A novel dehydroabietic acid-based multifunctional fluorescent probe for the detection and bioimaging of Cu2+/Zn2+/ClO.

A multifunctional dehydroabietic acid-based fluorescent probe (CPS) was designed and synthesized by introducing the 2,6-bis(1H-benzo[d]imidazol-2-yl)phenol fluorophore. The probe CPS could selectively recognize Cu2+, Zn2+ and ClO- ions from other analytes, and it showed fluorescence quenching behavior toward Cu2+ and a ratiometric response to Zn2+ and ClO- by changing from green fluorescence to blue and cyan, respectively. The detection limits toward Cu2+, Zn2+ and ClO- ions were 3.8 nM, 0.253 µM and 0.452 µM, respectively. In addition, CPS presented many fascinating merits, such as high selectivity, a short response time (15-20 s), a wide pH range (3-10) and high photostability. The sensing mechanisms of CPS were verified by 1H-NMR, ESI-MS, FT-IR and Job's plot methods. Meanwhile, CPS exhibited satisfactory detection performance in water samples. More importantly, the probe could be applied as a promising tool for visual bioimaging of three ions in living cells and zebrafishes.

Synthesis and Antifungal Activity of Novel L-Menthol Hydrazide Derivatives as Potential Laccase Inhibitors.

To discover novel laccase inhibitors as potential fungicides, twenty-six novel L-menthol hydrazide derivatives were designed and synthesized. In the in vitro antifungal assay, most of the target compounds displayed pronounced antifungal activity against Sclerotinia sclerotiorum, Fusarium graminearum, and Botryosphaeria dothidea. Especially, the EC50 of compounds 3 b and 3 q against B. dothidea was 0.465 and 0.622 mg/L, which was close to the positive compound fluxapyroxad (EC50 =0.322 mg/L). Scanning electron microscopy (SEM) analysis showed that compound 3 b could significantly damage the mycelial morphology of B. dothidea. In vivo antifungal experiments on apple fruits showed that 3 b exhibited excellent protective and curative effects. Furthermore, in the in vitro laccase inhibition assay, 3 b showed outstanding inhibitory activity with the IC50 value of 2.08 µM, which is much stronger than positive control cysteine and PMDD-5Y. These results indicated that this class of L-menthol derivatives could be promising leads for the discovery of laccase-targeting fungicides.

Synthesis and Anti-Fungal/Oomycete Activity of Novel Sulfonamide Derivatives Containing Camphor Scaffold.

Twelve novel camphor sulfonamide derivatives 2a-2l were synthesized and characterized by 1 H-NMR, 13 C-NMR and HRMS spectra. The anti-fungal/oomycete activity bioassay showed that some of the title compounds displayed moderate to good anti-fungal/oomycete activities against B. dothidea and P. capsici. Compound 2d exhibited the best in vitro antifungal activity toward B. dothidea. The in vivo experiment revealed that compound 2d possessed considerable anti-B. dothidea effect at 200 mg/L. Mechanism study showed that compound 2d could increase the cell membrane permeability. In addition, the in vitro enzyme inhibition assay and molecular docking results indicated that compound 2d could be a potential SDH inhibitor.

Synthesis, Antifungal Activity, DFT Study and Molecular Dynamics Simulation of Novel 4-(1,2,4-Oxadiazol-3-yl)-N-(4-phenoxyphenyl)benzamide Derivatives.

In order to find novel potential antifungal agrochemicals, a series of new 4-(1,2,4-oxadiazol-3-yl)-N-(4-phenoxyphenyl)benzamide derivatives 3a-j were designed, synthesized and characterized by their 1 H-, 13 C-NMR and HRMS spectra. The preliminary antifungal assay in vitro revealed that compounds 3a-j exhibited moderate to good antifungal activity against five plant pathogenic fungi. Especially, compound 3e presented significant antifungal activity against Alternaria solani, Botrytis cinerea and Sclerotinia sclerotiorum, superior to positive control boscalid. In the in vivo antifungal assay on tomato plants and cucumber leaves, compound 3e presented good inhibition rate against B. cinerea at 200 mg/L. Molecular dynamics simulation revealed that compound 3e could bind with the active site of class II histone deacetylase (HDAC).

Design, synthesis and antifungal evaluation of novel nopol derivatives as potent laccase inhibitors.

BACKGROUND: To explore further potential natural product-based antifungal agents, a series of novel nopol-based carboxamide and hydrazide derivatives containing a natural pinene structure were designed, synthesized, and evaluated for their inhibitory activities against seven phytopathogenic fungi and oomycetes. RESULTS: The bioassay results indicated that some compounds exhibited good inhibitory activities against Gibberella zeae, Sclerotinia sclerotiorum, and Phytophthora capsici. Among them, compound 3h displayed excellent in vitro activities against G. zeae, with EC50 values of 1.09 mg L-1 , which was comparable with the commercial fungicides bixafen and carbendazim (median effective concentration [EC50 ] = 1.21 and 0.89 mg L-1 , respectively). Notably, in vivo bioassay results suggested that compound 3h also showed prominent protective and curative effects (95.6% and 94.2%) at 200 mg L-1 against G. zeae. The scanning electron microscopy study indicated that compound 3h could destroy the morphological integrity of G. zeae hyphae. The in vitro enzyme inhibitory bioassay revealed that compound 3h exhibited potent inhibitory activity against laccase with median inhibitory concentration (IC50 ) values of 4.93 µm, superior to positive control cysteine (IC50  = 35.50 µm), and its binding modes with laccase were elucidated by molecular docking study. In addition, the fluorescent imaging of the dansylamide-labeled derivatives 8 on wheat leaf epidermal cells and the hyphae of G. zeae revealed that this class of hydrazide derivatives could readily permeate into wheat leaves and reached the laccase target in fungal cells. CONCLUSION: Some nopol-based hydrazide derivatives exhibited excellent anti-G. zeae activity and laccase inhibitory activity, which merits further development as a new fungicide candidate for controlling Fusarium head blight. © 2023 Society of Chemical Industry.

Novel Camphor Sulfonohydrazide and Sulfonamide Derivatives as Potential Succinate Dehydrogenase Inhibitors against Phytopathogenic Fungi/Oomycetes.

To discover novel fungicidal agrochemicals for treating wheat scab, 39 novel camphor sulfonohydrazide/sulfonamide derivatives 4a-4t and 6a-6s were designed and synthesized. In the in vitro antifungal/antioomycete assay, compounds 4g, 4n, and 4o displayed significant inhibitory activities against Fusarium graminearum, Botryosphaeria dothidea, and Phytophthora capsici. Among them, 4n exhibited the best antifungal activity against F. graminearum with an EC50 value of 0.41 mg/L, which was at the same level as that of pydiflumetofen. The in vivo experiment revealed that 4n presented excellent protective and curative efficacy toward F. graminearum. In the antifungal mechanism study, 4n could increase the cell membrane permeability and reduce the exopolysaccharide and ergosterol content of F. graminearum. Scanning electron microscopy (SEM) and transmission electron microscopy (TEM) analyses revealed that 4n could significantly damage the surface morphology and the cell ultrastructure of mycelia to interfere with the growth of F. graminearum. Furthermore, 4n exhibited potent succinate dehydrogenase (SDH) inhibitory activity in vitro with an IC50 value of 3.94 µM, which was equipotent to pydiflumetofen (IC50 = 4.07 µM). The molecular dynamics simulation and docking study suggested that compound 4n could well occupy the active site and form strong interactions with the key residues of SDH. The above-mentioned results demonstrated that the title camphor sulfonohydrazide/sulfonamide derivatives could be promising lead compounds for further succinate dehydrogenase inhibitor (SDHI) fungicide development.

Design, Synthesis, and Biological Evaluation of Novel Camphor-Based Hydrazide and Sulfonamide Derivatives as Laccase Inhibitors against Plant Pathogenic Fungi/Oomycetes.

To discover novel natural product-based fungicidal agrochemicals, 41 novel camphanic acid hydrazide and camphor sulfonamide derivatives were designed, synthesized, and tested for their antifungal profile against four plant pathogenic fungi and three oomycetes. As a result, some derivatives presented pronounced inhibitory activities toward Botryosphaeria dothidea, Fusarium graminearum, Phytophthora capsici, and Phytophthora nicotianae. Especially, compound 4b demonstrated the most potent anti-B. dothidea activity (EC50 = 1.28 mg/L), much stronger than positive control chlorthalonil. The in vivo assay showed that 4b displayed significant protective and curative effects on apple fruits infected by B. dothidea. The primary antifungal mechanism study revealed that 4b could obviously enhance the cell membrane permeability, destroy the mycelial surface morphology and the cell ultrastructure, and reduce the ergosterol and exopolysaccharide contents of B. dothidea. Further, 4b showed potent laccase inhibitory activity in vitro with an IC50 value of 11.3 µM, superior to positive control cysteine. The molecular docking study revealed that 4b could dock well into the active site of laccase by forming multiple interactions with the key residues in the pocket. The acute oral toxicity test in rats presented that 4b had slight toxicity with an LD50 value of 849.1 mg/kg bw (95% confidence limit: 403.9-1785.3 mg/kg bw). This research identified that the camphanic acid hydrazide derivatives could be promising leads for the development of novel laccase-targeting fungicides.

Design, Synthesis, and Fungicidal Evaluation of Novel 1,3-Benzodioxole-Pyrimidine Derivatives as Potential Succinate Dehydrogenase Inhibitors.

A series of novel 1,3-benzodioxole-pyrimidine derivatives were designed and synthesized. The in vitro bioassay indicated that compounds 4e, 4g, 4n, 5c, and 5e displayed excellent fungicidal activities against test fungal strains. Especially, in the in vitro experiments, 5c exhibited a broad spectrum of fungicidal activity against Botrytis cinerea, Rhizoctonia solani, Fusarium oxysporum, Alternaria solani, and Gibberella zeae with EC50 values of 0.44, 6.96, 6.99, 0.07, and 0.57 mg/L, respectively, which were significantly more potent than those of positive control boscalid (EC50: 5.02, >50, >50, 0.16, and 1.28 mg/L). In vivo testing on tomato fruits and leaves showed that 5c displayed considerable protective and curative efficacy against A. solani. Scanning electron microscopy analysis indicated that 5c possessed a strong ability to destroy the surface morphology of mycelia and seriously interfere with the growth of the fungal pathogen. In the in vitro enzyme inhibition assay, 5c exhibited pronounced succinate dehydrogenase (SDH) inhibitory activity with an IC50 value of 3.41 µM, equivalent to that of boscalid (IC50: 3.40 µM). In addition, fluorescence quenching experiment further confirmed the strong interaction of 5c with SDH. Through chiral resolution, 5c was separated into two enantiomers. Among them, (S)-5c exhibited stronger fungicidal activity (EC50: 0.06 mg/L) and SDH inhibitory (2.92 µM) activity than the R-enantiomer (EC50: 0.17 mg/L and SDH IC50: 3.68 µM), which was in accordance with the molecular docking study (CDOCKER Interaction Energy for (R)-5c and (S)-5c: -28.23 and -29.98 kcal/mol, respectively). These results presented a promising lead for the discovery of novel SDHIs as antifungal pesticides.