Dimethyl Sulfoxide Provides Three Different Units in Synthesis of Chroman-4-ones Containing Sulfur and a Quaternary Carbon Center under HOAc Conditions.

HOAc-promoted construction of chroman-4-ones with a sulfur atom and an α-carbonyl quaternary carbon center directly from ortho-hydroxyacetophenones and DMSO is described. In these unique reactions, DMSO is activated by HOAc and provides three different units (CH2, CH2OH, and CH2SMe) in the target molecules. This reaction displays good substrate scope and reaction yields with a series of substitutes. The mechanism showed that the three units were formed in sequential order.

Structural modification of (3E)-4,8-dimethyl-1,3,7-nontriene enhances its ability to kill Plutella xylostella insect pests.

BACKGROUND: Plant secondary metabolites and their modified derivatives play an important role in the discovery and development of novel insecticides. The natural plant product (3E)-4,8-dimethyl-1,3,7-nontriene (DMNT) has been proven to be able to effectively repel and kill the lepidopteran insect pest Plutella xylostella. RESULTS: In this study, four oxygenated derivatives of DMNT were synthesized by allylic hydroxylation and subsequent etherification or esterification. Bioassays on P. xylostella larvae showed that the compounds DMNT-OCH3 (2), DMNT-OCy (3) and DMNT-OAc (4) were more toxic to the larvae than DMNT alone. The most pronounced effect was observed for compound 2, which showed a 22.23% increase in lethality at a concentration of 0.25 µm. Moreover, the peritrophic matrix (PM) barrier in the insect midgut was more severely damaged by compounds 2, 3 and 4 than by DMNT. The median lethal concentration (LC50 , 48 h) of compounds 2, 3 and 4 on P. xylostella was determined to be 0.98, 1.13 and 1.11 mg mL-1 , respectively, which is much lower than the commercial insecticides eucalyptol (2.89 mg mL-1 ) and thymol (2.45 mg mL-1 ). CONCLUSION: These results suggested that oxygenated DMNT derivatives offer a significantly improved killing effect over DMNT on P. xylostella. This work has provided a basis for further design, structural modification and development of DMNT as botanical insecticides. © 2023 Society of Chemical Industry.

Structural characterization, derivatization and antibacterial activity of secondary metabolites produced by termite-associated Streptomyces showdoensis BYF17.

BACKGROUND: Insect-associated Streptomyces is a valuable resource for development of compounds with antibacterial potential. However, relatively little is known of the secondary metabolites produced by termite-associated Streptomyces. RESULTS: Here, seven compounds including o-acetaminophenol (1), phenazine-1,6-dicarboxylic acid (2), phenylacetic acid (3), phenazinolin D (4), izumiphenazine A (5), izumiphenazine B (6) and phenazinolin E (7) were obtained from the fermentation broth of a termite-associated Streptomyces showdoensis BYF17, which was isolated from the body surfaces of Odontotermes formosanus. Two additional novel derivative compounds (6a and 6b) were synthesized via acetylation and methylation, respectively. The structures of these compounds were elucidated by spectroscopic analyses. The antibacterial bioassay showed that compound 6a displayed strong inhibitory effects against Pseudomonas syringae pv. actinidiae (Psa), with a zone of inhibition (ZOI) diameter of 20.6 mm, which was comparable to that of positive gentamicin sulfate with a ZOI value of 25.6 mm. Furthermore, the Day 5 curative activities of both compounds 6 and 6a against kiwifruit bacterial canker were 71.5%, which was higher than those of referred oxine-copper (55.0%) and ethylicin (46.8%) at a concentration of 200 µg mL-1 . In addition, the mechanism analysis based on scanning electron microscopic observation revealed that both compounds 6 and 6a destroyed the integrity of the Psa cell membrane. CONCLUSION: The results of biological tests showed that these bioactive compounds exhibit potent antimicrobial activities, which have the potential to be developed into new antibacterial agents. © 2023 Society of Chemical Industry.

Bis-iodine-labeled Curcumin as a Potential CT Imaging Agent for ß-amyloid Plaques in the Brain.

BACKGROUND: Alzheimer's disease (AD) is one of the most common causes of dementia, affecting many old people. OBJECTIVES: By designing and synthesizing intracerebral imaging probes, we tried to provide a new solution for the early diagnosis of AD. METHODS: We designed and synthesized bis-iodine-labeled curcumin, and verified its performance through in vivo and in vitro experiments. RESULTS: In this study, bis-iodine-labeled curcumin (7, BICUR) was synthesized. In the in vitro mass spectrum binding assay, Kd values of BICUR with Aß1-40 and Aß1-42 aggregates were 46.29 nM and 64.29 nM, respectively. Aß plaques in AD brain adjacent sections were positively stained by BICUR, which was similar to some other curcumin derivatives. The Log P value of BICUR was 1.45. In the biodistribution experiment, BICUR showed the highest initial brain uptake (5.87% compared to the blood concentration) two minutes after the tail vein injection and rapid clearance from the mouse brain. In the acute toxicity experiment, BICUR showed low toxicity, and the LD50 was >100 mg/kg. Moreover, BICUR showed a high stability in vitro (86.68% unchanged BICUR after incubation for 120min in mouse brain homogenate). Besides, BICUR produced an enhanced CT imaging effect that could be sensitively detected in vitro, but it also showed an obvious differentiation from surrounding tissues after intracerebral injection. CONCLUSION: All results suggested that BICUR could probably act as a targeted CT imaging agent for Aß plaques in the brain.

Inactivation of RPX1 in Arabidopsis confers resistance to Plutella xylostella through the accumulation of the homoterpene DMNT.

The lepidopteran crop pest Plutella xylostella causes severe constraints on Brassica cultivation. Here, we report a novel role for RPX1 (resistance to P. xylostella) in resistance to this pest in Arabidopsis thaliana. The rpx1-1 mutant repels P. xylostella larvae, and feeding on the rpx1-1 mutant severely damages the peritrophic matrix structure in the midgut of the larvae, thereby negatively affecting larval growth and pupation. This resistance results from the accumulation of defence compounds, including the homoterpene (3E)-4,8-dimethyl-1,3,7-nonatriene (DMNT), due to the upregulation of PENTACYCLIC TRITERPENE SYNTHASE 1 (PEN1), which encodes a key DMNT biosynthetic enzyme. P. xylostella infestation and wounding induce RPX1 protein degradation, which may confer a rapid response to insect infestation. RPX1 inactivation and PEN1 overexpression are not associated with negative trade-offs for plant growth but have much higher seed production than the wild-type in the presence of P. xylostella infestation. This study offers a new strategy for plant molecular breeding against P. xylostella.

Multi-fluorine labeled indanone derivatives as potential MRI imaging probes for ß-Amyloid plaques.

In order to realize the early diagnosis of Alzheimer's disease (AD), we designed and synthesized a series of multi-fluorine labeled indanone derivatives based on indanone which could target ß-amyloid (Aß). Through the in vitro staining experiment and affinity experiment, we selected 7d out, and then evaluated it through other in vivo and in vitro experiments. The staining of AD human brain adjacent sections revealed that compound 7d could bind to Aß plaques with high affinity. In the in vitro binding assay, 7d showed a balanced affinity with Aß1-40 (Kd = 367 ± 13) and Aß1-42 (Kd = 384 ± 56). Also, 7d exhibited a low toxicity (LD50 > 50 mg/kg) and an excellent ability to pass through the blood-brain barrier (Log p = 3.87). The biodistribution experiment in mice showed that 7d reached the highest brain uptake after 1 h of tail vein injection and cleared after 24 h. A low concentration of 7d (1.875 mg/ml) showed a strong imaging ability (19F-weighted mode), and the imaging capability increased with the increasing of concentration. All the results showed that 7d could provide a feasible solution for the early diagnosis of AD under non-radioactive condition.

Diversity and antibacterial potential of the Actinobacteria associated with Apis mellifera ligustica.

Insect-associated Actinobacteria are a potentially rich source of novel natural products with antibacterial activity. Here, the community composition of Actinobacteria associated with Apis mellifera ligustica was investigated by integrated culture-dependent and independent methods. A total of 61 strains of Streptomyces genera were isolated from the honeycomb, larva, and different anatomical parts of the honeybee's body using the culture-dependent method. Amplicon sequencing analyses revealed that the actinobacterial communities were dominated by the family of Bifidobacteriaceae and Microbacteriaceae in the honeybee gut, and Nocardiaceae and Pseudonocardiaceae in the honeycomb, whereas only Streptomyces genera were isolated by the culture-dependent method. Culture-independent analyses showed more diverse actinobacterial communities than those of culture-dependent methods. The antibacterial bioassay showed that most crude extracts of representative isolates exhibited antibacterial activities. Among them, the crude extract of Streptomyces sp. FCF01 showed the best antibacterial activities against Staphylococcus aureus, Micrococcus tetragenus, and Pseudomonas syringae pv. actinidiae (Psa) with the disc diameter of inhibition zone diameter (IZD) of 23.00, 15.00, and 13.33 mm, respectively. Chemical analysis of Streptomyces sp. FCF01 led to the isolation of three secondary metabolites, including mayamycin (1), mayamycin B (2), and N-(2-Hydroxyphenyl) acetamide (3). Among them, compound 1 displayed strong antibacterial activity against S. aureus, M. tetragenus, and Psa with minimum inhibitory concentrations (MIC) values of 6.25, 12.5, and 6.25 µg/ml, respectively. In addition, two novel derivative compounds 1a and 1b were synthesized by acetylation of compound 1. Both compounds 1a and 1b displayed similar antibacterial activities with those of metabolite 1. These results indicated that Streptomyces species associated with honeybees had great potential in finding antibiotics.

Preference Test of Plutella xylostella Larvae upon DMNT Treatment.

We describe a method to test the preference of insects in response to (3E)-4,8-dimethyl-1,3,7-nonatriene (DMNT). We use a device that includes a horizontal glass tube, two grooves (with activated carbon), air flow, rubber stoppers/tubes, transparent glass containers (optional), and a holder for the glass tube (optional). Equal amounts of activated carbon in the groove (removable) are placed at both ends to avoid air contamination. The air flow is generated by an air pump. In the closed device, different samples are placed at each end of the glass tube. The air pump at the top of the glass tube forms an air flow that converges to the middle site of the glass tube. In each test, insect larvae are located in the middle of the glass test tube. If the test samples release DMNT that can be sensed by insects, the insects will selectively move to one specific end of the glass tube. The number of insects that move to each end will be recorded for further studies. This method can also be used to test the preference of insects in response to other volatile compounds.

Volatile DMNT directly protects plants against Plutella xylostella by disrupting the peritrophic matrix barrier in insect midgut.

Insect pests negatively affect crop quality and yield; identifying new methods to protect crops against insects therefore has important agricultural applications. Our analysis of transgenic Arabidopsis thaliana plants showed that overexpression of pentacyclic triterpene synthase 1, encoding the key biosynthetic enzyme for the natural plant product (3E)-4,8-dimethyl-1,3,7-nonatriene (DMNT), led to a significant resistance against a major insect pest, Plutella xylostella. DMNT treatment severely damaged the peritrophic matrix (PM), a physical barrier isolating food and pathogens from the midgut wall cells. DMNT repressed the expression of PxMucin in midgut cells, and knocking down PxMucin resulted in PM rupture and P. xylostella death. A 16S RNA survey revealed that DMNT significantly disrupted midgut microbiota populations and that midgut microbes were essential for DMNT-induced killing. Therefore, we propose that the midgut microbiota assists DMNT in killing P. xylostella. These findings may provide a novel approach for plant protection against P. xylostella.

Comparative genomic analysis of the PAL genes in five Rosaceae species and functional identification of Chinese white pear.

Phenylalanine ammonia lyase (PAL) plays an important role in the biosynthesis of secondary metabolites regulating plant growth response. To date, the evolutionary history of the PAL family in Rosaceae plants remains unclear. In this study, we identified 16 PAL homologous genes in five Rosaceae plants (Pyrus bretschneideri, Fragaria vesca, Prunus mume, Prunus persica, and Malus × domestica). We classified these PALs into three categories based on phylogenetic analysis, and all PALs were distributed on 13 chromosomes. We tracked gene duplication events and performed sliding window analysis. These results revealed the evolution of PALs in five Rosaceae plants. We predicted the promoter of the PbPALs by PLANT CARE online software, and found that the promoter region of both PbPAL1 and PbPAL3 have at least one AC element. The results of qRT-PCR analysis found that PbPAL1 and PbPAL2 were highly expressed in the stems and roots, while expression level of PbPAL3 was relatively low in different tissues. The expression of PbPAL1 and PbPAL2 increased firstly and then decreased at different developmental periods of pear fruit. Among them, the expression of PbPAL1 reached the highest level 55 days after flowering. Three PbPALs were induced by abiotic stress to varying degrees. We transfected PbPAL1 and PbPAL2 into Arabidopsis thaliana, which resulted in an increase in lignin content and thickening of the cell walls of intervascular fibres and xylem cells. In summary, this research laid a foundation for better understanding the molecular evolution of PALs in five Rosaceae plants. Furthermore, the present study revealed the role of PbPALs in lignin synthesis, and provided basic data for regulating lignin synthesis and stone cells development in pear plants.

In Silico Genome-Wide Analysis of the Pear (Pyrus bretschneideri) KNOX Family and the Functional Characterization of PbKNOX1, an Arabidopsis BREVIPEDICELLUS Orthologue Gene, Involved in Cell Wall and Lignin Biosynthesis.

Stone cells are a characteristic trait of pear fruit, but the contents and sizes of stone cells negatively correlate with fruit texture and flavor. Secondary cell wall thickening and lignification have been established as key steps of stone cell development. KNOTTED-LIKE HOMEOBOX (KNOX) proteins play important roles in plant cell growth and development, including cell wall formation and lignification. Although the characteristics and biological functions of KNOX proteins have been investigated in other plants, this gene family has not been functionally characterized in pear. Eighteen PbKNOX genes were identified in the present study, and all of the identified family members contained the KNOX I and/or KNOX II domains. Based on the phylogenetic tree and chromosomal localization, the 18 PbKNOX genes were divided into five subfamilies [SHOOT MERISTEMLESS (STM)-like, BREVIPEDICELLUS (BP)-like, KNOTTED ARABIDOPSIS THALIANA 2/6 (KNAT2/6)-like, KNAT7-like, and KNAT3-5-like] and were distributed among 10 chromosomes. In addition, we identified 9, 11, and 11 KNOX genes in the genomes of grape, mei, and strawberry, respectively, and the greatest number of collinear KNOX gene pairs formed between pears and peaches. Analyses of the spatiotemporal expression patterns showed that the tissue specificity of PbKNOX gene expression was not very significant and that the level of the PbKNOX1 transcript showed an opposite trend to the levels of stone cells and lignin accumulation. Furthermore, PbKNOX1 has high sequence identity and similarity with Arabidopsis BP. Compared with wild-type Arabidopsis, plants overexpressing PbKNOX1 not only showed an approximately 19% decrease in the secondary cell wall thickness of vessel cells but also exhibited an approximately 13% reduction in the lignin content of inflorescence stems. Moreover, the expression of several genes involved in lignin biosynthesis was downregulated in transgenic lines. Based on our results, PbKNOX1/BP participates in cell wall-thickening and lignin biosynthesis and represses the transcription of key structural genes involved in lignin synthesis, providing genetic evidence for the roles of KNOX in cell wall thickening and lignin biosynthesis in pear.

In Silico Genome-Wide Analysis of Respiratory Burst Oxidase Homolog (RBOH) Family Genes in Five Fruit-Producing Trees, and Potential Functional Analysis on Lignification of Stone Cells in Chinese White Pear.

: The accumulation of lignin in fruit has a significant negative impact on the quality of fruit-producing trees, and in particular the lignin formation stimulates the development of stone cells in pear fruit. Reactive oxygen species (ROS) are essential for lignin polymerization. However, knowledge of the RBOH family, a key enzyme in ROS metabolism, remains unknown in most fruit trees. In this study, a total of 40 RBOHs were identified from five fruit-producing trees (Pyrusbretschneideri, Prunuspersica, Citrussinensis, Vitisvinifera, and Prunusmume), and 10 of these sequences came from Pyrusbretschneideri. Multiple sequence alignments revealed that all 10 PbRBOHs contained the NADPH_Ox domain and the six alpha-helical transmembrane domains (TM-I to TM-VI). Chromosome localization and interspecies phylogenetic tree analysis showed that 10 PbRBOHs irregularly distributed on 8 chromosomes and 3 PbRBOHs (PbRBOHA, PbRBOHB, and PbRBOHD) are closely related to known lignification-related RBOHs. Furthermore, hormone response pattern analysis showed that the transcription of PbRBOHs is regulated by SA, ABA and MeJA. Reverse transcription-quantitative real-time polymerase chain reaction (qRT-PCR) and transcriptome sequencing analysis showed that PbRBOHA, PbRBOHB, and PbRBOHD accumulated high transcript abundance in pear fruit, and the transcriptional trends of PbRBOHA and PbRBOHD was consistent with the change of stone cell content during fruit development. In addition, subcellular localization revealed that PbRBOHA and PbRBOHD are distributed on the plasma membrane. Combining the changes of apoplastic superoxide (O2.-) content and spatio-temporal expression analysis, these results indicate that PbRBOHA and PbRBOHD, which are candidate genes, may play an important role in ROS metabolism during the lignification of pear stone cells. This study not only provided insight into the molecular characteristics of the RBOH family in fruit-producing trees, but also lays the foundation for studying the role of ROS in plant lignification.

Family-1 UDP glycosyltransferases in pear (Pyrus bretschneideri): Molecular identification, phylogenomic characterization and expression profiling during stone cell formation.

Stone cells are a characteristic trait of pear fruits, and excessive stone cell formation has a significant negative impact on the texture and flavour of the pulp. Lignin is one of the main components of stone cells. Family-1 uridine diphosphate-glycosyltransferases (UGTs) are responsible for the glycosylation modification of monolignols. However, information remains limited regarding the relationship between UGTs and stone cell formation. To address this problem, we identified 139 UGTs from the pear genome, which were distributed in 15 phylogenetic groups (A-M, O, and P). We also performed a collinearity analysis of UGTs among four Rosaceae plants (pear, peach, mei, and strawberry). Phylogenetic analysis suggested that 13 PbUGTs might be related to the glycosylation of monolignols. Analysis of expression patterns demonstrated that most putative monolignol glycosylation-related PbUGTs not only showed high expression levels in flowers and buds but were also induced by exogenous ABA, SA, and MeJA. In addition, the transcript level of Pbr005014.1 (named PbUGT72AJ2) was consistent with the changing trend of lignin content in pear fruit, and the transcript level was also higher in 'Dangshan Su' pear with higher lignin and stone cell contents. Subcellular localization results showed that PbUGT72AJ2 was located mainly in the cytomembrane and cytoplasm. Based on our study, PbUGT72AJ2 is considered to be a monolignol glycosylation-related UGT. Our results provide an important source for the identification of UGTs and a foundation for the future understanding and manipulation of lignin metabolism and stone cell formation in pear fruit.

Molecular identification, phylogenomic characterization and expression patterns analysis of the LIM (LIN-11, Isl1 and MEC-3 domains) gene family in pear (Pyrus bretschneideri) reveal its potential role in lignin metabolism.

Lignin is the main component of stone cells, which are a key factor in determining pear quality. Therefore, modification of lignin biosynthesis has important implications for regulating stone cell formation. LIMs are involved in plant development, stress response and metabolism. However, there is still a lack of knowledge about the pear LIM family and lignin-related LIMs. To address this problem, we identified 14 LIMs from the pear genome and named them. Phylogenomic and feature domain analysis showed that they were divided into CRP- and DA&DAR-LIM groups and five subclades. LIMs from the genomes of four rosids (Prunus mummer, Prunus persica, Fragaria vesca and Vitis vinifera) were also identified, and microsynteny analysis revealed the most orthologous gene pairs in the cross of pear/grape and pear/mei. The transcript levels of PbLIMs were significantly affected by SA, ABA and MeJA. Spatio-temporal expression analysis showed that PbLIMs of the δLIM2 subfamily were highly expressed in the flowers. Changes in the expression levels of PbWLIM1a and PbWLIM1b during fruit development was consistent with the changes in lignin content. Combining phylogenetic analyses, protein three-dimensional structure determination and sequence alignment analyses, these two genes were suggested as lignin-related PbLIMs. Subcellular localization results showed that PbWLIM1a and PbWLIM1b were located mainly in the chloroplast. This study lays the foundation for revealing the mechanism of LIM-mediated lignin metabolism to regulate stone cell formation.

Modular synthesis of (E)-cinnamaldehydes directly from allylarenes via a metal-free DDQ-mediated oxidative process.

An efficient synthesis of (E)-cinnamaldehydes by a metal-free DDQ-mediated oxidative transformation of allylarenes was developed. The protocol provides a practical method to prepare diverse (E)-cinnamaldehydes with broad functional group tolerance in good to excellent yields, including easy access to natural products randainal and geranyloxy sinapyl aldehyde from plant extracts. Finally, the mechanism of a single-electron transfer process was proposed.

Discovery of N-(5-((5-chloro-4-((2-(isopropylsulfonyl)phenyl)amino)pyrimidin-2-yl)amino)-4-methoxy-2-(4-methyl-1,4-diazepan-1-yl)phenyl)acrylamide (CHMFL-ALK/EGFR-050) as a potent ALK/EGFR dual kinase inhibitor capable of overcoming a variety of ALK/EGFR associated drug resistant mutants in NSCLC.

Recently, more and more concomitant EGFR mutations and ALK rearrangement are observed from the clinic, which still lacks effective single-agent therapy. Starting from ALK inhibitor 14 (TAE684), we have developed a highly potent EGFR/ALK dual kinase inhibitor compound 18 (CHMFL-ALK/EGFR-050), which potently inhibited EGFR L858R, del 19 and T790M mutants as well as EML4-ALK, R1275Q, L1196M, F1174L and C1156Y mutants biochemically. Compound 18 significantly inhibited the proliferation of EGFR mutant and EML4-ALK driven NSCLC cell lines. In the cellular context it strongly affected EGFR and ALK mediated signaling pathways, induced apoptosis and arrested cell cycle at G0/G1 phase. In the in vivo studies, 18 significantly suppressed the tumor growth in H1975 cell inoculated xenograft model (40 mg/kg/d, TGI: 99%) and H3122 cell inoculated xenograft model (40 mg/kg/d, TGI: 78%). Compound 18 might be a potential drug candidate for EGFR- or ALK-individual as well as concomitant EGFR/ALK NSCLC.

Synthesis of 3-acylindoles by palladium-catalyzed acylation of free (N-H) indoles with nitriles.

An efficient palladium-catalyzed synthesis of 3-acylindoles using free (N-H) indoles and nitriles has been developed. The acylation reaction proceeded well under the Pd(OAc)(2)/2,2'-bipyridine system and with D-(+)-camphorsulfonic acid as the additive. A possible mechanism involving carbopalladation of nitriles and subsequent hydrolysis of ketimines is proposed.

Palladium-catalyzed ortho-alkoxylation of anilides via C-H activation.

A palladium-catalyzed ortho-alkoxylation of anilides with both primary and secondary alcohols via ligand-directed C-H activation has been explored. This alkoxylation promoted by catalytic methanesulfonic acid proceeds well at room temperature in most cases and affords aryl alkyl ethers in moderate to good yields.

Palladium-catalyzed oxidative tandem reaction of allylamines with aryl halides leading to alpha,beta-unsaturated aldehydes.

A novel tandem protocol involving a Heck reaction process for the synthesis of alpha,beta-unsaturated aldehydes has been developed. In the presence of Pd(OAc), PPh3, NaOAc, TBAB and air, N-allylbenzenamines underwent the reaction with various aryl halides to afford the corresponding alpha,beta-unsaturated aldehydes selectively in moderate to good yields. The protocol can also be used as a valuable route for the deallylation of arylamines.

Palladium-catalyzed intramolecular 5-exo-dig hydroarylations of N-arylpropiolamides: thermodynamics-controlled stereoselective synthesis of 3-methyleneoxindoles.

Palladium-catalyzed intramolecular hydroarylation of N-arylpropiolamides has been developed for the stereoselective synthesis of 3-(monosubstituted methylene)oxindoles. In the presence of Pd(OAc)(2) and dppf, a variety of N-arylpropiolamides successfully underwent the intramolecular hydroarylation reaction to afford the corresponding 3-(monosubstituted-methylene)oxindoles in moderate to excellent yields. It is noteworthy that the stereoselectivity of the reaction can be controlled by varying the reaction temperature.