Mechanistic studies of the triggered release of liposomal contents by matrix metalloproteinase-9

Matrix metalloproteinases (MMPs) constitute a class of extracellular-matrix-degrading enzymes overexpressed in many cancers and contribute to the metastatic ability of the cancer cells. We have recently demonstrated that liposomal contents can be released when triggered by the enzyme MMP-9. Herein, we report the results of our mechanistic studies of the MMP-9-triggered release of liposomal contents. We synthesized peptides containing the cleavage site for MMP-9 and conjugated them with fatty acids to prepare the corresponding lipopeptides. By employing circular dichroism (CD) spectroscopy, we demonstrated that the lipopeptides, when incorporated into liposomes, are demixed in the lipid bilayers and generate triple-helical structures. MMP-9 cleaves the triple-helical peptides, leading to the release of the liposomal contents. Other MMPs, which cannot hydrolyze triple-helical peptides, fail to release the contents from the liposomes. We also observed that the rate and extent of release of the liposomal contents depend on the mismatch between the acyl chains of the synthesized lipopeptide and phospholipid components of the liposomes. CD spectroscopic studies imply that the observed differences in the release reflect the ability of the liposomal membrane to anneal the defects following the enzymatic cleavage of the liposome-incorporated lipopeptides.     

Oligonucleotide Analogues with Integrated Bases and Backbone

The thiomethylene‐linked U*[s]U⁽*⁾ dimers 9 – 14 were synthesized by substitution of the 6‐[(mesyloxy)methyl]uridine 6 by the thiolate derived from the uridine‐5′‐thioacetates 7 and 8 followed by O‐deprotection. Similarly, the thiomethylene‐linked A*[s]A⁽*⁾ dimers 9 – 14 were obtained from the 8‐(bromomethyl)adenosine 15 and the adenosine‐5′‐thioacetates 16 and 17 . The concentration dependence of both H? N(3) of the U*[s]U⁽*⁾ dimers 9 – 14 evidences the formation of linear and cyclic duplexes, and of linear higher associates, C(8 or 6)CH₂OH and/or C(5′/II)OH groups favouring the formation of cyclic duplexes. The concentration dependence of the chemical shift for both H₂N? C(6) of the A*[s]A⁽*⁾ dimers 18 – 23 evidences the formation of mainly linear associates. The heteroassociation of U*[s]U⁽*⁾ to A*[s]A⁽*⁾ dimers is stronger than the homoassociation of U*[s]U⁽*⁾ dimers, as evidenced by diluting equimolar mixtures of 11 / 20 and 13 / 22 . A 1 : 1 stoichiometry of the heteroassociation is evidenced by a Job's plot for 11 / 20 , and by mole ratio plots for 9 / 18, 10 / 19, 12 / 21, 13 / 22 , and 14 / 23 .     

Oligonucleotide Analogues with Integrated Bases and Backbones. Part 19.

The A*[s]U⁽*⁾ dinucleosides 1 and 2 form thermoreversible gels in organic solvents. The basis of the gelation is the formation of linear aggregates by base pairing following desolvation of the nucleobases. This is evidenced by the absence of gel formation by the C(6)‐deaminated analogue 3 of 1 , the correlation of gelation with the anti‐conformation, as preferred for 1 , and the temperature‐, concentration‐, and time‐dependent CD spectra. The gels were also characterized by the minimum gelation concentration, the gel–sol transition (melting) temperature, and rheological properties.     

Heteroleptic copper(I) complexes coupled with methano[60]fullerene: synthesis, electrochemistry, and photophysics

Heteroleptic copper(I) complexes CuPOP-F and CuFc-F have been prepared from a fullerene-substituted phenanthroline ligand and bis[2-(diphenylphosphino)phenyl] ether (POP) and 1,1'-bis(diphenylphosphino)ferrocene (dppFc), respectively. Electrochemical studies indicate that some ground-state electronic interaction between the fullerene subunit and the metal-complexed moiety are present in both CuPOP-F and CuFc-F. Their photophysical properties have been investigated by steady state and time-resolved UV-vis-NIR luminescence spectroscopy and nanosecond laser flash photolysis in a CH2Cl2 solution and compared to those of the corresponding model copper(I) complexes CuPOP and CuFc and of the fullerene model compound F. Selective excitation of the methanofullerene moiety in CuPOP-F results in regular deactivation of the lowest singlet and triplet states, indicating no intercomponent interactions. Conversely, excitation of the copper(I)-complexed unit (405 nm, 40% selectivity) shows that the strongly luminescent triplet metal-to-ligand charge-transfer ((3)MLCT) excited state located at 2.40 eV is quenched by the carbon sphere with a rate constant of 1.6 x 10(8) s(-1). Details on the mechanism of photodynamic processes in CuPOP-F via transient absorption are hampered by the rather unfavorable partition of light excitation between the two chromophores. By determination of the yield of formation of the lowest fullerene triplet level through sensitized singlet oxygen luminescence in the NIR region, it is shown that the final sink of photoinduced processes is always the fullerene triplet. This can be populated via a two-step charge-separation charge-recombination process and a less favored (3)MLCT --> (3)C60 triplet-triplet energy-transfer pathway. In CuFc-F, both of the photoexcited copper(I)-complexed and fullerene moieties are quenched by the presence of the ferrocene unit, most likely via ultrafast energy transfer.     

A 2D coordination polymer with canted ferromagnetism constructed from ferromagnetic [Ni(II)Co(II)] nodes

A bimetallic coordination polymer, infinity (2)[{LNi (II)Co (II)}(dca) 2], has been constructed from heterobinuclear [Ni (II)Co (II)] nodes and dicyanamido spacers [L (2-) is the dianion of the Schiff base resulting from the 2:1 condensation of 3-methoxysalicyladehyde with 1,3-propanediamine; L (2-) = N, N'-propylenebis(3-methoxysalycilideneiminato)]. The intranode Co (II)-Ni (II) interaction was found to be ferromagnetic because of the orthogonality of the magnetic orbitals. Below 12 K, the onset of the canted ferromagnetic ordering is observed.     

Interaction of mixed-donor macrocycles containing the 1,10-phenanthroline subunit with selected transition and post-transition metal ions: metal ion recognition in competitive liquid-liquid solvent extraction of Cu(II), Zn(II), Pb(II), Cd(II), Ag(I), and Hg(II)

Two new mixed aza-thia crowns 5-aza-2,8-dithia[9]-(2,9)-1,10-phenanthrolinophane (L(4)) and 2,8-diaza-5-thia[9]-(2,9)-1,10-phenanthrolinophane (L(7)) have been synthesized and characterized. The coordination behavior of L(4) and L(7) toward the metal ions Cu(II), Zn(II), Pb(II), Cd(II), Hg(II), and Ag(I) was studied in aqueous solution by potentiometric methods, in CD3CN/D2O 4:1 (v/v) by (1)H NMR titrations and in the solid state. The data obtained were compared with those available for the coordination behavior toward the same metal ions of structurally analogous mixed donor macrocyclic ligands L(1)-L(3), L(5), L(6): all these contain a phenanthroline subunit but have only S/O/N(aromatic) donor groups in the remaining portion of the ring and are, therefore, less water-soluble than L(4) and L(7). The complexes [Cd(NO3)2(L(5))], [Pb(L(7))](ClO4)2 x 1/2MeCN, [Pb(L(4))](ClO4)2 x MeCN, and [Cu(L(7))](ClO4)2 x 3/2MeNO2 were characterized by X-ray crystallography. The efficacy of L(1)-L(7) in competitive liquid-liquid metal ion extraction of Cu(II), Zn(II), Cd(II), Pb(II), Ag(I), and Hg(II) was assessed. In the absence of Hg(II), a clear extraction selectivity for Ag(I) was observed in all systems investigated.     

GaCl3-assisted cyclization reactions in hypersilyl(trimethylsilyl)aminodichlorophosphine

Hypersilyl(trimethylsilyl)aminodichlorophosphine, (hyp)N(SiMe3)PCl2, was treated with GaCl3, which resulted in the formation of an interesting novel bicycle, composed of a four-membered SiNP2 ring and a five-membered P2Ga2Cl ring. In the presence of Me3SiN3, the same reaction provided access to a cyclo-2-phospha-4-sila-1,3-diazenium tetrachlorogallate. The free chloro-cyclo-phosphasiladiazane was obtained by the addition of nucleophilic bases.     

Boraguanidinates: synthesis, X-ray structures, and reactions of {Li2[(i)Pr2NB(NDipp)2]}2 with p-block and group 12 element halides

The new Dipp-substituted boranes (Dipp = 2,6-diisopropylphenyl) (i)Pr2NB(NHDipp)2 (5), BrB(NHDipp)2 (7b), and B(NHDipp)3 (8) have been prepared in high yields and characterized in solution by multinuclear NMR spectroscopy ((1)H, (11)B and (13)C) and in the solid state by X-ray crystallography. Reaction of 5 with 2 equiv of (n)BuLi in n-hexane produces {Li2[(i)Pr2NB(NDipp)2]}2 (6) the first example of a stable dilithiated boraguanidinate. The unsolvated complex 6 has a dimeric structure in the solid state. A survey of the reactions of 6 with p-block and Group 12 element halides revealed various types of reactivity including (a) disproportionation (InCl), (b) reduction to the metal (PbCl2, CdCl2, TeBr4, TeI4, and TlCl), and (c) simple metathesis (GeCl2, MgCl2, and ZnCl2). The metathetical products were characterized by multinuclear NMR spectra but, in contrast to the dilithiated complex 6, they readily decompose in non-coordinating solvents to form the diprotonated compound 5.     

Perturbative calculation of Franck-Condon integrals: new hints for a rational implementation

Perturbative approaches to the mode mixing effects in the calculation of Franck-Condon integrals are analyzed and discussed. The zero order multidimensional Franck-Condon integrals are factorized into products of one-dimensional ones, so that recurrence relations can be used without need of storing a huge number of data. Calculations on model systems show that at the second order of perturbation, the method gives results in very good agreement with the exact ones, even in the case of significantly large Duschinsky effect. The accuracy of the results can be substantially improved by grouping together all those modes which are strongly mixed with each other, usually a few ones, for which Franck-Condon integrals can be computed exactly, and using the perturbative approach for treating the smaller mixing between all the other modes.