Dithiocarbamate Formation Research Articles

N-Fluoroalkylated Morpholinos - a New Class of Nucleoside Analogues.

The first concise and efficient synthesis of some fluorine-containing morpholino nucleosides has been developed. One synthetic strategy was based on the oxidative ring cleavage of the vicinal diol unit of uridine, cytidine adenosine and guanosine derivatives, followed by cyclisation of the dialdehyde intermediates by double reductive amination with fluorinated primary amines to obtain various N-fluoroalkylated morpholinos. Another approach involved cyclisation of the diformyl intermediates with ammonia source, followed by dithiocarbamate formation and desulfurization-fluorination with diethylaminosulfur trifluoride yielding the corresponding morpholine-based nucleoside analogues with a N-CF3 element in their structure.

Design, synthesis and biological evaluation of rhein-piperazine-dithiocarbamate hybrids as potential anticancer agents

A series of novel rhein-piperazine-dithiocarbamate hybrids 3 were efficiently synthesized from rhein through a catalyst-free and one-pot, three-step sequence involving chlorination and N-acylation followed by dithiocarbamate formation. Hybrids 3 were evaluated for their in vitro cytotoxic potency by MTT assay against several human cancer and non-cancer cells. Five of the hybrids were more cytotoxic to human lung cancer cell line A549 than the parent rhein and the reference, cytarabine (CAR). Structure-activity relationship (SAR) analysis indicated that cytotoxicity was significantly enhanced when ester groups were incorporated into the hybrids (3h–j). In particular, hybrid 3h (IC50 = 10.93 μg/mL), containing a long-chain alkyl ester, was the most potent compound toward A549 tumor cells, being 7- and 5-fold more toxic than rhein (IC50 = 77.11 μg/mL) and CAR (IC50 = 49.27 μg/mL), respectively. Additionally, hybrid 3h was less toxic to the corresponding normal human lung fibroblast cell line, WI-38, with a higher selectivity index (SI, WI-38/A549 ≈ 5) than doxorubicin (DOX, SI ≈ 0), CAR (SI ≈ 2) and rhein (SI ≈ 1). Furthermore, hybrid 3h displayed more toxicity against four types of lung cancer cells (A549, Calu-1, PC-9, and H460; IC50 = 10.81–23.78 μg/mL) than against six other types of cancer cells (Huh-7, 786-O, HCT116, Hela, SK-BR-3, and SK-OV-3; IC50 = 23.85–51.98 μg/mL). Further mechanistic studies showed that hybrid 3h induced apoptosis in a concentration-dependent manner in human lung adenocarcinoma cell line PC-9. In vivo safety studies showed that hybrid 3h had no acute toxicity to the major organs of mice and did not lead to blood biochemical index changes. Our results exhibit prominent anti-cancer cell inhibition ability and no obvious systemic toxicity to normal organs, indicating that hybrid 3h has promising potential for further applications in anti-lung cancer drug development.

Rational Design, Synthesis and Evaluation of Novel Rodanine Derivatives for Antihyperglycemic Activity

A new series of rhodanines with phenyl substitution at the fifth position of rhodanine ring were designed targeting the PPAR-γ receptor for their possible antihyperglycemic activity. The rational design and molecular modeling studies were performed against PPAR-γ (PDB ID: 2PRG). The compounds were docked and thereafter subjected for the synthesis via dithiocarbamate formation followed by Knoevenagel condensation. Structures were confirmed by IR, NMR and Mass spectral analysis. All synthesized compounds were screened for in vitro glucose uptake assay using isolated hepatocytes. The Compounds A5 and B5 having N-(p-methoxy benzyl) at third position of rhodanine ring showed good glucose uptake activity. These two compounds were further studied for the in vivo antihyperglycemic study against dexamethasone induced insulin resistance. The compounds were tested at single dose level of 10 mg/kg, per oral. The compound B5 showed significant antihyperglycemic activity when compared with the standard drug, pioglitazone. The illustration, about the molecular design, synthesis, analysis, glucose uptake study and in vivo antihyperglycemic activity is reported here along with the structure activity relationships.

Development of Novel Rhodanine Analogs as Anticancer Agents: Design, Synthesis, Evaluation and CoMSIA Study.

A series of novel 5-substituted benzylidene rhodanine derivatives using four different amines were designed based on our previously developed CoMSIA (Comparative molecular similarity indices analysis) model for the anticancer activity. The designed rhodanines were synthesized via dithiocarbamate formation, cyclization and Knoevenagel condensation. The structures of the synthesized compounds were confirmed and analyzed by spectral studies. The synthesized rhodanines were investigated for in vitro anticancer activities and the analogs have displayed mild to significant cytotoxicity against MCF-7 breast cancer cells. The compounds with benzyloxy substitution at the fifth position of rhodanine ring (Compounds 20, 33 and 38) system showed significant cytotoxic activity against MCF-7 cells. CoMSIA, a three-dimensional quantitative structureactivity relationship (3D-QSAR) technique was accomplished to elucidate structure-activity relationships. Based on the information derived from CoMSIA contour plots, some key features for increasing the activity of compounds have been identified and used to design new anti-cancer agents. The present developed CoMSIA model displayed good external predictability, r2pred of 0.841 and good statistical robustness.

Stabilization of Silver Nanoparticles with a Dithiocarbamate Ligand and Formation of Nanocomposites by Combination with Polythiophene Derivative Nanoparticles

Spherical morphology for silver nanoparticles (Ag NPs) stabilized with dithiocarbamate (DTC) by reducing silver nitrate with sodium borohydride was obtained, while the addition of sodium citrate and hydrogen peroxide allowed the formation of silver nanotriangles (Ag NTs). Solutions of bright yellow and blue colors characteristic of both morphologies were observed. UV-vis optical analysis of NPs stabilized with DTC showed a plasmonic absorption band at 393 nm characteristic for spherical morphology, while two bands were observed at 332 nm and 762 nm, and a shoulder around 500 nm for the triangular morphology; with these spectra each morphology was confirmed. In these spectra an absorption band between 250 and 260 nm confirms the presence of DTC ligand. The stability of the NPs was achieved using an 8.69 × 10-3 mM solution of 4-(ethylaminodithiocarbamate) methylpyridine di-n-butyltin (IV) through a transmetallation reaction. Silver nanoparticles (Ag NPs) with spherical morphology of average diameter of 12.7 ± 1.2 nm and triangular morphology with 28.9 ± 0.8 nm for each side of the triangles were analyzed by high resolution scanning electron microscopy (HR-SEM). UV-vis spectra also showed the stability of NPs with DTC for more than three months. A copolymer derived of 3-hexylthiophene with (E)-2-(ethyl(4-((4-nitrophenyl) diazenyl) phenyl) amino) ethyl 2-(thiophen-3-yl) acetate (PA) was tested to get polymer NPs by reprecipitation method using THF/water systems. PA Polymer NPs having average diameter of 9.0 ± 1.7 nm were found. By quick and easy procedure, the formation of nanocomposite (NC) of spherical Ag NPs and PA polymer NPs was reached. This NC could be used as imaging agent, electrochemical biosensor, and photonic and optoelectronic device materials.

Synthesis and Antifungal Activity against Fusarium oxysporum of Some Brassinin Analogs Derived from l-tryptophan: A DFT/B3LYP Study on the Reaction Mechanism.

An efficient methodology to obtain novel antifungal analogs of brassinin 1 is described. Starting from l-tryptophan 2, N,N′-dialkylthiourea 4, 4-[(1H-indol-3-yl)methylene]-2-sulfanylidene-1,3-thiazolidin-5-one 5 and alkyl (2S)-3-(1H-indol-3-yl)-2-{[(alkylsulfanyl)carbonothioyl]amino}propanoate 6 type compounds were obtained as main products in different ratios depending on the reaction conditions via a tandem dithiocarbamate formation and Michael addition reaction. In order to understand the dependence of the reaction conditions on the mechanism pathway, a DFT/B3LYP study was performed. The results suggested the existence of competitive mechanistic routes which involve the presence of an ionic dithiocarbamate intermediate 9. Antifungal activities of all products were then evaluated against Fusarium oxysporum through mycelial growth inhibition using a microscale amended-medium assay. IC50 values were thus determined for each compound. These results showed that 6-related compounds can be considered as promissory antifungal agents.

Carbon disulfide mediated self-assembly of Laccase and iron oxide nanoparticles on gold surfaces for biosensing applications

A simple one-step methodology was explored to prepare enzyme-modified nanostructured electrodes for the development of biosensing interfaces. Magnetite type nanoparticles conjugated with Laccase were immobilized on gold surfaces. This approach relies on the reaction between carbon disulfide and amine groups of biomolecules to form dithiocarbamate (DTC) moieties, as well as on the strong affinity between sulfur species and metals. Special emphasis was given to demonstrate DTC formation in aqueous solution and further attachment to iron oxide nanoparticles and to gold electrodes. UV–visible spectroscopy confirmed the functionalization of nanoparticles by DTC using a model secondary amine (N-hexylmethylamine). The direct attachment of modified iron oxide nanoparticles (with ca. 20 or 40nm mean sizes) to gold electrodes was investigated using the hormone epinephrine, with well-known electrochemical properties. A high amount of immobilized epinephrine and a facilitated redox conversion was observed for modified electrodes containing iron oxide nanoparticles. The success of this simple and robust method was confirmed by X-ray photoelectronic spectroscopy. Finally, the catalytic activity of modified gold with iron oxide nanoparticles and Laccase was evaluated toward 2,2′-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid diammonium salt (ABTS). Chronoamperometric studies revealed a significant catalytic activity of immobilized Laccase in the presence of the nanoparticles, in particular for the largest ones (40nm), with a sensitivity for ABTS oxidation of 100mAM−1cm−2.

One-pot approach to modify nanostructured gold surfaces through in situ dithiocarbamate linkages

This work describes a simple methodology to bio-functionalize flat Au(111) electrodes through the one-step reaction between gold nanoparticles (AuNPs), carbon disulfide and a secondary amine (epinephrine) and an aminoacid (tryptophan). The process relies on the in situ dithiocarbamate formation between carbon disulphide and amine groups and also on the strong linkage between sulfur and gold. The redox behavior of modified gold with epinephrine or tryptophan, prepared from both ethanolic and aqueous solutions confirms their covalent immobilization and reveals a significant increase of their amount on the electrode due to the presence of AuNPs. Electrochemical reductive desorption in basic solution provided qualitative information on the amount of sulfur linked the gold surface and complements the redox studies. The co-immobilization of an enzyme (glucose oxidase, GOx) and gold nanoparticles, using carbon disulfide has been also tested. The presence of GOx on modified Au(111) electrodes has been confirmed by electrochemical detection of the catalytic oxidation of glucose in the presence of a redox mediator and through the evaluation of H2O2 reduction, formed during the catalytic reaction. XPS analysis, topographic and phase imaging by atomic force microscopy (AFM) further confirmed surface modification by AuNPs and also their functionalization. The successful one-step amine adsorption, in the presence of AuNPs, from aqueous solutions reveals the potential of this method in the construction of nanostructured biosensing interfaces.

Mixed-surface, lipid-tethered quantum dots for targeting cells and tissues

Quantum dots (QDs), with their variable luminescent properties, are rapidly transcending traditional labeling techniques in biological imaging and hold vast potential for biosensing applications. An obstacle in any biosensor development is targeted specificity. Here we report a facile procedure for creating QDs targeted to the cell membrane with the goal of cell-surface protease biosensing. This procedure generates water-soluble QDs with variable coverage of lipid functional groups. The resulting hydrophobicity is quantitatively controlled by the molar ratio of lipids per QD. Appropriate tuning of the hydrophobicity ensures solubility in common aqueous cell culture media, while providing affinity to the lipid bilayer of cell membranes. The reaction and exchange process was directly evaluated by measuring UV absorption spectra associated with dithiocarbamate formation. Cell membrane binding was assessed using flow cytometry and total internal reflection fluorescence imaging with live cells, and tissue affinity was measured using histochemical staining and fluorescence imaging of frozen tissue sections. Increases in cell and tissue binding were found to be regulated by both QD hydrophobicity and surface charge, underlying the importance of QD surface properties in the optimization of both luminescence and targeting capability.

Controlled and Sustained Release of Drugs from Dendrimer–Nanoparticle Composite Films

A dendrimer-nanoparticle hybrid scaffold based on robust dithiocarbamate formation provides a controlled drug delivery system. These composite films are nontoxic and can incorporate a variety of guests, providing sustained drug release over multiple uses. The system is highly modular: the release process can be easily tuned by altering the dendrimer generation and the size of the AuNPs, generating a versatile delivery system.

Biocompatible Charged and Uncharged Surfaces Using Nanoparticle Films

Biocompatible surfaces: Cationic, anionic, and neutral nanoparticles (NPs) were immobilized via dithiocarbamate formation onto polyethylenimine (PEI) films to create biofouling resistant surfaces. These surfaces are designed to bind proteins without denaturation, hence preventing biofouling. Extended incubation of these surfaces with BSA and serum demonstrate minimal fouling, providing a new methodology for generating surfaces resistant towards protein fouling.

Nanoparticle Immobilization on Surfaces via Activatable Heterobifunctional Dithiocarbamate Bond Formation

A simple and reliable technique has been developed to deposit robust monolayers of different types of nanoparticles including gold, iron-platinum, and core/shell CdSe/ZnS, using in situ dithiocarbamate formation.

Controlling the Cellular Uptake of Gold Nanorods

Gold nanorods coated with cetyltrimethylammonium bromide (CTAB), a cationic micellar surfactant used in nanorod synthesis, were rapidly and irreversibly internalized by KB cells via a nonspecific uptake mechanism. Internalized nanorods near the cell surface were monitored by two-photon luminescence (TPL) microscopy and observed to migrate toward the nucleus with a quadratic rate of diffusion. The internalized nanorods were not excreted but formed permanent aggregates within the cells, which remained healthy and grew to confluence over a 5-day period. Nonspecific nanorod uptake could be greatly reduced by displacing the CTAB surfactant layer with chemisorptive surfactants, particularly by the conjugation of poly(ethylene glycol) chains onto nanorods using in situ dithiocarbamate formation.

Nuclear Factor κB Is a Molecular Target for Sulforaphane-mediated Anti-inflammatory Mechanisms

Sulforaphane (SFN), an aliphatic isothiocyanate, is a known cancer chemopreventive agent. Aiming to investigate anti-inflammatory mechanisms of SFN, we here report a potent decrease in lipopolysaccharide (LPS)-induced secretion of pro-inflammatory and pro-carcinogenic signaling factors in cultured Raw 264.7 macrophages after SFN treatment, i.e. NO, prostaglandin E(2), and tumor necrosis factor alpha. SFN did not directly interact with NO, nor did it inhibit inducible nitric-oxide synthase enzymatic activity. Western blot analyses revealed time- and dose-dependent reduction of LPS-induced inducible nitric-oxide synthase as well as Cox-2 protein expression, which was suppressed at the transcriptional level. To reveal the target of SFN beyond its anti-inflammatory action, we performed electrophoretic mobility shift assay analyses of transcription factor-DNA binding. Consequently, nuclear factor kappa B (NF-kappa B), a pivotal transcription factor in LPS-stimulated pro-inflammatory response, was identified as the key mediator. SFN selectively reduced DNA binding of NF-kappa B without interfering with LPS-induced degradation of the inhibitor of NF-kappa B nor with nuclear translocation of NF-kappa B. Because SFN can interact with thiol groups by dithiocarbamate formation, it may impair the redox-sensitive DNA binding and transactivation of NF-kappa B. Sulforaphane could either directly inactivate NF-kappa B subunits by binding to essential Cys residues or interact with glutathione or other redox regulators like thioredoxin and Ref-1 relevant for NF-kappa B function. Our data provide novel evidence that anti-inflammatory mechanisms contribute to sulforaphane-mediated cancer chemoprevention.

Carbon disulfide mediated protein cross-linking by N,N-diethyldithiocarbamate.

N,N-Diethyldithiocarbamate and its disulfide are used as pesticides, in industrial processes, and as therapeutic agents, providing numerous opportunities for human exposure. Animal studies and in vitro investigations have demonstrated adverse effects following exposure to dithiocarbamates. The ability of dithiocarbamates to decompose to parent amine and CS2 suggests that these adverse effects may be mediated through release of CS2. The toxicity of CS2 is well established, and covalent cross-linking of proteins has been presented as a potential molecular mechanism of CS2 induced neuropathy. In the present investigation the ability of N,N-diethyldithiocarbamate to effect covalent cross-linking of proteins under physiological conditions is examined. Using 13C NMR, cross-linking was observed to proceed through dithiocarbamate formation on protein amino groups followed by the production of bis(thiocarbamoyl) disulfide, dithiocarbamate ester, and thiourea cross-linking structures. The presence of bis(lysyl) thiourea cross-linking structures was verified by complete protein hydrolysis in conjunction with GC/MS. Generation of inter- and intramolecular cross-linking was established using denaturing polyacrylamide gel electrophoresis under reducing conditions and revealed that cross-linking proceeded more rapidly for N,N-diethyldithiocarbamate than for equimolar CS2 under similar conditions. Covalent cross-linking of solubilized neurofilament triplet proteins, the putative neurotoxic targets, was examined. Both N,N-diethyldithiocarbamate and CS2 were able to covalently cross-link the low molecular weight component of the neurofilament triplet proteins, but neither produced intermolecular cross-linking of the medium or high molecular weight component. These results establish that N,N-diethyldithiocarbamate promoted protein cross-linking occurs under physiological conditions and proceeds through liberation of CS2.(ABSTRACT TRUNCATED AT 250 WORDS)

Characterization of isothiocyanates, thioureas, and other lysine adduction products in carbon disulfide-treated peptides and protein.

Carbon disulfide (CS2) is an industrial solvent used in rayon production and as an organic synthetic precursor. It is also a member of the class of neuropathy-inducing xenobiotics known as the "neurofilament (NF) neurotoxicants". Current hypotheses propose direct reaction of CS2 with NF lysine epsilon-amine moieties as a step in the mechanism of this neuropathy. In this study, covalent CS2 binding in a lysine-containing dipeptide and in bovine serum albumin (BSA) in vitro was characterized. Dipeptide and BSA, incubated with 14CS2, exhibited stable incorporation of radioactivity after removal of unbound CS2 and reincubation in physiological buffer for up to 10 days. In contrast, free thiol levels decreased from a maximum immediately following CS2 exposure to near-base-line levels after 10 days, consistent with time-dependent conversion of initially formed N-substituted dithiocarbamate adducts into secondary products. HPLC/thermospray-MS and HPLC/UV photodiode-array analysis of CS2-dipeptide adducts confirmed dithiocarbamate formation and demonstrated their conversion into N-alkylisothiocyanates and, ultimately, N,N'-disubstituted thioureas and ureas. The results of UV spectrophotometry of CS2-treated BSA were also consistent with loss of dithiocarbamate and appearance of thioureas. Similar time-dependent formation of these products, in addition to N,N'-disubstituted thiuram disulfides, was demonstrated in CS2-treated BSA by means of 13C-NMR spectroscopy. SDS-PAGE analysis of adducted protein revealed a discrete, higher mobility band, likely representing a specific intramolecular cross-link. In contrast, no evidence for intermolecular protein cross-linking was obtained. Identical results were obtained with cysteinyl-blocked BSA, indicating the lack of formation of N,S-dialkyldithiocarbamate (dithiourethane) cross-links in these preparations.(ABSTRACT TRUNCATED AT 250 WORDS)

Studies in the dithiocarbamate series. Part II. The reaction of substituted N-benzylpiperidines with carbon disulphide

A series of substituted N-benzylpiperidines has been prepared, and their reaction with carbon disulphide studied. When the benzyl group possesses a 4-hydroxy-substituent together with 3,5-substituents such as alkyl and methoxy-the reaction leads to dithiocarbamate formation by ‘ insertion’ of carbon disulphide between the benzylic carbon atom and nitrogen. A three-stage mechanism is proposed, one stage involving the formation of a p-quinone methide as a transient intermediate. An alternative two-stage mechanism is considered but shown to be less likely.

Vulcanization of Elastomers. 24. The Effect of Fillers on the Course of Vulcanization Reactions. I

Abstract The effect of Aerosil on the chemical reactions and the kinetics observed in thiuram vulcanizations was subjected to closer scrutiny. Results now available show: 1. Aerosil does not alter the chemical mechanism of thiuram vulcanizations. It also has no qualitative effect on the kinetics of the various reactions involved. Thiuram disappearance and dithiocarbamate formation over by far the greatest range of conversions are first order reactions. Nevertheless, the limiting value of dithiocarbamate formation is somewhat higher than in the absence of Aerosil. 2. Quantitatively speaking, essential differences are involved. The rate constants for both thiuram loss and dithiocarbamate formation rise considerably when Aerosil is used; in both cases there is a dependence on the Aerosil content of the rubber compound. Yet the activation energies of thiuram disappearance and dithiocarbamate formation are practically the same as in Aerosil-free thiuram vulcanizations, amounting to about 21 kcal/mole. 3. Measurements of the limiting equilibrium swelling reveal that adding Aerosil does not result in additional crosslinking, so that in thiuram vulcanizations the catalytic action displayed by silica gel is the outstanding feature.

Vulcanization of Elastomers. 23. The Chemical Kinetics of Vulcanization Reactions and The Physical Properties of The Vulcanizates. I

Abstract As a complement to earlier investigations, and in order to examine more closely the connection between the chemical kinetics and the changes with vulcanization time of the physical properties in the case of vulcanization reactions, we used thiuram vulcanizations as an example, and concerned ourselves with the dependence of stress values (moduli) at different degrees of elongation and different vulcanization temperatures. We found: 1. Stress values attain a limiting value, dependent on the degree of elongation, but independent of the vulcanization temperature at constant elongation. 2. The rise in stress values with the vulcanization time is characterized by an initial delay, which, however, is practically nonexistent at higher temperatures. 3. The kinetics of the increase in stress values with vulcanization time are both qualitatively and quantitatively in accord with the dependence of the reciprocal equilibrium swelling on the vulcanization time; both processes, after a retardation, go according to the first order law and at the same rate. 4. From the temperature dependence of the rate constants of reciprocal equilibrium swelling, as well as of the increase in stress, an activation energy of 22 kcal/mole can be calculated, in good agreement with the activation energy of dithiocarbamate formation in thiuram vulcanizations.

Vulcanization of Elastomers. 21. Vulcanization of Natural Rubber with Thiuram Disulfides. VI

Abstract Bound sulfur in a pure thiuram vulcanizate increases relatively rapidly at first at all temperatures, reaches a poorly defined maximum at about 27 to 30%, independent of temperature, and then recedes slightly to reach a limiting value of 25% also independent of temperature, based on the original thiuram disulfide. The rise in sulfur content at the start points to a temperature-independent limiting value of 33%. It is shown that the combination of sulfur in this region initially follows a first order reaction, and goes at the same rate as the reduction in concentration of thiuram disulfide. It can be seen from the above that sulfur may be combined in thiuram vulcanization without simultaneous crosslinking. The dithiocarbamate formation increases rapidly in the region of longer vulcanization times, after the maximum in bound sulfur has been reached, without further combination of sulfur with the vulcanizate. The rate constants for thiuram decrease, for dithiocarbamate increase and for sulfur combination were calculated. The temperature dependence of each of these reactions has practically the same activation energy, 23 kcal/mole. The bound sulfur content of the vulcanizates in pure thiuram vulcanizations is no criterion of the state of vulcanization.