Synthesis, characterisation and antimicrobial activity of supramolecular cobalt-peptide conjugates.

Herein, we describe the synthesis and characterisation of four new supramolecular cobalt conjugates of antimicrobial peptides functionalised with terpyridine ligands (L). Peptides were chosen based on the well-established arginine-tryptophan (RW)3 motif, with terpyridine-derivatized lysine (Lys(tpy)) added to the sequence, or replacing tryptophan residues. Self-assembly of the antimicrobial peptides with Co(BF4)2·6H2O formed exclusively CoL2 dimers (for peptides with one tpy ligand each) and Co2L4 metallo-macrocycles (for peptides with two tpy ligands for each peptide), which could be 'locked' by oxidation of Co(+II) to Co(+III) with ammonium ceric nitrate. The Co-peptide complexes were characterised by mass spectrometry and in solution by NMR spectroscopy, including 2D diffusion ordered NMR spectroscopy (DOSY) which confirmed the proposed stoichiometries. The antimicrobial activity of the novel peptides and their metallo-supramolecular assemblies was investigated by determination of their minimal inhibitory concentration (MIC) against a panel of Gram-positive and Gram-negative bacteria. Complexation with cobalt increases the activity of the peptides in almost every case. Most of the new metal-peptide conjugates showed good activity against Gram-positive bacteria, including a multi-resistant S. aureus strain and the opportunistic pathogenic yeast C. albicans (down to 7 µmol l-1 for the most active Co2L4 derivate), a value that is increased five-fold compared to the lysine-derivatized peptide ligand alone. Interestingly, conjugates of the CoL2 type also showed decent activity against Gram-negative bacteria including the WHO-flagged problematic A. baumannii strain (down to 18 µmol l-1 for the most active derivative).

Synthesis and DNA interaction studies of Ru(II) cell penetrating peptide (CPP) bioconjugates.

The synthesis of the first bioconjugates of a set of ruthenium(II) dipyridophenazine complexes with two different cell penetrating peptides (CPPs) is described. The CPPs, an arginine rich TAT-9 (RKKRRQRRR) sequence and the Xentry peptide (LCLRPVG), were synthesized using standard SPPS protocols, and the bioconjugates were obtained by the microwave-assisted coupling of the HOBt/TBTU preactivated metal complexes with the respective peptides on Wang resin. The racemic metal complexes were obtained by modified literature procedures. The bioconjugates were cleaved from the resin, purified by semi-preparative HPLC and characterized by analytical HPLC, high resolution mass spectrometry (HR-MS), and NMR spectroscopy. Despite the bioconjugation of the peptides to the dppz ligand, DNA intercalation was observed by CD spectroscopy, viscometry and the characteristic switch-on fluorescence of this class of compounds. Furthermore, the cellular uptake of the Xentry bioconjugates was confirmed by live cell imaging. Like the parent metal complexes, the bioconjugates show low in vitro cytotoxicity (IC50 > 80 µM), which is similar to the respective metal complexes alone.

Comparison of Proteomic Responses as Global Approach to Antibiotic Mechanism of Action Elucidation.

New antibiotics are urgently needed to address the mounting resistance challenge. In early drug discovery, one of the bottlenecks is the elucidation of targets and mechanisms. To accelerate antibiotic research, we provide a proteomic approach for the rapid classification of compounds into those with precedented and unprecedented modes of action. We established a proteomic response library of Bacillus subtilis covering 91 antibiotics and comparator compounds, and a mathematical approach was developed to aid data analysis. Comparison of proteomic responses (CoPR) allows the rapid identification of antibiotics with dual mechanisms of action as shown for atypical tetracyclines. It also aids in generating hypotheses on mechanisms of action as presented for salvarsan (arsphenamine) and the antirheumatic agent auranofin, which is under consideration for repurposing. Proteomic profiling also provides insights into the impact of antibiotics on bacterial physiology through analysis of marker proteins indicative of the impairment of cellular processes and structures. As demonstrated for trans-translation, a promising target not yet exploited clinically, proteomic profiling supports chemical biology approaches to investigating bacterial physiology.

Probing CO Generation through Metal-Assisted Alcohol Dehydrogenation in Metal-2-(arylazo)phenol Complexes Using Isotopic Labeling (Metal = Ru, Ir): Synthesis, Characterization, and Cytotoxicity Studies.

The reaction of 2-{2-(benzo[1,3]dioxol-5-yl)- diazo}-4-methylphenol (HL) with [Ru(PPh3)3Cl2] in ethanol resulted in the carbonylated ruthenium complex [RuL(PPh3)2(CO)] (1), wherein metal-assisted decarbonylation via in situ ethanol dehydrogenation is observed. When the reaction was performed in acetonitrile, however, the complex [RuL(PPh3)2(CH3CN)] (2) was obtained as the main product, probably by trapping of a common intermediate through coordination of CH3CN to the Ru(II) center. The analogous reaction of HL with [Ir(PPh3)3Cl] in ethanol did not result in ethanol decarbonylation and instead gave the organoiridium hydride complex [IrL(PPh3)2(H)] (3). Unambiguous evidence for the generation of CO via ruthenium-assisted ethanol oxidation is provided by the synthesis of the 13C-labeled complex, [Ru(PPh3)2L(13CO)] (1A) using isotopically labeled ethanol, CH313CH2OH. To summarize all the evidence, a ruthenium-assisted mechanistic pathway for the decarbonylation and generation of alkane via alcohol dehydrogenation is proposed. In addition, the in vitro antiproliferative activity of complexes 1-3 was tested against human cervical (HeLa) and human colorectal adenocarcinoma (HT-29) cell lines. Complexes 1-3 showed impressive cytotoxicity against both HeLa (half-maximal inhibitory concentration (IC50) value of 3.84-4.22 µM) and HT-29 cancer cells (IC50 values between 3.3 and 4.5 µM). Moreover, the complexes were comparatively less toxic to noncancerous NIH-3T3 cells.

Direct Crystallographic Observation of CO2 Captured in Zig Zag Channels of a Copper(I) Metal-Organic Framework.

Single crystal X-ray diffraction has been used to study the CO2 absorption sites in a microporous Cu-MOF, [CuI2(py-pzpypz)2(µ-CN)2]n (1) (where py-pzpypz = 4-(4-pyridyl)-2,5-dipyrazyl-pyridine), which features zigzag-shaped channels, at a range of CO2 pressures (1, 5, and 10 bar) and at two temperatures (240 and 298 K). Unlike the acetonitrile molecules in the as-synthesized MOF, 1·MeCN, the CO2 molecules in 1·nCO2 (n = 0.8, 0.7, 0.45) are preferentially centered on the vertices of each zig and zag, which allows for weak (azine) C-H···OCO interactions with the H atoms on the electron-deficient pyrazine and pyridine rings of the MOF.

A CuAAC Click Approach for the Introduction of Bidentate Metal Complexes to a Sulfanilamide-Derived Antibiotic Fragment.

A simple "click-chemistry" approach was employed in order to functionalize the known antibiotic fragment sulfanilamide with a bidentate pyridyl-triazole pocket, which allowed for the synthesis of ruthenium(II) and rhenium(I) carbonyl chloride complexes. Six new complexes were prepared and comprehensively characterized, including five single crystal X-ray structures, photophysical characterization, and testing for antimicrobial activity. Interestingly, functionalization of the pyridine ring with an ortho-hydroxymethyl group resulted in a greater than 100-fold increase in the rate of ligand release in a dimethylsulfoxide solution. Subsequent studies indicated this process could be further accelerated by irradiation with 265 nm light. Structural characterization of four of the complexes indicates that this is the result of a lengthening and weakening of the Re-NPyridine bond (average (Ltri) = 2.19 Å vs LtriOH = 2.25 Å) due to the steric influence of the hydroxymethyl group. The organometallic rhenium(I) pyridyl-triazole functionality maintains its characteristic fluorescent properties despite the presence of the sulfonamide moiety. Two of the compounds showed modest antimicrobial activity against methicillin-resistant Staphylococcus aureus, whereas the structurally similar sulfamethoxazole alone showed no activity under the same conditions.

From Enzymes to Functional Materials-Towards Activation of Small Molecules.

The design of non-noble metal-containing heterogeneous catalysts for the activation of small molecules is of utmost importance for our society. While nature possesses very sophisticated machineries to perform such conversions, rationally designed catalytic materials are rare. Herein, we aim to raise the awareness of the overall common design and working principles of catalysts incorporating aspects of biology, chemistry, and material sciences.

Commensurate CO2 Capture, and Shape Selectivity for HCCH over H2CCH2, in Zigzag Channels of a Robust Cu(I)(CN)(L) Metal-Organic Framework.

A novel copper(I) metal-organic framework (MOF), {[Cu(I)2(py-pzpypz)2(µ-CN)2]·MeCN}n (1·MeCN), with an unusual topology is shown to be robust, retaining crystallinity during desolvation to give 1, which has also been structurally characterized [py-pzpypz is 4-(4-pyridyl)-2,5-dipyrazylpyridine)]. Zigzag-shaped channels, which in 1·MeCN were occupied by disordered MeCN molecules, run along the c axis of 1, resulting in a significant solvent-accessible void space (9.6% of the unit cell volume). These tight zigzags, bordered by (Cu(I)CN)n chains, make 1 an ideal candidate for investigations into shape-based selectivity. MOF 1 shows a moderate enthalpy of adsorption for binding CO2 (-32 kJ mol(-1) at moderate loadings), which results in a good selectivity for CO2 over N2 of 4.8:1 under real-world operating conditions of a 15:85 CO2/N2 mixture at 1 bar. Furthermore, 1 was investigated for shape-based selectivity of small hydrocarbons, revealing preferential uptake of linear acetylene gas over ethylene and methane, partially due to kinetic trapping of the guests with larger kinetic diameters.

Spin Crossover in Dinuclear N4S2 Iron(II) Thioether-Triazole Complexes: Access to [HS-HS], [HS-LS], and [LS-LS] States.

Access to a new family of thioether-linked PSRT ligands, 4-substituted-3,5-bis{[(2-pyridylmethyl)sulfanyl]methyl}-4H-1,2,4-triazoles (analogues of the previously studied amino-linked PMRT ligands), has been established. Four such ligands have been prepared, PSPhT, PS(i)BuT, PS(t-Bu)PhT, and PS(Me)PhT, with R = Ph, (i)Bu, (t-Bu)Ph, and (Me)Ph, respectively. Three dinuclear colorless to pale green iron(II) complexes, [Fe(II)2(PSRT)2](BF4)4·solvent, featuring N4S2 donor sets, were prepared. Single-crystal structure determinations on [Fe(II)2(PSPhT)2](BF4)4·2MeCN·H2O, [Fe(II)2(PSPhT)2](BF4)4·2(1)/2MeCN·(1)/2H2O·THF, [Fe(II)2(PS(Me)PhT)2](BF4)4·2MeCN, and [Fe(II)2(PS(i)BuT)2](BF4)4·4MeCN reveal that all four are stabilized in the [HS-HS] state to 100 K and that both possible binding modes of the bis-terdentate ligands, cis- and trans-axial, are observed. Variable-temperature magnetic susceptibility studies of air-dried crystals (solvatomorphs of the single crystal samples) reveal the first examples of spin crossover (SCO) for a dinuclear iron(II) complex with N4S2 coordination. Specifically, [Fe(II)2(PSPhT)2](BF4)4·2(1)/2H2O undergoes a multistep but complete SCO from [HS-HS] to [LS-LS], whereas [Fe(II)2(PS(Me)PhT)2](BF4)4·1(1)/2MeCN·2H2O exhibits a half-SCO from [HS-HS] to [HS-LS]. In contrast, [Fe(II)2(PS(i)BuT)2](BF4)4·MeCN·H2O remains [HS-HS] down to 50 K. The reflectance spectrum of pale green [Fe(II)2(PSPhT)2](BF4)4·(1)/2CHCl3·2(1)/2H2O (solvatomorph A) reveals a trace of LS character (572 nm band (1)A1g → (1)T1g). Evans' (1)H NMR method and UV-vis spectroscopy studies revealed that on cooling dark green acetonitrile solutions of these complexes from 313 to 233 K, all three undergo SCO centered at or near room temperature. The tendency of the complexes to go LS in solution reflects the electronic impact of R on the σ-donor strength of the PSRT ligand, whereas the opposite trend in stabilization of the LS state is seen in the solid state, where crystal packing effects, of the R group and solvent content, dominate the SCO behavior.

Reversible quantitative guest sensing via spin crossover of an iron(ii) triazole.

A new phenyl-triazole-pyrazine ligand, 4-p-tolyl-3-(phenyl)-5-(2-pyrazinyl)-1,2,4-triazole (tolpzph), was prepared in order to enforce pyrazine coordination of the iron(ii) centre in the resulting complex, [FeII(tolpzph)2(NCS)2]·THF (1·THF). Structure determinations carried out on this discrete mononuclear complex, 1·THF, at 273 K (mostly high spin) and 100 K (mostly low spin) demonstrate this was successful, and that spin crossover (SCO) occurred on cooling. Subsequent magnetic measurements on 1·THF revealed that it shows highly sensitive and reversible solvent-dependent SCO, with T1/2(1·THF) = 255 K vs. T1/2(1) = 212 K (with SCO of 1 more abrupt and occurring with a 4 K hysteresis loop), a drop of 43 K due to THF loss. This is reversible over at least 10 cycles of re-solvating with THF followed by re-drying, so 1 ↔ 1·THF can be considered an 'on-off' THF sensor, monitored by the T1/2 reversibly shifting (by 43 K). Furthermore, quantitative sensing of the fractional amount of THF present in 1·nTHF, 0 ≤ n ≤ 1, is demonstrated. Monitoring the T1/2 and using TGA to quantify n(THF) revealed a linear dependence (25 data points; Pearson r2 = 0.93): T1/2 = 41.1n(THF) + 219. Finally, 1 is also shown to take up CHCl3 [T1/2(1·CHCl3) = 248 K], with a logarithmic T1/2 dependence on the fractional amount of CHCl3 present (10 data points; Pearson r2 = 0.98): T1/2 = 27.0 log10[n(CHCl3)] + 243. This study is a proof of principle that a (multi-use) quantitative sensor material based on spin crossover is feasible.

Pressure induced separation of phase-transition-triggered-abrupt vs. gradual components of spin crossover.

The application of pressure on [Co(II)(dpzca)2], which at ambient pressure undergoes abrupt spin crossover (SCO) with thermal hysteresis, gives unique insights into SCO. It reversibly separates the crystallographic phase transition (I41/a↔P21/c) and associated abrupt SCO from the underlying gradual SCO, as shown by detailed room temperature (RT) X-ray crystallography and temperature dependent magnetic susceptibility studies, both under a range of 10 different pressures. The pressure effects are shown to be reversible. The crystal structure of the pressure-induced low-spin state is determined at RT at 0.42(2) and 1.78(9) GPa. At the highest pressure [1.78(9) GPa] the Co-N bond lengths are consistent with the complex being fully LS, and the conjugated terdentate ligands are significantly distorted out of plane. The abrupt SCO event can be shifted up to RT by application of a hydrostatic pressure of ∼0.4 GPa. These magnetic susceptibility (vs. temperature) and X-ray crystallography (at RT) studies, under a range of pressures, show that the SCO can be tuned over a wide range of temperature and pressure space, including RT SCO.

Spin Crossover, Reversible Redox, and Supramolecular Interactions in 3d Complexes of 4-(4-Pyridyl)-2,5-dipyrazyl-pyridine.

A new terpyridine-inspired terdentate ligand, 4-(4-pyridyl)-2,5-dipyrazyl-pyridine (py-pzpypz), featuring three "spare" nitrogen donors "out the back", has been used to synthesize five bis-ligand complexes, [M(II)(py-pzpypz)2]X2, where M = Mn with X = ClO4, or M = Fe, Co, Ni, and Zn with X = BF4. In contrast, when M = Cu(II), regardless of the M:L ratio employed, 1:1 M:L products were obtained: for X = BF4 a 1D chain {[Cu(II)(py-pzpypz)(DMF)2](BF4)2}n, and for X = Cl a monometallic complex [Cu(py-pzpypz)Cl2]. All seven complexes were structurally characterized, confirming the expected N6 coordination of the M(II) centers in all cases except Cu(II). Notably, a Jahn-Teller elongation is observed in the Co(II) complex, consistent with it being low spin at 100 K. The Cu(II) 1D chain complex has an N4O2 coordination sphere as in this case the "spare" pyridine donor out the back of the py-pzpypz ligand bridges to the next Cu(II) center in the chain, hence providing both a terdentate site and a monodentate pyridine to the next Cu(II) center, and the coordination sphere is completed by weak axial coordination by two DMF solvent molecules. The Cu(II) center in the monometallic complex has an N3Cl2 square pyramidal coordination sphere. In all cases, the noncoordinating, "spare", pyrazine nitrogen atoms are involved in interesting intermolecular interactions, including NPz-π interactions and nonclassical C-H···NPz hydrogen bonding. The Fe(II) complex is low spin as expected. Two polymorphs of the Co(II) complex were obtained, both of which showed gradual spin crossover, with a room temperature T1/2. Two reversible redox processes are observed for [Co(II)(py-pzpypz)2](BF4)2, with Em(M(I)/M(II)) = -0.63 V and Em(M(II)/M(III)) = +0.37 V, and a quasireversible redox process for [Fe(II)(py-pzpypz)2](BF4)2, with Em(M(II)/M(III)) = +1.26 V, versus 0.01 M AgNO3/Ag in MeCN. These potentials are shifted to significantly higher potentials (by ∼0.45 V) than the literature values for the corresponding Fe(II) and Co(II) complexes of the equivalent all-pyridine ligand, consistent with replacement of the two pyridine rings by two pyrazine rings significantly stabilizing the lower oxidation states.

Pyrazine-imide complexes: reversible redox and MOF building blocks.

The synthesis of the symmetric pyrazine imide ligand, N-(2-pyrazylcarbonyl)-2-pyrazinecarboxamide, (Hdpzca) and five new first row transition metal complexes of it are reported: [M(II)(dpzca)(2)], M(II) = Fe, Cu, Zn; [Cu(II)(dpzca)(H(2)O)(2)]BF(4), [Cu(II)(dpzca)(H(2)O)(3)](2)SiF(6). The crystal structures of Hdpzca, [Co(II)(dpzca)(2)], [Cu(II)(dpzca)(2)], {[Co(III)(dpzca)(2)](BF(4))}(2)·5CH(3)CN and [Cu(II)(dpzca)(H(2)O)(3))](2)SiF(6)·2H(2)O were determined and reveal an orthogonal positioning of the 'spare' pyrazine nitrogen atoms and 'spare' pairs of imide oxygen atoms. The [M(II)(dpzca)(2)] complexes are therefore useful six-coordinate building blocks for producing larger supramolecular assemblies. Two examples of secondary assembly of [M(II)(dpzca)(2)] complexes, with M = Co and Ni, with silver nitrate gave single crystals; {[Co(III)(dpzca)(2)Ag](NO(3))(2)·2H(2)O}n and {([Ni(II)(dpzca)(2)Ag(I)(1/2)](1/2NO(3))(xH(2)O}n were structurally characterised. The redox processes of [M(II)(dpzca)(2)], with M(II) = Fe, Ni, Cu and Zn, are reported and, as seen for M(II) = Co, reversible metal- and ligand-based redox processes are observed, with E(m)(M(II)/M(III)) values 0.15-0.24 V higher than for the analogous complexes of Hpypzca (non-symmetric pyridine/pyrazine imide ligand), and 0.35-0.36 V higher than for the complexes of Hbpca (symmetric pyridine imide ligand).

Selective gas adsorption in a pair of robust isostructural MOFs differing in framework charge and anion loading.

Activation of the secondary assembly instructions in the mononuclear pyrazine imide complexes [Co(III)(dpzca)2](BF4) or [Co(II)(dpzca)2] and [Ni(II)(dpzca)2] has facilitated the construction of two robust nanoporous three-dimensional coordination polymers, [Co(III)(dpzca)2Ag](BF4)2·2(H2O) [1·2(H2O)] and [Ni(II)(dpzca)2Ag]BF4·0.5(acetone) [2·0.5(acetone)]. Despite the difference in charge distribution and anion loading, the framework structures of 1·2(H2O) and 2·0.5(acetone) are isostructural. One dimensional channels along the b-axis permeate the structures and contain the tetrafluoroborate counterions (the Co(III)-based MOF has twice as many BF4(-) anions as the Ni(II)-based MOF) and guest solvent molecules. These anions are not readily exchanged whereas the solvent molecules can be reversibly removed and replaced. The H2, N2, CO2, CH4, H2O, CH3OH, and CH3CN sorption behaviors of the evacuated frameworks 1 and 2 at 298 K have been studied, and modeled, and both show very high selectivity for CO2 over N2. The increased anion loading in the channels of Co(III)-based MOF 1 relative to Ni(II)-based MOF 2 results in increased selectivity for CO2 over N2 but a decrease in the sorption kinetics and storage capacity of the framework.

Hysteretic spin crossover in iron(II) complexes of a new pyridine-triazole-pyrazine ligand is tuned by choice of NCE co-ligand.

A family of three new mononuclear complexes of the general form [Fe(L(pz))2(NCE)2] has been prepared (L(pz) = 4-p-tolyl-3-(2-pyrazinyl)-5-(2-pyridyl)-1,2,4-triazole; E = S, Se, BH3). All three exhibit spin crossover, in two cases with hysteresis, with T1/2 being predictably tuned by varying the coordinated anion.