Discovering monoacylglycerol lipase inhibitors by a combination of fluorogenic substrate assay and activity-based protein profiling
The endocannabinoid 2-arachidonoylglycerol (2-AG) is predominantly metabolized by monoacylglycerol lipase (MAGL) in the brain. Selective inhibitors of MAGL provide valuable insights into the role of 2-AG in a variety of (patho)physiological processes and are potential therapeutics for the treatment...
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2022-08-01
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| Series: | Frontiers in Pharmacology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fphar.2022.941522/full |
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| author | Hui Deng Hui Deng Qianwen Zhang Qian Lei Qian Lei Na Yang Na Yang Kai Yang Jianbing Jiang Zhiyi Yu |
| author_facet | Hui Deng Hui Deng Qianwen Zhang Qian Lei Qian Lei Na Yang Na Yang Kai Yang Jianbing Jiang Zhiyi Yu |
| author_sort | Hui Deng |
| collection | DOAJ |
| description | The endocannabinoid 2-arachidonoylglycerol (2-AG) is predominantly metabolized by monoacylglycerol lipase (MAGL) in the brain. Selective inhibitors of MAGL provide valuable insights into the role of 2-AG in a variety of (patho)physiological processes and are potential therapeutics for the treatment of diseases such as neurodegenerative disease and inflammation, pain, as well as cancer. Despite a number of MAGL inhibitors been reported, inhibitors with new chemotypes are still required. Here, we developed a substrate-based fluorescence assay by using a new fluorogenic probe AA-HNA and successfully screened a focused library containing 320 natural organic compounds. Furthermore, we applied activity-based protein profiling (ABPP) as an orthogonal method to confirm the inhibitory activity against MAGL in the primary substrate-based screening. Our investigations culminated in the identification of two major compound classes, including quinoid diterpene (23, cryptotanshinone) and β-carbolines (82 and 93, cis- and trans-isomers), with significant potency towards MAGL and good selectivity over other 2-AG hydrolases (ABHD6 and ABHD12). Moreover, these compounds also showed antiproliferative activities against multiple cancer cells, including A431, H1975, B16-F10, OVCAR-3, and A549. Remarkably, 23 achieved complete inhibition towards endogenous MAGL in most cancer cells determined by ABPP. Our results demonstrate the potential utility of the substrate-based fluorescence assay in combination with ABPP for rapidly discovering MAGL inhibitors, as well as providing an effective approach to identify potential targets for compounds with significant biological activities. |
| first_indexed | 2024-04-14T02:17:53Z |
| format | Article |
| id | doaj.art-908871b0b41d41d88cc1c93e87e959cd |
| institution | Directory Open Access Journal |
| issn | 1663-9812 |
| language | English |
| last_indexed | 2024-04-14T02:17:53Z |
| publishDate | 2022-08-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Pharmacology |
| spelling | doaj.art-908871b0b41d41d88cc1c93e87e959cd2022-12-22T02:18:08ZengFrontiers Media S.A.Frontiers in Pharmacology1663-98122022-08-011310.3389/fphar.2022.941522941522Discovering monoacylglycerol lipase inhibitors by a combination of fluorogenic substrate assay and activity-based protein profilingHui Deng0Hui Deng1Qianwen Zhang2Qian Lei3Qian Lei4Na Yang5Na Yang6Kai Yang7Jianbing Jiang8Zhiyi Yu9Department of Respiratory and Critical Care Medicine, West China Hospital, Sichuan University, Chengdu, Sichuan, ChinaTargeted Tracer Research and Development Laboratory, Precision Medicine Key Laboratory of Sichuan Province & Precision Medicine Center, West China Hospital, Sichuan University, Chengdu, Sichuan, ChinaDepartment of Respiratory and Critical Care Medicine, West China Hospital, Sichuan University, Chengdu, Sichuan, ChinaDepartment of Respiratory and Critical Care Medicine, West China Hospital, Sichuan University, Chengdu, Sichuan, ChinaTargeted Tracer Research and Development Laboratory, Precision Medicine Key Laboratory of Sichuan Province & Precision Medicine Center, West China Hospital, Sichuan University, Chengdu, Sichuan, ChinaDepartment of Respiratory and Critical Care Medicine, West China Hospital, Sichuan University, Chengdu, Sichuan, ChinaTargeted Tracer Research and Development Laboratory, Precision Medicine Key Laboratory of Sichuan Province & Precision Medicine Center, West China Hospital, Sichuan University, Chengdu, Sichuan, ChinaDepartment of Obstetrics and Gynecology, West China Second University Hospital, Sichuan University, Chengdu, Sichuan, ChinaDepartment of Medicinal Chemistry, School of Pharmaceutical Sciences, Cheeloo College of Medicine, Shandong University, Jinan, Shandong, ChinaDepartment of Obstetrics and Gynecology, West China Second University Hospital, Sichuan University, Chengdu, Sichuan, ChinaThe endocannabinoid 2-arachidonoylglycerol (2-AG) is predominantly metabolized by monoacylglycerol lipase (MAGL) in the brain. Selective inhibitors of MAGL provide valuable insights into the role of 2-AG in a variety of (patho)physiological processes and are potential therapeutics for the treatment of diseases such as neurodegenerative disease and inflammation, pain, as well as cancer. Despite a number of MAGL inhibitors been reported, inhibitors with new chemotypes are still required. Here, we developed a substrate-based fluorescence assay by using a new fluorogenic probe AA-HNA and successfully screened a focused library containing 320 natural organic compounds. Furthermore, we applied activity-based protein profiling (ABPP) as an orthogonal method to confirm the inhibitory activity against MAGL in the primary substrate-based screening. Our investigations culminated in the identification of two major compound classes, including quinoid diterpene (23, cryptotanshinone) and β-carbolines (82 and 93, cis- and trans-isomers), with significant potency towards MAGL and good selectivity over other 2-AG hydrolases (ABHD6 and ABHD12). Moreover, these compounds also showed antiproliferative activities against multiple cancer cells, including A431, H1975, B16-F10, OVCAR-3, and A549. Remarkably, 23 achieved complete inhibition towards endogenous MAGL in most cancer cells determined by ABPP. Our results demonstrate the potential utility of the substrate-based fluorescence assay in combination with ABPP for rapidly discovering MAGL inhibitors, as well as providing an effective approach to identify potential targets for compounds with significant biological activities.https://www.frontiersin.org/articles/10.3389/fphar.2022.941522/fullmonoacylglycerol lipaseinhibitor discoveryfluorogenic substrate assayactivity-based protein profilinganticancer activity |
| spellingShingle | Hui Deng Hui Deng Qianwen Zhang Qian Lei Qian Lei Na Yang Na Yang Kai Yang Jianbing Jiang Zhiyi Yu Discovering monoacylglycerol lipase inhibitors by a combination of fluorogenic substrate assay and activity-based protein profiling Frontiers in Pharmacology monoacylglycerol lipase inhibitor discovery fluorogenic substrate assay activity-based protein profiling anticancer activity |
| title | Discovering monoacylglycerol lipase inhibitors by a combination of fluorogenic substrate assay and activity-based protein profiling |
| title_full | Discovering monoacylglycerol lipase inhibitors by a combination of fluorogenic substrate assay and activity-based protein profiling |
| title_fullStr | Discovering monoacylglycerol lipase inhibitors by a combination of fluorogenic substrate assay and activity-based protein profiling |
| title_full_unstemmed | Discovering monoacylglycerol lipase inhibitors by a combination of fluorogenic substrate assay and activity-based protein profiling |
| title_short | Discovering monoacylglycerol lipase inhibitors by a combination of fluorogenic substrate assay and activity-based protein profiling |
| title_sort | discovering monoacylglycerol lipase inhibitors by a combination of fluorogenic substrate assay and activity based protein profiling |
| topic | monoacylglycerol lipase inhibitor discovery fluorogenic substrate assay activity-based protein profiling anticancer activity |
| url | https://www.frontiersin.org/articles/10.3389/fphar.2022.941522/full |
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