Synthesis and evaluation of novel 7- and 8-aminophenoxazinones for the detection of β-alanine aminopeptidase activity and the rapid identification of Pseudomonas aeruginosa in clinical samples

A series of novel 8-aminophenoxazin-3-one and 7-aminophenoxazin-3-one chromogens and their corresponding β-alanine derivatives were synthesized and evaluated for their ability to detect β-alanyl aminopeptidase activity in bacteria known to hydrolyse β-alanine derivatized substrates. The results provided insight into the structural requirements for effective visualization of enzymatic activity and the mechanism of formation of phenoxazinon-3-ones. 8-Aminophenoxazin-3-one substrates 23c, 23d and 23e were prepared in good to high overall yield and were selective for β-alanyl aminopeptidase activity in bacteria, producing a lighter agar background coloration facilitating visualization of colored colonies, with variable localization to the colonies, but had lower sensitivities for the detection of Pseudomonas aeruginosa in comparison to the analogous 7-aminophenoxazin-3-one substrates. The synthetic methodology employed here allows the preparation of a range of substrates for evaluation and the establishment of structure-activity relationships. For example, the 2-pentyl substituted aminophenoxazin-3-one 22b performed with analogous sensitivity to the corresponding 1-pentyl-7-aminophenoxazin-3-one substrate 1 used commercially, highlighting that the position of the pentyl substituent can be varied while maintaining detection sensitivity

Pharmacological Activities of Aminophenoxazinones

Aminophenoxazinones are degradation products resulting from the metabolism of different plant species, which comprise a family of natural products well known for their pharmacological activities. This review provides an overview of the pharmacological properties and applications proved by these compounds and their structural derivatives during 2000-2021. The bibliography was selected according to our purpose from the references obtained in a SciFinder database search for the Phx-3 structure (the base molecule of the aminophenoxazinones). Compounds Phx-1 and Phx-3 are among the most studied, especially as anticancer drugs for the treatment of gastric and colon cancer, glioblastoma and melanoma, among others types of relevant cancers. The main information available in the literature about their mechanisms is also described. Similarly, antibacterial, antifungal, antiviral and antiparasitic activities are presented, including species related directly or indirectly to significant diseases. Therefore, we present diverse compounds based on aminophenoxazinones with high potential as drugs, considering their levels of activity and few adverse effects

Synthesis, structure and diverse coordination chemistry of cobalt(III) complexes derived from a Schiff base ligand and their biomimetic catalytic oxidation of o-aminophenols

This paper deals with the syntheses and structural characterizations of four new cobalt(III) compounds (1–4) derived from a N3O donor Schiff base ligand and their catalytic activity towards the aerobic oxidation of o-aminophenols. Both counter ions and solvents used for the synthesis have significant influence on structural diversity of the resulting complexes. X-ray crystallography reveals that although the geometry of cobalt(III) centres are octahedral in all the cases but the coordination environments are significantly different in them. All these complexes show diverse reactivity towards the catalytic oxidation of o-aminophenols in which availability of substitutionally labile sites at the metal centre for substrate o-aminophenols binding is the main reason for higher catalytic activity in 2 and 3 than others. Furthermore, we have examined the detailed kinetic studies of the aerobic oxidation of one substituted o-aminophenol, namely 2-amino-5-methylphenol, using 2 and 3 as catalysts in which facile oxidation of the substituted o-aminophenol was noticed. ESI mass spectral study has been carried out in order to get insight into mechanistic pathway of functioning such catalytic activity.publishe

Stability and pKa Modulation of Aminophenoxazinones and Their Disulfide Mimics by Host-Guest Interaction with Cucurbit[7]uril. Direct Applications in Agrochemical Wheat Models

Aqueous solubility and stability often limit the application of aminophenoxazinones and their sulfur mimics as promising agrochemicals in a sustainable agriculture inspired by allelopathy. This paper presents a solution to the problem using host-guest complexation with cucurbiturils (CBn). Computational studies show that CB7 is the most suitably sized homologue due to its strong affinity for guest molecules and its high water solubility. Complex formation has been studied by direct titrations monitored using UV-vis spectroscopy, finding a preferential interaction with protonated aminophenoxazinone species with high binding affinities (CB7 center dot APOH+ , Ka = (1.85 +/- 0.37) x 106 M-1; CB7 center dot DiS-NH3+ , Ka = (3.91 +/- 0.53) x 104 M-1; and DiS-(NH3+)2 , Ka= (1.27 +/- 0.42) x 105M-1). NMR characterization and stability analysis were also performed and revealed an interesting pKa modulation and stabilization by cucurbiturils (2-amino-3H-phenoxazin-3-one (APO), pKa = 2.94 +/- 0.30, and CB7 center dot APO, pKa = 4.12 +/- 0.15; 2,2 '-disulfanediyldianiline (DiS-NH2), pKa = 2.14 +/- 0.09, and CB7 center dot DiS-NH2 , pKa = 3.26 +/- 0.09), thus favoring applications in different kinds of crop soils. Kinetic studies have demonstrated the stability of the CB7 center dot APO complex at different pH media for more than 90 min. An in vitro bioassay with etiolated wheat coleoptiles showed that the bioactivity of APO and DiS-NH2 is enhanced upon complexation

Pharmaceutically relevant (hetero)cyclic compounds and natural products from lignin-derived monomers:Present and perspectives

Lignin, the richest source of renewable aromatics on the planet, is an intriguing raw material for the construction of value-added aromatics. In the past decade, much progress has been made regarding the development of efficient lignin depolymerization methods, able to produce specific monophenol derivatives in high-enough selectivity and yields. This now serves as an excellent basis for developing powerful downstream conversion strategies toward a wide range of products, including fine chemical building blocks. The inherent structural features of lignin-derived platform chemicals undoubtedly inspire the development of novel, creative, atom-economic synthetic routes toward biologically active molecules or natural products. In this perspective we attempt to bridge the structural features of lignin-derived platform chemicals with existing synthetic strategies toward the construction of heterocycles and provide a summary of efforts for the production of natural products from aromatics that can be, in principle, obtained from lignin. Last, we comment on the latest efforts that present entire value-chains from wood to valuable pharmaceutically relevant compounds

Bacterial tolerance to host-exuded specialized metabolites structures the maize root microbiome.

Plants exude specialized metabolites from their roots, and these compounds are known to structure the root microbiome. However, the underlying mechanisms are poorly understood. We established a representative collection of maize root bacteria and tested their tolerance against benzoxazinoids (BXs), the dominant specialized and bioactive metabolites in the root exudates of maize plants. In vitro experiments revealed that BXs inhibited bacterial growth in a strain- and compound-dependent manner. Tolerance against these selective antimicrobial compounds depended on bacterial cell wall structure. Further, we found that native root bacteria isolated from maize tolerated the BXs better compared to nonhost Arabidopsis bacteria. This finding suggests the adaptation of the root bacteria to the specialized metabolites of their host plant. Bacterial tolerance to 6-methoxy-benzoxazolin-2-one (MBOA), the most abundant and selective antimicrobial metabolite in the maize rhizosphere, correlated significantly with the abundance of these bacteria on BX-exuding maize roots. Thus, strain-dependent tolerance to BXs largely explained the abundance pattern of bacteria on maize roots. Abundant bacteria generally tolerated MBOA, while low abundant root microbiome members were sensitive to this compound. Our findings reveal that tolerance to plant specialized metabolites is an important competence determinant for root colonization. We propose that bacterial tolerance to root-derived antimicrobial compounds is an underlying mechanism determining the structure of host-specific microbial communities

Maize root bacteria cooperate to tolerate and metabolise host-secreted plant specialized metabolites

Plant roots are colonized by microbial communities which can promote growth, provide nutrients and protection against pathogens for the host. Plant specialized metabolites exuded by roots, like benzoxazinoids (BX) produced by maize, can structure root-associated microbial communities. However, the underlying mechanisms are poorly understood. The present thesis aimed to uncover the contribution of i) bacterial tolerance to benzoxazinoids, ii) bacterial metabolisation of benzoxazinoids and iii) the interactions of both for structuring root bacterial communities. First, we established a collection of maize root bacteria and in an extensive high-throughput in vitro screening we uncovered that benzoxazinoids inhibit bacterial growth in a strain-dependent and compound-dependent manner. Among the benzoxazinoids tested, 6-methoxybenzoxazolin-2(3H)-one (MBOA), which is the dominant metabolite structuring maize rhizosphere microbiomes, is the most selective compound. Tolerance to MBOA correlated positively with the benzoxazinoid-dependent colonisation of maize roots by the bacteria. We propose that tolerance to secreted antimicrobial compounds presents an important mechanism for structuring the microbial community on plant roots. Second, we identified maize root bacteria that metabolise the abundant benzoxazinoid in the rhizosphere, MBOA to 2-amino-7-methoxyphenoxazin-3-one (AMPO). The characteristic red colour of AMPO enabled us to develop a simple plate assay to screen the maize root bacteria strain collection for their ability to metabolise MBOA to AMPO. Few bacterial lineages including Sphingobium and Microbacterium convert MBOA to AMPO. AMPO-forming bacteria were enriched on roots of BX-producing but not of BX-deficient plants. We utilized the phenotypic diversity within the genus of Microbacteria to identify an N-acyl homoserine lactonase (BxdA) as the key enzyme for converting MBOA to AMPO. This study demonstrated the specific recruitment of adapted bacteria on BX-producing roots that can metabolise the host secondary metabolites. Third, we studied benzoxazinoid tolerance and metabolisation in a community context. We found that bacteria cooperate to tolerate and metabolise benzoxazinoids in synthetic microbial communities. Benzoxazinoid metabolisation conferred MBOA tolerance to the SynComs. Cooperation lead to the formation of an alternative MBOA conversion product, N-(2-hydroxy-methoxyphenyl)acetamide (HMPAA), a dominant metabolite that is not formed by single strains. We discovered that HMPAA is formed by the combined activity of an MBOA-degrading Microbacterium with a Pseudomonas. This study demonstrated that bacteria on maize roots cooperate to metabolise benzoxazinoids and as a community, they benefitted from an enhanced tolerance to these compounds. This research reveals microbiological and biochemical mechanisms of how plant specialized metabolites contribute to shape the root microbiota. The ability of maize root bacteria to tolerate and metabolise benzoxazinoids are important traits defining their abundance on BX-producing maize roots. We demonstrate that strains when combined in synthetic communities, they divide labour and cooperate to metabolise and tolerate benzoxazinoids. The deepened understanding of how plant specialized metabolites sculpt community composition provides a tool to selectively steer microbiome structure. Further work is required to understand how bacterial BX-mediated mechanisms affect microbiomes to harness the functions of microbial communities for sustainable agriculture

Differential Impact of Plant Secondary Metabolites on the Soil Microbiota

Plant metabolites can shape the microbial community composition in the soil. Two indole metabolites, benzoxazolinone (BOA) and gramine, produced by different Gramineae species, and quercetin, a flavonoid synthesized by many dicot species, were studied for their impacts on the community structure of field soil bacteria. The three plant metabolites were directly added to agricultural soil over a period of 28 days. Alterations in bacterial composition were monitored by next generation sequencing of 16S rRNA gene PCR products and phospholipid fatty acid analysis. Treatment of the soil with the plant metabolites altered the community composition from phylum to amplicon sequence variant (ASV) level. Alpha diversity was significantly reduced by BOA or quercetin, but not by gramine. BOA treatment caused a decrease of the relative abundance of 11 ASVs, while only 10 ASVs were increased. Gramine or quercetin treatment resulted in the increase in relative abundance of many more ASVs (33 or 38, respectively), most of them belonging to the Proteobacteria. Isolation and characterization of cultivable bacteria indicated an enrichment in Pseudarthrobacter or Pseudomonas strains under BOA/quercetin or BOA/gramine treatments, respectively. Therefore, the effects of the treatments on soil bacteria were characteristic for each metabolite, with BOA exerting a predominantly inhibitory effect, with only few genera being able to proliferate, while gramine and quercetin caused the proliferation of many potentially beneficial strains. As a consequence, BOA or gramine biosynthesis, which have evolved in different barley species, is accompanied with the association of distinct bacterial communities in the soil, presumably after mutual adaptation during evolution

On the formulation of disulfide herbicides based on aminophenoxazinones: polymeric nanoparticle formulation and cyclodextrin complexation to combat crop yield losses

BackgroundThe resistance of weeds to herbicides is a significant issue in ensuring future food supply. Specific examples are Plantago lanceolata, Portulaca oleracea and Lolium rigidum, which mainly infect rice, wheat, barley and pastures, and cause high yield losses every year. In this regard, natural products and their mimics have provided new hope as a result of their different modes-of-action, activity at low concentrations and reduced pollution effects relative to conventional herbicides. However, the poor water solubility and physicochemical properties of these compounds limit their broad application. These problems can be addressed by formulation techniques, and encapsulation appears to be of great interest. ResultsDisulfide herbicides inspired by aminophenoxazinones have been formulated with 2-hydroxypropyl-beta-cyclodextrin (HP-beta-CD), gamma-CD and polymeric nanoparticles (NPs). In silico studies were employed to identify which complexes would be generated and complex formation was confirmed by nuclear magnetic resonance spectroscopy. Solubility diagrams were generated to assess any improvement in water solubility, which was enhanced 2-13-fold. Scanning electron microscopy and energy-dispersive X-ray spectra confirmed the success of the formulation process for the nanoparticles. Formulated compounds were evaluated in an in vitro wheat coleoptile bioassay, with almost 100% elongation inhibition achieved using only water for the bioassay. Specific in vitro testing on weed phytotoxicity showed that the application of core/shell NPs is highly effective in the fight against P. lanceolata seed germination. ConclusionsThe formulation of disulfide herbicides with CD complexes and NPs led to an enhancement in water solubility and bioactivity. These systems can be applied in pre-emergent mode against P. lanceolata, using only water to prepare the sample, and they showed better activity than the positive controls. (c) 2022 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industr

Pigments of aminophenoxazinones and viridomycins produced by termite-associated Streptomyces tanashiensis BYF-112

Termite-associated Streptomyces tanashiensis BYF-112 was found as a potential source for yellow and green pigments, which were stable under the tested temperature, light and metal ions. Eight metabolites (1–8), including four new natural yellow pigments aminophenoxazinones (1–4), and two rarely iron dependent green pigments viridomycin A and F (9–10) were isolated from BYF-112 cultured in YMS and YMS treated with FeSO4, respectively. The metabolites 2–4 displayed a significant safety performance on the normal liver cell line L-02, while the metabolite 1 showed weak cytotoxicity against the L-02 and several cancer cells. Especially, in the filter paper disc tests, the compound 1 possessed strong antibacterial activity against methicillin-resistant Staphylococcus aureus (MRSA) with the zone of inhibition (ZOI) of 15.3 mm, which was equal to that of referenced levofloxacin (ZOI = 15.2 mm). And the metabolite 1 also showed moderate antibacterial activities against Micrococcus teragenus and S. aureus, with the ZOI values of 15.3 and 17.2 mm. In addition, by the minimum inhibitory concentration (MIC) assay, the compound 1 displayed potential antibacterial activities against M. teragenus, S. aureus and MRSA, with the MIC values of 12.5, 12.5, and 25.0 μg/ml, respectively. The present results indicate that BYF-112 may be a promising source for safe and bioactive pigments, which can be used for further development and industrial applications