Synthesis and Biological Activity Evaluation of Benzoxazinone-Pyrimidinedione Hybrids as Potent Protoporphyrinogen IX Oxidase Inhibitor.

Protoporphyrinogen IX oxidase (PPO/Protox, E.C. 1.3.3.4) is recognized as one of the most important targets for herbicide discovery. In this study, we report our ongoing research efforts toward the discovery of novel PPO inhibitors. Specifically, we identified a highly potent new compound series containing a pyrimidinedione moiety and bearing a versatile building block-benzoxazinone scaffold. Systematic bioassays resulted in the discovery of compound 7af, ethyl 4-(7-fluoro-6-(3-methyl-2,6-dioxo-4-(trifluoromethyl)-3,6-dihydropyrimidin-1(2H)-yl)-3-oxo-2,3-dihydro-4H-benzo[b][1,4]oxazin-4-yl)butanoate, which exhibited broad-spectrum and excellent herbicidal activity at the dosage of 37.5 g a.i./ha through postemergence application. The inhibition constant (Ki) value of 7af to Nicotiana tabacum PPO (NtPPO) was 14 nM, while to human PPO (hPPO), it was 44.8 µM, indicating a selective factor of 3200, making it the most selective PPO inhibitor to date. Moreover, molecular simulations further demonstrated the selectivity and the binding mechanism of 7af to NtPPO and hPPO. This study not only identifies a candidate that showed excellent in vivo bioactivity and high safety toward humans but also provides a paradigm for discovering PPO inhibitors with improved performance through molecular simulation and structure-guided optimization.

Discovery of a Subnanomolar Inhibitor of Protoporphyrinogen IX Oxidase via Fragment-Based Virtual Screening.

Protoporphyrinogen IX oxidase (PPO, E.C. 1.3.3.4), a key functional enzyme existing in various organisms, is acknowledged to be one of the most important action targets in the development of herbicides due to its pivotal roles in chlorophyll and heme biosynthesis pathways. As our persistent research work on the discovery of novel PPO-inhibiting herbicides, a new compound methyl 2-((5-(3-chloro-4,5,6,7-tetrahydro-2H-indazol-2-yl)-6-fluorobenzo[d]thiazol-2-yl)thio)acetate (8aj, Ki = 16 nM) was screened out as a hit compound via a fragment-based virtual screening method performed in the Auto Core Fragment in silico Screening web server. Subsequently, through a fused process of "hit-to-lead" optimization guided by molecular simulation, a total of 30 3-chloro-4,5,6,7-tetrahydro-2H-indazol-benzo[d]thiazole derivatives were synthesized and characterized. The results of the enzymatic inhibition bioassay showed that more than half of the newly synthesized compounds displayed higher activity against Nicotiana tabacum PPO (NtPPO) than oxadiazon, a commercial PPO-inhibiting herbicide. In particular, compound 8ab, a subnanomolar inhibitor with a Ki value of 380 pM against NtPPO, was discovered, which showed to be 71-fold more active than the commercial control oxadiazon (Ki = 27 nM), and was proven to be the most potent PPO inhibitor so far. Furthermore, the greenhouse assay demonstrated that most of the synthetic compounds showed good herbicidal activity toward the tested weeds. Especially, compound 8ad (Ki = 670 pM) showed the most promising post-emergence herbicidal activity with a broad spectrum of weed control even at a concentration as low as 37.5 g a.i./ha and relatively safe to rice at a dosage of 150 g a.i./ha, indicating that 8ad has the greatest potential to be developed as a new herbicide for weed control in paddy fields. This work provides a paradigm for the rational design and discovery of a novel PPO-inhibiting herbicide guided by the fragment-based drug design.

Surface acoustic wave ammonia sensor based on ZnO/SiO2 composite film.

A surface acoustic wave (SAW) resonator with ZnO/SiO2 (ZS) composite film was used as an ammonia sensor in this study. ZS composite films were deposited on the surface of SAW devices using the sol-gel method, and were characterized using SEM, AFM, and XRD. The performance of the sensors under ammonia gas was optimized by adjusting the molar ratio of ZnO:SiO2 to 1:1, 1:2 and 1:3, and the sensor with the ratio of ZnO to SiO2 equaling to 1:2 was found to have the best performance. The response of sensor was 1.132 kHz under 10 ppm NH3, which was much higher than that of the sensor based on a pristine ZnO film. Moreover, the sensor has good selectivity, reversibility and stability at room temperature. These can be attributed to the enhanced absorption of ammonia and unique surface reaction on composite films due to the existence of silica.

Highly sensitive room-temperature surface acoustic wave (SAW) ammonia sensors based on Co3O4/SiO2 composite films.

Surface acoustic wave (SAW) sensors based on Co3O4/SiO2 composite sensing films for ammonia detection were investigated at room temperature. The Co3O4/SiO2 composite films were deposited onto ST-cut quartz SAW resonators by a sol-gel method. SEM and AFM characterizations showed that the films had porous structures. The existence of SiO2 was found to enhance the ammonia sensing property of the sensor significantly. The sensor based on a Co3O4/SiO2 composite film, with 50% Co3O4 loading, which had the highest RMS value (3.72), showed the best sensing property. It exhibited a positive frequency shift of 3500 Hz to 1 ppm ammonia as well as excellent selectivity, stability and reproducibility at room temperature. Moreover, a 37% decrease in the conductance of the composite film as well as a positive frequency shift of 12,500 Hz were observed when the sensor was exposed to 20 ppm ammonia, indicating the positive frequency shift was derived from the decrease in film conductance.