10.1002/anie.201806854
Angewandte Chemie International Edition
COMMUNICATION
µg/mL) to induce an inflammatory response. This response is
accompanied by an increase in NO production, which we
monitored by measuring nitrite (NO2 ) accumulation. Our
we also demonstrate that γ-KetoTCM-1 releases COS/H2S in live
cells and reduces LPS-induced NO formation, which is consistent
with anti-inflammatory activities. Taken together, γ-KetoTCM
compounds provide a promising new platform for H2S donation
and readily enables colorimetric measurement of H2S donation,
making them as key research tools in H2S investigations.
–
expectation was that the COS/H2S donor would decrease LPS-
–
induced NO2 formation, indicating anti-inflammatory activity of
the donor. To determine whether the observed effects were due
to COS/H2S release, we also performed control experiments with
γ-KetoTCM-3, γ-KetoCM-1, and GYY4137 under the identical
condition. We used GYY4137 as a positive control because it has
shown anti-inflammatory effects previously,[21] and because it
generates a slow, continuous release of H2S. We chose to use 25
µM of each compound in this study because this concentration
did not provide significant cytotoxicity (Figure S7).
Acknowledgements
We acknowledge financial support from the NIH (R01GM113030),
Dreyfus Foundation, and NSF (AKS; DGE-1309047). NMR, MS,
microscopy instrumentation in the CAMCOR facility is supported
by the NSF (CHE-1427987, CHE-1531189, CHE-1625529).
In comparison to the control group, in which cells were only
incubated in FBS-free DMEM, the LPS-treated cells showed a
–
significant NO2 increase. γ-KetoTCM-1 pretreatment, however,
Keywords: hydrogen sulfide • γ-ketothiocarbamate • carbonyl
–
significantly reduced LPS-induced NO2 production. Control
sulfide • colorimetric • anti-inflammation
–
experiments using γ-KetoCM-1 also reduced NO2 levels,
although to a lesser extent than γ-KetoTCM-1. We attribute the
observed effects from KetoCM-1 to MVK release, which we
confirmed independently (Figure S8). We also observed a modest
[1]
[2]
K. Abe, H. Kimura, J. Neurosci. 1996, 16, 1066-1071.
(a) R. Wang, Physiol. Rev. 2012, 92, 791-896; (b) C. Szabo, Nat. Rev.
Drug. Discov. 2007, 6, 917-935.
–
reduction of LPS-induced NO2 production from γ-KetoTCM-3,
[3]
(a) J. M. Fukuto, S. J. Carrington, D. J. Tantillo, J. G. Harrison, L. J.
Ignarro, B. A. Freeman, A. Chen, D. A. Wink, Chem. Res. Toxicol. 2012,
25, 769-793; (b) O. Kabil, R. Banerjee, Antioxid. Redox. Signal. 2014, 20,
770-782.
although this effect was significantly attenuated from that of γ-
KetoTCM-1. GYY4137 exhibited a less pronounced effect on
LPS-induced NO2– production at the same concentration (25 M),
which supports the increased efficiency of γ-KetoTCM-1 (Figure
5). Taken together, these investigations demonstrate that γ-
KetoTCM-1 can deliver H2S in complex environment and provide
protection against LPS-induced inflammation, suggesting
potential therapeutic applications of γ-KetoTCM-based H2S
donors. In addition these experiments highlight the benefits of
having access to key control compounds that enable specific
contributions to cellular protections to be analysed.
[4]
[5]
(a) L. Li, P. Rose, P. K. Moore, Annu. Rev. Pharmacol. Toxicol. 2011, 51,
169-187; (b) K. R. Olson, Antioxid. Redox. Signal. 2012, 17, 32-44; (c) C.
Szabo, Antioxid. Redox. Signal. 2012, 17, 68-80.
(a) M. D. Pluth, T. S. Bailey, M. D. Hammers, M. D. Hartle, H. A. Henthorn,
A. K. Steiger, Synlett 2015, 26, 2633-2643; (b) Y. Zhao, T. D. Biggs, M.
Xian, Chem. Commun. 2014, 50, 11788-11805; (c) Y. Zhao, A. Pacheco,
M. Xian, Handb. Exp. Pharmacol. 2015, 230, 365-388; (d) Y. Zheng, B.
Yu, L. K. De La Cruz, M. Roy Choudhury, A. Anifowose, B. Wang, Med.
Res. Rev. 2018, 38, 57-100; (e) C. Szabo, A. Papapetropoulos,
Pharmacol. Rev. 2017, 69, 497-564.
[6]
[7]
G. A. Benavides, G. L. Squadrito, R. W. Mills, H. D. Patel, T. S. Isbell, R.
P. Patel, V. M. Darley-Usmar, J. E. Doeller, D. W. Kraus, Proc. Natl. Acad.
Sci. U. S. A. 2007, 104, 17977-17982.
(a) Y. Zhao, H. Wang, M. Xian, J. Am. Chem. Soc. 2011, 133, 15-17; (b)
Y. Zhao, S. Bhushan, C. Yang, H. Otsuka, J. D. Stein, A. Pacheco, B.
Peng, N. O. Devarie-Baez, H. C. Aguilar, D. J. Lefer, M. Xian, ACS Chem.
Biol. 2013, 8, 1283-1290; (c) Y. Zhao, J. Kang, C. M. Park, P. E. Bagdon,
B. Peng, M. Xian, Org. Lett. 2014, 16, 4536-4539; (d) Y. Zhao, C. Yang,
C. Organ, Z. Li, S. Bhushan, H. Otsuka, A. Pacheco, J. Kang, H. C.
Aguilar, D. J. Lefer, M. Xian, J. Med. Chem. 2015, 58, 7501-7511; (e) J.
C. Foster, C. R. Powell, S. C. Radzinski, J. B. Matson, Org. Lett. 2014,
16, 1558-1561; (f) A. Martelli, L. Testai, V. Citi, A. Marino, I. Pugliesi, E.
Barresi, G. Nesi, S. Rapposelli, S. Taliani, F. Da Settimo, M. C. Breschi,
V. Calderone, ACS Med. Chem. Lett. 2013, 4, 904-908; (g) T. Roger, F.
Raynaud, F. Bouillaud, C. Ransy, S. Simonet, C. Crespo, M. P.
Bourguignon, N. Villeneuve, J. P. Vilaine, I. Artaud, E. Galardon,
Chembiochem 2013, 14, 2268-2271.
Figure 5. Effects of γ-KetoTCM-1 on LPS-induced NO2– formation. RAW 264.7
cells were pretreated with γ-KetoTCM-1 (25 µM) or control compounds for 6 h,
followed by LPS (1.0 µg/mL, 18-h). Results are expressed as mean ± SD (n =
4). ***P < 0.001 vs the control group; ###P < 0.001 vs vehicle-treated group; and
$$$P < 0.001 between γ-KetoTCM-1-treated and γ-KetoCM-1-treated groups.
[8]
[9]
(a) Y. Zheng, B. Yu, K. Ji, Z. Pan, V. Chittavong, B. Wang, Angew. Chem.
Int. Ed. 2016, 55, 4514-4518; (b) P. Shukla, V. S. Khodade, M.
SharathChandra, P. Chauhan, S. Mishra, S. Siddaramappa, B. E.
Pradeep, A. Singh, H. Chakrapani, Chem Sci 2017, 8, 4967-4972.
J. Kang, Z. Li, C. L. Organ, C. M. Park, C. T. Yang, A. Pacheco, D. Wang,
D. J. Lefer, M. Xian, J. Am. Chem. Soc. 2016, 138, 6336-6339.
In summary, we prepared and evaluated a series of γ-
ketothiocarbamate compounds that function as COS/H2S donors
and provide a colorimetric response upon donor activation. The
PNA generated upon donor activation provides an optical readout,
which allows for the COS/H2S release to be monitored and
quantified directly during the course of an experiment. In addition,
[10] (a) N. O. Devarie-Baez, P. E. Bagdon, B. Peng, Y. Zhao, C. M. Park, M.
Xian, Org. Lett. 2013, 15, 2786-2789; (b) N. Fukushima, N. Ieda, K.
Sasakura, T. Nagano, K. Hanaoka, T. Suzuki, N. Miyata, H. Nakagawa,
Chem. Commun. 2014, 50, 587-589.
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