A new class of macrocyclic lanthanide complexes for cell labeling and magnetic resonance imaging applications

Lanthanide complexes have wide applications in biochemical research and biomedical imaging. We have designed and synthesized a new class of macrocyclic lanthanide chelates, Ln/DTPA-PDA-C(n), for cell labeling and magnetic resonance imaging (MRI) applications. Two lipophilic Gd3+ complexes, Gd/DTPA-PDA-C(n) (n = 10, 12), labeled a number of cultured mammalian cells noninvasively at concentrations as low as a few micromolar. Cells took up these agents rapidly and showed robust intensity increases in T1-weighed MR images. Labeled cells showed normal morphology and doubling time as control cells. In addition to cultured cells, these agents also labeled primary cells in tissues such as dissected pancreatic islets. To study the mechanism of cellular uptake, we applied the technique of diffusion enhanced fluorescence resonance energy transfer (DEFRET) to determine the cellular localization of these lipophilic lanthanide complexes. After loading cells with a luminescent complex, Tb/DTPA-PDA-C10, we observed DEFRET between the Tb3+ complex and extracellular, but not intracellular, calcein. We concluded that these cyclic lanthanide complexes label cells by inserting two hydrophobic alkyl chains into cell membranes with the hydrophilic metal binding site facing the extracellular medium. As the first imaging application of these macrocyclic lanthanide chelates, we labeled insulin secreting beta-cells with Gd/DTPA-PDA-C12. Labeled cells were encapsulated in hollow fibers and were implanted in a nude mouse. MR imaging of implanted beta-cells showed that these cells could be followed in vivo for up to two weeks. The combined advantages of this new class of macrocyclic contrast agents ensure future imaging applications to track cell movement and localization in different biological systems.     

The synthesis of di- and tetramethyl substituted macrocyclic polyether-diester ligands

Three series of macrocyclic polyether-diester ligands have been prepared from dimethyl triethylene glycol ( 20 ), two dimethyl tetraethylene glycols ( 21,23 ), dimethyl pentaethylene glycol ( 22 ) and tetramethyl tetraethylene glycol ( 24 ) and diglycolyl chloride (products 5–9 ), thiadiglycolyl chloride (products 10–14 ) and 2,6-pyridine dicarbonyl chloride (products 15–19 ). The eighteen-membered rings ( 6 and 16 ) formed solid potassium thiocyanate complexes. The eighteen- and twenty-one-membered ring compounds 6–8 and 16–18 complexed with benzylammonium perchlorate in methylene chloride-d₂ as shown by significant chemical shift changes in the ¹H nmr spectra.     

Photo-induced DNA cleavage activity and remarkable photocytotoxicity of lanthanide(III) complexes of a polypyridyl ligand

Lanthanide(III) complexes [Ln(pyphen)(acac)(2)(NO(3))] (1, 2), [Ln(pydppz)(acac)(2)(NO(3))] (3, 4) and [La(pydppz)(anacac)(2)(NO(3))] (5), where Ln is La(III) (in 1, 3, 5) and Gd(III) (in 2, 4), pyphen is 6-(2-pyridyl)-1,10-phenanthroline, pydppz is 6-(2-pyridyl)-dipyrido[3,2-a:2',3'-c]phenazine, anacac is anthracenylacetylacetonate and acac is acetylacetonate, were prepared, characterized and their DNA photocleavage activity and photocytotoxicity studied. The crystal structure of complex 2 displays a GdO(6)N(3) coordination. The pydppz complexes 3-5 show an electronic spectral band at ~390 nm in DMF. The La(III) complexes are diamagnetic, while the Gd(III) complexes are paramagnetic with seven unpaired electrons. The molar conductivity data suggest 1 : 1 electrolytic nature of the complexes in aqueous DMF. They are avid binders to calf thymus DNA giving K(b) in the range of 5.4 × 10(4)-1.2 × 10(6) M(-1). Complexes 3-5 efficiently cleave supercoiled DNA to its nicked circular form in UV-A light of 365 nm via formation of singlet oxygen ((1)O(2)) and hydroxyl radical (HO˙) species. Complexes 3-5 also exhibit significant photocytotoxic effect in HeLa cancer cells giving respective IC(50) value of 0.16(±0.01), 0.15(±0.01) and 0.26±(0.02) μM in UV-A light of 365 nm, while they are less toxic in dark with an IC(50) value of >3 μM. The presence of an additional pyridyl group makes the pydppz complexes more photocytotoxic than their dppz analogues. FACS analysis of the HeLa cells treated with complex 4 shows apoptosis as the major pathway of cell death. Nuclear localization of complex 5 having an anthracenyl moiety as a fluorophore is evidenced from the confocal microscopic studies.     

Template synthesis, spectroscopic studies and biological screening of macrocyclic complexes derived from thiocarbohydrazide and benzil

A novel series of complexes of the type [M(TML)X₂]; where TML is a tetradentate macrocyclic ligand; M = Co(II), Ni(II), Cu(II) or Zn(II); X = Cl⁻, CH₃COO⁻ or NO ₃ ⁻ have been synthesized by template condensation of benzil and thiocarbohydrazide in the presence of divalent metal salts in methanolic medium. The complexes have been characterized with the help of elemental analyses, conductance measurements, molecular weight determination, magnetic measurements, electronic, NMR, infrared and far infrared spectral studies. Electronic spectra along with magnetic moments suggest the six coordinate octahedral geometry for these complexes. The low value of molar conductance indicates them to be non-electrolytes. The biological activities of metal complexes have been tested in vitro against a number of pathogenic bacteria to assess their inhibiting potential.     

DNA cleavage and antimicrobial studies of 17-membered schiff base macrocyclic triazoles: synthesis and spectroscopic approach

A novel series of 17-membered complexes [MLCl₂] (M = Co²⁺, Ni²⁺ and Cu²⁺) have been synthesized with newly derived biologically active ligands (L^I–L^IV). These ligands were synthesized by the condensation of 3-subtituted-4-amino-5-hydrazino-1,2,4-triazole with bis(phthalaldehyde)ethylenediamine precursor. The structure of the complexes has been proposed by elemental analyses, IR, EPR, electronic spectral studies, conductivity, magnetic, thermal and electrochemical studies. All the complexes are soluble in DMF and DMSO and are non-electrolytes. All these Schiff bases and their complexes have been screened for their antibacterial (Escherichia coli, Staphylococus aureus, Salmonella typhi, Pseudomonas aeruginosa) and antifungal activities (Aspergillus niger, Aspergillus flavus and Cladosporium) by the Agar and Potato dextrose agar diffusion method. The DNA cleavage study was done by Agarose gel electrophoresis technique.     

Rapid hydrolytic cleavage of the mRNA model compound HPNP by glycine based macrocyclic lanthanide ribonuclease mimics

The lanthanide ion based macrocyclic complexes 1.Ln mimic the hydrophobic nature of ribonucleases, where the lanthanide ions induce the formation of a hydrophobic cavity for 1, giving rise to a large order of magnitude enhancement in the hydrolytic cleavage of HPNP.     

Spectroscopy: the study of DNA cleavage by newly synthesized polydentate macrocyclic ligand and its copper(II) complexes

A novel hexadentate nitrogen donor [N6] macrocyclic ligand, i.e. 2,6,12,16,21,22-hexaaza-3,5,13,15-tetramethyl-4,14-diethyl-tricyclo-[15.3.1.1(7-11)]docosane-1(21),2,5,7(22),8,10,12,15,17,19-decaene (L), has been synthesized. Copper(II) complexes with this ligand have been prepared and subjected to elemental analyses, molar conductance measurements, magnetic susceptibility measurements, mass, 1H NMR (ligand), IR, electronic, and EPR spectral studies. On the basis of molar conductance the complexes may be formulated as [Cu(L)X2] [X = Cl(-), Br(-), NO3(-) and CH3COO(-)] due to their nonelectrolytic nature in N,N'-dimethylformamide (DMF). All the complexes are of the high spin type and are six coordinated. On the basis of IR, electronic and EPR spectral studies tetragonal geometry has been assigned to the Cu(II) complexes. The interaction of these complexes with calf thymus DNA has been explored by using absorption, emission, viscosity measurements, electrochemical studies and DNA cleavage. All the experimental results suggest that the complexes bind to DNA and also promote the cleavage plasmid pBR 322, in the presence of H2O2 and ascorbic acid.     

Controlling the variation of axial water exchange rates in macrocyclic lanthanide(III) complexes

The rate of axial water exchange in well-defined series of lanthanide complexes depends on the extent of second sphere hydration which is determined by complex hydrophobicity and the nature of the lanthanide ion and its counter-ion.     

Synthesis,DNA-binding and cleavage studies of macrocyclic copper(II) complexes

Two macrocyclic copper(II) complexes, [CuL¹](ClO₄)₂ (L¹ = 2,6,9,13-tetraparacyclophane, a Schiff base) and [CuL²]Cl₂ [L² = 3,10-bis(2-benzyl)-1,3,5,8,10,13-hexaazacyclotetradecane] have been prepared and characterized by elemental analysis, u.v.–vis., i.r. and mass spectra. Absorption, fluorescence, circular dichroic spectra and viscosity experiments have been carried out on the interaction of the two complexes with calf thymus CT DNA. The results suggest that both complexes can bind to CT DNA by intercalation via the aromatic moiety ring in the macrocycle into the base pairs of DNA. [CuL¹](ClO₄)₂ binds to CT DNA more strongly than [CuL²]Cl₂. The position of the aromatic ring in the macrocycle plays an important role in deciding the extent of binding of the complexes to DNA. Significantly, the complexes have been found to be single-strand DNA cleavers in the presence of H₂O₂ or/and 2-mercaptoethanol.     

Studies on organolanthanide complexes: VII. Formation and cleavage of carbon-metal bonds of dicyclopentadienyl-early lanthanide complexes

Four new 1,10-phenanthroline-coordinated early lanthanide complexes containing a σ-carbon-metal bond 1–4 were synthesized by the reaction of alkynylsodium or alkyllithium with (η⁵-C₅H₅)₂LnCl·nPhen in THF at 0 or −78°C. The complexes (η⁵-C₅H₅)₂LnCl·nPhen were prepared from LnCl₃·nPhen and C₅H₅Na. 1,1′-Trimethylenedicyclopentadienyl(phenylacetylenylneodymium). THF 5 was also prepared. These complexes were identified by elemental analysis, IR, ¹H NMR spectroscopy and thermogravimetry. Protolysis reactions of these complexes with H₂O. CH₃OH and t-C₄H₉OH show that different protolytic reagents give the products with different cleavage extents of σ- and π-bonds. The ligands in the complexes also affect the cleavage of π-bonds. β-Hydrogen elimination of complex 3 takes place with thermal decomposition.     

The synthesis of the zero-valent nickel complexes via di-π-allylnickel

A new route to zero-valent nickel complexes, (cycloocta-1,5-diene)duroquinonenickel, (endo-dicyclopentadiene)duroquinonenickel, dicycloocta-1,5-dienenickel and bis(tetracyclone)nickel, as described. The synthetic procedure involves the reaction between di-π-allylnickel and duroquinone, cyclic dienes, or tetraphenylcyclopentadienone. The replacement of the π-allyl ligand with duroquinone or a cyclic diene is suggested to follow the associative mechanism. The ¹³C NMR and mass spectra of the compounds isolated are reported.     

Dissociation kinetics of macrocyclic trivalent lanthanide complexes of 1,4,7,10-tetraazacyclododecane-1,7-diacetic acid (DO2A)

The [H(+)]-catalyzed dissociation rate constants of several trivalent lanthanide (Ln) complexes of 1,4,7,10-tetraazacyclododecane-1,7-diacetic acid (LnDO2A(+), Ln = La, Pr, Eu, Er and Lu) have been determined in two pH ranges: 3.73-5.11 and 1.75-2.65 at four different temperatures (19-41.0 °C) in aqueous media at a constant ionic strength of 0.1 mol dm(-3) (LiClO(4)). For the study in the higher pH range, i.e. pH 3.73-5.11, copper(II) ion was used as the scavenger for the free ligand DO2A in acetate/acetic acid buffer medium. The rates of Ln(III) complex dissociation have been found to be independent of [Cu(2+)] and all the Ln(III) complexes studied show [H(+)]-dependence at low acid concentrations but become [H(+)]-independent at high acid concentrations. Influence of the acetate ion content in the buffer on the dissociation rate has also been investigated and all the complexes exhibit a first-order dependence on [Acetate]. The dissociation reactions follow the rate law: k(obs) = k(Ac)[Acetate] + K'k(lim)[H(+)]/(1 + K'[H(+)]) where k(AC) is the dissociation rate constant for the [Acetate]-dependent pathway, k(lim) is the limiting rate constant, and K' is the equilibrium constant for the reaction LnDO2A(+) + H(+) ? LnDO2AH(2+). In the lower pH range, i.e. pH 1.75-2.65, the dye indicator, cresol red, was used to monitor the dissociation rate, and all the Ln(III) complexes also show [H(+)]-dependence dissociation pathways but without the rate saturation observed at higher pH range. The dissociation reactions follow the simple rate law: k(obs) = k(H)[H(+)], where k(H) is the dissociation rate constant for the pathway involving monoprotonated species. The absence of an [H(+)]-independent pathway in both pH ranges indicates that LnDO2A(+) complexes are kinetically rather inert. The obtained k(AC) values follow the order: LaDO2A(+) > PrDO2A(+) > EuDO2A(+) > ErDO2A(+) > LuDO2A(+), whereas the k(lim) and k(H) values follow the order: LaDO2A(+) > PrDO2A(+) > ErDO2A(+) > EuDO2A(+) > LuDO2A(+), mostly consistent with their thermodynamic stability order, i.e. the more thermodynamically stable the more kinetically inert. In both pH ranges, activation parameters, ΔH*, ΔS* and ΔG*, for both acetate-dependent and proton-catalyzed dissociation pathways have been obtained for most of the La(III), Pr(III), Eu(III), Er(III) and Lu(III) complexes, from the temperature dependence measurements of the rate constants in the 19-41 °C range. An isokinetic (linear) relationship is found between ΔH* and ΔS* values, which supports a common reaction mechanism.     

Synthesis and characterisation of dimeric eight-coordinate lanthanide(III) complexes of a macrocyclic tribenzylphosphinate ligand

The macrocyclic ligand 1,4,7,10-tetraazacyclododecane-1,4,7-triyl(methylenebenzyl-phosphinic acid) H3L3, has been prepared and its complexes with Eu, Gd and Tb(III) studied by NMR, relaxometry, luminescence and single crystal X-ray crystallography. In solution and in the crystal, the complexes have eight-coordinate metal centres with bridging phosphinate groups linking the two twisted square antiprismatic coordination polyhedra. A single stereoisomer crystallises from solution with an RRR and SSS configuration at the P centres in each sub-unit. The relaxivity of [GdL3]2 is low (1.9 mM-1 s-1, 298 K, 20 MHz), consistent with the absence of any proximate water molecules. The terbium dimer possesses a relatively long excited state lifetime (2.47 ms, 298 K).     

A new method for the synthesis of macrocyclic ketones

Summary We have described the synthesis of macrocyclic diacetylene ketones by oxidative condensation of aliphatic ,-diacetyiene ketones.     

A high-throughput approach to lanthanide complexes and their rapid screening in the ring opening polymerisation of caprolactone

Libraries of lanthanide complexes supported by nitrogen and oxygen containing ligands have been synthesised using a high-throughput approach. The complexes were employed in the ring-opening polymerisation of epsilon-caprolactone, in some cases giving polycaprolactone of controlled molecular weight and narrow polydispersity. The libraries, based on twenty-one ligands and eight lanthanide reagents, were developed in order to determine the best combination of lanthanide metal and ligand. They were prepared via transamination reactions of [Ln[N(SiMe(3))(2)](3)] complexes with tetradentate dianionic ligands containing oxygen and nitrogen donors. 1H NMR spectroscopy was used to screen polymerisation activity. The steric demand of the ligand has a significant effect on the polymerisation process, as do the type of nitrogen donor and the size of the central Ln(3+) ion. Ligands containing aryl rings with bulky substituents such as tert-pentyl groups afforded species capable of performing controlled polymerisation of caprolactone, whereas less bulky groups such as methyl were not effective. Yttrium and mid-sized lanthanides such as samarium showed increased activity compared with the larger lanthanides, lanthanum and praseodymium, and the smaller lanthanides like ytterbium. X-ray crystal structures of a sterically demanding chelating amine-bis((2-hydroxyaryl)methyl) ligand and a chloride bridged dinuclear gadolinium complex are reported. The centrosymmetric molecule contains gadolinium in distorted capped trigonal prismatic environments bonded to two amine, two phenolate, one THF and two chloride donors.     

New chiral lanthanide amide ate complexes for the catalysed synthesis of scalemic nitrogen-containing heterocycles

New chiral binaphthylamido yttrium and ytterbium ate complexes with lithium and potassium counterions have been synthesised and characterised. X-ray structures have been obtained for [Li(thf)4][Ln{(R)-C20H12(NC5H9)2}2] (Ln=Yb, Y) and [K(thf)5][Yb{(R)-C20H12(NCH2CMe3)2}2] as isostructural complexes. The efficiency of these complexes for the enantioselective intramolecular hydroamination was examined. [Li(thf)4][Yb{(R)-C20H12(NC5H9)2}2] afforded the highest enantiomeric excess (up to 87 %) for the synthesis of a spiropyrrolidine, while [Li(thf)4][Y{(R)-C20H12(NC5H9)2}2] proved to be slightly more active. The role of the counter cation in the active catalytic species was evidenced by the comparison between lithium and potassium ate complexes. The most active catalyst of this series, [Li(thf)4][Yb{(R)-C20H12(NCH2CMe3)2}2], was successfully used for the cyclisation of aminopentenes with internal double bonds.     

Organoruthenium(II) thiosemicarbazone complexes: synthesis, spectral characterization, DNA binding and DNA cleavage studies

A series of new ruthenium(II) complexes were synthesized with Schiff bases derived from salicylaldehyde / o-hydroxyacetophenone/ o-vanillin / 2-hydroxy-1-naphthaldehyde with thiosemicarbazide and acetyl furan. They are characterized by elemental analysis, IR, electronic, ¹H NMR, ¹³C NMR and mass spectral studies. The elemental analysis suggests the stoichiometry to be 1:1 (metal:ligand). Four of these complexes were tested for its binding with CT-DNA using absorption spectroscopic studies and two of these complexes exhibit efficient DNA cleavage activity.