Insect ryanodine receptors constitute calcium ion channels, and are the target of novel diamide insecticides containing two types of phthalic and anthranilic diamides. Ryanodine receptors in Periplaneta americana leg muscle were extracted through homogenization and centrifugation. Based on the potential usefulness of a fluorescent probe identified in a previous study, a fluorescence polarization assay was developed with extracted Periplaneta ryanodine receptors, and this technique was used to test the affinity binding of two types of active diamide compounds. Results showed that anthranilic diamides could replace the binding of the fluorescent probe on the ryanodine receptor, but phthalic analogs, including flubendiamide, did not affect the binding of fluorescent probe. Therefore, the developed fluorescence polarization (FP) assay is an easier and more efficient technique to study the affinity binding of small molecules on fluorescent probe binding sites on ryanodine receptors than the [3H] labeled-based assay.

Chlorantraniliprole (CAP) is a new anthranilic diamide insecticide acting with high efficacy on ryanodine receptors of most of the species within the Lepidoptera order and has been used worldwide. To monitor its residue in food and environmental samples, we developed an indirect competitive enzyme-linked immunosorbent assay (icELISA), based on monoclonal antibodies. The established icELISA possesses a concentration of producing 50% inhibition (IC50) of 1.60 ng/mL. In addition, the assay had the average recoveries of 79–119% when CAP was fortified in tap water, soil, spinach (Spinacia oleracea), and rape (Brassica campestris). Moreover, the icELISA results of spinach samples were confirmed by high performance liquid chromatography–mass spectrometry. Overall, we demonstrate that the developed immunoassay could be used as the quick and convenient determination method of CAP residues in environmental and agricultural samples.

Six optically active α-hydroxyl-β,γ-unsaturated acid esters 1a to 1f were synthesised, and they are significant moieties of the cerebrosides. The chiral intermediate alkynol 4 prepared by catalytic asymmetric addition had 99% ee, and which was converted into the target compounds 1a to 1f with high enantiomeric purity.(R,E)-Methyl 2-acetoxytetradec-3-enoateC17H30O4[α]D20=+30.0 (c 0.5, CH2Cl2)Source of chirality: asymmetric catalysisAbsolute configuration: (2R)(R,E)-Methyl 2-acetoxypentadec-3-enoateC18H32O4[α]D20=+33.6 (c 0.5, CH2Cl2)Source of chirality: asymmetric catalysisAbsolute configuration: (2R)(R,E)-Methyl 2-acetoxyhexadec-3-enoateC19H34O4[α]D25=+64.6 (c 0.5, CH2Cl2)Source of chirality: asymmetric catalysisAbsolute configuration: (2R)(R,E)-Methyl 2-acetoxyheptadec-3-enoateC20H36O4[α]D20=+37 (c 0.5, CH2Cl2)Source of chirality: asymmetric catalysisAbsolute configuration: (2R)(R,E)-Methyl 2-acetoxyoctadec-3-enoateC21H38O4[α]D20=+64.5 (c 0.5, CH2Cl2)Source of chirality: asymmetric catalysisAbsolute configuration: (2R)(R,E)-Methyl 2-acetoxynonadec-3-enoateC22H40O4[α]D20=+88.2 (c 0.5, CH2Cl2)Source of chirality: asymmetric catalysisAbsolute configuration: (2R)

The discovery of new safe and effective pesticides is one of the main means of providing eco-friendly agricultural agents for modern crop protection. To identify new biological molecules based of the anthranilic diamide skeleton of the novel pesticide chlorantraniliprole, which acts on the ryanodine receptor and functional groups in acyl hydrazine insect growth regulators, more than 40 new compounds of 2-acylamino-substituted N′-benzylbenzohydrazide derivatives were designed and synthesized. The structures of the new compounds were characterized using 1H nuclear magnetic resonance (NMR), high-resolution mass spectrometry (HRMS), or electron impact mass spectrometry (EI-MS), and their biological activities at a concentration of 600 mg L–1 were determined against cotton aphid (Aphis gossypii Glover), carmine spider mite (Tetranychus cinnabarinus), and diamondback moth (Plutella xylostella). The results of a preliminary assay showed that compounds 6a-I-2 and 6d-III-4 maintained the lethal activity of anthranilic diamide against P. xylostella; compounds 6c-II-4, 6d-I-7, 6d-II-1, and 6d-III-5 exhibited good lethal activity against A. gossypii; and compounds 6a-II-1, 6a-III-1, 6b-I-7, 6c-I-1, and 6c-III-5 retained promising larvicidal activities against T. cinnabarinus. In subsequent further tests against T. cinnabarinus, compounds 6a-II-1, 6a-III-1, 6c-I-1, and 6c-III-5 showed an LC50 value of <90 mg L–1; especially, the LC50 of compound 6a-III-1 was only 27.9 mg L–1. In conclusion, the introduction of the functional fragment-substituted acyl hydrazine improved the acaricidal activity of the anthranilic diamide skeleton, and the halogen atom at X position and the methyl group at R1 play crucial roles in the biological activities of the compounds.

Fluobendiamide is the first commercialized artificial synthetic insecticide acting on the ryanodine receptor. This new molecule possesses a combination of excellent insecticidal activity and eco-friendly characteristics with a skeleton structure of phthalamide. In this study, we incorporated hydrazone, present in many pesticidal compounds reported during the last two decades, into phthalamide derivatives via Schiff-base condensation and aminolysis to obtain 21 new compounds; these compounds were characterized by proton nuclear magnetic resonance (1H NMR), infrared spectroscopy (IR), and high-resolution mass spectrometry (HRMS) or elemental analysis. A preliminary bioassay against peach aphids (Myzus persicae) revealed that the title compounds exhibited good stomach toxicity at 600 mg/L. Twelve new compounds were found to display higher activity than postive control flubendiamide (LC50 = 184.099 mg/L), however, LC50 was less than 100 mg/L only for compounds 4e, 4o, 4s, 4t (59−77 mg/L). That is, combinations of a p-fluorophenyl or (methyl)thienyl group at the Ar position with an isopropyl or cyclohexyl group at the R position might improve the lethality of the designed phthalamide derivative. Preliminary results of a bioassay at 600 mg/L against diamondback moth (Plutella xylostella, Linnaeus) showed that only the title compound 4e possessed good larvicidal activity. On comparison of the bioassay results of stomach toxicity and larvicidal activity, it is noteworthy that the compound incorporating phenylpyrazolyl exhibited good larvicidal activity and poor stomach activity.