Qiangen Wu, Jia-Long Fang, Suresh K. Nagumalli, Xiaoqing Guo, Frederick A. Beland
{"title":"NAD(P)+依赖的醇氧化还原酶将7-羟基大麻二酚氧化为活性甲酰基代谢物。","authors":"Qiangen Wu, Jia-Long Fang, Suresh K. Nagumalli, Xiaoqing Guo, Frederick A. Beland","doi":"10.1007/s00204-025-04140-x","DOIUrl":null,"url":null,"abstract":"<div><p>Cannabidiol (CBD) undergoes oxidation to 7-hydroxy-CBD in the liver via cytochrome P450 enzymes. 7-Hydroxy-CBD can be further oxidized to 7-carboxy-CBD, the principal circulating metabolite in humans. An aldehyde intermediate, 7-formyl-CBD, is hypothesized to be the precursor of 7-carboxy-CBD; however, the formation of 7-formyl-CBD and the enzymes leading to its formation and metabolism have not been thoroughly investigated. Upon incubating 7-hydroxy-CBD with human liver S9 or microsomes, and using <i>O</i>-(2,3,4,5,6-pentafluorobenzyl)hydroxylamine (PFBHA) as a trapping agent, we demonstrated the formation of 7-formyl-CBD as its oxime derivative 7-pentafluorobenzyl oxime-CBD (7-PFBO-CBD). The transformation of 7-hydroxy-CBD to 7-formyl-CBD in S9 or microsomes required NAD<sup>+</sup> or NADP<sup>+</sup>. Trapping 7-formyl-CBD with PFBHA decreased the formation of 7-carboxy-CBD, indicating that the formation of 7-carboxy-CBD depends on the availability of 7-formyl-CBD. The flavonoid kaempferol, which inhibits xanthine oxidoreductase and hydroxysteroid dehydrogenases, suppressed the formation of 7-PFBO-CBD and 7-carboxy-CBD in human liver S9 and microsomes incubated with 7-hydroxy-CBD. The xanthine oxidoreductase inhibitor allopurinol and its substrate xanthine did not affect the metabolism of 7-hydroxy-CBD. The hydroxysteroids estradiol and dehydroepiandrosterone reduced the formation of 7-carboxy-CBD in liver microsomes and S9. These results suggest that alcohol oxidoreductases associated with hydroxysteroid metabolism may play a role in the conversion of 7-hydroxy-CBD to 7-formyl-CBD. The irreversible aldehyde dehydrogenase inhibitors, disulfiram and WIN 18,446, inhibited the formation of 7-carboxy-CBD and led to a significant accumulation of 7-PFBO-CBD in the S9 and microsomal incubations. The accumulation of the aldehyde intermediate may play a role in the enhanced cytotoxicity of 7-hydroxy-CBD in HepG2 cells when co-treated with disulfiram. These findings indicate that NAD(P)<sup>+</sup>-dependent alcohol oxidoreductases may catalyze the conversion of 7-hydroxy-CBD to 7-formyl-CBD in human liver S9 and microsomes. The potentially toxic aldehyde intermediate is further oxidized by aldehyde dehydrogenase to 7-carboxy-CBD.</p></div>","PeriodicalId":8329,"journal":{"name":"Archives of Toxicology","volume":"99 11","pages":"4411 - 4421"},"PeriodicalIF":6.9000,"publicationDate":"2025-07-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"NAD(P)+-dependent alcohol oxidoreductases oxidize 7-hydroxycannabidiol to a reactive formyl metabolite\",\"authors\":\"Qiangen Wu, Jia-Long Fang, Suresh K. Nagumalli, Xiaoqing Guo, Frederick A. Beland\",\"doi\":\"10.1007/s00204-025-04140-x\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<div><p>Cannabidiol (CBD) undergoes oxidation to 7-hydroxy-CBD in the liver via cytochrome P450 enzymes. 7-Hydroxy-CBD can be further oxidized to 7-carboxy-CBD, the principal circulating metabolite in humans. An aldehyde intermediate, 7-formyl-CBD, is hypothesized to be the precursor of 7-carboxy-CBD; however, the formation of 7-formyl-CBD and the enzymes leading to its formation and metabolism have not been thoroughly investigated. Upon incubating 7-hydroxy-CBD with human liver S9 or microsomes, and using <i>O</i>-(2,3,4,5,6-pentafluorobenzyl)hydroxylamine (PFBHA) as a trapping agent, we demonstrated the formation of 7-formyl-CBD as its oxime derivative 7-pentafluorobenzyl oxime-CBD (7-PFBO-CBD). The transformation of 7-hydroxy-CBD to 7-formyl-CBD in S9 or microsomes required NAD<sup>+</sup> or NADP<sup>+</sup>. Trapping 7-formyl-CBD with PFBHA decreased the formation of 7-carboxy-CBD, indicating that the formation of 7-carboxy-CBD depends on the availability of 7-formyl-CBD. The flavonoid kaempferol, which inhibits xanthine oxidoreductase and hydroxysteroid dehydrogenases, suppressed the formation of 7-PFBO-CBD and 7-carboxy-CBD in human liver S9 and microsomes incubated with 7-hydroxy-CBD. The xanthine oxidoreductase inhibitor allopurinol and its substrate xanthine did not affect the metabolism of 7-hydroxy-CBD. The hydroxysteroids estradiol and dehydroepiandrosterone reduced the formation of 7-carboxy-CBD in liver microsomes and S9. These results suggest that alcohol oxidoreductases associated with hydroxysteroid metabolism may play a role in the conversion of 7-hydroxy-CBD to 7-formyl-CBD. The irreversible aldehyde dehydrogenase inhibitors, disulfiram and WIN 18,446, inhibited the formation of 7-carboxy-CBD and led to a significant accumulation of 7-PFBO-CBD in the S9 and microsomal incubations. The accumulation of the aldehyde intermediate may play a role in the enhanced cytotoxicity of 7-hydroxy-CBD in HepG2 cells when co-treated with disulfiram. These findings indicate that NAD(P)<sup>+</sup>-dependent alcohol oxidoreductases may catalyze the conversion of 7-hydroxy-CBD to 7-formyl-CBD in human liver S9 and microsomes. The potentially toxic aldehyde intermediate is further oxidized by aldehyde dehydrogenase to 7-carboxy-CBD.</p></div>\",\"PeriodicalId\":8329,\"journal\":{\"name\":\"Archives of Toxicology\",\"volume\":\"99 11\",\"pages\":\"4411 - 4421\"},\"PeriodicalIF\":6.9000,\"publicationDate\":\"2025-07-30\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Archives of Toxicology\",\"FirstCategoryId\":\"3\",\"ListUrlMain\":\"https://link.springer.com/article/10.1007/s00204-025-04140-x\",\"RegionNum\":2,\"RegionCategory\":\"医学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q1\",\"JCRName\":\"TOXICOLOGY\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Archives of Toxicology","FirstCategoryId":"3","ListUrlMain":"https://link.springer.com/article/10.1007/s00204-025-04140-x","RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q1","JCRName":"TOXICOLOGY","Score":null,"Total":0}
NAD(P)+-dependent alcohol oxidoreductases oxidize 7-hydroxycannabidiol to a reactive formyl metabolite
Cannabidiol (CBD) undergoes oxidation to 7-hydroxy-CBD in the liver via cytochrome P450 enzymes. 7-Hydroxy-CBD can be further oxidized to 7-carboxy-CBD, the principal circulating metabolite in humans. An aldehyde intermediate, 7-formyl-CBD, is hypothesized to be the precursor of 7-carboxy-CBD; however, the formation of 7-formyl-CBD and the enzymes leading to its formation and metabolism have not been thoroughly investigated. Upon incubating 7-hydroxy-CBD with human liver S9 or microsomes, and using O-(2,3,4,5,6-pentafluorobenzyl)hydroxylamine (PFBHA) as a trapping agent, we demonstrated the formation of 7-formyl-CBD as its oxime derivative 7-pentafluorobenzyl oxime-CBD (7-PFBO-CBD). The transformation of 7-hydroxy-CBD to 7-formyl-CBD in S9 or microsomes required NAD+ or NADP+. Trapping 7-formyl-CBD with PFBHA decreased the formation of 7-carboxy-CBD, indicating that the formation of 7-carboxy-CBD depends on the availability of 7-formyl-CBD. The flavonoid kaempferol, which inhibits xanthine oxidoreductase and hydroxysteroid dehydrogenases, suppressed the formation of 7-PFBO-CBD and 7-carboxy-CBD in human liver S9 and microsomes incubated with 7-hydroxy-CBD. The xanthine oxidoreductase inhibitor allopurinol and its substrate xanthine did not affect the metabolism of 7-hydroxy-CBD. The hydroxysteroids estradiol and dehydroepiandrosterone reduced the formation of 7-carboxy-CBD in liver microsomes and S9. These results suggest that alcohol oxidoreductases associated with hydroxysteroid metabolism may play a role in the conversion of 7-hydroxy-CBD to 7-formyl-CBD. The irreversible aldehyde dehydrogenase inhibitors, disulfiram and WIN 18,446, inhibited the formation of 7-carboxy-CBD and led to a significant accumulation of 7-PFBO-CBD in the S9 and microsomal incubations. The accumulation of the aldehyde intermediate may play a role in the enhanced cytotoxicity of 7-hydroxy-CBD in HepG2 cells when co-treated with disulfiram. These findings indicate that NAD(P)+-dependent alcohol oxidoreductases may catalyze the conversion of 7-hydroxy-CBD to 7-formyl-CBD in human liver S9 and microsomes. The potentially toxic aldehyde intermediate is further oxidized by aldehyde dehydrogenase to 7-carboxy-CBD.
期刊介绍：
Archives of Toxicology provides up-to-date information on the latest advances in toxicology. The journal places particular emphasis on studies relating to defined effects of chemicals and mechanisms of toxicity, including toxic activities at the molecular level, in humans and experimental animals. Coverage includes new insights into analysis and toxicokinetics and into forensic toxicology. Review articles of general interest to toxicologists are an additional important feature of the journal.
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