PEST MANAGEMENT SCIENCE
·
2018
·Vol. 75
·期 3
·718-725
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IF 3.249
Lu Xu,Jun Zhao,Yang Sun,Dejin Xu,Guangchun Xu,Xiaolong Xu,Yueliang Zhang,Shuijin Huang,Zhaojun Han,Zhongyan Gu
BACKGROUND The rice striped stem borer (SSB), Chilo suppressalis (Walker), which is one of the most economically important phytophagous pests, has developed resistance to multiple insecticides. The resistance of SSB against chlorantraniliprole has been investigated in detail. However, the mechanism of its metabolic resistance has rarely been studied. RESULTS A field population from Wuhu City, China was used to establish chlorantraniliprole resistant and susceptible strains (WHR and WHS) by laboratory continuous selection. Enzyme activities data suggested the potential involvement of cytochrome P450 monooxygenase in WHR. CYP6CV5 , CYP9A68 , CYP321F3 , and CYP324A12 were significantly overexpressed in WHR (from 4.48 to 44.88-fold). These four P450 genes were expressed in the late developmental stages of WHR; however, they were almost absent during the egg stage. In addition, their expressions were much more sensitive to chlorantraniliprole induction in WHR than in WHS. Injection of individual and mixture dsRNAs reduced the expression of the four target genes (55.2–73.2% and 43.2–50.2%, respectively) and caused significant larvae mortality (55.1–65.1% and 88.2%, respectively). CONCLUSION Multiple overexpressed P450 genes were potentially associated with chlorantraniliprole resistance, as confirmed by the RNA interference (RNAi) assay. Our findings suggested that metabolic resistance to chlorantraniliprole might be mediated by P450s. © 2018 Society of Chemical Industry
研究领域:
昆虫学
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分子生物学
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害虫管理
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DOI
PubMed
高影响因子
Frontiers in Veterinary Science
·
2025
·
IF 2.9
Jianping Liang,Can Cui,Wenge Ren,Linglin Li,Linyi Lv,Xianhui Huang,Xiangmei Li
Introduction Tenvermectin (TVM) is a novel avermectin-class drug that has attracted attention for its superior antiparasitic potency, low toxicity, and broad-spectrum activity. However, uncertainty about its interaction with cytochrome P450 enzymes (CYPs) has raised concerns about potential therapeutic failure, increased risk of toxicity, dangerous drug combinations, and prolonged discontinuation periods. Method To address these critical safety concerns, we conducted a systematic comparative study using a highly selective and quantitatively accurate substrate conversion assay to assess and compare the effects of TVM and ivermectin (IVM) on the activities of key CYPs (CYP1A1/2, 2B1, 2C6, 2D2, and 3A1/2). Results TVM induced CYP1A, 2C, 2D and 3A activities only at high therapeutic doses (2.5 mg/kg), and its induction was significantly weaker than that of IVM at all doses tested, with the most pronounced difference for CYP3A1/2. Although TVM had weak inhibitory effects on CYP2B1 and 2D2, at therapeutic concentrations these effects are presumably unlikely to cause clinically significant CYP-mediated drug interactions. Conclusion As the first study to report the effects of TVM on CYP enzyme activity, these findings provide important experimental evidence and a theoretical framework for its clinical safety assessment, development of optimal dosing regimens, and rational polypharmacy strategies.
研究领域:
药理学
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毒理学
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药物代谢
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临床药理学
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寄生虫学
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DOI
PubMed
高影响因子
TOXICOLOGY IN VITRO
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2021
·
IF 3.5
Jing Gao,Yuanjin Zhang,Xueqin Lei,Yuan Xu,Zhenliang Sun,Xin Wang
Hydroxygenkwanin (HGK), a natural flavonoid extracted from the buds of Daphne genkwa Sieb.et Zucc. (Thymelaeaceae), possesses a wide range of pharmacological activities, including anti-inflammatory, antibacterial and anticancer. However, the inhibitory effect of HGK on cytochrome P450 (CYP) remains unclear. This study investigated the potential inhibitory effects of HGK on CYP1A2, 2B1/6, 2C9/11, 2D1/6, 2E1 and 3A2/4 enzymes in human and rat liver microsomes (HLMs and RLMs) by the cocktail approach. HGK exhibited no time-dependent inhibition of CYP activities in HLMs and RLMs. Enzyme inhibition kinetics indicated that HGK was not only a competitive inhibitor of human CYP1A2 and 2C9, but also competitively inhibited rat CYP1A2 and 2C11 activities, with K i value at 0.84 ± 0.03, 8.09 ± 0.44, 2.68 ± 0.32 and 8.35 ± 0.31 μM, respectively. Further studies showed that the inhibitory effect of HGK on CYP enzymes was weaker than that of diosmetin, which may be related to the substitution of hydroxyl and methoxy in the A and B rings of the flavone skeleton. Therefore, the low K i values of HGK for CYP1A2 and 2C may lead to potential drug-drug interactions and toxicity.
研究领域:
药理学
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酶学
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毒理学
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DOI
PubMed
高影响因子
JOURNAL OF ORGANIC CHEMISTRY
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2021
·
IF 4.354
Jiawei Guo,Fengwei Li,Fangyuan Cheng,Li Ma,Xiaohui Liu,Pradeepraj Durairaj,Gang Zhang,Dandan Tang,Xiangtian Long,Wei Zhang,Lei Du,Xingwang Zhang,Shengying Li
Human drug metabolites (HDMs) are important chemicals widely used in drug-related studies. However, acquiring these enzyme-derived and regio-/stereo-selectively modified compounds through chemical approaches is complicated. PikC is a biosynthetic P450 enzyme involved in pikromycin biosynthesis from the bacterium Streptomyces venezuelae. Here, we identify the mutant PikCD50N as a potential biocatalyst, with a broad substrate scope, diversified product profile, and high catalytic efficiency, for preparation of HDMs. Remarkably, PikCD50N can mediate the drug-metabolizing reactions using the low-cost H2O2 as a direct electron and oxygen donor.
研究领域:
生物化学
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药理学
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酶学
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产品关联
DOI
PubMed
高影响因子
Frontiers in Physiology
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2022
·
IF 4.755
Hafeez Muhammad,Li Xiaowei,Ullah Farman,Zhang Zhijun,Zhang Jinming,Huang Jun,Fernández-Grandon G. Mandela,Khan Muhammad Musa,Siddiqui Junaid Ali,Chen Limin,Ren Xiao Yun,Zhou Shuxing,Lou Yonggen,Lu Yaobin
The fall armyworm (FAW), Spodoptera frugiperda (J.E. Smith), is a pest of many important crops globally. Effective control is challenging, with the pest exhibiting resistance to different synthetic pesticides across various groups. However, the mechanisms employed by resistant insects for overexpression of relevant detoxification genes remain unclear. The activity of detoxification enzymes was investigated in this study. Additionally, using RNA interference (RNAi), a functional analysis was completed of two P450s genes in an indoxacarb resistant population of fall armyworms. Elevated resistance levels (resistance ratio = 31.37-fold) in indoxacarb-selected populations of FAW were observed after 14 generations. The qRT-PCR showed higher expression of two cytochrome P450 genes, CYP321A7 and CYP6AE43, in this selected population compared to the control population. RNAi was applied to knock down the P450 dsCYP321A7 and dsCYP6AE43 genes in the FAW larvae. Droplet feeding of the dsRNAs (CYP321A7 and CYP6AE43) via an artificial diet significantly increased mortality rates in the indoxacarb treated population. A shorter larval developmental time of FAW was detected in all dsRNAs-fed larvae. Correspondingly, larval mass was reduced by dsRNAs in indoxacarb resistant populations of fall armyworm. Larval feeding assays demonstrate that dsRNAs targeting, specifically of CYP321A7 and CYP6AE43 enzymes, could be a beneficial technique in the management of indoxacarb resistant populations. Further study on the potential use of dsRNA and its application should be conducted in efforts to counter the development of resistance in FAW against various insecticides in the field.
研究领域:
昆虫学
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分子生物学
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害虫管理
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DOI
PubMed
高影响因子
PHYTOMEDICINE
·
2019
·
IF 4.18
Shuai Ji,Dan-dan He,Zhen-yu Su,Yan Du,Yu-jie Wang,Shi-kai Gao,Meng-zhe Guo,Dao-quan Tang
Background Herbal compatibility of compound formulas can enhance therapeutic effects or reduce side effects of the monarch drugs , but majority of compatibility mechanisms are still unknown. Sangju-Yin, a well-known Chinese compound formula, is currently used to treat common cold in clinical. Purpose In this study, we proposed a strategy to explore the compatibility mechanism of Sangju-Yin by investigating P450 enzymes-based metabolic interactions between monarch drugs and the other constituent herbs. Methods Under the guidance of traditional Chinese medicine theory, the constituent herbs of Sangju-Yin were divided into four groups, including monarch drugs, monarch drugs with addition of minister drugs, monarch drugs with addition of minister and adjuvant drugs, as well as the whole recipe, namely monarch drugs with addition of minister, adjuvant and conductant drugs. Their effects on rats in vivo P450 (CYP1A2, CYP2A3, CYP2C6, CYP2C11 and CYP3A1) activities after oral administration were evaluated using probe drug assay based on LC-MS/MS. Moreover, effects of the four groups of herbs on mRNA expression of P450 enzymes after oral administration, as well as in vitro P450 activities after co-incubation, were investigated to explore the underlying mechanisms. Results Comparing with monarch drugs, addition of different constituent herbs significantly enhanced CYP1A2 and CYP2C6 activities, and inhibited CYP2A3 and CYP3A1 activities, indicating their possible influences on plasma concentrations of active constituents in the monarch drugs. Mechanism study suggested that these herbs affected P450 activities by transcriptional regulation and/or direct interaction with the enzymes. Conclusion This study clarified the compatibility mechanism of Sangju-Yin from the aspect of P450 enzymes-based metabolic interactions, which would benefit better understanding of the therapeutic basis of Sangju-Yin.
研究领域:
中药学
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药物代谢
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分子生物学
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产品关联
DOI
PubMed
高影响因子
PHYTOTHERAPY RESEARCH
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2022
·
IF 6.388
Yuanjin Zhang,Jing Gao,Yuan Xu,Jie Liu,Shengbo Huang,Guihong Li,Bingyi Yao,Zhenliang Sun,Xin Wang
Deoxyshikonin, a natural naphthoquinone compound extracted from Lithospermum erythrorhizon Sieb. et Z ucc (Boraginaceae), has a wide range of pharmacological activities, including anti-tumor, anti-bacterial and wound healing effects. However, the inhibitory effect of deoxyshikonin on cytochrome P450 (CYP) remains unclear. This study investigated the potential inhibitory effects of deoxyshikonin on CYP1A2, 2B1/6, 2C9/11, 2D1/6, 2E1 and 3A2/4 enzymes in human and rat liver microsomes (HLMs and RLMs) by the cocktail approach in vitro . The single-point inactivation experiment showed that deoxyshikonin presented no time-dependent inhibition on CYP activities in HLMs and RLMs. Enzyme inhibition kinetics indicated that in HLMs, deoxyshikonin was not only a competitive inhibitor of CYP1A2 and 2E1, but also a mixed inhibitor of CYP2B6, 2C9, 2D6 and 3A4, with K i of 2.21, 1.78, 1.68, 0.20, 4.08 and 0.44 μM, respectively. In RLMs, deoxyshikonin not only competitively inhibited CYP2B1 and 2E1, but also exhibited mixed inhibition on CYP1A2, 2C11, 2D1 and 3A2, with K i values of no more than 18.66 μM. In conclusion, due to the low K i values of deoxythiokonin on CYP enzymes in HLMs, this may lead to drug–drug interactions (DDI) and potential toxicity.
研究领域:
药理学
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酶学
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毒理学
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DOI
PubMed
高影响因子
JOURNAL OF HAZARDOUS MATERIALS
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2023
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IF 13.6
Shu-Ting Zhang,Tao Li,Shi-Kai Deng,Jim C. Spain,Ning-Yi Zhou
Nitroanisoles are used widely as synthetic intermediates and explosives. Although bacteria have been reported to degrade 4-nitroanisole (4NA) under aerobic conditions , the key enzymes and the catalytic mechanism have remained elusive. Rhodococcus sp. strain JS3073 was isolated for its ability to grow on 4NA as the sole carbon and energy source. In this study, whole cell biotransformation experiments indicated that 4NA degradation is initiated by O -demethylation to form 4-nitrophenol (PNP), which undergoes subsequent degradation by a previously established pathway involving formation of 1,2,4-benzenetriol and release of nitrite. Based on comparative transcriptomics and heterologous expression, a novel three-component cytochrome P450 system encoded by pnaABC initiates the O -demethylation of 4NA to yield formaldehyde and PNP. The pnaABC genes encode a phthalate dioxygenase type reductase (PnaA), a cytochrome P450 monooxygenase (PnaB), and an EthD family protein (PnaC) with putative function similar to ferredoxins. This unusual P450 system also has a broad substrate specificity for nitroanisole derivatives. Sequence analysis of PnaAB revealed high identity with multiple self-sufficient P450s of the CYP116B subfamily. The findings revealed the molecular basis of the catabolic pathway for 4NA initiated by an unusual O -demethylase PnaABC and extends the understanding of the diversity among P450s and their electron transport chains.
研究领域:
微生物学
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环境科学
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分子生物学
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DOI
PubMed
高影响因子
CHEMICAL RESEARCH IN TOXICOLOGY
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2019
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IF 3.274
Xiucai Guo,Wei Li,Qingmei Li,Yan Chen,Guode Zhao,Ying Peng,Jiang Zheng
Tofacitinib (TFT) is an oral JAK inhibitor which has been approved for the treatment of moderately and severely active rheumatoid arthritis. TFT was found to show concentration-, time-, and NADPH-dependent inhibition of CYP3A4, and irreversibility of the inactivation was also observed. Incubation (40 min, 37 °C) of recombinant CYP3A4 with TFT at 200 μM resulted in >70% loss of CYP3A4 activity. Estimated kinact and KI were 0.037 min–1 and 93.2 μM, respectively. GSH and superoxide dismutase/catalase revealed minor or little protection against the CYP3A4 inactivation. Furthermore, ketoconazole attenuated TFT-mediated CYP3A4 inactivation. Epoxide and α-keto-aldehyde intermediates of TFT were trapped and characterized in microsomal incubations, respectively. The aldehyde intermediate is believed to be the key for the enzyme inactivation. Multiple P450 enzymes, including CYPs2C19, 3A4, 2D6, and 1A2, participated in the metabolism of TFT to the epoxide, while the formation of the aldehyde was mainly catalyzed by CYP3A4. In conclusion, TFT was proven to be a mechanism-based inactivator of CYP3A4.
研究领域:
药理学
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药物代谢
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酶学
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产品关联
DOI
PubMed
高影响因子
BIOTECHNOLOGY AND BIOENGINEERING
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2024
·Vol. 122
·期 3
·724-735
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IF 3.5
Xia Ke,Hong-Duo Dong,Xi-Man Zhao,Xin-Xin Wang,Zhi-Qiang Liu,Yu-Guo Zheng
C14-functionalized steroids enabled diverse biological activities in anti-gonadotropin and anticancer therapy. However, access to C14-functionalized steroids was impeded by the deficiency of chemical synthetic methods. Recently, several membrane-bound fungal cytochrome P450s (CYPs) have been identified with steroid C14α-hydroxylation activity. However, the lack of efficient heterologous overexpression strategy hampered their further characterization and molecular engineering. In the present study, sequences of fungi-derived CYP genes encoding putative 14α-hydroxylase were selected and bioinformatically analyzed. Substitution of the N-terminal hydrophobic helix by a soluble maltose binding protein tag significantly enhanced the soluble expression level in Escherichia coli . A novel CYP originated from Bipolaris oryzae was discovered with high steroidal C14α-hydroxylation activity when coupled with the redox partner CPR lun . A catalytically self-sufficient chimeric CYP-CPR was built by intramolecular fusion, and the electronic transfer rate was improved. A coenzyme NADPH regeneration system was finally constructed by the co-expression of glucose dehydrogenase. The developed soluble multi-enzyme cascade biotransformation system supported the selective C14α-hydroxylation toward progesterone with a final titer of 34.54 mg/L, the highest level achieved in E. coli -based heterologous expression system. This study provides insightful ideas on the functional expression of fungi-derived CYPs and promises an efficient C14α-hydroxylation system for steroidal drugs through protein engineering.
研究领域:
生物化学
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分子生物学
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药物开发
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DOI
高影响因子
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
·
2013
·Vol. 14
·期 12
·24211-24229
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IF 2.464
Xueqing Yang,Xianchun Li,Yalin Zhang
Cytochrome P450 monooxygenases (CYPs or P450s) play paramount roles in detoxification of insecticides in a number of insect pests. However, little is known about the roles of P450s and their responses to insecticide exposure in the codling mothCydia pomonella(L.), an economically important fruit pest. Here we report the characterization and expression analysis of the first P450 gene, designated asCYP9A61, from this pest. The full-length cDNA sequence ofCYP9A61is 2071 bp long and its open reading frame (ORF) encodes 538 amino acids. Sequence analysis shows that CYP9A61 shares 51%–60% identity with other known CYP9s and contains the highly conserved substrate recognition site SRS1, SRS4 and SRS5. Quantitative real-time PCR showed thatCYP9A61were 67-fold higher in the fifth instar larvae than in the first instar, and more abundant in the silk gland and fat body than other tissues. Exposure of the 3rd instar larvae to 12.5 mg L−1of chlorpyrifos-ethyl for 60 h and 0.19 mg L−1of lambda-cyhalothrin for 36 h resulted in 2.20-and 3.47-fold induction ofCYP9A61, respectively. Exposure of the 3rd instar larvae to these two insecticides also significantly enhanced the total P450 activity. The results suggested thatCYP9A61is an insecticide-detoxifying P450.Keywords:Cydia pomonella;cytochrome P450;induction;response;insecticides
研究领域:
昆虫学
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分子生物学
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毒理学
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产品关联
DOI
PubMed
高影响因子
PEST MANAGEMENT SCIENCE
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2021
·Vol. 77
·期 9
·4128-4137
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IF 4.845
Li-Xiang Wang,Sha Tao,Yan Zhang,Ya-Long Jia,Shun-Fan Wu,Cong-Fen Gao
BACKGROUND Pymetrozine is commonly used for the control of Nilaparvata lugens , and resistance to pymetrozine has been frequently reported in the field populations in recent years. However, the mechanism of brown planthopper resistance to pymetrozine is still unknown. RESULTS In this study, a pymetrozine-resistant strain (PMR) was established, and the potential biochemical resistance mechanism of N. lugens to pymetrozine was investigated. Pymetrozine was synergized by the inhibitor piperonyl butoxide (PBO) in the PMR with 2.83-fold relative synergistic ratios compared with the susceptible strain (Sus). Compared with the Sus, the cytochrome P450 monooxygenase activity of PMR was increased by 1.7 times, and two P450 genes ( NlCYP6CS1 and NlCYP301B1 ) were found to be significantly overexpressed more than 6.0-fold in the PMR. Pymetrozine exposure induced upregulation of NlCYP6CS1 expression in the Sus, but the expression of NlCYP301B1 did not change significantly. In addition, RNA interference (RNAi)-mediated suppression of NlCYP6CS1 gene expression dramatically increased the toxicity of pymetrozine against N. lugens . Moreover, transgenic lines of Drosophila melanogaster expressing NlCYP6CS1 were less susceptible to pymetrozine, and had a stronger ability to metabolize pymetrozine. CONCLUSIONS Taken together, our findings indicate that the overexpression of NlCYP6CS1 is one of the key factors contributing to pymetrozine resistance in N. lugens . And this result is helpful in proposing a management strategy for pymetrozine resistance.
研究领域:
昆虫学
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分子生物学
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害虫管理
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产品关联
DOI
PubMed
高影响因子
危险品化学品经营许可证（带存储）