-releasing
properties are still lacking.[7] Recently, Liang and co-workers clarified
the misunderstanding of diallyl disulfide (DADS) as a rapid H2S-
releasing donor, which was attributed to its DATS impurity.[8] The
results showed that H2S was released in fact via a sluggish reaction
of DADS with GSH through an α-carbon nucleophilic substitution
pathway within hours. Thus, we envisioned the core structure of
DADS (diallyl disulfide) to be valuable template for the design of H2S
slow-releasing donors (Scheme 1, Step c).
Introduction
H2S has been recognized as an important signaling molecule owing
to its significant functions in many aspects of human health and
disease.[1] Many studies have shown various therapeutic effects of
H2S, including vasodilation, anti-inflammation, anti-oxidation, and
cardiovascular protection.[2] In order to explore the physiological and
pathological properties of H2S, researchers used inorganic sulfide
salts such as Na2S and NaHS to mimic endogenous H2S generation.[3]
However, the uncontrollable and rapid release of H2S from sulfide
salts does not mimic the slow and controllable H2S production in the
living system, which greatly limits their usage.[4] Considering these
disadvantages, small H2S-releasing organic molecules (H2S donors)
have been developed and used as the primary tools, mainly including
In addition, due to the ubiquitous targets of H2S, localized
generation of H2S is highly desirable but remains a great challenge.
Site-directed delivery of H2S by donors ideally shall be triggered by
specific cellular species or events.[9] H2S has been found to improve
cardiac functions and serve as a cardioprotectant for the treatment
of heart failure and ischemic heart diseases.[10] Under the condition
of cardiovascular injury, oxidative stress triggered by reactive oxygen
species (ROS) including superoxide, hydroxyl radical and hydrogen
peroxide plays a vital role.[11] As it is well known that the sulfide can
be easily converted into disulfide under oxidative conditions, the
oxidative stress in the injured cells could be slickly used to trigger the
localized generation of H2S within injured cells (Scheme 1, Step b). [12]
a. State Key Laboratory of Natural Medicines and Department of Medicinal Chemistry,
China Pharmaceutical University, 24 Tong Jia Xiang, Nanjing 210009, P. R. China. E-
mail: jinyixu@china.com ; cpuxst@cpu.edu.cn
Based on the above researches, we envision that allyl mercaptan
is a valuable template for the design of H2S slow-releasing donors
with cardioprotective effect. However, the simplicity and instability
of allyl mercaptan limit its direct application. Since
transthioesterification is a common strategy to transfer thioester to
thiol in the presence of Cys or GSH (Scheme 1, Step a).[13] We
b. Department of Pharmacology, School of Pharmacy, Nanjing Medical University,
Nanjing 211166, P. R. China
c. Division of Molecular Therapeutics & Formulation, School of Pharmacy, the
University of Nottingham, University Park Campus, Nottingham NG7 2RD, U.K.
†Electronic Supplementary Information (ESI) available: [details of any
supplementary information available should be included here]. See
DOI: 10.1039/x0xx00000x
‡These authors contributed equally to this work.
This journal is © The Royal Society of Chemistry 20xx
J. Name., 2018, 00, 1-3 | 1
Please do not adjust margins

ChemComm
Please do not adjust margins
Page 2 of 5
ARTICLE
Journal Name
designed a series of thioester-based hydrogen sulfide donors, which
could release H2S in a controllable and slow manner.
Lys, couldn’t trigger the H2S release of 5a, we supposeVdietwhAartticitlemOinglihnet
DOI: 10.1039/C9CC02829C
be due to the weaker nucleophilicity of those amino acids. We then
systematically compared the H2S-releasing capabilities of these
donors, and the data are summarized in Table 1. In contrast to DATS
(48.8%, 0.35 h), the results of our H2S donors showed that up to 52.2
% of the H2S can be detected (compound 5f) and the release time
lasts up to 8.5 h (compound 5k). All those thioester-based H2S donors
Herein, we reported a class of thioester-based H2S donors with
novel H2S-releasing mechanism and in vitro H2S-releasing kinetics.
Also their protective effects on H9c2 cells subjected with H2O2
challenge were evaluated, and the cardioprotective effects of
selected donors in a mouse model of MI were presented.
O
could release H2S in a much slower manner than DATS.
O
S
OH
PBS/THF(9:1), 37oC
NH2
S
H2S
Cys
O
5a 100 M
(
)
alpha-carbon
nucleophilic
substitution
HS
OH
SSH
NH2
Cys
allyl mercaptan
SH
(DADS)
O
S
R
S
H2S
S
c
a
b
O
S
Tansthioesterification
Site-directed delivery
Slow generation
O
OH
NH2
R
O
Scheme 1. Proposed controllable thioester-based hydrogen sulfide slow release donors.
Result and discussion
To test this idea, a series of thioester derivatives 5a-l were prepared
and the synthesis route is illustrated in Scheme 2. Various readily
available benzoic acids were used as the starting materials. The
treatment of acids with thionyl chloride afforded acyl chlorides 2 as
the reactive intermediates, which were further reacted with
thioacetamide in toluene to give compounds 3 in almost quantitive
yields. After hydrolysis, various thiobenzoic acids 4 were obtained,
which were treated with alkyl bromides or benzyl bromides directly
in the presence of triethylamine (TEA) to provide the desired
compounds 5a-l in 72-88% total yields. We expected that different
substituents would affect the reaction rates of compounds 5 with Cys
Figure 2. Cysteine mediated H2S release from compound 5a.
Table 1. H2S-releasing profiles of thioester-based hydrogen sulfide donors.
O
Cys (10 equiv)
H2S
R1
S
R2
to generate thiols, thereby regulating the rate of H2S generation.
PBS (pH 7.4, 50 mM)/THF
(9:1), 37oC
100 M
S
O
NH
O
O
1) aq.10% NaOH
SOCl2
NH2
S
OH
R1
Cl
R1
R1
DCM, rt.
Toluene
Tpeak
(h)
[H2S]peak
(μM)
41.5
17.5
38.3
34.5
50.6
52.2
42.4
Tpeak
[H2S]peak
(μM)
19.4
15.6
24.3
15.8
15.3
48.8
2) 10% HCl
Donors
Donors
1
2
3
(h)
3.5
5.0
3.0
8.5
7.5
0.27
5a
5b
5c
5d
5e
5f
8.0
7.0
6.5
5.0
8.0
6.0
7.0
5h
5i
5j
5k
5l
O
O
R2 Br
TEA
S
R2
SH
R1
R1
5
4
O
O
O
O
DATS
O
S
S
S
S
S
5g
O
5a
5b
5c
5d
O
O
O
O
S
S
S
To better understand the mechanism of H2S generation from these
thioester-based hydrogen sulfide donors, we analyzed the reaction
between 5a and Cys (10 equiv). As shown in Scheme 3, the
decomposition of 5a to 8 was monitored by the formation of 9 using
mass spectrometric analysis (Supporting Information, Fig. S1). The
formation of disulfide 10 from allyl mercaptan (8) was also confirmed
by mass spectra and NMR (Fig. S2). Besides, a great decline in the
release of H2S was observed under anaerobic conditions. On the
basis of the products observed and the previous knowledge,[8] we
proposed the following mechanism: The reaction is initiated by
reversible thiol exchange between 5a and Cys to generate the allyl
mercaptan (8), which suffers oxidation to form DADS (10).
Meanwhile, the resultant S-benzoyl cysteine 7 should undergo a
native chemical ligation (NCL)-type acyl transfer to form a more
stable N-benzoyl cystein 9. Compound 10 then undergoes α-carbon
nucleophilic substitution with Cys (Path A) or allyl mercaptan (Path
F
F3
C
O
5e
O
OCH3
5f
5g
5h
O
O
O
S
S
S
S
CF3
5l
5k
5i
F
5j
Scheme 2. Synthesis of thioester-based hydrogen sulfide donors 5a-l.
With these compounds in hand, we tested their H2S-releasing
kinetics in aqueous buffers firstly. These thioesters were stable in
PBS buffers, but a time- and dose-dependent H2S release was
detected when Cys was added into the solutions using the standard
methylene blue (MB) method.[14] Finally, the optimized condition
was determined to be 100 µM donors in PBS buffer (pH 7.4, 50 mM)
containing 10% THF in the presence of 10 equiv of Cys. As shown in
Figure 2, taking compound 5a as the example, a maximum of 41.5
µM of H2S at 8 h was detected from 100 µM donor in PBS buffer
containing 1 mM Cys. To our surprise, other amino acid, like Gly or
2 | J. Name., 2012, 00, 1-3
This journal is © The Royal Society of Chemistry 20xx
Please do not adjust margins

Page 3 of 5
ChemComm
Please do not adjust margins
Journal NameARTICLE
B) to generate perthiol intermediate (12), which releases H2S in the cytoplasmic enzyme found in all cells.[18] Thus, in tVhiieswsAtrtuicdleyO, ntlhinee
DOI: 10.1039/C9CC02829C
presence of Cys. However, it is worth mentioning that allyl sulfides contents of LDH in medium were measured to indicate the extent of
are substantially active because of the presence of allylic system, the cellular injury. As shown in Figure 3, the exposure of H9c2 cells to
precise and comprehensive H2S-releaseing mechanism of these H2O2 (400 µM, 1 h) caused the damage of cell membrane, leading to
donors was not clear yet, additional pathways to generate the H2S the release of LDH to the medium. The LDH release was significantly
might exist in this complex system.[15]
reduced when cells were pretreated with different concentrations
(10, 20, and 50 μM) of donors for 5 h in the presence of Cys (10
equiv), indicating the protective effects of these compounds. Among
these donors, 5e exhibited the most potent cardioprotective effect.
Changes in the mitochondrial membrane potential (∆ψm, MMP)
have been postulated to be early events in H2O2-induced apoptosis
signaling pathway.[19] Thus, fluorescent probe JC-1 was used to
measure the changes of ∆ψm of cells treated with 5e. As shown in
Figure S5, normal H9c2 cells had bright red fluorescence, and cells
treated with H2O2 appeared green fluorescence, indicating that H2O2
caused severe damage to cells and reduced ∆ψm. The pre-incubation
of cells with 20 or 50 μM of 5e greatly impeded this ∆ψm loss, as
indicated by a decrease in green emission and an increase in red
emission. These findings further confirmed that these H2S donors
exhibited potent cellular protection against oxidative injury.
O
O
O
+
S
HO
SH
S
HO
S
HO
S
NH2
O
NH2
NH2
6
(Cys)
13
5a
6
(Cys)
6
(Cys)
14
Path A
Path B
S
O
SH
S
H2
S
SH
S
12
10
8
O
O
S
HO
S
SH
11
NH2
7
NCL
8
O
H
N
HO
HS
O
9
Scheme 3. Proposed Mechanism of H2S generation.
H2S has been widely proposed to protect the cardiovascular
system through its antioxidative role.[16] In order to test the
protective effects of our newly synthesized H2S donors in cells, the
H2S releasing abilities of these donors in cells were firstly verified in
H9c2 cells by a selective H2S fluorescent probe WSP-1 (Fig. S3).[17] The
results showed that the typical donor 5a released H2S well in H2O2-
stimulated H9c2 cells. Thus, in vitro model of cardiomyoblasts were
established in H2O2-stimulated H9c2 cells. The cell viability was
analyzed by MTT colorimetry and the results were exhibited in Table
2. Compounds 5a, 5c-g significantly attenuated H2O2-induced cell
damage at concentrations of both 20 and 50 µM. The weaker
protective effects of these donors at higher concentrations (> 100
μM) might be attributed to their poor solubility (data not shown).
Overall, the protective effects of these donors were similar to that of
NaHS, DADS and DATS, and were consistent with their H2S release
abilities, suggesting that their protective effects are likely due to H2S
release.
Figure 3. LDH assay of compounds 5a-g on H9c2 cells. Cell viability was analyzed by LDH
assay from culture media of indicated cells which were pretreated with different
concentrations (10, 20, and 50 μM) of compounds for 5 h or NaHS (100 μM, 1 h). The
data are expressed as percentages of H2O2-stimulated cells (model group).
Table 2. Protective effect of compounds on H2O2-stimulated H9c2 cells.
Cell viability (%)
NO
Comp.
1 μM
58.1
59.6
64.6
62.8
65.0
66.0
63.1
62.8
63.3
63.7
63.3
62.6
58.6
60.3
59.6
51.9
10 μM
61.1
64.2
69.3
65.8
71.9
65.7
66.2
66.0
59.5
55.4
62.9
59.6
67.0
67.5
64.8
20 μM
74.5
66.3
77.9
68.9
71.3
64.4
72.1
69.9
59.0
57.1
64.7
57.1
68.5
71.7
71.2
50 μM
68.3
66.9
65.5
71.1
77.7
70.5
63.3
61.7
54.8
55.2
57.1
53.8
69.6
70.8
69.6
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
5a
5b
5c
5d
5e
5f
5g
5h
5i
5j
5k
5l
NaHS
DATS
DADS
Model
aValues are expressed as means from three independent experiments, the bar graph
expressed as mean ± SD was included in Supporting Information Fig. S4.
Cell viability is most often defined based on the integrity of the cell
membrane. A common method for detecting cell viability is based on
measuring the activity of cytoplasmic enzymes released by damaged
cells, such as lactate dehydrogenase (LDH), which is a stable
Figure 4. Isolated hearts from mice treated with different concentrations of 5e (15
mg/kg/day, 30 mg/kg/day) were sliced into 2 mm sections and stained with TTC to
determine the infarction size. (n=6 for each group). *p<0.05 versus model group,
**p<0.01 versus model group.
This journal is © The Royal Society of Chemistry 20xx
J. Name., 2013, 00, 1-3 | 3
Please do not adjust margins

ChemComm
Please do not adjust margins
Page 4 of 5
ARTICLE
Journal Name
The cardioprotective effects of 5e were further verified in vivo in
Notes and references
View Article Online
the MI mice model.[20] Mice treated with 5e for 7 days were
subjected to coronary occlusion and the hearts were harvested 72 h
post MI. Infarct size was assessed with TTC staining (2,3,5-
triphenyltetrazolium chloride), this white compound was
enzymatically reduced to red TPF (1,3,5-triphenylformazan) in living
tissues, while it remains as white TTC in areas of infarction since
these enzymes have been degraded due to apoptosis. For this reason,
healthy viable heart muscle will be stained in deep red, while areas
of potential infarctions will be more pale. Infarction size was
calculated as the ratio of scar average circumferences to left
ventricular average inner circumferences. The results showed that
infarct size was significantly larger in post MI mice (Model group)
than that in sham-operated mice (Normal group) (Figure 4a).
Remarkably, donor 5e treatment relived myocardial infarction in a
dose-dependent manner. Low (15 mg/kg/day) and high (30
mg/kg/day) dose of 5e reduce infarction size to 74.75% and 63.97%
of the model group, respectively (Figures 4b-c).
1
a) J.L. Wallace, R. Wang, Nat. Rev. DruDgODI:is1c0o.1v0.3290/C195C, C1042, 832299C-
345; b) Y. Zhao, T. D. Biggs, M. Xian, Chem. Commun. 2014, 50,
11788-11805.
2
a) J.L. Greaney, J.L. Kutz, S.W.Shank, S. Jandu, L. Santhanam, L.M.
Alexander, Hypertension 2017, 69, 902-909; b) N.L. Kanagy, C.
Szabo, A. Papapetropoulos, Am. J. Physiol. Cell. Physio. 2017,
312, C537-C549.
3
D. Yoo, R.C. Jupiter, E.A. Pankey, V.G. Reddy, J.A. Edward, K.W.
Swan, T.C. Reak, R. Mostany, P.J. Kadowitz, Am. J. Physiol. Heart
Circ. Physiol. 2015, 309, 605-614.
4
5
M.D. Hartle, M.D. Pluth, Chem. Soc. Rev. 2016, 45, 6108-6117.
a) C.Y. Tsai, S.Y. Wen, M.A. Shibu, Y.C. Yang, H. Peng, B. Wang,
Y.M. Wei, H.Y. Chang, C.Y. Lee, C.Y. Huang, W.W. Kuo, Int. J.
Cardiol. 2015, 195. 300-310; b) M. Lv, Y. Liu, T.H. Xiao, W. Jiang,
B.W. Lin, X.M. Zhang, Y.M. Lin, Z.S. Xu, Am. J. Transl. Res. 2017,
9, 1183-1192; c) U. Hasegawa, A.J. van der Vlies, Bioconjugate
Chem. 2014, 25, 1290-1300; d) Y. Zhao, J.M. Kang, C.M. Park, P.E.
Bagdon, B. Peng, M. Xian, Org. Lett. 2014, 16, 4536-4539.
a) Y. Zhao, S.G. Bolton, M.D. Pluth, Org. Lett. 2017, 19, 2278-
2281; b) J.M. Kang, Z. Li, C. L. Organ, C. M. Park, C. T. Yang, A.
Pacheco, D. F. Wang, D. J. Lefer, M. Xian, J. Am. Chem. Soc. 2016,
138, 6336-6339; c) Y. Zhao, M.D. Pluth, Angew. Chem. Int. Ed.
2016, 55, 14638-14642; d) Y. Zhao; A.K. Steiger, M.D. Pluth,
Angew. Chem. Int. Ed. 2018, 57, 1-6; e) Y. Zhao. A.K. Steiger, M.D.
Pluth, Chem. Commun. 2018, 54, 4951-4954.
H&E staining was then applied to evaluate histological feature of
infarcted hearts. As shown in Figure 5b, in peri-infarct heart,
myofilaments condensed, leaving damaged gap junction with
infiltrated nucleus from necrotic and apoptotic cells. However, donor
5e significantly restrained the damage within epimyocardium and
alleviated the morphological damage of cardiomyocytes (Figure 5c).
Furthermore, TUNEL staining was used to determine apoptosis of
cardiomyocytes in peri-infarction area. As shown in Figure S6, TUNEL
positive staining cells were hardly found in Sham operated mice,
while massive red fluorescent dots indicating fragments of DNA were
found in heart section of model group. Donor 5e displayed an
obvious benefit in reducing the number of TUNEL-positive cells.
6
7
8
9
Y. Zhao, A. Pacheco, M. Xian, Exp. Pharmacol. 2015, 230, 365-
388.
D. Liang, H.X. Wu, M.W. Wong, D.J. Huang, Org. Lett. 2015, 17,
4196-4199.
a) Y. Zheng, B. Yu, L.K. De La Cruz, M. Roy Choudhury, A.
Anifowose, B. Wang, Med. Res. Rev. 2018, 38, 57-100; b) S. Feng,
Y. Zhao, M. Xian, Q. Wang, Acta Biomater. 2015, 27, 205-213.
10 Y.D. Bai, Y.R. Yang, X.P. Mu, G. Lin, Y.P. Wang, S. Jin, Y. Chen, M.J.
Wang, Y.C. Zhu, Oxid. Med. Cell. Longev. 2018, 2018, 3402809.
11 C.A. Papaharalambus, K.K. Griendling, Trends Cardiovasc. Med.
2007, 17, 48-54.
12 a) M. Jereb, D. Dolenc, RSC Adv. 2015, 5, 58292-58306; b) V.
Kesavan, D. Bonnet-Delpon, J. Bégué, Synthesis 2000, 2, 223-225.
13 a) Y. Zhao, H. Wang, M. Xian, J. Am. Chem. Soc. 2011, 133, 15-17;
b) X.F. Yang, Q. Huang, Y. Zhong, Z. Li, H. Li, M. Lowry, J.O.
Escobedo, R. M. Strongin, Chem. Sci. 2014, 5, 2177-2183.
14 Z.G. Li, Methods Enzymol. 2015, 554, 101-110.
15 a) K.V. Ruddraraju, Z.D. Parsons, C.D. Lewis, K.S. Gates, J. Org.
Chem. 2017, 82, 776-780; b) G.A. Benavides, G.L. Squadrito, R.W.
Mills, R.P. Patel, V.M. Darley-Usmar, J.E. Doeller, D.W. Kraus, T.S.
Isbell, T.P. Patel, Proc. Natl. Acad. Sci. U.S.A. 2007, 104, 17977-
17982.
16 a) Z. Zhang, S. Jin, X. Teng, X. Duan, Y. Chen, Y. Wu, Nitric Oxide.
2017, 67, 10-25; b) S.Y. Wen, C.Y. Tsai, P.Y. Pai, Y.W. Chen, Y.C.
Yang, R. Aneja, C.Y. Huang, W.W. Kuo, Environ. Toxicol. 2018, 33,
93-103.
17 B. Peng, W. Chen, C. Liu, E.W. Rosser, A. Pacheco, Y. Zhao, H.C.
Aguilar, M. Xian, Chem-Eur. J. 2014, 20, 1010-1016.
18 N. Uchide, K. Ohyama, T. Bessho, H. Toyoda, Intervirology 2009,
52,164-173.
Figure 5. H&E staining was performed using heart sections to determine the
morphological changes. (a) Normal group; (b) Model group; (c) Treated with 5e, 30
mg/kg/day. Scale bar: 10 μm.
In summary, a series of thioester-based H2S donors were
developed. These donors are stable in aqueous solution, and release
significant amounts of H2S in a slow and controllable manner in the
presence of Cys. The most potent donor 5e exhibited promising
cardioprotective activity against H2O2-induced oxidative stress by
decreasing the mitochondrial membrane potential loss and lactate
dehydrogenase release in H9c2 cells. More importantly, donor 5e
also exhibited potent cardioprotective effects in an in vivo
myocardial infarction mice model by reducing myocardial infarct size
and cardiomyoblasts apoptosis. Taken together, our study
demonstrated that the new allyl thioesters may have potential as
cardioprotective agents by releasing H2S.
Acknowledgements
19 L. Wang, H. Huang, Y. Fan, B. Kong, H. Hu, K. Hu, J. Guo, Y. Mei,
W. L. Liu, Oxid. Med. Cell. Longev. 2014, 2014, 960362.
20 S.S. Luo, X.F. Gu, F.F. Ma, C.H. Liu, Y.Q. Shen, R.W. Ge, Y.Z. Zhu,
Rree Radic. Biol. Med. 2016, 92, 1-14.
We gratefully acknowledge financial support from the National
Natural Science Foundation of China (81673306, 81703348), and
"Double First-Class" University project CPU2018GY04, CPU2018GY35,
China Pharmaceutical University.
4 | J. Name., 2012, 00, 1-3
This journal is © The Royal Society of Chemistry 20xx
Please do not adjust margins

Page 5 of 5
ChemComm
View Article Online
DOI: 10.1039/C9CC02829C
199x98mm (300 x 300 DPI)