ANALYTICAL AND BIOANALYTICAL CHEMISTRY
·
2014
·Vol. 406
·期 28
·7379-7387
·
IF 3.578
Wang Jing,Liu Shu,Ma Bing,Chen Lina,Song Fengrui,Liu Zhiqiang,Liu Chun-ming
Xanthine oxidase (XOD) catalyzes the metabolism of hypoxanthine and xanthine to uric acid, the overproduction of which could cause hyperuricemia, a risk factor for gout. Inhibition of XOD is a major treatment for gout, and biflavonoids have been found to act as XOD-inhibitory compounds. In this study, ultrafiltration liquid chromatography with photodiode-array detection coupled to electrospray-ionization tandem mass spectrometry (UF-LC-PDA–ESI-MS) was used to screen and identify XOD inhibitors from S. tamariscina . High-performance counter-current chromatography (HPCCC) was used to separate and isolate the active constituents of these XOD inhibitors. Furthermore, ultrahigh-performance liquid chromatography (UPLC) and triple-quadrupole mass spectrometry (TQ-MS) was used to determine the XOD-inhibitory activity of the obtained XOD inhibitors, and enzyme kinetics was performed with Lineweaver–Burk (LB) plots using xanthine as the substrate. As a result, two compounds in S. tamariscina were screened as XOD inhibitors: 65.31 mg amentoflavone and 0.76 mg robustaflavone were isolated from approximately 2.5 g S. tamariscina by use of HPCCC. The purities of the two compounds obtained were over 98 % and 95 %, respectively, as determined by high-performance liquid chromatography (HPLC). Lineweaver–Burk plot analysis indicated that amentoflavone and robustaflavone were non-competitive inhibitors of XOD, and the IC
50 values of amentoflavone and robustaflavone for XOD inhibition were 16.26 μg mL −1 (30.22 μmol L −1 ) and 11.98 μg mL −1 (22.27 μmol L −1 ), respectively. The IC
50 value of allopurinol, used as the standard, was 7.49 μg mL −1 (46.23 μmol L −1 ). The results reveal that the method for systematic screening, identification, and isolation of bioactive components in S. tamariscina and for detecting their inhibitory activity using ultrafiltration LC–ESI-MS, HPCCC, and UPLC–TQ-MS is feasible and efficient, and could be expected to extend to screening and separation of other enzyme inhibitors.
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研究领域:
药理学
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分析化学
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DOI
PubMed
高影响因子
BIOMEDICINE & PHARMACOTHERAPY
·
2022
·
IF 7.419
Tianqiao Yong,Danling Liang,Chun Xiao,Longhua Huang,Shaodan Chen,Yizhen Xie,Xiong Gao,Qingping Wu,Huiping Hu,Xiangmin Li,Yuancao Liu,Manjun Cai
In this paper, we reported the hypouricemic effect of 2,4-dihydroxybenzoic acid methyl ester (DAE), a component of Ganoderma applanatum , in hyperuricemic mice through inhibiting XOD and down-regulating URAT1. Computationally, DAE showed a high similarity to allopurinol and depicted a high affinity in docking to XOD. In vitro , DAE exhibited an inhibitory effect against XOD. Importantly, DAE demonstrated a remarkable hypouricemic effect, decreasing serum uric acids (SUAs) of hyperuricemic mice (407 ± 31 μmol/L) to 195 ± 23, 145 ± 33 and 134 ± 16 μmol/L ( P < 0.01) at the doses of 20, 40, and 80 mg/kg with a dose-dependent manner and showing efficacies at 54–68 %, which were close to the efficacies of allopurinol (61 %) and benzbromarone (57 %). DAE depicted higher and negatively dose-independent urinary uric acids in comparison with that of the hyperuricemic control, implying DAE exerted an uricosuric effect and also a reduction effect on uric acid production. Unlike toxic allopurinol and benzbromarone, no general toxicity on body weights and no negative influence on liver, kidney, spleen and thymus were observed for DAE. Mechanistically, DAE inhibited XOD activities in vivo . Moreover, DAE up-regulated OAT1 and down-regulated GLUT9, URAT1 and CNT2. Overall, DAE may present a hypouricemic effect through inhibiting XOD and up-regulating OAT1 and down-regulating GLUT9, URAT1 and CNT2. This work provided novel insights into the hypouricemic effect of DAE and G. applanatum.
研究领域:
药理学
·
生物化学
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分子生物学
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医学
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DOI
PubMed
高影响因子
Food Bioscience
·
2022
·
IF 5.318
Mengfan Hou,Huan Xiang,Xiao Hu,Shengjun Chen,Yanyan Wu,Jiachao Xu,Xianqing Yang
This study aimed to isolate and identify the novel potential Xanthine oxidase (XOD) inhibitory peptides derived from Trachinotus ovatus hydrolysate (TOH) (hydrolysis by neutral protease), and investigate their structure-function relationship. Initially, TOH was separated to four fractions (F1–F4) by gel filtration chromatography and F4 was chosen with higher XOD inhibitory rate (85.16% ± 0.566, 25 mg/mL) and lower molecular weight (95.73%, <1000 Da). F4 was further characterized by reversed-phase high performance liquid chromatography (RP-HPLC) coupled with Nano-HPLC-MS/MS and four peptides (FPAW, LLPW, WLLP and FHLP) were identified with IC 50 value of 3.81 ± 0.18 mM, 4.17 ± 0.12 mM, 43.06 ± 0.73 mM, and over 50.00 mM respectively. It can be concluded that Trp-containing peptides especially located in C-terminal could inhibit XOD effectively. Moreover, the XOD-inhibiting actions of these peptides involved π-π stacking (Phe914 and Phe1009 in XOD), hydrogen bonding via molecular docking . Lineweaver-Burk equation showed the inhibition type of FPAW was a mixed inhibition. Circular dichroism (CD) spectra indicated FPAW could combine and change the secondary structure of XOD. Therefore, two peptides of FPAW and LLPW might have potential to be used to alleviate hyperuricemia in the future.
研究领域:
生物化学
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药理学
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蛋白质组学
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DOI
高影响因子
MOLECULES
·
2024
·Vol. 29
·期 18
·4288
·
IF 4.2
Mingyu Yang,Yitang Xu,Qihua Yu,Mengyu Li,Liyong Yang,Ye Yang
Xanthine oxidase (XOD) is a key enzyme that promotes the oxidation of xanthine/hypoxanthine to form uric acid, and the accumulation of uric acid leads to hyperuricaemia. The prevalence of gout caused by hyperuricaemia is increasing year by year. TAOZHI (TZ) can be used for the treatment of rheumatic arthralgia due to qi stagnation and blood stasis and contains a large number of polyphenolic components. The aim of this study was to investigate the relationship between chromatograms and XOD inhibition of 21 batches of TZ total polyphenol extract samples. Chemometric methods such as grey correlation analysis, bivariate correlation analysis, and partial least squares regression were used to identify the active ingredient groups in the total polyphenol extracts of TZ, which were validated using molecular docking techniques. The total polyphenol content contained in the 21 batches did not differ significantly, and all batches showed inhibitory effects on XOD. Spectroeffect correlation analysis showed that the inhibitory effect of TZ on XOD activity was the result of the synergistic effect of multiple components, and the active component groups screened to inhibit XOD were F2 (4-O-Caffeoylquinic acid), F4, and F10 (naringenin). The molecular docking results showed that the binding energies of all nine dockings were lower than −7.5 kcal/mol, and the binding modes included hydrogen bonding, hydrophobic forces, salt bridges, and π-staking, and the small molecules might exert their pharmacological effects by binding to XOD through the residue sites of the amino acids, such as threonine, arginine, and leucine. This study provides some theoretical basis for the development and utilisation of TZ total polyphenols.
研究领域:
药理学
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生物化学
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计算生物学
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DOI
PubMed
高影响因子
BIOORGANIC CHEMISTRY
·
2024
·
IF 4.5
Fengxin Zheng,Suiqing Mai,Xiaolin Cen,Pei Zhao,Wenjie Ye,Jiale Ke,Shiqin Lin,Huazhong Hu,Zitao Guo,Shuqin Zhang,Hui Liao,Ting Wu,Yuanxin Tian,Qun Zhang,Jianxin Pang,Zean Zhao
The development of XOD/URAT1 dual target inhibitors has emerged as a promising therapeutic strategy for the management of hyperuricemia. Here, through virtual screening, we have identified digallic acid as a novel dual target inhibitor of XOD/URAT1 and subsequently evaluated its pharmacological properties, pharmacokinetics, and toxicities. Digallic acid inhibited URAT1 with an IC 50 of 5.34 ± 0.65 μM, which is less potent than benzbromarone (2.01 ± 0.36 μM) but more potent than lesinurad (10.36 ± 1.23 μM). Docking and mutation analysis indicated that residues S35, F241 and R477 of URAT1 confer a high affinity for digallic acid. Digallic acid inhibited XOD with an IC 50 of 1.04 ± 0.23 μM. Its metabolic product, gallic acid, inhibited XOD with an IC 50 of 0.91 ± 0.14 μM. Enzyme kinetic studies indicated that both digallic acid and gallic acid act as mixed-type XOD inhibitors. It shares the same binding mode as digallic acid, and residues E802, R880, F914, T1010, N768 and F1009 contribute to their high affinity. The anion group (carboxyl) of digallic acid contribute significantly to its inhibition activity on both XOD and URAT1 as indicated by docking analysis. Remarkably, at a dosage of 10 mg/kg in vivo, digallic acid exhibited a stronger urate-lowering and uricosuric effect compared to the positive drug benzbromarone and lesinurad. Pharmacokinetic study indicated that digallic acid can be hydrolyzed into gallic acid in vivo and has a t 1/2 of 0.77 ± 0.10 h. Further toxicity evaluation indicated that digallic acid exhibited no obvious renal toxicity, as reflected by CCK-8, biochemical analysis (CR and BUN) and HE examination. The findings of our study can provide valuable insights for the development of XOD/URAT1 dual target inhibitors, and digallic acid deserves further investigation as a potential anti-hyperuricemic drug.
研究领域:
药理学
·
药物化学
·
药物发现
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DOI
PubMed
高影响因子
PHYTOMEDICINE
·
2022
·
IF 6.656
Tianqiao Yong,Danling Liang,Shaodan Chen,Chun Xiao,Xiong Gao,Qingping Wu,Yizhen Xie,Longhua Huang,Huiping Hu,Xiangmin Li,Yuancao Liu,Manjun Cai
Background Hyperuricemia is characterized with high serum uric acids (SUAs) and directly causes suffering gout. Caffeic acid phenethyl ester (CAPE) is widely included in dietary plants and especially propolis of honey hives. Hypothesis/Purpose Since CAPE exerts a property resembling a redox shuttle, the hypothesis is that it may suppress xanthine oxidase (XOD) and alleviate hyperuricemia. The aim is to unveil the hypouricemic effect of CAPE and the underlying mechanisms. Methods By establishing a hyperuricemic model with potassium oxonate (PO) and hypoxanthine (HX) together, we investigated the hypouricecmic effect of CAPE. On this model, the expressions of key mRNAs and proteins, including glucose transporter 9 (GLUT9) and urate transporter 1 (URAT1), and the activity of XOD were assayed in vivo . Also, the inhibitory effect of CAPE against XOD was assayed in vitro through enzymatic activity tests and by molecular docking. Results CAPE demonstrated a remarkable hypouricemic effect, which reduced the SUAs of hyperuricemic mice (401 ± 111 µmol/l) to 209 ± 56, 204 ± 65 and 154 ± 40 µmol/l ( p < 0.01) at the doses of 15, 30 and 60 mg/kg respectively, depicting efficacies between 48 and 62% and approaching allopurinol's efficacy (52%). Serum parameters, body weights, inner organ coefficients, and H&E staining suggested that CAPE displayed no general toxicity and it alleviated the liver and kidney injuries caused by hyperuricemia. Mechanistically, CAPE decreased XOD activities significantly in vivo , presented an IC 50 at 214.57 µM in vitro and depicted a favorable binding to XOD in molecular simulation, indicating that inhibiting XOD may be an underlying mechanism of CAPE against hyperuricemia. CAPE did decreased GLUT9 protein and down-regulated URAT1 mRNA and protein. In addition, CAPE up-regulated ATP binding cassette subfamily G member 2 (ABCG2) and organic anion transporter 3 (OAT3) mRNA and proteins in comparison with that of the hyperuricemic control. All above, CAPE may alleviate hyperuricmia through inhibiting XOD, decreasing GLUT9 and URAT1 and increasing ABCG2 and OAT3. Conclusion CAPE presented potent hypouricemic effect in hyperuricemic mice through inhibiting XOD activity and up-regulating OAT3. CAPE may be a promising treatment against hyperuricemia.
研究领域:
药理学
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生物化学
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分子生物学
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医学
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DOI
PubMed
高影响因子
COMPOSITES COMMUNICATIONS
·
2023
·
IF 8
Jiangjuan Yuan,Huan Kan,Han Wang,Ningli Wang,Yun Liu,Dong Pei,Qingli Qu
Enzymatic microreactors have gradually become a research hotspot as they can provide an ideal environment for the enzymatic reaction to ensure that the enzymatic reaction can be carried out in an organized manner. Here, inspired by the cellular structure of eukaryotic organisms, we developed Alginate@Chitosan (Alg@CS) microparticles for enzymatic reactions by oil-free gas-shearing method, allowing free access to substrate and product while shielding the enzyme from its surroundings. With the interpenetrating network crosslinked by Alg, CS and Ca 2+ as the substrate, XOD can be encapsulated in particles at the same time of crosslinking. The microreactors are more stable and could provide more active sites for XOD with the addition of CS. The microreactors show excellent recyclability and 4 °C storage stability. This work opens up an intriguing avenue for enzyme microreactor preparation, which is important for the construction of artificial biomimetic catalytic systems.
研究领域:
生物技术
·
材料科学
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化学工程
·
生物化学
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DOI
高影响因子
CHEMICO-BIOLOGICAL INTERACTIONS
·
2024
·
IF 4.7
Yong-jun Chen,Zi-tao Guo,Hai-qiao Chen,Shi-fan Zhang,Ying-xia Bao,Zhoufan Xie,Jia-le Ke,Wen-jie Ye,Jia-cheng Liang,Jia-chen Chen,Ning Li,Feng-xin Zheng,Hui Liao,Ting Wu,Jian-xin Pang
Long-term hyperuricemia can induce kidney damage, clinically referred to as hyperuricemic nephropathy (HN), which is characterized by renal fibrosis, inflammation, and oxidative stress. However, currently used uric acid-lowering drugs are not capable of protecting the kidneys from damage. Therefore, uric acid-lowering drugs that can also protect the kidneys are urgently needed. In this study, we first discovered that salinomycin, an antibiotic, can regulate uric acid homeostasis and ameliorate kidney damage in mice with HN. Mechanistically, salinomycin inhibited serum and hepatic xanthine oxidase (XOD) activities and downregulated renal urate transporter 1 (URAT1) expression and transport activity, thus exerting uric acid-lowering effects in mice with HN. Furthermore, we found that salinomycin promoted p-NRF2 Ser40 expression, resulting in increased nuclear translocation of NRF2 and activation of NRF2. More importantly, salinomycin affected the gut microbiota and promoted the generation of short-chain fatty acids (SCFAs) in mice with HN. In conclusion, our results revealed that salinomycin maintains uric acid homeostasis and alleviates kidney injury in mice with HN by multiple mechanisms, suggesting that salinomycin might be a desirable candidate for HN treatment in the clinic.
研究领域:
药理学
·
肾病学
·
微生物学
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分子生物学
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DOI
PubMed
高影响因子
Journal of Functional Foods
·
2026
·
IF 3.9
Xiqian Tan,Zhiqi Xing,Bolin Zhou,Lijun You,Fangchao Cui,Xuepeng Li,Jianrong Li
Hyperuricemia (HUA) is a metabolic disorder that can trigger gout and cause renal dysfunction. In this study, microbial fermentation with Bacillus amyloliquefaciens Y11 was employed to ferment salmon dark meat. Subsequently, novel peptides with high xanthine oxidase (XOD) inhibitory activity were derived from the fermented samples, purified, and identified. The hypouricemic effect of these peptides was evaluated using an in vitro HK-2 cell HUA model. Five fractions (G1–G5) were isolated via ultrafiltration and Sephadex G15 gel chromatography, and the fractions with higher XOD inhibition rates (G1 and G5) were combined to prepare a mixed fraction. Subsequently, 4122 peptides were identified from the mixed fraction using liquid chromatography with tandem mass spectrometry. Based on predictive bioactivity, toxicity, allergenicity, and solubility scores, five peptides and seven corresponding truncated peptides were selected for synthesis. Their XOD inhibitory activity was assessed, and molecular docking analysis was performed. Ultimately, GEADFMDYGR and its truncated peptide GEADF, which showed high XOD inhibitory activity in vitro (inhibition rates of 90.72 ± 2.85% and 94.06 ± 1.30%, respectively), were screened out. In the HK-2 HUA model, both peptides significantly reduced intracellular uric acid levels. Specifically, 0.5 mg/mL GEADFMDYGR and GEADF decreased the uric acid levels in these cells by 39.70 ± 8.75 and 27.43 ± 3.31 μmol/L, respectively. Further analyses revealed that GEADFMDYGR and GEADF are reversible mixed-type XOD inhibitors. These findings provide new insights and experimental evidence guiding the development of novel therapeutic peptides for HUA.
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高影响因子
BIOMEDICAL CHROMATOGRAPHY
·
2020
·Vol. 34
·期 8
·e4852
·
IF 1.728
Jing Wang,Yong Zhang,Meiqi Zhou,Meizhu Zheng,Jing Cui,Zhiqiang Liu,Chunming Liu,Shu Liu
Superoxide anion radical scavenger and xanthine oxidase inhibitor play an important role in the treatment of several relevant human diseases. In the present study, ultrafiltration liquid chromatography–mass spectrometry coupled to microplate reader was applied to screen and identify superoxide anion radical scavengers and xanthine oxidase inhibitors from total flavonoids of Ginkgo biloba leaves. As a result, four compounds (quercetin, apigenin, kaempferol and isorhamnetin) were screened as xanthine oxidase inhibitors by ultrafiltration LC–MS, and the 50% scavenging concentration values of the screened flavonoids were lower than those for allopurinol. Lineweaver–Burk plot results indicated that kaempferol was a competitive xanthine oxidase inhibitor; the other flavonoids were all anticompetitive inhibitors. Four flavonoids—rutin, quercetin, kaempferol and isorhamnetin—were screened as superoxide anion radical scavengers by LC–MS. The results demonstrate that the method for screening and evaluation of superoxide anion radical scavenger and xanthine oxidase inhibitor from a complex mixture system is feasible and efficient.
研究领域:
药理学
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分析化学
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产品关联
DOI
PubMed
高影响因子
Foods
·
2025
·Vol. 14
·期 3
·524
·
IF 5.1
Zexuan Xu,Miaoyu Gan,Weiliang Guan,Fang Tian,Yuxi Wang,Jinjie Zhang,Luyun Cai
Hyperuricemia arises from imbalanced uric acid metabolism, contributing to gout and related chronic diseases. When traditional drugs are used to treat hyperuricemia, side effects are inevitable, which promotes the exploration of new bioactive compounds. Protein hydrolysates and peptides are gradually showing potential in the treatment of hyperuricemia. This study investigated the uric acid inhibitory activity of peptides extracted fromTrachurus japonicususing in silico and in vitro methods. We employed in silico virtual enzymolysis and experimental validation to identify bioactive peptides fromTrachurus japonicusproteins. Four peptides (DF, AGF, QPSF, and AGDDAPR) were comprehensively screened by molecular docking and database analysis. After solid-phase synthesis, the inhibitory effects of these peptides on hyperuricemia were further verified in vitro and at the cellular level. The results showed that all four peptides have good hyperuricemia-inhibiting activities. Molecular docking and molecular dynamics revealed that peptides DF and AGDDAPR affect the production of uric acid by binding to the active sites of urate transporter 1 (URAT1), glucose transporter 9 (GLUT9), and xanthine oxidase (XOD), while peptides QPSF and AGF mainly influence the XOD active site, confirming that it is feasible to rapidly screen hyperuricemia-inhibiting peptides by molecular docking.
研究领域:
药理学
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生物化学
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分子生物学
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食品科学
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产品关联
DOI
PubMed
高影响因子
Journal of Future Foods
·
2026
·
IF 10.6
Zhi Wei,Pengfei Meng,Yanxin Wang,Xiaolong Chen,Wenjun Li,Jiajie Xu,Kai Liao,Xing Zhang,Chenyang Lu
With rising global hyperuricemia due to lifestyle changes, conventional assays and mouse models faced limitations in technical complexity and cost. In this study, we established a zebrafish model of hyperuricemia through a combined genetic and dietary induction approach. Wild-type zebrafish were treated with the uricase inhibitor potassium oxonate (PO) and the uric acid precursor xanthine, resulting in uric acid levels increasing to approximately 2-fold that of wild-type. To reduce reliance on PO and create a more stable model, we knocked out the uox gene encoding uricase using CRISPR/Cas9. The resulting mutant uox -/- increased the uric acid level to 2.83-fold compared to wild-type zebrafish. Treatment with xanthine further increased the uric acid to 3.79-fold compared to wild-type zebrafish, thereby generating a zebrafish hyperuricemia model with highly elevated uric acid. Subsequently, high-throughput screening of tuna protein hydrolysate identified a dominant peptide, Lys-Asn-Arg-Ala-Lys (KNRAK), predicted to target xanthine oxidase via molecular docking. KNRAK effectively reduced uric acid levels by 74.4% and 29.8% in hyperuricemia models derived from wild-type and uox -/- zebrafish, respectively. Transcriptomic analysis revealed that KNRAK reduced uric acid accumulation in the wild-type derived model by modulating purine metabolism, oxidative stress responses, and urate transport–related pathways, whereas in the uox -/- derived model, KNRAK attenuated NF-κB/MAPK-mediated inflammatory signaling. In summary, this study developed a zebrafish hyperuricemia model with high uric acid levels through combined genetic and dietary induction, and identifies KNRAK as a promising therapeutic peptide with dual mechanisms of action.
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高影响因子
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