ABSTRACT
Vitamin B12 deficiency causes significant changes in cellular metabolism leading to various clinical symptoms, such as hematological, psychiatric, and neurological disorders. We hypothesize that skin pigmentation disorders may be a diagnostically important manifestation of vitamin B12 deficiency, however the cellular and molecular mechanisms underlying these effects remain unknown. The aim of this study was to examine the effect of vitamin B12 deficiency on melanocytes homeostasis. Hypocobalaminemia in vitro model was developed by treating epidermal melanocytes with synthesized vitamin B12 antagonist-hydroxycobalamin(c-lactam). The cells were examined using immunoenzymatic, spectrophotometric, and fluorimetric assays as well as image cytometry. Significant melanogenesis stimulation-the increase of relative melanin content and tyrosinase activity up to 131% and 135%, respectively-has been indicated. Cobalamin-deficient cells displayed the elevation (by 120%) in reactive oxygen species level. Moreover, the redox status imbalance was stated. The study provided a scientific evidence for melanocytes homeostasis disturbance under hypocobalaminemia, thus indicating a significant element of the hyperpigmentation mechanism due to vitamin B12 deficiency. Furthermore, the implication between pigmentary and hematological and/or neuropsychiatric symptoms in cobalamin-deficient patients may be an important issue.
Subject(s)
Homeostasis , Melanins/metabolism , Melanocytes/metabolism , Vitamin B 12 Deficiency/metabolism , Cells, Cultured , Humans , Monophenol Monooxygenase/metabolism , Reactive Oxygen Species/metabolism , Vitamin B 12/metabolismABSTRACT
Vitamin B12 is a natural cobalt complex that, while reduced to the "supernucleophilic" Co(I) form, can easily react with electrophiles via an SN2 mechanism. It is also shown to react via an SN2' mechanism with allylic compounds allowing for photochemical deprotection of (allyloxy)arenes. A sustainable alternative to commonly used noble metal-catalyzed deprotection reactions is presented.
ABSTRACT
Vitaminâ B12 is a cofactor for many enzymes, but it also functions as a catalyst in C-C bond-forming reactions. Herein, the impact of corrin structural modifications on their catalytic efficacy was examined. Derivatives with various substituents at c-, d-, and meso-positions were synthesised by using traditional and new microwave methodologies, and then tested in the model reaction of 1,1-diphenylethylene with ethyl diazoacetate. To complement the experimental data, cyclic voltammetry and DFT calculations were performed. Mainly alterations at the c- or d-positions influence both the reaction yield and selectivity.
Subject(s)
Vitamin B 12/chemistry , Biological Products/chemistry , Catalysis , Diazonium Compounds/chemistry , Models, Molecular , Molecular Conformation , Structure-Activity RelationshipABSTRACT
The binding of vitaminâ B12 derivatives to human B12 transporter proteins is strongly influenced by the type and site of modification of the cobalamin original structure. We have prepared the first cobalamin derivative modified at the phosphate moiety. The reaction conditions were fully optimized and its limitations examined. The resulting derivatives, particularly those bearing terminal alkyne and azide groups, were isolated and used in copper-catalyzed alkyne-azide cycloaddition reactions (CuAAC). Their sensitivity towards light revealed their potential as photocleavable molecules. The binding abilities of selected derivatives were examined and compared with cyanocobalamin. The interaction of the alkylated derivatives with haptocorrin was less affected than the interaction with intrinsic factor. Furthermore, the configuration of the phosphate moiety was irrelevant to the binding process.
Subject(s)
Intrinsic Factor/metabolism , Transcobalamins/metabolism , Vitamin B 12/analogs & derivatives , Alkynes/chemistry , Azides/chemistry , Catalysis , Copper/chemistry , Cycloaddition Reaction , Humans , Intrinsic Factor/chemistry , Light , Phosphates/metabolism , Photolysis/radiation effects , Protein Binding , Transcobalamins/chemistry , Ultraviolet Rays , Vitamin B 12/chemical synthesis , Vitamin B 12/metabolismABSTRACT
Cobalester, a natural nontoxic vitamin B12 derivative, was found to catalyse unusual olefinic sp(2) C-H alkylation with diazo reagents as a carbene source instead of the expected cyclopropanation.
Subject(s)
Alkenes/chemistry , Cobalt/chemistry , Alkylation , CatalysisABSTRACT
Specially designed B-ring-modified cobalamin derivatives were synthesized and tested as potential activators of soluble guanylyl cyclase (sGC). Herein, we disclose the influence of substituents at the c- and d-positions in hydrophilic and hydrophobic cobyrinic acid derivatives on their capacities to activate sGC. The presence of the amide group at c-/d-position in cobyrinic acid derivatives strongly influence the level of sGC activation. Removal of the d-position altogether has a profound effect for hydrophobic compounds. In contrast, little differences were observed in hydrophilic ones.
Subject(s)
Cobamides/chemistry , Guanylate Cyclase/metabolism , Enzyme Activation , Molecular StructureABSTRACT
We present a new method for the preparation of cobinamide (CN)2Cbi, a vitamin B12 precursor, that should allow its broader utility. Treatment of vitamin B12 with only NaCN and heating in a microwave reactor affords (CN)2Cbi as the sole product. The purification procedure was greatly simplified, allowing for easy isolation of the product in 94% yield. The use of microwave heating proved beneficial also for (CN)2Cbi(c-lactone) synthesis. Treatment of (CN)2Cbi with triethanolamine led to (CN)2Cbi(c-lactam).
Subject(s)
Cobamides/chemical synthesis , Ethanolamines/chemistry , Lactams, Macrocyclic/chemical synthesis , Vitamin B 12/chemistry , Cobamides/chemistry , Lactams, Macrocyclic/chemistry , MicrowavesABSTRACT
Hybrid molecules composed of PpIX and cobyrinic acid derivatives conjugated through linkers of varying length and composition were prepared via 1,3-dipolar cycloaddition (CuAAC) or amidation/esteryfication reactions. They were tested for activation of soluble guanylyl cyclase (sGC), a key enzyme in the NO/cGMP signaling pathway, by an in vitro GTPâcGMP conversion assay. Using purified heme-deficient sGC and truncated sGC variants lacking a heme-binding domain, we demonstrated that such hybrid molecules may activate sGC by targeting heme-binding and/or catalytic domain. While all conjugates activated sGC, only selected compounds served as bifunctional regulators and were capable of simultaneous targeting both heme and catalytic domains of sGC. The length and type of a linker connecting both components had a profound effect on the extent of sGC activation, indicating that the linker's type is crucial for their binding affinities with regulatory and catalytic domains. Only hybrids with the conjugated linker of 13-16 atom length synergistically target both domains and displayed the lowest EC50 and highest activating potency. Compounds with shorter connecting linkers were much less potent and were no more active than the cobyrinic acid component alone. The most active conjugate, which showed a 60-fold activation of sGC, was compound 11, in which PpIX and cobyrinic acid components are separated by 11 atoms chain with the triazole moiety in between.
Subject(s)
Guanylate Cyclase/metabolism , Organometallic Compounds/chemical synthesis , Organometallic Compounds/pharmacology , Receptors, Cytoplasmic and Nuclear/metabolism , Chemistry Techniques, Synthetic , Copper/chemistry , Enzyme Activation/drug effects , Guanylate Cyclase/chemistry , Humans , Models, Molecular , Organometallic Compounds/chemistry , Protein Conformation , Protoporphyrins/chemistry , Receptors, Cytoplasmic and Nuclear/chemistry , Soluble Guanylyl Cyclase , Structure-Activity RelationshipABSTRACT
Various newly prepared and previously known vitamin B12 derivatives have been studied as potential soluble guanylyl cyclase (sGC) activators. All compounds tested were found to activate the sGC enzyme, although to differing extents. The best results were obtained with the derivatives synthesized from c-lactone and possessing aliphatic amides in the c- and d-positions.
Subject(s)
Enzyme Activators/chemical synthesis , Guanylate Cyclase/chemistry , Lactones/chemical synthesis , Receptors, Cytoplasmic and Nuclear/chemistry , Vitamin B 12/analogs & derivatives , Vitamin B 12/chemical synthesis , Amides/chemical synthesis , Amides/chemistry , Enzyme Activators/chemistry , Humans , Lactones/chemistry , Soluble Guanylyl Cyclase , Structure-Activity Relationship , Vitamin B 12/chemistryABSTRACT
The acid-sensitivity of vitamin B(12) derived mono- and diamides was studied. It was found that the use of reductive ring-opening of the lactone moiety deactivated undesired decomposition of c-mono- and c,d-diamides under acidic conditions. As a result, reactions gave respectively c- or d-acids which were further functionalized via coupling with amino acids. Though mono- and diamides exhibited acid sensitivity, they were used for the preparation of several highly functionalized molecules showing their stability under various reaction conditions.
Subject(s)
Diamide/chemistry , Lactones/chemistry , Vitamin B 12/chemistry , Amino Acids/chemistry , Catalysis , Crystallography, X-Ray , Magnetic Resonance Spectroscopy , Molecular Structure , Sensitivity and Specificity , StereoisomerismABSTRACT
A selective synthesis of new hydrophobic cobalamin derivatives bearing two different spacers has been accomplished via ring-opening reaction of c-lactone. The reaction of c-lactone with various amines afforded three types of amides (a, b, and c) depending on the reaction conditions. The structure of lactone b was determined by the X-ray analysis confirming the position of ring closure. It also reveals the presence of a hydrogen bond between the terminal hydroxy group and one of the axial cyanide ligands.
