Novel phosphate–phosphonate hybrid nanomaterials applied to biology

A new process for preparing oligonucleotide arrays is described that uses surface grafting chemistry which is fundamentally different from the electrostatic adsorption and organic covalent binding methods normally employed. Solid supports are modified with a mixed organic/inorganic zirconium phosphonate monolayer film providing a stable, well-defined interface. Oligonucleotide probes terminated with phosphate are spotted directly to the zirconated surface forming a covalent linkage. Specific binding of terminal phosphate groups with minimal binding of the internal phosphate diesters has been demonstrated. On the other hand, the reaction of a bisphosphonate bone resorption inhibitor (Zoledronate) with calcium deficient apatites (CDAs) was studied as a potential route to local drug delivery systems active against bone resorption disorders. A simple mathematical model of the Zoledronate/CDA interaction was designed that correctly described the adsorption of Zoledronate onto CDAs. The resulting Zoledronate-loaded materials were found to release the drug in different phosphate-containing media, with a satisfactory agreement between experimental data and the values predicted from the model.     

苯甲酰亚胺动态化学键在构筑pH响应药物载体中的应用

介绍了作者近期利用苯甲酰亚胺化学结构构筑对生理环境pH值响应的药物输送体系的工作进展.亚胺(Schiff碱)作为动态共价键早已为人们所熟知,但其在pH敏感药物输送载体领域的应用并不多.苯甲酰亚胺由于苯环与碳氮双键共轭作用使其在生理pH值下的稳定性更好,同时具有在弱酸环境下(如固态肿瘤细胞外环境、内涵体环境等)的水解特性...     

Biological activity of functionalized SiO2 thin films prepared by sol-gel method

In this work we investigated the biological properties of sol-gel films in aqueous medium. Functionalized silica films were prepared by the sol-gel process, from organically modified silicon alkoxides with amino or thiol groups. Covalent binding of proteins with different orientations according to the hydrophilic or hydrophobic character of the surface was studied. This binding occurred via a covalent coupling agent providing a very stable linkage. No denaturation was detected and a good detection of the antigen was observed. Immunoassays have demonstrated the biological activity of grafted antibodies.     

Metal phosphonates applied to biotechnologies: a novel approach to oligonucleotide microarrays

A new process for preparing oligonucleotide arrays is described that uses surface grafting chemistry which is fundamentally different from the electrostatic adsorption and organic covalent binding methods normally employed. Solid supports are modified with a mixed organic/inorganic zirconium phosphonate monolayer film providing a stable, well-defined interface. Oligonucleotide probes terminated with phosphate are spotted directly on to the zirconated surface forming a covalent linkage. Specific binding of terminal phosphate groups with minimal binding of the internal phosphate diesters has been demonstrated. The mixed organic/inorganic thin films have also been extended for use arraying DNA duplex probes, and therefore represent a viable general approach to DNA-based bioarrays. Ideas for interfacing mixed organic/inorganic interfaces to other bioapplications are also discussed.     

EQUILIBRIUM AMONG HEMATOPORPHYRIN DERIVATIVE COMPONENTSs—II. EFFECT OF ESTERASE ACTIVITY

Photofrin II is the hematoporphyrin-derivative fraction enriched in covalently-linked oligomers, characterized by a high degree of folding. Interaction with hydrophobic structures, such as biomolecules and cell structures, results in a modification of the equilibria among the different species, as a consequence of an unfolding effect exerted towards the electrostatic aggregates. The effect of esterase activity was evaluated, taking into account the nature suggested for the covalent linkage of the oligomers (ether and/or ester). The study was performed in Photofrin II aqueous solution by means of absorption and fluorescence spectral analysis. The results showed that the esterase is active only towards the unfold oligomers: that is, in Photofrin II solution supplemented with albumin. In these conditions, spectral analysis revealed the presence of a monomerization process, which is clearly evident during the first four hours of incubation. The monomerization effect induced by the enzyme was also proven by both equilibrium-dialysis measurements and zinc ion complexation. Zinc ion complexes with high affinity for monomeric species, giving rise to a very distinct emission band at 580 nm. The amount of ester linkage shown in the oligomers through enzyme hydrolysis appeared to be less than might have been expected, owing to the inhibiting effect of the monomer produced on the enzyme. The results are a step toward clarifying the intracellular and intratissue turnover of the drug observed after administration.     

Glutaraldehyde-modified electrode for nonlabeling voltammetric detection of <Emphasis Type="Italic">p16</Emphasis><Superscript><Emphasis Type="Italic">INK4A</Emphasis></Superscript> gene

A nonlabeling electrochemical detection method for analyzing the polymerase-chain-reaction-amplified sequence-specific p16 ^INK4A gene, in which the basis for the covalent immobilization of deoxyribonucleic acid (DNA) probe is described, has been developed. The self-assembly process was based on the covalent coupling of glutaraldehyde (GA) as an arm molecule onto an amino-functional surface. The p16 ^INK4A gene was used as the model target for the methylation detection of early cancer diagnosis. An amino-modified DNA probe was successfully assembled on the GA-coupling surface through the formation of Schiff base under potential control. The hybridization of amino-modified DNA probes with the target was investigated by means of electrochemical measurements, including cyclic voltammetry and square wave voltammetry. Furthermore, the functions of GA coupling for sequence-specific detection were compared with those obtained based on mercaptopropionic acid. Hybridization experiments indicated that the covalent coupling of GA was suitable for the immobilization of DNA probe and was sensitive to the electrochemical detection of single-base mismatches of label-free DNA targets in hybridization. Moreover, reported probe-modified surfaces exhibited excellent stability, and the hybridization reactions were found to be completely reversible and highly specific for recognition in subsequent hybridization processes. The strategy provided the potential for taking full advantage of existing modified electrode technologies and was verified in microarray technology, which could be applied as a useful and powerful tool in electrochemical biosensor and microarray technology.     

布洛芬高分子前体药物及纳米微球的合成和表征

将非甾体消炎药布洛芬以共价键连接到舍双键的甲基丙烯酸-2-羟基乙酯(HEMA)上,制成含布洛芬药物的单体,进而通过自聚或共聚,合成了含布洛芬的高分子药物;采用乳液聚合方法制备了高分子药物蚋米微球。研究了影响聚合反应的有关因素,并对所合成的产物用^1H-NMR,GPC和TEM等检测手段进行了表征。     

Supramolecular Polymerizations

Unimers of both natural and synthetic origin self‐assemble into linear, helical, columnar, planar and three‐dimensional structures depending upon the functionality of supramolecular interactions. Recent reports describing the mechanism of formation, properties and possible applications of these systems are critically reviewed. The assembling of one‐dimensional systems produces equilibrium polymers showing a length distribution and a degree of polymerization that may far exceed that of typical condensation polymers. Their growth may occur by a step‐by‐step process akin to polycondensation, and by cooperative processes such as helical growth or growth coupled to liquid crystallinity. Of particular interest are functional systems based on the coupling of a chemical reaction to supramolecular polymerization, and systems based on a covalent polymer hosted within the cavity of a supramolecular one. The assembly of two and three‐dimensional systems occurs through a process akin to crystallization. The supramolecular organization of amphiphiles such as block copolymers is currently well described by the mean‐field theory of unstable modes in homogeneous melts. An alternative, less sophisticated approach considers the growth of specifically designed building blocks. Possible applications are in areas that expand the uses of covalent polymers, electrochemical and photonic devices, ion‐selective channels, separation processes, microengines mimicking the performance of biological systems, storage of sequential information, biocompatible and patterned surfaces, sensors. A classification including additional systems that have been described as supramolecular polymers is presented.     

苯佐卡因高分子载体药物及其纳米微球的合成

将苯佐卡因通过酯键和甲基丙烯酸相连,合成了含苯佐卡因的可聚合单体(BM),BM自聚得到苯佐卡因高分子载体药物(PBM),并采用乳液聚合法制备了PBM纳米微球。PBM的结构经1H NMR和TEM表征。     

Versatile Bispidine-Based Bifunctional Chelators for 64CuII-Labelling of Biomolecules

Bifunctional chelators as parts of modular metal-based radiopharmaceuticals are responsible for stable complexation of the radiometal ion and for covalent linkage between the complex and the targeting vector. To avoid loss of complex stability, the bioconjugation strategy should not interfere with the radiometal chelation by occupying coordinating groups. The C9 position of the very stable Cu^II chelator 3,7-diazabicyclo[3.3.1]nonane (bispidine) is virtually predestined to introduce functional groups for facile bioconjugation as this functionalisation does not disturb the metal binding centre. We describe the preparation and characterisation of a set of novel bispidine derivatives equipped with suitable functional groups for diverse bioconjugation reactions, including common amine coupling strategies (bispidine-isothiocyanate) and the Cu-free strain-promoted alkyne–azide cycloaddition. We demonstrate their functionality and versatility in an exemplary way by conjugation to an antibody-based biomolecule and validate the obtained conjugate in vitro and in vivo.     

Immobilized sucrose phosphorylase fromLeuconostoc mesenteroides

Sucrose phosphorylase fromLeuconostoc mesenteroides was immobilbilized by covalent linkage to several supports, and the specific activity recovery was 2-11%. The enzyme adsorbed onto DEAE-cellulose re tained about 18% specific activity and was stable over eight months. The optimum pH (7.0) and temperature (30°C) did not change after immobilization. Also there was no improvement of thermal stability, and Km for sucrose and phosphate was lower compared to the soluble enzyme.     

Reticular synthesis of covalent organic borosilicate frameworks

This paper reports the synthesis and characterization of a new crystalline 3D covalent organic framework, COF-202: [C(C6H4)4]3[B3O6 (tBuSi)2]4, formed from condensation of a divergent boronic acid, tetra(4-dihydroxyborylphenyl)methane, and tert-butylsilane triol, tBuSi(OH)3. This framework is constructed through strong covalent bonds (Si-O, B-O) that link triangular and tetrahedral building units to form a structure based on the carbon nitride topology. COF-202 demonstrates high thermal stability, low density, and high porosity with a surface area of 2690 m2 g-1. The design and synthesis of COF-202 expand the type of linkage that could be used to crystallize new materials with extended covalent organic frameworks.     

Synthesis and physicochemical characteristics of a liver-targeted conjugate of fluorodeoxyuridine monophosphate with lactosaminated human albumin

In previous experiments fluorodeoxyuridine monophosphate (FUdRMP) was conjugated with lactosaminated human albumin (L-HSA). Fluorodeoxyuridine (FUdR) is an anticancer agent and L-HSA is a hepatotropic carrier of drugs obtained by the covalent linkage of lactose residues to the albumin molecule. The conjugate was synthesised via the imidazolide of FUdRMP at alkaline pH. Peripheral venous administration of L-HSA-FUdRMP produced enhanced FUdR levels in hepatic blood and might accomplish a non-invasive loco-regional chemotherapy of liver micrometastases. In the present paper some physicochemical characteristics of L-HSA-FUdRMP are reported. Polyacrylamide gel electrophoresis indicated that the coupling reaction did not cause covalent aggregation of the L-HSA molecules. 31P NMR spectra of the conjugate showed that FUdRMP was linked to L-HSA by phosphoamide bonds to lysine and histidine residues, and the area of the peak due to the lysine bond represented more than 80% of the spectrum of L-HSA-FUdRMP. MALDI analysis revealed a partial degradation of the peptide backbone of the conjugate which could not be detected using other methods of analysis. The degradation was not caused by the coupling of lactose molecules to albumin, but rather a consequence of FUdRMP conjugation with L-HSA. This fragmentation was dependent on the pH of the medium used for the FUdRMP coupling reaction. By decreasing the pH to 7.5, conjugates were obtained with a lower drug load but with a substantially reduced fragmentation, which should be preferred for a clinical use of L-HSA-FUdRMP.     

α-SrMnO3电子结构的第一性原理研究

采用平面波赝势方法对钙钛矿型锰酸盐氧化合物α-SrMnO3的电子结构进行了第一性原理研究. 六方钙钛矿型结构α-SrMnO3化合物为磁性绝缘体, 磁基态对应于共面八面体及共顶点八面体间的磁性交换作用均为反铁磁性(AFM), 其禁带宽度为1.6 eV; 费米能级附近的Mn3d态与O2p态存在很强的杂化作用, 属于共价绝缘体, 这种强共价性使得Mn4+的自旋磁矩偏离理想值. 采用Noodleman的对称性破缺方法, 根据α-SrMnO3不同磁有序态的总能量拟合出α-SrMnO3中的自旋交换耦合常数. α-SrMnO3的局部微结构(Mn—O—Mn)决定了整个体系的特殊磁性交换作用. 共面及共顶点的八面体间均存在AFM交换作用, 并且共顶点八面体间的AFM作用比较强.     

A highly stable short alpha-helix constrained by a main-chain hydrogen-bond surrogate

Herein we describe a strategy for the preparation of artificial alpha-helices involving replacement of one of the main-chain hydrogen bonds with a covalent linkage. To mimic the C=O...H-N hydrogen bond as closely as possible, we envisioned a covalent bond of the type C=X-Y-N, where X and Y are two carbon atoms connected through an olefin metathesis reaction. Our results demonstrate that the replacement of a hydrogen bond between the i and i + 4 residues at the N-terminus of a short peptide with a carbon-carbon bond results in a highly stable constrained alpha-helix at physiological conditions as indicated by CD and NMR spectroscopies. The advantage of this strategy is that it allows access to short alpha-helices with strict preservation of molecular recognition surfaces required for biomolecular interactions.     

Elongated multiple electronic cascade and cyclization spacer systems in activatible anticancer prodrugs for enhanced drug release.

The design and synthesis of several novel elongated self-elimination spacer systems for application in prodrugs is described. These elongated spacer systems can be incorporated between a cleavable specifier and the parent drug. Naphthalene- and biphenyl-containing spacers were synthesized but did not eliminate. Prodrugs of the anticancer agents doxorubicin and paclitaxel are reported that contain two or three electronic cascade spacers. A novel catalytic application of HOBt was found for the synthesis of N-aryl carbamates through reacting a 4-nitrophenyl carbonate with an aniline derivative, to connect the 1,6-elimination spacers via a carbamate linkage. In addition, a double spacer-containing paclitaxel prodrug was synthesized, comprising a 1,6-elimination spacer and a bis-amine linker connected to paclitaxel via a 2'-carbamate linkage. Prodrugs in which the novel spacer systems were incorporated between a specific tripeptide specifier and the parent drug doxorubicin or paclitaxel proved to be significantly faster activated by plasmin in comparison with prodrugs containing conventional spacer systems. It is expected that the generally applicable novel spacer systems reported herein will contribute to future development of improved enzymatically activated prodrugs.     

Cyclization-activated prodrugs

Many drugs suffer from an extensive first-pass metabolism leading to drug inactivation and/or production of toxic metabolites, which makes them attractive targets for prodrug design. The classical prodrug approach, which involves enzyme-sensitive covalent linkage between the parent drug and a carrier moiety, is a well established strategy to overcome bioavailability/toxicity issues. However, the development of prodrugs that can regenerate the parent drug through non-enzymatic pathways has emerged as an alternative approach in which prodrug activation is not influenced by inter- and intraindividual variability that affects enzymatic activity. Cyclization-activated prodrugs have been capturing the attention of medicinal chemists since the middle-1980s, and reached maturity in prodrug design in the late 1990 s. Many different strategies have been exploited in recent years concerning the development of intramoleculary-activated prodrugs spanning from analgesics to anti-HIV therapeutic agents. Intramolecular pathways have also a key role in two-step prodrug activation, where an initial enzymatic cleavage step is followed by a cyclization-elimination reaction that releases the active drug. This work is a brief overview of research on cyclization-activated prodrugs from the last two decades.     

Surface functionalization of ultrananocrystalline diamond films by electrochemical reduction of aryldiazonium salts

The surface functionalization of ultrananocrystalline diamond (UNCD) thin films via the electrochemical reduction of aryl diazonium cations is described. The one-electron-transfer reaction leads to the formation of solution-based aryl radicals, which in turn react with the UNCD surface forming stable covalent C-C bonds. Cyclic voltammetry (CV), X-ray photoelectron spectroscopy (XPS), ac impedance spectroscopy, and contact angle measurements have been employed to characterize the organic overlayer and estimate the surface coverage. The grafting of 3,5-dichlorophenyl groups renders the UNCD surface hydrophobic, whereas the attachment of 4-aminophenyl groups makes the surface relatively hydrophilic. The surface coverage, estimated from the electrochemical and XPS measurements, is as high as 70% of a compact monolayer. The aminophenyl terminated surface was obtained by electrochemical reduction of the tethered nitrophenyl groups. This two-step approach yields a UNCD surface with functional moieties available for the potential covalent coupling of a wide variety of biomolecules (e.g., DNA and proteins).     

A comparative study on the interaction with calf thymus DNA of a Ni(II) complex of the anticancer drug adriamycin and a Ni(II) complex of sodium 1,4-dihydroxy-9,10-anthraquinone-2-sulphonate

The anthracycline drug adriamycin and its metal complexes are efficient in treating several forms of human cancers with recognized antineoplastic activity attributed to strong interactions with DNA within the target cells. The hydroxy-9,10-anthraquinone unit present in the molecule controls and regulates drug action. Metal ions when linked to adriamycin help to reduce the generation of radicals responsible for toxic side effects. A complex of adriamycin with Ni(II) was prepared and its physicochemical characteristics and DNA-binding ability were compared to a Ni(II) complex of sodium-1,4-dihydroxy-9,10-anthraquinone-2-sulphonate (NaLH₂), an analog of adriamycin. Interactions with calf thymus DNA of both complexes were studied by UV-Vis and fluorescence spectroscopy. Binding parameters determined for both complexes agree with each other. Binding of the Ni(II)-adriamycin complex to DNA was five to eight times stronger than for the Ni(II) complex of the hydroxy-9,10-anthraquinone analog, Na₂[Ni(NaLH)₂Cl₂]?·?2H₂O, i.e., Ni(NaLH)₂. The difference in binding was attributed to the presence of sugar units in adriamycin and to its absence in NaLH₂. Although the Ni(II) complex of the hydroxy-9,10-anthraquinone analog of adriamycin [Ni(NaLH)₂] was slightly weaker in binding DNA than the drug and its Ni(II) complex, a much lower cost of the former justifies its consideration as a substitute for the anthracycline drugs that are now in use.