New applications of clioquinol in the treatment of inflammation disease by directly targeting arginine 335 of NLRP3

Peipei Chen; Yunshu Wang; Huaiping Tang; Chao Zhou; Zhuo Liu; Shenghan Gao; Tingting Wang; Yun Xu; Sen-Lin Ji

doi:10.1016/j.jpha.2024.101069

Volume 15 Issue 1

Feb. 2025

Turn off MathJax

Article Contents

Article Navigation > Journal of Pharmaceutical Analysis > 2025 > 15(1): 101069

Peipei Chen, Yunshu Wang, Huaiping Tang, Chao Zhou, Zhuo Liu, Shenghan Gao, Tingting Wang, Yun Xu, Sen-Lin Ji. New applications of clioquinol in the treatment of inflammation disease by directly targeting arginine 335 of NLRP3[J]. Journal of Pharmaceutical Analysis, 2025, 15(1): 101069. doi: 10.1016/j.jpha.2024.101069

Citation:

Peipei Chen, Yunshu Wang, Huaiping Tang, Chao Zhou, Zhuo Liu, Shenghan Gao, Tingting Wang, Yun Xu, Sen-Lin Ji. New applications of clioquinol in the treatment of inflammation disease by directly targeting arginine 335 of NLRP3[J]. Journal of Pharmaceutical Analysis, 2025, 15(1): 101069. doi: 10.1016/j.jpha.2024.101069

Citation:

Peipei Chen, Yunshu Wang, Huaiping Tang, Chao Zhou, Zhuo Liu, Shenghan Gao, Tingting Wang, Yun Xu, Sen-Lin Ji. New applications of clioquinol in the treatment of inflammation disease by directly targeting arginine 335 of NLRP3[J]. Journal of Pharmaceutical Analysis, 2025, 15(1): 101069. doi: 10.1016/j.jpha.2024.101069

PDF( 9258 KB)

New applications of clioquinol in the treatment of inflammation disease by directly targeting arginine 335 of NLRP3

doi: 10.1016/j.jpha.2024.101069

Peipei Chen^a,b,c,d,
Yunshu Wang^a,b,c,d,
Huaiping Tang^a,b,c,d,
Chao Zhou^a,b,c,d,
Zhuo Liu^a,b,c,d,
Shenghan Gao^a,b,c,d,
Tingting Wang^{e
,
,},
Yun Xu^{a,b,c,d
,
,},
Sen-Lin Ji^{a,b,c,d
,
,}

a. Department of Neurology, Nanjing Drum Tower Hospital, Affiliated Hospital of Medical School, Nanjing University, Nanjing, 210008, China;
b. State Key Laboratory of Pharmaceutical Biotechnology and Institute of Translational Medicine for Brain Critical Diseases, Nanjing University, Nanjing, 210093, China;
c. Jiangsu Key Laboratory for Molecular Medicine, Medical School of Nanjing University, Nanjing, 210008, China;
d. Nanjing Neurology Clinical Medical Center, Nanjing, 210000, China;
e. State Key Laboratory of Pharmaceutical Biotechnology, Chemistry and Biomedicine Innovation Center (ChemBIC), Jiangsu Key Laboratory of Molecular Medicine, Division of Immunology, Medical School, Nanjing University, Nanjing, 210008, China

Funds:

This research was supported by the National Natural Science Foundation of China (Grant Nos.: 82101417, 81920108017, and 82130036), the STI2030-Major Projects (Project No.: 2022ZD0211800), Jiangsu Province Key Medical Discipline (Grant No.: ZDXK202216), the Key Research and Development Program of Jiangsu Province of China (Program No.: BE2020620), and Nanjing Medical Science and technology development Foundation, China (Grant No.: YKK20061).

Received Date: May 09, 2024
Accepted Date: Aug. 10, 2024
Rev Recd Date: Aug. 05, 2024
Publish Date: Aug. 18, 2024

Abstract

Abstract

The NOD-like receptor protein 3 (NLRP3) inflammasome is essential in innate immune-mediated inflammation, with its overactivation implicated in various autoinflammatory, metabolic, and neurodegenerative diseases. Pharmacological inhibition of NLRP3 offers a promising treatment strategy for inflammatory conditions, although no medications targeting the NLRP3 inflammasome are currently available. This study demonstrates that clioquinol (CQ), a clinical drug with chelating properties, effectively inhibits NLRP3 activation, resulting in reduced cytokine secretion and cell pyroptosis in both human and mouse macrophages, with a half maximal inhibitory concentration (IC₅₀) of 0.478 μM. Additionally, CQ mitigates experimental acute peritonitis, gouty arthritis, sepsis, and colitis by lowering serum levels of interleukin-1β (IL-1β), IL-6, and tumor necrosis factor-α (TNF-α). Mechanistically, CQ covalently binds to Arginine 335 (R335) in the NACHT domain, inhibiting NLRP3 inflammasome assembly and blocking the interaction between NLRP3 and its component protein. Collectively, this study identifies CQ as an effective natural NLRP3 inhibitor and a potential therapeutic agent for NLRP3-driven diseases.
- Clioquinol,
- NLRP3 inflammasome,
- NACHT domain,
- Sepsis,
- Peritonitis,
- Colitis

FullText(HTML)

References(44)

References

[1]	E.I. Elliott, F.S. Sutterwala, Initiation and perpetuation of NLRP3 inflammasome activation and assembly, Immunol. Rev. 265 (2015) 35-52.
[2]	K.V. Swanson, M. Deng, J.P.Y. Ting, The NLRP3 inflammasome: Molecular activation and regulation to therapeutics, Nat. Rev. Immunol. 19 (2019) 477-489.
[3]	Y. Huang, W. Xu, R. Zhou, NLRP3 inflammasome activation and cell death, Cell. Mol. Immunol. 18 (2021) 2114-2127.
[4]	X. Zhan, Q. Li, G. Xu, et al., The mechanism of NLRP3 inflammasome activation and its pharmacological inhibitors, Front. Immunol. 13 (2023), 1109938.
[5]	P. Duewell, H. Kono, K.J. Rayner, et al., NLRP3 inflammasomes are required for atherogenesis and activated by cholesterol crystals, Nature 464 (2010) 1357-1361.
[6]	R.A. Cherny, C.S. Atwood, M.E. Xilinas, et al., Treatment with a copper-zinc chelator markedly and rapidly inhibits β-amyloid accumulation in Alzheimer’s disease transgenic mice, Neuron 30 (2001) 665-676.
[7]	J. Li, L. Zhuang, X. Luo, et al., Protection of MCC950 against Alzheimer’s disease via inhibiting neuronal pyroptosis in SAMP8 mice, Exp. Brain Res. 238 (2020) 2603-2614.
[8]	A.P. Perera, R. Fernando, T. Shinde, et al., MCC950, a specific small molecule inhibitor of NLRP3 inflammasome attenuates colonic inflammation in spontaneous colitis mice, Sci. Rep. 8 (2018), 8618.
[9]	R. Wykowski, A.M. Fuentefria, S.F. de Andrade, Antimicrobial activity of clioquinol and nitroxoline: A scoping review, Arch. Microbiol. 204 (2022), 535.
[10]	D. Kaur, F. Yantiri, S. Rajagopalan, et al., Genetic or pharmacological iron chelation prevents MPTP-induced neurotoxicity in vivo, Neuron 37 (2003) 899-909.
[11]	T. Nguyen, A. Hamby, S.M. Massa, Clioquinol down-regulates mutant huntingtin expression in vitro and mitigates pathology in a Huntington’s disease mouse model, Proc. Natl. Acad. Sci. USA 102 (2005) 11840-11845.
[12]	S. Lu, Y. Ke, C. Wu, et al., Radio sensitization of clioquinol and zinc in human cancer cell lines, BMC Cancer 18 (2018), 448.
[13]	X. Lv, Z. Fan, F. Cao, et al., Clioquinol induces autophagy by down-regulation of calreticulin in human neurotypic SH-SY5Y cells, Chem. Biol. Interact. 369 (2023), 110268.
[14]	A.I. Bush, Drug development based on the metal’s hypothesis of Alzheimer’s disease, J. Alzheimers Dis. 15 (2008) 223-240.
[15]	C. Chen, Y. Zhou, X. Ning, et al., Directly targeting ASC by lonidamine alleviates inflammasome-driven diseases, J. Neuroinflammation 19 (2022), 315.
[16]	R. Yuan, W. Zhao, Q. Wang, et al., Cucurbitacin B inhibits non-small cell lung cancer in vivo and in vitro by triggering TLR4/NLRP3/GSDMD-dependent pyroptosis, Pharmacol. Res. 170 (2021), 105748.
[17]	Y. Huang, H. Jiang, Y. Chen, et al., Tranilast directly targets NLRP3 to treat inflammasome-driven diseases, EMBO Mol. Med. 10 (2018), e8689.
[18]	J. Fu, H. Wu, Structural mechanisms of NLRP3 inflammasome assembly and activation, Annu. Rev. Immunol. 41 (2023) 301-316.
[19]	S. Li, Y. Sun, M. Song, et al., NLRP3/caspase-1/GSDMD-mediated pyroptosis exerts a crucial role in astrocyte pathological injury in mouse model of depression, JCI Insight 6 (2021), e146852.
[20]	M.Y. Pai, B. Lomenick, H. Hwang, et al., Drug affinity responsive target stability (DARTS) for small-molecule target identification, Methods Mol. Biol. 1263 (2015) 287-298.
[21]	V.G. Magupalli, R. Negro, Y. Tian, et al., HDAC6 mediates an aggresome-like mechanism for NLRP3 and pyrin inflammasome activation, Science 369 (2020), eaas8995.
[22]	L.G. Danielski, A.D. Giustina, S. Bonfante, et al., The NLRP3 inflammasome and its role in sepsis development, Inflammation 43 (2020) 24-31.
[23]	R. Ungaro, S. Mehandru, P.B. Allen, et al., Ulcerative colitis, The Lancet 389 (2017) 1756-1770.
[24]	C. Bauer, P. Duewell, C. Mayer, et al., Colitis induced in mice with dextran sulfate sodium (DSS) is mediated by the NLRP3 inflammasome, Gut 59 (2010) 1192-1199.
[25]	H.M. Lee, J.J. Kim, H.J. Kim, et al., Upregulated NLRP3 inflammasome activation in patients with type 2 diabetes, Diabetes 62 (2013) 194-204.
[26]	M.T. Heneka, M.P. Kummer, A. Stutz, et al., NLRP3 is activated in Alzheimer’s disease and contributes to pathology in APP/PS1 mice, Nature 493 (2013) 674-678.
[27]	W. Barclay, M.L. Shinohara, Inflammasome activation in multiple sclerosis and experimental autoimmune encephalomyelitis (EAE), Brain Pathol. 27 (2017) 213-219.
[28]	Y. Dong, S. Li, Y. Lu, et al., Stress-induced NLRP3 inflammasome activation negatively regulates fear memory in mice, J. Neuroinflammation 17 (2020), 205.
[29]	H.S.G.R. Investigators, Safety, tolerability, and efficacy of PBT2 in Huntington’s disease: A phase 2, randomised, double-blind, placebo-controlled trial, Lancet Neurol. 14 (2015) 39-47.
[30]	X. Han, S. Sun, Y. Sun, et al., Small molecule-driven NLRP3 inflammation inhibition via interplay between ubiquitination and autophagy: Implications for Parkinson disease, Autophagy 15 (2019) 1860-1881.
[31]	Q. Huang, P. Yang, Y. Liu, et al., The interplay between alpha-Synuclein and NLRP3 inflammasome in Parkinson’s disease, Biomed. Pharmacother. 168 (2023), 115735.
[32]	Z. Huang, L. Wang, L. Chen, et al., Induction of cell cycle arrest via the p21, p27-cyclin E, A/Cdk2 pathway in SMMC-7721 hepatoma cells by clioquinol, Acta Pharm. 65 (2015) 463-471.
[33]	I. Tanida, T. Ueno, E. Kominami, LC3 and autophagy, Methods Mol. Biol. 445 (2008) 77-88.
[34]	L. Zhang, Y. Gai, Y. Liu, et al., Tau induces inflammasome activation and microgliosis through acetylating NLRP3, Clin. Transl. Med. 14 (2024), e1623.
[35]	S. Zheng, X. Que, S. Wang, et al., ZDHHC5-mediated NLRP3 palmitoylation promotes NLRP3-NEK7 interaction and inflammasome activation, Mol. Cell 83 (2023) 4570-4585.e7.
[36]	K. Lee, B.K. Hong, S. Lee, et al., Nuclear receptor coactivator 6 is a critical regulator of NLRP3 inflammasome activation and gouty arthritis, Cell. Mol. Immunol. 21 (2024) 227-244.
[37]	S. Chen, Y. Li, Y. You, et al., Theaflavin mitigates acute gouty peritonitis and septic organ injury in mice by suppressing NLRP3 inflammasome assembly, Acta Pharmacol. Sin. 44 (2023) 2019-2036.
[38]	R.C. Coll, A.A.B. Robertson, J.J. Chae, et al., A small-molecule inhibitor of the NLRP3 inflammasome for the treatment of inflammatory diseases, Nat. Med. 21 (2015) 248-255.
[39]	H. He, H. Jiang, Y. Chen, et al., Oridonin is a covalent NLRP3 inhibitor with strong anti-inflammasome activity, Nat. Commun. 9 (2018), 2550.
[40]	J. Shao, W. Li, J. Sun, et al., Britannin as a novel NLRP3 inhibitor, suppresses inflammasome activation in macrophages and alleviates NLRP3-related diseases in mice, Acta Pharmacol. Sin. 45 (2024) 803-814.
[41]	G. Xu, S. Fu, X. Zhan, et al., Echinatin effectively protects against NLRP3 inflammasome-driven diseases by targeting HSP90, JCI Insight 6 (2021), e134601.
[42]	J. Zhao, H. Liu, Z. Hong, et al., Tanshinone I specifically suppresses NLRP3 inflammasome activation by disrupting the association of NLRP3 and ASC, Mol. Med. 29 (2023), 84.
[43]	X. Wu, P. Xu, X. Shi, et al., Intra-articular injection of clioquinol ameliorates osteoarthritis in a rabbit model, Front. Med. 9 (2022), 1028575.
[44]	L. Mao, A. Kitani, W. Strober, et al., The role of NLRP3 and IL-1β in the pathogenesis of inflammatory bowel disease, Front. Immunol. 9 (2018), 2566.