Benzisothiazolinone derivatives as potent allosteric monoacylglycerol lipase inhibitors that functionally mimic sulfenylation of regulatory cysteines

Riccardo Castelli; Laura Scalvini; Federica Vacondio; Alessio Lodola; Mattia Anselmi; Stefano Vezzosi; Caterina Carmi; Michele Bassi; Francesca Ferlenghi; Silvia Rivara; Ingvar R Möller; Kasper D Rand; Jennifer Daglian; Don Wei; Emmanuel Yaw Dotsey; Faizy Ahmed; Kwang-Mook Jung; Nephi Stella; Simar Singh; Marco Mor; Daniele Piomelli

doi:10.1021/acs.jmedchem.9b01679

Benzisothiazolinone derivatives as potent allosteric monoacylglycerol lipase inhibitors that functionally mimic sulfenylation of regulatory cysteines

Riccardo Castelli, Laura Scalvini, Federica Vacondio, Alessio Lodola, Mattia Anselmi, Stefano Vezzosi, Caterina Carmi, Michele Bassi, Francesca Ferlenghi, Silvia Rivara, Ingvar R Möller, Kasper D Rand, Jennifer Daglian, Don Wei, Emmanuel Yaw Dotsey, Faizy Ahmed, Kwang-Mook Jung, Nephi Stella, Simar Singh, Marco MorDaniele Piomelli

3 Citations (Scopus)

Abstract

We describe a set of benzisothiazolinone (BTZ) derivatives that are potent inhibitors of monoacylglycerol lipase (MGL), the primary degrading enzyme for the endocannabinoid 2-arachidonoyl-sn-glycerol (2-AG). Structure-activity relationship studies evaluated various substitutions on the nitrogen atom and the benzene ring of the BTZ nucleus. Optimized derivatives with nanomolar potency allowed us to investigate the mechanism of MGL inhibition. Site-directed mutagenesis and mass spectrometry experiments showed that BTZs interact in a covalent reversible manner with regulatory cysteines, Cys201 and Cys208, causing a reversible sulfenylation known to modulate MGL activity. Metadynamics simulations revealed that BTZ adducts favor a closed conformation of MGL that occludes substrate recruitment. The BTZ derivative 13 protected neuronal cells from oxidative stimuli and increased 2-AG levels in the mouse brain. The results identify Cys201 and Cys208 as key regulators of MGL function and point to the BTZ scaffold as a useful starting point for the discovery of allosteric MGL inhibitors.

Original language	English
Journal	Journal of Medicinal Chemistry
ISSN	0022-2623
DOIs	https://doi.org/10.1021/acs.jmedchem.9b01679
Publication status	Published - 13 Feb 2020

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Castelli, R., Scalvini, L., Vacondio, F., Lodola, A., Anselmi, M., Vezzosi, S., Carmi, C., Bassi, M., Ferlenghi, F., Rivara, S., Möller, I. R., Rand, K. D., Daglian, J., Wei, D., Dotsey, E. Y., Ahmed, F., Jung, K.-M., Stella, N., Singh, S., ... Piomelli, D. (2020). Benzisothiazolinone derivatives as potent allosteric monoacylglycerol lipase inhibitors that functionally mimic sulfenylation of regulatory cysteines. Journal of Medicinal Chemistry. https://doi.org/10.1021/acs.jmedchem.9b01679

Castelli, R, Scalvini, L, Vacondio, F, Lodola, A, Anselmi, M, Vezzosi, S, Carmi, C, Bassi, M, Ferlenghi, F, Rivara, S, Möller, IR, Rand, KD, Daglian, J, Wei, D, Dotsey, EY, Ahmed, F, Jung, K-M, Stella, N, Singh, S, Mor, M & Piomelli, D 2020, 'Benzisothiazolinone derivatives as potent allosteric monoacylglycerol lipase inhibitors that functionally mimic sulfenylation of regulatory cysteines', Journal of Medicinal Chemistry. https://doi.org/10.1021/acs.jmedchem.9b01679

@article{d3c113958c4c480f98d9d17e64584326,

title = "Benzisothiazolinone derivatives as potent allosteric monoacylglycerol lipase inhibitors that functionally mimic sulfenylation of regulatory cysteines",

abstract = "We describe a set of benzisothiazolinone (BTZ) derivatives that are potent inhibitors of monoacylglycerol lipase (MGL), the primary degrading enzyme for the endocannabinoid 2-arachidonoyl-sn-glycerol (2-AG). Structure-activity relationship studies evaluated various substitutions on the nitrogen atom and the benzene ring of the BTZ nucleus. Optimized derivatives with nanomolar potency allowed us to investigate the mechanism of MGL inhibition. Site-directed mutagenesis and mass spectrometry experiments showed that BTZs interact in a covalent reversible manner with regulatory cysteines, Cys201 and Cys208, causing a reversible sulfenylation known to modulate MGL activity. Metadynamics simulations revealed that BTZ adducts favor a closed conformation of MGL that occludes substrate recruitment. The BTZ derivative 13 protected neuronal cells from oxidative stimuli and increased 2-AG levels in the mouse brain. The results identify Cys201 and Cys208 as key regulators of MGL function and point to the BTZ scaffold as a useful starting point for the discovery of allosteric MGL inhibitors.",

author = "Riccardo Castelli and Laura Scalvini and Federica Vacondio and Alessio Lodola and Mattia Anselmi and Stefano Vezzosi and Caterina Carmi and Michele Bassi and Francesca Ferlenghi and Silvia Rivara and M{\"o}ller, {Ingvar R} and Rand, {Kasper D} and Jennifer Daglian and Don Wei and Dotsey, {Emmanuel Yaw} and Faizy Ahmed and Kwang-Mook Jung and Nephi Stella and Simar Singh and Marco Mor and Daniele Piomelli",

year = "2020",

month = feb,

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doi = "10.1021/acs.jmedchem.9b01679",

language = "English",

journal = "Journal of Medicinal Chemistry",

issn = "0022-2623",

publisher = "American Chemical Society",

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T1 - Benzisothiazolinone derivatives as potent allosteric monoacylglycerol lipase inhibitors that functionally mimic sulfenylation of regulatory cysteines

AU - Castelli, Riccardo

AU - Scalvini, Laura

AU - Vacondio, Federica

AU - Lodola, Alessio

AU - Anselmi, Mattia

AU - Vezzosi, Stefano

AU - Carmi, Caterina

AU - Bassi, Michele

AU - Ferlenghi, Francesca

AU - Rivara, Silvia

AU - Möller, Ingvar R

AU - Rand, Kasper D

AU - Daglian, Jennifer

AU - Wei, Don

AU - Dotsey, Emmanuel Yaw

AU - Ahmed, Faizy

AU - Jung, Kwang-Mook

AU - Stella, Nephi

AU - Singh, Simar

AU - Mor, Marco

AU - Piomelli, Daniele

PY - 2020/2/13

Y1 - 2020/2/13

N2 - We describe a set of benzisothiazolinone (BTZ) derivatives that are potent inhibitors of monoacylglycerol lipase (MGL), the primary degrading enzyme for the endocannabinoid 2-arachidonoyl-sn-glycerol (2-AG). Structure-activity relationship studies evaluated various substitutions on the nitrogen atom and the benzene ring of the BTZ nucleus. Optimized derivatives with nanomolar potency allowed us to investigate the mechanism of MGL inhibition. Site-directed mutagenesis and mass spectrometry experiments showed that BTZs interact in a covalent reversible manner with regulatory cysteines, Cys201 and Cys208, causing a reversible sulfenylation known to modulate MGL activity. Metadynamics simulations revealed that BTZ adducts favor a closed conformation of MGL that occludes substrate recruitment. The BTZ derivative 13 protected neuronal cells from oxidative stimuli and increased 2-AG levels in the mouse brain. The results identify Cys201 and Cys208 as key regulators of MGL function and point to the BTZ scaffold as a useful starting point for the discovery of allosteric MGL inhibitors.

AB - We describe a set of benzisothiazolinone (BTZ) derivatives that are potent inhibitors of monoacylglycerol lipase (MGL), the primary degrading enzyme for the endocannabinoid 2-arachidonoyl-sn-glycerol (2-AG). Structure-activity relationship studies evaluated various substitutions on the nitrogen atom and the benzene ring of the BTZ nucleus. Optimized derivatives with nanomolar potency allowed us to investigate the mechanism of MGL inhibition. Site-directed mutagenesis and mass spectrometry experiments showed that BTZs interact in a covalent reversible manner with regulatory cysteines, Cys201 and Cys208, causing a reversible sulfenylation known to modulate MGL activity. Metadynamics simulations revealed that BTZ adducts favor a closed conformation of MGL that occludes substrate recruitment. The BTZ derivative 13 protected neuronal cells from oxidative stimuli and increased 2-AG levels in the mouse brain. The results identify Cys201 and Cys208 as key regulators of MGL function and point to the BTZ scaffold as a useful starting point for the discovery of allosteric MGL inhibitors.

U2 - 10.1021/acs.jmedchem.9b01679

DO - 10.1021/acs.jmedchem.9b01679

M3 - Journal article

C2 - 31714779

SN - 0022-2623

JO - Journal of Medicinal Chemistry

JF - Journal of Medicinal Chemistry

ER -

Benzisothiazolinone derivatives as potent allosteric monoacylglycerol lipase inhibitors that functionally mimic sulfenylation of regulatory cysteines

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