ABSTRACT
While thiocarbonyl-stabilised phosphonium ylides generally react upon flash vacuum pyrolysis by the extrusion of Ph3PS to give alkynes in an analogous way to their carbonyl-stabilised analogues, two examples with a hydrogen atom on the ylidic carbon are found to undergo a quite different process. The net transfer of a phenyl group from P to S gives (Z)-configured 1-diphenylphosphino-2-(phenylsulfenyl)alkenes in a novel isomerisation process via intermediate λ5-1,2-thiaphosphetes. These prove to be versatile hemilabile ligands with a total of seven complexes prepared involving five different transition metals. Four of these are characterised by X-ray diffraction with two involving the bidentate ligand forming a five-membered ring metallacycle and two with the ligand coordinating to the metal only through phosphorus.
ABSTRACT
The prevalent occurrence of herbicide resistant weeds increases the necessity for new site of action herbicides for effective control as well as to relax selection pressure on the known sites of action. As a consequence, interest increased in the unexploited molecule cinmethylin as a new solution for the control of weedy grasses in cereals. Therefore, the mechanism of action of cinmethylin was reevaluated. We applied the chemoproteomic approach cellular Target Profiling™ from Evotec to identify the cinmethylin target in Lemna paucicostata protein extracts. We found three potential targets belonging to the same protein family of fatty acid thioesterases (FAT) to bind to cinmethylin with high affinity. Binding of cinmethylin to FAT proteins from Lemna and Arabidopsis was confirmed by fluorescence-based thermal shift assay. The plastid localized enzyme FAT plays a crucial role in plant lipid biosynthesis, by mediating the release of fatty acids (FA) from its acyl carrier protein (ACP) which is necessary for FA export to the endoplasmic reticulum. GC-MS analysis of free FA composition in Lemna extracts revealed strong reduction of unsaturated C18 as well as saturated C14, and C16 FAs upon treatment with cinmethylin, indicating that FA release for subsequent lipid biosynthesis is the primary target of cinmethylin. Lipid biosynthesis is a prominent target of different herbicide classes. To assess whether FAT inhibition constitutes a new mechanism of action within this complex pathway, we compared physiological effects of cinmethylin to different ACCase and VLCFA synthesis inhibitors and identified characteristic differences in plant symptomology and free FA composition upon treatment with the three herbicide classes. Also, principal component analysis of total metabolic profiling of treated Lemna plants showed strong differences in overall metabolic changes after cinmethylin, ACCase or VLCFA inhibitor treatments. Our results identified and confirmed FAT as the cinmethylin target and validate FAT inhibition as a new site of action different from other lipid biosynthesis inhibitor classes.
Subject(s)
Arabidopsis/drug effects , Araceae/drug effects , Fatty Acids/antagonists & inhibitors , Herbicides/metabolism , Plant Proteins/metabolism , Thiolester Hydrolases/metabolism , Arabidopsis/metabolism , Araceae/metabolism , Biological Transport , Crystallography, X-Ray , Endoplasmic Reticulum/metabolism , Fatty Acid Synthesis Inhibitors/metabolism , Fatty Acid Synthesis Inhibitors/pharmacology , Fatty Acids/biosynthesis , Fluorescence , Gas Chromatography-Mass Spectrometry , Herbicide Resistance , Herbicides/pharmacology , Principal Component Analysis , Protein Conformation , Thiolester Hydrolases/chemistryABSTRACT
While thermolysis of thiocarbonyl-stabilised phosphonium ylides generally results in extrusion of Ph(3)PS to give alkynes, those with a P=CH function instead undergo a novel P to S transfer of a phenyl group to give (Z)-configured 1-phosphino-2-sulfenylalkenes of interest as bidentate ligands.
Subject(s)
Alkynes/chemical synthesis , Organophosphorus Compounds/chemical synthesis , Sulfhydryl Compounds/chemistry , Sulfur Compounds/chemical synthesis , Crystallography, X-Ray , Models, Molecular , Molecular Structure , TemperatureABSTRACT
Azeotropic reflux chromatography (in which the eluent is continuously recycled by means of refluxing) was used to separate a mixture of spiroketal intermediates in the scale-up synthesis of spongistatin 1, leading to an improved separation and an approximately 35-fold reduction in the amount of solvent used.
ABSTRACT
A series of small molecule orally bioavailable ghrelin receptor agonists have been identified through systematic optimisation of a high throughput screening hit.
