Disclosing the Antifungal Mechanisms of the Cyclam Salt H4[H2(4-CF3PhCH2)2Cyclam]Cl4 against Candida albicans and Candida krusei.

Mycoses are one of the major causes of morbidity/mortality among immunocompromised individuals. Considering the importance of these infections, the World Health Organization (WHO) defined a priority list of fungi for health in 2022 that include Candida albicans as belonging to the critical priority group and Pichia kudriavzevii (Candida krusei) to the medium priority group. The existence of few available antifungal drugs, their high toxicity, the acquired fungal resistance, and the appearance of new species with a broader spectrum of resistance, points out the need for searching for new antifungals, preferably with new and multiple mechanisms of action. The cyclam salt H4[H2(4-CF3PhCH2)2Cyclam]Cl4 was previously tested against several fungi and revealed an interesting activity, with minimal inhibitory concentration (MIC) values of 8 µg/mL for C. krusei and of 128 µg/mL for C. albicans. The main objective of the present work was to deeply understand the mechanisms involved in its antifungal activity. The effects of the cyclam salt on yeast metabolic viability (resazurin reduction assay), yeast mitochondrial function (JC-1 probe), production of reactive oxygen species (DCFH-DA probe) and on intracellular ATP levels (luciferin/luciferase assay) were evaluated. H4[H2(4-CF3PhCH2)2Cyclam]Cl4 induced a significant decrease in the metabolic activity of both C. albicans and C. krusei, an increase in Reactive Oxygen Species (ROS) production, and an impaired mitochondrial function. The latter was observed by the depolarization of the mitochondrial membrane and decrease in ATP intracellular levels, mechanisms that seems to be involved in the antifungal activity of H4[H2(4-CF3PhCH2)2Cyclam]Cl4. The interference of the cyclam salt with human cells revealed a CC50 value against HEK-293 embryonic kidney cells of 1.1 µg/mL and a HC10 value against human red blood cells of 0.8 µg/mL.

Use of Limestone Sludge in the Preparation of É©-Carrageenan/Alginate-Based Films.

The use of processed limestone sludge as a crosslinking agent for films based on Na-alginate and É©-carrageenan/Na-alginate blends was studied. Sorbitol was tested as a plasticizer. The produced gel formulations included alginate/sorbitol and carrageenan/alginate/sorbitol mixtures, with tested sorbitol concentrations of 0.0, 0.5 and 1.0 wt%. The limestone sludge waste obtained from the processing of quarried limestone was converted into an aqueous solution of Ca2+ by dissolution with mineral acid. This solution was then diluted in water and used to induce gel crosslinking. The necessity of using sorbitol as a component of the crosslinking solution was also assessed. The resulting films were characterized regarding their dimensional stability, microstructure, chemical structure, mechanical performance and antifungal properties. Alginate/sorbitol films displayed poor dimensional stability and were deemed not viable. Carrageenan/alginate/sorbitol films exhibited higher dimensional stability and smooth and flat surfaces, especially in compositions with 0.5 wt% sorbitol. However, an increasing amount of plasticizer appears to result in severe surface cracking, the development of a segregation phenomenon affecting carrageenan and an overall decrease in films' mechanical resistance. Although further studies regarding film composition-including plasticizer fraction, film optimal thickness and film/mold material interaction-are mandatory, the attained results show the potential of the reported É©-carrageenan/alginate/sorbitol films to be used towards the development of viable films derived from algal polysaccharides.

A Cyclam Salt as an Antifungal Agent: Interference with Candida spp. and Cryptococcus neoformans Mechanisms of Virulence.

The importance of fungal infections, particularly those caused by yeasts, is increasing among the medical community. Candida albicans and Cryptococcus neoformans are amongst the high-priority fungal species identified by the World Health Organization (WHO) and are considered in the critical group, while Candida krusei is included in the medium-priority group. The cyclam salt H4[H2(4-CF3PhCH2)2Cyclam]Cl4 proved to be active against the growth of these three yeasts, and the aim of this work was to verify its interference with their virulence mechanisms, whether shared or unique. H4[H2(4-CF3PhCH2)2Cyclam]Cl4 significantly inhibited biofilm production and catalase activity, being able to interfere with C. albicans dimorphic transition and C. neoformans melanin production. At the minimal inhibitory concentration (MIC) values, H4[H2(4-CF3PhCH2)2Cyclam]Cl4 had no antioxidant effect, as determined by the DPPH method. When using the RAW264.7 macrophage cell line, H4[H2(4-CF3PhCH2)2Cyclam]Cl4 reduced nitric oxide (NO) detection (the Griess reaction), but this effect was associated with a significant toxic effect on the cells.

Reactions of Heteroallenes with Salan-based Ti(IV) Complexes: A Joint Experimental and Computational Study.

The reaction of Ti(NMe2)4 with the salan ligand precursor H2N2O2H2 led to the formation of [(L*)Ti(NHMe2)2] (L*=N2O2 4-) that forms [(H2N2O2)TiCl2] upon reaction with two equiv. of Me3SiCl. [(L*)Ti(py)2] was obtained from the reaction of [Ti(NtBu)Cl2(py)3] with the sodium salt H2N2O2Na2. Treatment of [(L*)Ti(NHMe2)2] with two equiv. of tBuNCO led to the insertion of the isocyanate molecules into the Ti-Nsalan bonds with the formation of [{L*(N(tBu)CO)2}Ti]. Conversely, the reaction of [(H2N2O2)Ti(OiPr)2] with two equiv. of tBuNCO led to the insertion of one isocyanate molecule into a Ti-Nsalan bond with the formation of [{(HN2O2)(N(tBu)CO)}Ti(OiPr)]. Computational studies were performed to gain insight into the reactivity of isocyanates with salan-based Ti(IV) complexes.

Synthesis, Characterization, and Reactivity Studies of New Cyclam-Based Y(III) Complexes.

[(Bn2Cyclam)Y(N(SiMe3)2)] was prepared by reaction of H2Bn2Cyclam with Y[N(SiMe3)2]3. The protonation of the macrocycle ligand in [(Bn2Cyclam)Y(N(SiMe3)2)] is observed upon reaction with [HNMe3][BPh4] leading to the formation of [(HBn2Cyclam)Y(N(SiMe3)2)][BPh4]. DFT analysis of [(Bn2Cyclam)Y(N(SiMe3)2)] showed that the HOMO is located on the anionic nitrogen atoms of the cyclam ring indicating that protonation follows orbital control. Addition of H2Bn2Cyclam and H2(3,5-tBu2Bn)2Cyclam to a 1:3 mixture of YCl3 and LiCH2SiMe3 in THF resulted in the formation of [((C6H4CH2)BnCyclam)Y(THF)(µ-Cl)Li(THF)2] and [Y{(η3-3,5-tBu2Bn)2Cyclam}Li(THF)], respectively. The reaction of H23,5-tBu2Bn2Cyclam with Y(CH2SiMe3)3(THF)2 was studied and monitored by a temperature variation NMR experiment revealing the formation of [(3,5-tBu2Bn2Cyclam)Y(CH2SiMe3)]. Preliminary catalytic assays have shown that [Y{(η3-3,5-tBu2Bn)2Cyclam}Li(THF)] is a very efficient catalyst for the intramolecular hydroamination of 2,2-diphenyl-pent-4-enylamine.

Synthesis and Application of New Salan Titanium Complexes in the Catalytic Reduction of Aldehydes.

Complexes of formula [(H2N2O2)TiCl2] and [(H2N2O2)Ti(OiPr)2] (H2N2O2H2 = HOPh'CH2NH(CH2)2NHCH2Ph'OH, where Ph' = 2,4-(CMe2Ph)C6H2) were synthesized by the reaction of the salan ligand precursor H2N2O2H2 with TiCl4 and Ti(OiPr)4, respectively, in high yields. The dichlorido complex [(H2N2O2)TiCl2] revealed to be an efficient catalyst for the reduction of benzaldehyde in toluene. Full conversion was observed after 24 h at 55 °C in THF. The same catalyst also converted phenylacetaldehyde and hydrocinnamaldehyde into the corresponding alkanes quantitatively.

Bacterial Nosocomial Infections: Multidrug Resistance as a Trigger for the Development of Novel Antimicrobials.

Nosocomial bacterial infections are associated with high morbidity and mortality, posing a huge burden to healthcare systems worldwide. The ongoing COVID-19 pandemic, with the raised hospitalization of patients and the increased use of antimicrobial agents, boosted the emergence of difficult-to-treat multidrug-resistant (MDR) bacteria in hospital settings. Therefore, current available antibiotic treatments often have limited or no efficacy against nosocomial bacterial infections, and novel therapeutic approaches need to be considered. In this review, we analyze current antibacterial alternatives under investigation, focusing on metal-based complexes, antimicrobial peptides, and antisense antimicrobial therapeutics. The association of new compounds with older, commercially available antibiotics and the repurposing of existing drugs are also revised in this work.

Investigations into the Structure/Antibacterial Activity Relationships of Cyclam and Cyclen Derivatives.

A series of cyclam- and cyclen-derived salts are described in the present work; they were designed specifically to gain insights into their structure and antibacterial activity towards Staphylococcus aureus and Escherichia coli, used respectively, as Gram-positive and Gram-negative model organisms. The newly synthesized compounds are monosubstituted and trans-disubstituted tetraazamacrocycles that display benzyl, methylbenzyl, trifluoromethylbenzyl, or trifluoroethylbenzyl substituents appended on the nitrogen atoms of the macrocyclic ring. The results obtained show that the chemical nature, polarity, and substitution patterns of the benzyl groups, as well as the number of pendant arms, are critical parameters for the antibacterial activity of the cyclam-based salts. The most active compounds against both bacterial strains were the trans-disubstituted cyclam salts displaying CF3 groups in the para-position of the aromatic rings of the macrocyclic pendant arms. The analogous cyclen species presents a lower activity, revealing that the size of the macrocyclic backbone is an important requirement for the antibacterial activity of the tetraazamacrocycles. The nature of the anionic counterparts present on the salts was found to play a minor role in the antibacterial activity.

CoII Cryptates Convert CO2 into CO and CH4 under Visible Light.

Three CoII octaazacryptates, with different substituents on the aromatic rings (Br, NO2 , CCH), were synthesised and characterised. These and the already published non-substituted cryptate catalysed CO2 photoreduction to CO and CH4 under blue visible light at room temperature. Although CO was observed after short irradiation times and a large range of catalyst concentrations, CH4 was only observed after longer irradiation periods, such as 30 h, but with a small catalyst concentration (25 nm). Experiments with 13 C labelled CO2 showed that CO is formed and reacts further when the reaction time is long. The CCH catalyst is deactivated faster than the others and the more efficient catalyst for CH4 production is the one with Br. This reactivity trend was explained by an energy decomposition analysis based on DFT calculations.

New Cyclams and Their Copper(II) and Iron(III) Complexes: Synthesis and Potential Application as Anticancer Agents.

New cyclam derivatives (HOCH2 CH2 CH2 )2 (PhCH2 )2 Cyclam and (HOCH2 CH2 CH2 )2 ( 4-CF3 PhCH2 )2 Cyclam, as well as their CuII and FeIII complexes, were synthesized and characterized and their stability in cellular media was assessed. The cytotoxic effect of all compounds was examined on human cervical cancer (HeLa) cells, revealing strong anticancer activity. After 24 h, only complexes with the (HOCH2 CH2 CH2 )2 ( 4-CF3 PhCH2 )2 Cyclam ligand are cytotoxic, whereas after incubation for 72 h all compounds show significant antiproliferative effects. Notably, compounds containing 4-CF3 PhCH2 pendant arms on the cyclam ring revealed the most activity, with cytotoxicity values up to 12 times higher than those of cisplatin. All metal complexes seem to induce cell death through the formation of reactive oxygen species.

Cooperative Metal-Ligand Hydroamination Catalysis Supported by C-H Activation in Cyclam Zr(IV) Complexes.

Complexes of the type (R2Cyclam)ZrCl2 (where R = CH2═C(H)CH2 (All), CH2═C(Me)CH2 (MeAll), and PhCH2 (Bn)) react with suitable Grignard reagents to produce the corresponding alkyl derivatives (R2Cyclam)ZrR'2 (R' = Me, CH2Ph). Thermally induced double metalation of the pending arms of the cyclam ligand led to the formation of complexes ((CH═C(H)CH2)2Cyclam)Zr, 14, ((CH═C(Me)CH2)2Cyclam)Zr, 15, or ((C6H4CH2)2Cyclam)Zr, 16. These reactions proceed through C(sp2)-H bond activation and R'H elimination and convert the original dianionic tetracoordinated cyclam-based ligands in tetraanionic hexacoordinated ligands that establish two new Zr-C bonds. The cleavage of the Zr-C bonds may be readily achieved by treatment of the bis( ortho-metalated) species 16 with protic substrates ( tert-butanol, phenol, thiophenol, aniline, benzophenone hydrazone, pyrazole, and N, N'-diphenylhydrazine), to give rise to (Bn2Cyclam)ZrX2 complexes (X = OtBu, OPh, SPh, NHPh, NHNCPh2, C3H3N2, N, N'-PhNNPh). In catalytic conditions, complexes (All2Cyclam)Zr(NMe2)2, 14, 15, or 16 convert 2,2-diphenyl-pent-4-enylamine to 2-methyl-4,4-diphenylpyrrolidine with 100% selectivity and conversion values varying between 61 and 88% in 4.5 h, at 115 °C. Complexes 14 and 15, which display metalated allyl and methallyl pending groups on the cyclam ring, are the most active species (1.7 < TOF < 2.0 h-1). The mechanism of this reaction was studied by density functional theory that revealed two competitive paths, one proceeding through an imido intermediate and another that occurs via an amido ligand. Both cases represent cooperative mechanisms with active participation of the cyclam, as proton exchange between the coordinated substrate and the ligand side arm with reversible C-H activation is a crucial feature of the mechanism.

Stable, Yet Highly Reactive Nonclassical Iron(II) Polyhydride Pincer Complexes: Z-Selective Dimerization and Hydroboration of Terminal Alkynes.

The synthesis, characterization, and catalytic activity of nonclassical iron(II) polyhydride complexes containing tridentate PNP pincer-type ligands is described. These compounds of the general formula [Fe(PNP)(H)2(η2-H2)] exhibit remarkable reactivity toward terminal alkynes. They efficiently promote the catalytic dimerization of aryl acetylenes giving the corresponding conjugated 1,3-enynes in excellent yields with low catalyst loadings. When the reaction is carried out in the presence of pinacolborane, vinyl boronates are obtained. Both reactions take place under mild conditions and are highly chemo-, regio-, and stereoselective with up to 99% Z-selectivity.

Synthesis, antimicrobial activity and toxicity to nematodes of cyclam derivatives.

The antimicrobial activity and toxicity to nematodes of the cyclam salt [H2{H2(4-CF3PhCH2)2Cyclam}](CH3COO)2⋅(CH3COOH)2 were evaluated. Estimated minimum inhibitory concentrations (MICs) of 9, 261 and 15 µg/mL were obtained for Escherichia coli, Pseudomonas aeruginosa and Staphylococcus aureus, respectively. For selected Candida spp., the estimated MICs obtained ranged from 32 µg/mL to 63 µg/mL. Bactericidal activity was demonstrated but the compound was not reliably fungicidal. Concentrations of the cyclam salt up to 32 µg/mL did not significantly affect survival of the nematode Caenorhabditis elegans; however, concentrations equal or above this value significantly affected nematode survival in a dose-dependent manner.

Reactions of heteroallenes with cyclam-based Zr(IV) complexes.

This work describes reactions of heteroallenes with diamido-diamine cyclam-based Zr(iv) complexes of the general formula (Bn2Cyclam)ZrX2 (X = O(t)Bu, , O(i)Pr, , SPh, , NH(t)Bu, ) as well as the di-orthometallated species ((C6H4CH2)2Cyclam)Zr, . The reactions of isocyanates or isothiocyanates with , or resulted in the formation of N-bonded ureate or thioureate cyclam complexes upon [2 + 2] cycloaddition of the Zr-Namido bonds of the cyclam to the heteroallene (). DFT calculations showed that κ(2)-N,N'-ureate bonding is favoured over κ(2)-N,O-ureates, which in turn may be formed in reactions with bulky isocyanates as 1-naphthyl isocyanate (NpN[double bond, length as m-dash]C[double bond, length as m-dash]O). The reactions of with N,N'-cyclohexylcarbodiimide (CyN[double bond, length as m-dash]C[double bond, length as m-dash]NCy) and carbon disulfide afforded guanidinate and dithiocarbamate fragments, respectively, appended to one of the nitrogen atoms of the cyclam ligand. These reactions represent a reliable method for the synthesis of asymmetrically N-functionalized cyclams giving rise to C1 symmetry Zr(iv) species by addition of one equivalent of heteroallenes. The reaction of (Bn2Cyclam)Zr(NH(t)Bu)2, , with one equivalent of mesityl isocyanate (MesN[double bond, length as m-dash]C[double bond, length as m-dash]O) also proceeds through insertion, involving one Zr-NH(t)Bu bond. However, it was observed that the reaction of (Bn2Cyclam)Zr(NH(t)Bu)2, , with MesN[double bond, length as m-dash]C[double bond, length as m-dash]O follows a different path if the reaction is carried out at 60 °C. In this case the reaction leads to [2 + 2] addition of the Zr-Ncyclam bond to the isocyanate, with a concomitant occurrence of orthometallation of the one benzyl pending group of the cyclam ring. The reaction of (t)BuN[double bond, length as m-dash]C[double bond, length as m-dash]O with the di-orthometallated complex ((C6H4CH2)2Cyclam)Zr, , also gave a κ(2)-N,N'-ureate fragment, by isocyanate addition to the macrocycle. DFT calculations on these systems were conducted in an attempt to rationalise the reactivity patterns observed.

Synthesis and structural characterization of novel cyclam-based zirconium complexes and their use in the controlled ROP of rac-lactide: access to cyclam-functionalized polylactide materials.

Novel Bn(2)Cyclam-based zirconium complexes of the type (Bn(2)Cyclam)Zr(X)(X') (3, X = X' = OPh; 4, X = X' = SPh; 5, X = Cl, X' = O(i)Pr) were synthesized in good yields via metathesis routes involving the reaction of the dichloro precursor (Bn(2)Cyclam)Zr(Cl)(2) and the appropriate lithium salts. The molecular structures of compounds 3, 4 and 5, as determined by X-ray crystallographic studies, all confirmed the effective chelation of the Bn(2)Cyclam ligand in a κ(4)-N(2)N(2)' fashion, with the hexa-coordinated Zr center adopting a trigonal prismatic geometry. Complexes 3-5 as well as the diisopropoxide derivative (Bn(2)Cyclam)Zr(O(i)Pr)(2) (2) were all found to initiate the ring-opening polymerization (ROP) of rac-lactide in a controlled manner, as deduced from SEC data and linear correlations between molecular weight numbers (M(n)) and monomer conversion as the ROP proceeds. While initiator 2 polymerizes rac-lactide to afford, as expected, an O(i)Pr-ester-end PLA, the ROP of rac-lactide by species 3 or 4 affords an unusual cyclam-end group PLA, as deduced from MALDI-TOF data. The bonding and the electronic structures of the dialkoxides 2 and 3 were assessed by DFT and their possible influence on the polymerization mechanism is discussed.

Cyclam functionalization through isocyanate insertion in Zr-N bonds.

The insertion of isocyanates in (Bn(2)Cyclam)ZrX(2) is regioselective; (Bn(2)Cyclam)Zr(OR)(2) produces urea-like moieties by the insertion of RN═C═O in the Zr-N(amido) bonds of the cyclam ring. Depending on the bulkiness of the isocyanate R groups, O- and N-bound ureates are formed. (Bn(2)Cyclam)Zr(NH(t)Bu)(2) reacts with MesN═C═O at the terminal Zr-N bonds.