ABSTRACT
4-Hydroxyphenylpyruvate dioxygenase inhibiting herbicides (HIHs) represent a recent class (HRAC group 27) of herbicides that offer many advantages, such as broad-spectrum activity, crop selectivity, and low resistance rates. However, emerging studies have highlighted the potential toxicity of HIHs in the environment. This review aims to provide a comprehensive summary of the toxicity of HIHs toward nontarget organisms, including plants, microorganisms, animals, and humans. Furthermore, the present work discusses the ecological roles of these organisms in the environment and their significance in agriculture. By shedding light on the toxicity of HIHs, this study seeks to raise awareness among end users, including environmentalists, researchers, and farmers, regarding the potential ecological implications of these herbicides. Hopefully, this knowledge can contribute to informed decision-making and sustainable practices in green agriculture and environmental management.
Subject(s)
4-Hydroxyphenylpyruvate Dioxygenase , Herbicides , Herbicides/toxicity , 4-Hydroxyphenylpyruvate Dioxygenase/antagonists & inhibitors , 4-Hydroxyphenylpyruvate Dioxygenase/metabolism , Humans , Animals , Enzyme Inhibitors/toxicity , Plants/drug effectsABSTRACT
Weed resistance is a critical issue in crop production. Among the known herbicides, 4-hydroxyphenylpyruvate dioxygenase (HPPD) inhibitors are crucial for addressing weed resistance. HPPD inhibitors constitute a pivotal aspect of contemporary crop protection strategies. The advantages of these herbicides are their broad weed spectrum, flexible application, and excellent compatibility with other herbicides. They also exhibit satisfactory crop selectivity and low toxicity and are environmentally friendly. An increasing number of new HPPD inhibitors have been designed by combining computer-aided drug design with conventional design approaches. Herein, the molecular design and structural features of innovative HPPD inhibitors are reviewed to guide the development of new HPPD inhibitors possessing an enhanced biological efficacy.
Subject(s)
4-Hydroxyphenylpyruvate Dioxygenase , Drug Design , Enzyme Inhibitors , Herbicides , Plant Weeds , 4-Hydroxyphenylpyruvate Dioxygenase/antagonists & inhibitors , 4-Hydroxyphenylpyruvate Dioxygenase/chemistry , 4-Hydroxyphenylpyruvate Dioxygenase/metabolism , Herbicides/chemistry , Herbicides/pharmacology , Herbicides/chemical synthesis , Enzyme Inhibitors/chemistry , Enzyme Inhibitors/chemical synthesis , Enzyme Inhibitors/pharmacology , Plant Weeds/drug effects , Plant Weeds/enzymology , Plant Proteins/chemistry , Plant Proteins/antagonists & inhibitors , Weed Control , Herbicide Resistance , Structure-Activity Relationship , Molecular StructureABSTRACT
In the risk assessment of agrochemicals, there has been a historical paucity of using data to refine the default adjustment factors, even though large datasets are available to support this. The current state of the science for addressing uncertainty regarding animal to human extrapolation (AFA) is to develop a "data-derived" adjustment factor (DDEF) to quantify such differences, if data are available. Toxicokinetic (TK) and toxicodynamic (TD) differences between species can be utilized for the DDEF, with human datasets being ideal yet rare. We identified a case for a currently registered herbicide, mesotrione, in which human TK and TD are available. This case study outlines an approach for the development of DDEFs using comparative human and animal data and based on an adverse outcome pathway (AOP) for inhibition of 4-hydroxyphenol pyruvate dioxygenase (HHPD). The calculated DDEF for rat to human extrapolation (AFA) for kinetics (AFAK = 2.5) was multiplied by the AFA for dynamics (AFAD = 0.3) resulting in a composite DDEF of â¼1 (AFA = 0.75). This reflects the AOP and available scientific evidence that humans are less sensitive than rats to the effects of HPPD inhibitors. Further analyses were conducted utilizing in vitro datasets from hepatocytes and liver cytosols and extrapolated to whole animal using in vitro to in vivo extrapolation (IVIVE) to support toxicodynamic extrapolation. The in vitro datasets resulted in the same AFAD as derived for in vivo data (AFAD = 0.3). These analyses demonstrate that a majority of the species differences are related to toxicodynamics. Future work with additional in vitro/in vivo datasets for other HPPD inhibitors and cell types will further support this result. This work demonstrates utilization of all available toxicokinetic and toxicodynamic data to replace default uncertainty factors for agrochemical human health risk assessment.
Subject(s)
4-Hydroxyphenylpyruvate Dioxygenase , Cyclohexanones , Humans , Animals , Rats , Cyclohexanones/toxicity , Risk Assessment , 4-Hydroxyphenylpyruvate Dioxygenase/antagonists & inhibitors , Species Specificity , Herbicides/toxicity , Toxicokinetics , Adverse Outcome PathwaysABSTRACT
Tripyrasulfone is currently the only HPPD-inhibiting herbicide that possesses outstanding selectivity even for direct-seeded rice (Oryza sativa) when applied POST to control grass weeds; however, the underlying mechanisms remain unclear. In this study, the inhibitory effects of the real active HDT of tripyrasulfone on recombinant 4-hydroxyphenylpyruvate dioxygenase (HPPDs) from rice and barnyard grass (Echinochloa crus-galli) were similar, with consistent structural interactions and similar binding energies predicted by molecular docking. However, the HPPD expression level in rice was significantly greater than that in barnyard grass after tripyrasulfone treatment. Tripyrasulfone was rapidly taken up and hydrolyzed into HDT, which was similarly distributed within the whole plants of rice and barnyard grass at 24 h after treatment. Compared with barnyard grass, rice has more uniform epicuticular wax in the cuticle of its leaves, absorbing less tripyrasulfone and metabolizing much more tripyrasulfone. Overall, to a greater extent, the different sensitivities to tripyrasulfone between barnyard grass and rice resulted from metabolic variations.
Subject(s)
4-Hydroxyphenylpyruvate Dioxygenase , Echinochloa , Herbicides , Molecular Docking Simulation , Oryza , Plant Proteins , Oryza/metabolism , Oryza/chemistry , Echinochloa/drug effects , Echinochloa/genetics , Echinochloa/metabolism , Echinochloa/growth & development , Echinochloa/chemistry , Herbicides/pharmacology , Herbicides/chemistry , Herbicides/metabolism , Plant Proteins/metabolism , Plant Proteins/genetics , Plant Proteins/chemistry , 4-Hydroxyphenylpyruvate Dioxygenase/metabolism , 4-Hydroxyphenylpyruvate Dioxygenase/antagonists & inhibitors , 4-Hydroxyphenylpyruvate Dioxygenase/genetics , 4-Hydroxyphenylpyruvate Dioxygenase/chemistry , Plant Weeds/drug effects , Plant Weeds/metabolism , Enzyme Inhibitors/pharmacology , Enzyme Inhibitors/chemistryABSTRACT
4-Hydroxyphenylpyruvate dioxygenase (HPPD) is a crucial target enzyme in albino herbicides. The inhibition of HPPD activity interferes with the synthesis of carotenoids, blocking photosynthesis and resulting in bleaching and necrosis. To develop herbicides with excellent activity, a series of 3-hydroxy-2-(6-substituted phenoxynicotinoyl)-2-cyclohexen-1-one derivatives were designed via active substructure combination. The title compounds were characterized via infrared spectroscopy, 1H and 13C nuclear magnetic resonance spectroscopies, and high-resolution mass spectrometry. The structure of compound III-17 was confirmed via single-crystal X-ray diffraction. Preliminary tests demonstrated that some compounds had good herbicidal activity. Crop safety tests revealed that compound III-29 was safer than the commercial herbicide mesotrione in wheat and peanuts. Moreover, the compound exhibited the highest inhibitory activity against Arabidopsis thaliana HPPD (AtHPPD), with a half-maximal inhibitory concentration of 0.19 µM, demonstrating superior activity compared with mesotrione (0.28 µM) in vitro. A three-dimensional quantitative structure-activity relationship study revealed that the introduction of smaller groups to the 5-position of cyclohexanedione and negative charges to the 3-position of the benzene ring enhanced the herbicidal activity. A molecular structure comparison demonstrated that compound III-29 was beneficial to plant absorption and conduction. Molecular docking and molecular dynamics simulations further verified the stability of the complex formed by compound III-29 and AtHPPD. Thus, this study may provide insights into the development of green and efficient herbicides.
Subject(s)
4-Hydroxyphenylpyruvate Dioxygenase , Arabidopsis , Drug Design , Enzyme Inhibitors , Herbicides , 4-Hydroxyphenylpyruvate Dioxygenase/antagonists & inhibitors , Arabidopsis/drug effects , Arabidopsis/enzymology , Arabidopsis Proteins/antagonists & inhibitors , Arabidopsis Proteins/chemistry , Arabidopsis Proteins/metabolism , Cyclohexanones/chemistry , Cyclohexanones/pharmacology , Cyclohexanones/chemical synthesis , Enzyme Inhibitors/chemical synthesis , Enzyme Inhibitors/pharmacology , Herbicides/chemical synthesis , Herbicides/pharmacology , Ketones/chemical synthesis , Ketones/pharmacology , Molecular Docking Simulation , Molecular Structure , Structure-Activity Relationship , Triticum/drug effectsABSTRACT
4-Hydroxyphenylpyruvate dioxygenase (HPPD) inhibitor herbicides are widely used in modern agriculture. Plant root exudates (REs) play an important role in the adsorption, degradation, migration and transformation of pesticides in soil. In the present study, the structural affinity and interaction mechanism between four HPPD inhibitors (HPPDi) and soybean REs were investigated via multispectral technologies and two-dimensional correlation analysis (2D-COS). UV-vis absorption and fluorescence spectra showed that mesotrione, tembotrione, sulcotrione and topramezone effectively quench the intrinsic fluorescence of soybean REs through static quenching. The binding constant Ka revealed that the binding ability of HPPDi to soybean REs takes the following order: mesotrione > tembotrione > sulcotrione > topramezone. According to the thermodynamic parameters, the main interaction force between tembotrione, sulcotrione, topramezone and soybean REs is electrostatic interaction, while the main interaction force is a hydrogen bond or van der Waals force between mesotrione and soybean REs. The conformational changes of REs were attributed to HPPDi by 3D spectral evaluation. FTIR spectroscopy and 2D-COS analysis suggested that soybean REs mainly formed stable complexes with HPPDi through functional groups such as carbonyl, carboxyl, methoxy and nitrate, and the first binding groups were carbonyl and carboxyl. These results provide helpful information for the adsorption and desorption process of environmental pollutants on the surface of plants and soil.
Subject(s)
Herbicides , 4-Hydroxyphenylpyruvate Dioxygenase/antagonists & inhibitors , 4-Hydroxyphenylpyruvate Dioxygenase/metabolism , Exudates and Transudates/metabolism , Herbicides/pharmacology , Herbicides/metabolism , Glycine maxABSTRACT
BACKGROUND: Blood-sucking insects incorporate many times their body weight of blood in a single meal. Because proteins are the major component of vertebrate blood, its digestion in the gut generates extremely high concentrations of free amino acids. Previous reports showed that the tyrosine degradation pathway plays an essential role in adapting these animals to blood feeding. Inhibition of 4-hydroxyphenylpyruvate dioxygenase (HPPD), the rate-limiting step of tyrosine degradation, results in the death of insects after a blood meal. Therefore, it has been suggested that compounds that block the catabolism of tyrosine could act selectively on blood-feeding insects. Here, we evaluated the toxicity against mosquitoes of three HPPD inhibitors currently used as herbicides and in human health. RESULTS: Of the compounds tested, nitisinone (NTBC) proved to be more potent than mesotrione (MES) and isoxaflutole (IFT) in Aedes aegypti. NTBC was lethal to Ae. aegypti in artificial feeding assays [median lethal dose (LD50 ): 4.53 µm] and in topical application (LD50 : 0.012 nmol/mosquito). NTBC was also lethal to Ae. aegypti populations that were resistant to neurotoxic insecticides, and to other mosquito species (Anopheles and Culex). CONCLUSION: HPPD inhibitors, particularly NTBC, represent promising new drugs for mosquito control. Because they affect only blood-feeding organisms, they represent a safer and more environmentally friendly alternative to conventional neurotoxic insecticides. © 2021 Society of Chemical Industry.
Subject(s)
4-Hydroxyphenylpyruvate Dioxygenase , Aedes , Culex , Insecticides , 4-Hydroxyphenylpyruvate Dioxygenase/antagonists & inhibitors , Animals , Humans , Mosquito Control , Mosquito VectorsABSTRACT
The liver plays central roles in coordinating different metabolic processes, such as the catabolism of amino acids. In this study, we identify a loss of tyrosine catabolism and a concomitant increase in serum tyrosine levels during liver cancer development. Liver cells with disordered tyrosine catabolism, as exemplified by the suppression of a tyrosine catabolic enzyme 4-hydroxyphenylpyruvate dioxygenase (HPD), display augmented tumorigenic and proliferative potentials. Metabolomics profiling and isotope tracing reveal the metabolic reliance of HPD-silenced cells on glutamine, coupled with increased tricarboxylic acid cycle metabolites and their associated amino acid pools. Mechanistically, HPD silencing reduces ketone bodies, which regulate the proliferative and metabolic phenotypes via the AMPK/mTOR/p70S6 kinase pathway and mTOR-dependent glutaminase (GLS) activation. Collectively, our results demonstrate a metabolic link between tyrosine and glutamine metabolism, which could be exploited as a potentially promising anticancer therapy for liver cancer.
Subject(s)
4-Hydroxyphenylpyruvate Dioxygenase/antagonists & inhibitors , Glutamine/metabolism , Liver Neoplasms/metabolism , TOR Serine-Threonine Kinases/drug effects , Tyrosine/metabolism , Animals , Cell Line, Tumor , Humans , Liver/metabolism , Liver Neoplasms/drug therapy , Mice, Inbred BALB C , TOR Serine-Threonine Kinases/metabolismABSTRACT
Tsetse transmit African trypanosomiasis, which is a disease fatal to both humans and animals. A vaccine to protect against this disease does not exist so transmission control relies on eliminating tsetse populations. Although neurotoxic insecticides are the gold standard for insect control, they negatively impact the environment and reduce populations of insect pollinator species. Here we present a promising, environment-friendly alternative to current insecticides that targets the insect tyrosine metabolism pathway. A bloodmeal contains high levels of tyrosine, which is toxic to haematophagous insects if it is not degraded and eliminated. RNA interference (RNAi) of either the first two enzymes in the tyrosine degradation pathway (tyrosine aminotransferase (TAT) and 4-hydroxyphenylpyruvate dioxygenase (HPPD)) was lethal to tsetse. Furthermore, nitisinone (NTBC), an FDA-approved tyrosine catabolism inhibitor, killed tsetse regardless if the drug was orally or topically applied. However, oral administration of NTBC to bumblebees did not affect their survival. Using a novel mathematical model, we show that NTBC could reduce the transmission of African trypanosomiasis in sub-Saharan Africa, thus accelerating current disease elimination programmes.
Subject(s)
Cyclohexanones/therapeutic use , Drug Repositioning , Infection Control/methods , Nitrobenzoates/therapeutic use , Trypanosomiasis, African/prevention & control , 4-Hydroxyphenylpyruvate Dioxygenase/antagonists & inhibitors , 4-Hydroxyphenylpyruvate Dioxygenase/metabolism , Animals , Bees/drug effects , Female , Humans , Insecticides/therapeutic use , Male , Metabolome/drug effects , Mice , Models, Theoretical , Neglected Diseases/prevention & control , Orphan Drug Production , Rats , Rats, Wistar , Toxicity Tests , Trypanosomiasis, African/transmission , Tsetse Flies/drug effects , Tsetse Flies/metabolism , Tyrosine/metabolismABSTRACT
4-Hydroxyphenylpyruvate dioxygenase (HPPD, EC 1.13.11.27) has been recognized as one of the most promising targets in the field of herbicide innovation considering the severity of weed resistance currently. In a persistent effort to develop effective HPPD-inhibiting herbicides, a structure-guided strategy was carried out to perform the structural optimization for triketone-quinazoline-2,4-diones, a novel HPPD inhibitor scaffold first discovered in our lab. Herein, starting from the crystal structure of Arabidopsis thaliana (At)HPPD complexed with 6-(2-hydroxy-6-oxocyclohex-1-ene-1-carbonyl)-1,5-dimethyl-3-(o-tolyl)quinazoline-2,4(1H,3H)-dione (MBQ), three subseries of quinazoline-2,4-dione derivatives were designed and prepared by optimizing the hydrophobic interactions between the side chain of the core structure at the R1 position and the hydrophobic pocket at the active site entrance of AtHPPD. 6-(2-Hydroxy-6-oxocyclohex-1-ene-1-carbonyl)-1,5-dimethyl-3-(3-(trimethylsilyl)prop-2-yn-1-yl)quinazoline-2,4(1H,3H)-dione (60) with the best inhibitory activity against AtHPPD was identified to be the first subnanomolar-range AtHPPD inhibitor (Ki = 0.86 nM), which significantly outperformed that of the lead compound MBQ (Ki = 8.2 nM). Further determination of the crystal structure of AtHPPD in complex with compound 60 (1.85 Å) and the binding energy calculation provided a molecular basis for the understanding of its high efficiency. Additionally, the greenhouse assay indicated that 6-(2-hydroxy-6-oxocyclohex-1-ene-1-carbonyl)-1,5-dimethyl-3-propylquinazoline-2,4(1H,3H)-dione (28) and compound 60 showed acceptable crop safety against peanut and good herbicidal activity with a broad spectrum. Moreover, compound 28 also showed superior selectivity for wheat at the dosage of 120 g ai/ha and favorable herbicidal efficacy toward the gramineous weeds at the dosage of as low as 30 g ai/ha. We believe that compounds 28 and 60 have promising prospects as new herbicide candidates for wheat and peanut fields.
Subject(s)
4-Hydroxyphenylpyruvate Dioxygenase/antagonists & inhibitors , Enzyme Inhibitors/chemistry , Enzyme Inhibitors/pharmacology , Herbicides/chemistry , Herbicides/pharmacology , Silicon/chemistry , Silicon/pharmacology , 4-Hydroxyphenylpyruvate Dioxygenase/chemistry , Arabidopsis/chemistry , Arabidopsis/drug effects , Arabidopsis/enzymology , Kinetics , Plant Weeds/drug effects , Plant Weeds/growth & development , Structure-Activity Relationship , Weed ControlABSTRACT
QYM201 is a 4-hydroxyphenylpyruvate dioxygenase (HPPD) inhibiting herbicide recently registered in China for controlling grass and broadleaf weeds in wheat. It is a novel herbicide, and its potential harm to soil ecosystems has not yet been reported. This study investigates the influence of QYM201 on soil enzyme activity and microorganism quantities in two different soils at concentrations of 0.1, 1, and 5 mg kg-1 soil. Results indicate that QYM201 initially inhibited soil protease, urease, and sucrase activity and this effect increased with concentration. During the later stages of incubation, inhibitory effects gradually weakened and by the end of the experiment (45 days), enzyme activity was restored to control levels. Catalase activity was stimulated by QYM201, with significant differences observed between the QYM201-treated groups and the control at the onset of exposure. This stimulation effect decreased during the later stages of the experiment. However, catalase activity was still significantly higher at the end of the experiment compared to the control. The effects of QYM201 on soil microorganisms differed. Initially, bacteria and actinomycetes quantities were decreased by QYM201 (10 days). As the incubation progressed, microorganism quantities in the lower concentration groups (0.1 and 1 mg kg-1 soil) were restored to control levels, while those of the high concentration group (5 mg kg-1 soil) did not fully recover. QYM201 did not significantly impact the quantity of fungi. The half-life and degradation rate constant (k) of QYM201 for the two studied soil types were 23.1 days and 16.1 days, and 0.030 and 0.043 day-1, respectively.
Subject(s)
Enzyme Inhibitors/toxicity , Herbicides , Piperidines/toxicity , Pyrazoles/toxicity , Soil Pollutants , 4-Hydroxyphenylpyruvate Dioxygenase/antagonists & inhibitors , China , Ecosystem , Herbicides/analysis , Herbicides/toxicity , Soil , Soil Microbiology , Soil Pollutants/analysis , Soil Pollutants/toxicityABSTRACT
4-Hydroxyphenylpyruvate dioxygenase (HPPD) has attracted extensive interest as a promising target for the genetic engineering of herbicide-resistant crops. However, naturally occurring HPPDs are generally very sensitive to HPPD inhibitors. In this study, random mutagenesis was performed to increase the HPPD inhibitors' resistance of Sphingobium sp. HPPD (SpHPPD). Two mutants, Q258M and Y333F, with improved resistance were obtained. Subsequently, a double-mutant (Q258M/Y333F) was generated through combined mutation. Q258M/Y333F exhibited the highest resistance to four HPPD inhibitors [topramezone, mesotrione, tembotrione, and diketonitrile (DKN)]. The enzyme fitness of Q258M/Y333F to topramezone, mesotrione, tembotrione, and DKN was increased by 4.0-, 4.1-, 4.2-, and 3.2-folds, respectively, in comparison with that of the wild-type. Molecular modeling and docking revealed that Q258M mutation leads to the decrease of enzyme-inhibitor-binding strength by breaking the hydrogen bond between the enzyme and the inhibitor, and Y333F mutation changes the conformational balance of the C-terminal helix H11, which hinders the binding of the inhibitor to the enzyme and thus would contribute to improved herbicide resistance. This study helps to further elucidate the structural basis for herbicide resistance and provides better genetic resources for the genetic engineering of herbicide-resistant crops.
Subject(s)
4-Hydroxyphenylpyruvate Dioxygenase/chemistry , 4-Hydroxyphenylpyruvate Dioxygenase/genetics , Bacterial Proteins/chemistry , Bacterial Proteins/genetics , Herbicides/chemistry , Sphingomonadaceae/enzymology , 4-Hydroxyphenylpyruvate Dioxygenase/antagonists & inhibitors , 4-Hydroxyphenylpyruvate Dioxygenase/metabolism , Bacterial Proteins/antagonists & inhibitors , Bacterial Proteins/metabolism , Directed Molecular Evolution , Enzyme Inhibitors/chemistry , Herbicide Resistance , Molecular Docking Simulation , Sphingomonadaceae/chemistry , Sphingomonadaceae/geneticsABSTRACT
Topramezone is a new 4-hydroxyphenylpyruvate dioxygenase (HPPD) inhibitor herbicide that is widely used on corn to control annual grass weeds and broadleaf weeds. Due to its broad-spectrum weed control capacity, improved activity, excellent crop selectivity, low mammalian toxicity and high environmental safety, topramezone is considered an ideal target herbicide for transgenic engineering of herbicide tolerance. In this study, a topramezone-resistant strain, Burkholderia sp. BW-1, was screened from soil, and a novel topramezone-resistant HPPD gene (Bkhppd) was cloned from this strain. Purified BkHPPD displayed relatively high HPPD activity and topramezone resistance with a half-maximal inhibitory concentration (IC50) of 572.0â¯nM. Two BkHPPD mutants designated as BkHPPDt31 and BkHPPDt76 were screened through pressure acclimation. BkHPPDt31 contained three amino acid substitutions (H65D, N160â¯T and N258S), whereas BkHPPDt76 contained four amino acid substitutions (H65D, N160â¯T, N258S and N343â¯T). The topramezone IC50 values of BkHPPDt31 and BkHPPDt76 were 1.1- and 2.3-fold higher, respectively, than that of wild-type BkHPPD. In addition, site-directed mutagenesis indicated that the increased resistance conferred by BkHPPDt31 resulted from the synergistic effects of the three site mutations rather than a single site mutation, and that substitution of asparagine 343 with threonine significantly decreased catalytic efficiency and affinity but increased topramezone resistance. In summary, this study provides a novel topramezone-resistant HPPD gene for the engineering of genetically modified herbicide-resistant crops and facilitates further elucidation of the resistance mechanism of BkHPPD and improvement of resistance through directed evolution.
Subject(s)
4-Hydroxyphenylpyruvate Dioxygenase/genetics , Herbicide Resistance/genetics , Herbicides/metabolism , Pyrazoles/metabolism , 4-Hydroxyphenylpyruvate Dioxygenase/antagonists & inhibitors , 4-Hydroxyphenylpyruvate Dioxygenase/metabolism , Acclimatization , Bacterial Proteins/antagonists & inhibitors , Bacterial Proteins/genetics , Bacterial Proteins/metabolism , Burkholderia/drug effects , Burkholderia/enzymology , Burkholderia/genetics , Catalysis , Enzyme Inhibitors/metabolism , Enzyme Inhibitors/pharmacology , Herbicides/pharmacology , Mutagenesis, Site-Directed , Mutation , Pyrazoles/pharmacology , Recombinant Proteins/genetics , Recombinant Proteins/metabolismABSTRACT
4-Hydroxyphenylpyruvate dioxygenase (HPPD) is an iron-dependent non-heme oxygenase involved in the catabolic pathway of tyrosine, which is an important enzyme in the transformation of 4-hydroxyphenylpyruvic acid to homogentisic acid, and thus being considered as herbicide target. Within this study, a set of multiple structure-based pharmacophore models for HPPD inhibitors were developed. The ZINC and natural product database were virtually screened, and 29 compounds were obtained. The binding mode of HPPD and its inhibitors obtained through molecular docking study showed that the residues of Phe424, Phe381, His308, His226, Gln307 and Glu394 were crucial for activity. Molecular-mechanics-generalized born surface area (MM/GBSA) results showed that the coulomb force, lipophilic and van der Waals (vdW) interactions made major contributions to the binding affinity. These efforts will greatly contribute to design novel and effective HPPD inhibitory herbicides.
Subject(s)
4-Hydroxyphenylpyruvate Dioxygenase/antagonists & inhibitors , Enzyme Inhibitors/chemistry , Molecular Structure , User-Computer Interface , 4-Hydroxyphenylpyruvate Dioxygenase/chemistry , Binding Sites , Enzyme Inhibitors/isolation & purification , Enzyme Inhibitors/pharmacology , Herbicides/chemistry , Herbicides/pharmacology , Molecular Docking Simulation , Molecular Dynamics Simulation , Protein Binding , Quantitative Structure-Activity RelationshipABSTRACT
4-Hydroxyphenylpyruvate dioxygenase (HPPD, EC 1.13.11.27) has been identified as one of the most significant targets in herbicide discovery for resistant weed control. In a continuing effort to discover potent novel HPPD inhibitors, we adopted a ring-expansion strategy to design a series of novel pyrazole-quinazoline-2,4-dione hybrids based on the previously discovered pyrazole-isoindoline-1,3-dione scaffold. One compound, 3-(2-chlorophenyl)-6-(5-hydroxy-1,3-dimethyl-1H-pyrazole-4-carbonyl)-1,5-dimethylquinazoline-2,4(1H,3H)-dione (9bj), displayed excellent potency against AtHPPD, with an IC50 value of 84 nM, which is approximately 16-fold more potent than pyrasulfotole (IC50 = 1359 nM) and 2.7-fold more potent than mesotrione (IC50 = 226 nM). Furthermore, the co-crystal structure of the AtHPPD-9bj complex (PDB ID 6LGT) was determined at a resolution of 1.75 Å. Similar to the existing HPPD inhibitors, compound 9bj formed a bidentate chelating interaction with the metal ion and a π-π stacking interaction with Phe381 and Phe424. In contrast, o-chlorophenyl at the N3 position of quinazoline-2,4-dione with a double conformation was surrounded by hydrophobic residues (Met335, Leu368, Leu427, Phe424, Phe392, and Phe381). Remarkably, the greenhouse assay indicated that most compounds displayed excellent herbicidal activity (complete inhibition) against at least one of the tested weeds at the application rate of 150 g of active ingredient (ai)/ha. Most promisingly, compounds 9aj and 9bi not only exhibited prominent weed control effects with a broad spectrum but also showed very good crop safety to cotton, peanuts, and corn at the dose of 150 g of ai/ha.
Subject(s)
4-Hydroxyphenylpyruvate Dioxygenase/antagonists & inhibitors , Enzyme Inhibitors/chemistry , Plant Proteins/antagonists & inhibitors , Plant Weeds/enzymology , Pyrazoles/chemistry , Quinazolines/chemistry , 4-Hydroxyphenylpyruvate Dioxygenase/chemistry , 4-Hydroxyphenylpyruvate Dioxygenase/metabolism , Enzyme Inhibitors/pharmacology , Herbicides/chemistry , Herbicides/pharmacology , Plant Proteins/chemistry , Plant Proteins/metabolism , Plant Weeds/chemistry , Plant Weeds/drug effects , Pyrazoles/pharmacology , Quinazolines/pharmacology , Structure-Activity Relationship , Weed ControlABSTRACT
Aim: 4-Hydroxyphenylpyruvate dioxygenase (HPPD) has attracted increasing attention as an important target against tyrosinemia type I. This paper aimed to explore the structure-activity relationship of HPPD inhibitors with pyrazole scaffolds and to design novel HPPD inhibitors. Methodology & results: The best 3D-quantitative structure-activity relationships model was established by two different strategies based on 40 pyrazole scaffold-based analogs. Screening of molecular fragments by topomer technology, combined with molecular docking, 14 structures were identified for potential human HPPD inhibitory activity. Molecular dynamics results demonstrated that all the compounds obtained bound to the enzyme and possessed a satisfactory binding free energy. Conclusion: The quantitative structure-activity relationship of HPPD inhibitors of pyrazole scaffolds was clarified and 14 original structures with potential human HPPD inhibitory activity were obtained.
Subject(s)
4-Hydroxyphenylpyruvate Dioxygenase/antagonists & inhibitors , Enzyme Inhibitors/pharmacology , Pyrazoles/pharmacology , Quantitative Structure-Activity Relationship , 4-Hydroxyphenylpyruvate Dioxygenase/metabolism , Enzyme Inhibitors/chemical synthesis , Enzyme Inhibitors/chemistry , Humans , Models, Molecular , Molecular Structure , Pyrazoles/chemical synthesis , Pyrazoles/chemistryABSTRACT
Bipyrazone, 1,3-dimethyl-4-(2-(methylsulfonyl)-4-(trifluoromethyl) benzoyl)-1H-pyrazol-5-yl 1,3-dimethyl-1H-pyrazole- 4-carboxylate, is a 4-hydroxyphenylpyaunate dioxygenase (HPPD)-inhibiting herbicide. Greenhouse and field experiments were conducted to explore the potential of post-emergence (POST) application of bipyrazone in wheat fields in China. In the greenhouse study, bipyrazone at 10 and 20 g active ingredient (a.i.) ha-1 effectively controlled Descurainia sophia L., Capsella bursa-pastoris (L.) Medic., Lithospermum arvense L. and Myosoton aquaticum L. Whereas, all tested 16 wheat cultivars showed high degree of tolerance to bipyrazone at 375 and 750 g a.i. ha-1. In a dose-response experiment carried on the Shannong 6 wheat cultivar and five weed biotypes, bipyrazone was safe to the wheat cultivar, and C. bursa-pastoris, M. aquaticum and D. sophia were sensitive to this herbicide. The selectivity index (SI) between the Shannong 6 and weeds ranged from 34 to 39. The field experiments confirmed that a mixture of bipyrazone and fluroxypyr-mepthyl is practical for controlling broadleaf weeds, and bipyrazone applied alone at 30 to 40 g a.i. ha-1 can also provide satisfactory control of sensitive broadleaf weeds. These findings suggest that bipyrazone POST application has good potential for broadleaf weed management in wheat fields.
Subject(s)
4-Hydroxyphenylpyruvate Dioxygenase/antagonists & inhibitors , Herbicides/pharmacology , Pyrazoles/pharmacology , Triticum/growth & development , Crops, Agricultural/drug effects , Crops, Agricultural/enzymology , Crops, Agricultural/growth & development , Greenhouse Effect , Herbicide Resistance , Herbicides/chemistry , Molecular Structure , Plant Proteins/antagonists & inhibitors , Plant Weeds/drug effects , Pyrazoles/chemistry , Triticum/drug effects , Triticum/enzymologyABSTRACT
4-Hydroxyphenylpyruvate dioxygenase (HPPD, EC 1.13.11.27) is an important target site for discovering new bleaching herbicides. To explore novel HPPD inhibitors with excellent herbicidal activity, a series of novel N-aroyl diketone/triketone derivatives were rationally designed by splicing active groups and bioisosterism. Bioassays revealed that most of these derivatives displayed preferable herbicidal activity against Echinochloa crus-galli (EC) at 0.045 mmol/m2 and Abutilon juncea (AJ) at 0.090 mmol/m2. In particular, compound I-f was more potent compared to the commercialized compound mesotrione. Molecular docking indicated that the corresponding active molecules of target compounds and mesotrione shared similar interplay with surrounding residues, which led to a perfect interaction with the active site of Arabidopsis thaliana HPPD.
Subject(s)
4-Hydroxyphenylpyruvate Dioxygenase/antagonists & inhibitors , Enzyme Inhibitors/chemistry , Herbicides/chemistry , Ketones/chemistry , Plant Proteins/antagonists & inhibitors , 4-Hydroxyphenylpyruvate Dioxygenase/chemistry , 4-Hydroxyphenylpyruvate Dioxygenase/metabolism , Catalytic Domain , Echinochloa/drug effects , Echinochloa/enzymology , Enzyme Inhibitors/chemical synthesis , Enzyme Inhibitors/pharmacology , Herbicides/chemical synthesis , Herbicides/pharmacology , Ketones/pharmacology , Malvaceae/drug effects , Malvaceae/enzymology , Molecular Docking Simulation , Plant Proteins/chemistry , Plant Proteins/metabolism , Plant Weeds/drug effects , Plant Weeds/enzymology , Structure-Activity RelationshipABSTRACT
Hereditary tyrosinemia type 1 (HT1) and alkaptonuria (AKU) are inherited metabolic disorders caused by defective enzymes involved in tyrosine catabolism. Nitisinone, an ex-herbicide and member of the ß-triketone family, is therapeutically applied to prevent accumulation of toxic metabolites in patients by inhibiting the enzyme 4-hydroxyphenylpyruvate dioxygenase (HPD). Here, we developed a colorimetric bacterial whole-cell screening system that allows quantifying the inhibitory effects of human HPD inhibitors in a high-throughput and a robust fashion. The principle of our screening system is based on the degradation of tyrosine through 4-hydroxyphenylpyruvate into homogentisate by human HPD expressed in E. coli and subsequent production of a soluble melanin-like pigment. With the aim to optimise the assay, we tested different E. coli strains, expression and reaction temperatures, and time-points for supplementing the substrate. We found that in our assay the addition of prototypical ß-triketone HPD inhibitors decreases pigment production in a dose-dependent manner with increasing inhibitor concentrations. In addition, plate uniformity, signal variability and spatial uniformity assessment showed that we have developed a robust high-throughput screening assay that is simple to use, cost-effective and enables identification and evaluation of novel therapeutic human HPD inhibitors for the treatment of tyrosine-related metabolic disorders.
Subject(s)
4-Hydroxyphenylpyruvate Dioxygenase/antagonists & inhibitors , Enzyme Inhibitors/pharmacology , High-Throughput Screening Assays/methods , Calorimetry/methods , Drug Discovery/methods , Enzyme Inhibitors/chemistry , Escherichia coli , Humans , Melanins/metabolism , Tyrosine/metabolismABSTRACT
Fumarylacetoacetate hydrolase (FAH) is the last enzyme in tyrosine catabolism, and mutations in the FAH gene are associated with hereditary tyrosinemia type I (HT1 or TYRSN1) in humans. In a behavioral screen of N-ethyl-N-nitrosourea mutagenized mice we identified a mutant line which we named "swingshift" (swst, MGI:3611216) with a nonsynonymous point mutation (N68S) in Fah that caused age-dependent disruption of sleep-wake patterns. Mice homozygous for the mutation had an earlier onset of activity (several hours before lights off) and a reduction in total activity and body weight when compared with wild-type or heterozygous mice. Despite abnormal behavioral entrainment to light-dark cycles, there were no differences in the period or phase of the central clock in mutant mice, indicating a defect downstream of the suprachiasmatic nucleus. Interestingly, these behavioral phenotypes became milder as the mice grew older and were completely rescued by the administration of NTBC [2-(2-nitro-4-trifluoromethylbenzoyl)-1,3-cyclohexanedione], an inhibitor of 4-hydroxyphenylpyruvate dioxygenase, which is upstream of FAH. Mechanistically, the swst mutation had no effect on the enzymatic activity of FAH, but rather promoted the degradation of the mutant protein. This led to reduced FAH protein levels and enzymatic activity in the liver and kidney (but not the brain or fibroblasts) of homozygous mice. In addition, plasma tyrosine-but not methionine, phenylalanine, or succinylacetone-increased in homozygous mice, suggesting that swst mutants provide a model of mild, chronic HT1.