Glycodendrimers are a novel group of dendrimers (DDMs) characterized by surface modifications with various types of glycosides. It has been shown previously that such modifications significantly decrease the cytotoxicity of DDMs. Here, we present an investigation of glucose-modified carbosilane DDMs (first-third-generation, DDM1-3Glu) interactions with two models of biological structures: lipid membranes (liposomes) and serum protein (human serum albumin, HSA). The changes in lipid membrane fluidity with increasing concentration of DDMs was monitored by spectrofluorimetry and calorimetry methods. The influence of glycodendrimers on serum protein was investigated by monitoring changes in protein fluorescence intensity (fluorescence quenching) and as protein secondary structure alterations by circular dichroism spectrometry. Generally, all generations of DDMGlu induced a decrease of membrane fluidity and interacted weakly with HSA. Interestingly, in contrast to other dendritic type polymers, the extent of the DDM interaction with both biological models was not related to DDM generation. The most significant interaction with protein was shown in the case of DDM2Glu, whereas DDM1Glu induced the highest number of changes in membrane fluidity. In conclusion, our results suggest that the flexibility of a DDM molecule, as well as its typical structure (hydrophobic interior and hydrophilic surface) along with the formation of larger aggregates of DDM2-3Glu, significantly affect the type and extent of interaction with biological structures.

• MeSH
• antitumorózní látky aplikace a dávkování   MeSH
• cirkulární dichroismus MeSH
• dendrimery chemie   farmakologie   MeSH
• fluidita membrány účinky léků   MeSH
• fluorescenční spektrometrie MeSH
• glukosa chemie   farmakologie   MeSH
• hydrofobní a hydrofilní interakce MeSH
• lidé MeSH
• lidský sérový albumin metabolismus   MeSH
• liposomy MeSH
• nádory farmakoterapie   MeSH
• nosiče léků chemie   farmakologie   MeSH
• silany chemie   farmakologie   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH

A recently described bangle lectin (PHL) from the bacterium Photorhabdus asymbiotica was identified as a mainly fucose-binding protein that could play an important role in the host-pathogen interaction and in the modulation of host immune response. Structural studies showed that PHL is a homo-dimer that contains up to seven L-fucose-specific binding sites per monomer. For these reasons, potential ligands of the PHL lectin: α-L-fucopyranosyl-containing mono-, di-, tetra-, hexa- and dodecavalent ligands were tested. Two types of polyvalent structures were investigated - calix[4]arenes and dendrimers. The shared feature of all these structures was a C-glycosidic bond instead of the more common but physiologically unstable O-glycosidic bond. The inhibition potential of the tested structures was assessed using different techniques - hemagglutination, surface plasmon resonance, isothermal titration calorimetry, and cell cross-linking. All the ligands proved to be better than free L-fucose. The most active hexavalent dendrimer exhibited affinity three orders of magnitude higher than that of standard L-fucose. To determine the binding mode of some ligands, crystal complex PHL/fucosides 2 - 4 were prepared and studied using X-ray crystallography. The electron density in complexes proved the presence of the compounds in 6 out of 7 fucose-binding sites.

• MeSH
• antibakteriální látky chemie   farmakologie   terapeutické užití   MeSH
• bakteriální infekce farmakoterapie   mikrobiologie   MeSH
• bakteriální proteiny antagonisté a inhibitory   chemie   izolace a purifikace   metabolismus   MeSH
• dendrimery chemie   farmakologie   terapeutické užití   MeSH
• erytrocyty MeSH
• fukosa analogy a deriváty   farmakologie   terapeutické užití   MeSH
• hemaglutinace účinky léků   MeSH
• interakce hostitele a patogenu účinky léků   MeSH
• krystalografie rentgenová MeSH
• lektiny antagonisté a inhibitory   chemie   izolace a purifikace   metabolismus   MeSH
• lidé MeSH
• ligandy MeSH
• molekulární modely MeSH
• Photorhabdus metabolismus   MeSH
• povrchová plasmonová rezonance MeSH
• rekombinantní proteiny chemie   izolace a purifikace   metabolismus   MeSH
• vazebná místa MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

For the design of a biohybrid structure as a ligand-tailored drug delivery system (DDS), it is highly sophisticated to fabricate a DDS based on smoothly controllable conjugation steps. This article reports on the synthesis and the characterization of biohybrid conjugates based on noncovalent conjugation between a multivalent biotinylated and PEGylated poly(amido amine) (PAMAM) dendrimer and a tetrameric streptavidin-small protein binding scaffold. This protein binding scaffold (SA-ABDwt) possesses nM affinity toward human serum albumin (HSA). Thus, well-defined biohybrid structures, finalized by binding of one or two HSA molecules, are available at each conjugation step in a controlled molar ratio. Overall, these biohybrid assemblies can be used for (i) a controlled modification of dendrimers with the HSA molecules to increase their blood-circulation half-life and passive accumulation in tumor; (ii) rendering dendrimers a specific affinity to various ligands based on mutated ABD domain, thus replacing tedious dendrimer-antibody covalent coupling and purification procedures.

• MeSH
• bakteriální proteiny genetika   metabolismus   MeSH
• biotin chemie   MeSH
• biotinylace MeSH
• buněčné linie MeSH
• dendrimery chemická syntéza   farmakologie   MeSH
• epitelové buňky cytologie   účinky léků   MeSH
• Escherichia coli genetika   metabolismus   MeSH
• exprese genu MeSH
• krysa rodu rattus MeSH
• lidé MeSH
• ligandy MeSH
• molekulární modely MeSH
• nanočástice chemie   ultrastruktura   MeSH
• polyaminy chemie   MeSH
• polyethylenglykoly chemie   MeSH
• rekombinantní fúzní proteiny genetika   metabolismus   MeSH
• sekvence aminokyselin MeSH
• sérový albumin chemie   MeSH
• streptavidin chemie   MeSH
• Streptomyces genetika   metabolismus   MeSH
• systémy cílené aplikace léků * MeSH
• vazba proteinů MeSH
• zvířata MeSH
• Check Tag
• krysa rodu rattus MeSH
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Research Support, N.I.H., Extramural MeSH

Dendrimers are well-defined macromolecules whose highly branched structure is reminiscent of many natural structures, such as trees, dendritic cells, neurons or the networks of kidneys and lungs. Nature has privileged such branched structures for increasing the efficiency of exchanges with the external medium; thus, the whole structure is of pivotal importance for these natural networks. On the contrary, it is generally believed that the properties of dendrimers are essentially related to their terminal groups, and that the internal structure plays the minor role of an 'innocent' scaffold. Here we show that such an assertion is misleading, using convergent information from biological data (human monocytes activation) and all-atom molecular dynamics simulations on seven families of dendrimers (13 compounds) that we have synthesized, possessing identical terminal groups, but different internal structures. This work demonstrates that the scaffold of nanodrugs strongly influences their properties, somewhat reminiscent of the backbone of proteins.

• MeSH
• aza sloučeniny chemie   farmakologie   MeSH
• biokompatibilní materiály chemie   farmakologie   MeSH
• bisfosfonáty chemie   farmakologie   MeSH
• dendrimery chemie   farmakologie   MeSH
• inhibitory kostní resorpce chemie   farmakologie   MeSH
• lidé MeSH
• molekulární struktura MeSH
• monocyty účinky léků   MeSH
• nanočástice chemie   MeSH
• polylysin chemie   farmakologie   MeSH
• polypropyleny chemie   farmakologie   MeSH
• průtoková cytometrie MeSH
• silany chemie   farmakologie   MeSH
• simulace molekulární dynamiky MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

High-molecular-weight star polymer drug nanocarriers intended for the treatment and/or visualisation of solid tumours were synthesised, and their physico-chemical and preliminary in vitro biological properties were determined. The water-soluble star polymer carriers were prepared by the grafting of poly(amido amine) (PAMAM) dendrimers by hetero-telechelic N-(2-hydroxypropyl)methacrylamide (HPMA) copolymers, synthesised by the controlled radical Reversible Addition Fragmentation chain Transfer (RAFT) polymerisation. The well-defined star copolymers with Mw values ranging from 2 · 10(5) to 6 · 10(5) showing a low dispersity (approximately 1.2) were prepared in a high yield. A model anticancer drug, doxorubicin, was bound to the star polymer through a hydrazone bond, enabling the pH-controlled drug release in the target tumour tissue. The activated polymer arm ends of the star copolymer carrier enable a one-point attachment for the targeting ligands and/or a labelling moiety. In this study, the model TAMRA fluorescent dye was used to prove the feasibility of the polymer carrier visualisation by optical imaging in vitro. The tailor-made structure of the star polymer carriers should facilitate the synthesis of targeted polymer-drug conjugates, even polymer theranostics, for simultaneous tumour drug delivery and imaging.

• MeSH
• dendrimery * chemická syntéza   chemie   farmakokinetika   farmakologie   MeSH
• doxorubicin * chemie   farmakokinetika   farmakologie   MeSH
• léky s prodlouženým účinkem chemická syntéza   chemie   farmakokinetika   farmakologie   MeSH
• lidé MeSH
• methakryláty * chemie   farmakokinetika   farmakologie   MeSH
• myši MeSH
• nádorové buněčné linie MeSH
• nádory * farmakoterapie   metabolismus   patologie   MeSH
• nanočástice chemie   ultrastruktura   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

N-acetyl-D-glucosamine-coated polyamidoamine dendrimer (GN8P), exerting high binding affinity to rodent recombinant NKR-P1A and NKR-P1C activating proteins, was shown previously to delay the development of rat colorectal carcinoma as well as mouse B16F10 melanoma, and to potentiate antigen-specific antibody formation in healthy C57BL/6 mice via NK cell stimulation. In this study, we investigated whether GN8P also modulates tumor-specific B cell responses. Serum anti-B16F10 melanoma IgG levels, IgG2a mRNA expression, antibody dependent cell-mediated cytotoxicity (ADCC), and counts of plasma as well as antigen presenting B cells were evaluated in tumor-bearing C57BL/6 mice treated with GN8P and in respective controls. To reveal the mechanism of GN8P effects, the synthesis of interferon-gamma (IFN-γ) and interleukin-4 (IL-4), cytokines involved in regulation of immunoglobulin class switch, was determined. The GN8P treatment significantly elevated IgG, and particularly IgG2a, response against B16F10 melanoma, which led to augmented ADCC reaction. The significant increase in production of IFN-γ, which is known to support IgG2a secretion, was observed solely in NK1.1 expressing cell populations, predominantly in NK cells. Moreover, GN8P raised the number of plasma cells, and promoted antigen presenting capacity of I-A/I-E-positive B lymphocytes by up-regulation of their CD80 and CD86 co-stimulatory molecule expression. These results indicate that GN8P-induced enhancement of tumor-specific antibody formation is triggered by NK cell activation, and contributes to complexity of anticancer immune response involving lectin-saccharide interaction.

• MeSH
• acetylglukosamin chemie   farmakologie   MeSH
• antigeny CD80 biosyntéza   genetika   MeSH
• antigeny CD86 biosyntéza   genetika   MeSH
• B-lymfocyty účinky léků   imunologie   metabolismus   MeSH
• buněčná cytotoxicita závislá na protilátkách účinky léků   imunologie   MeSH
• buňky NK účinky léků   imunologie   metabolismus   MeSH
• dendrimery chemie   farmakologie   MeSH
• imunoglobulin G biosyntéza   krev   genetika   imunologie   MeSH
• interferon gama biosyntéza   genetika   MeSH
• interleukin-4 biosyntéza   genetika   MeSH
• melanom experimentální genetika   imunologie   metabolismus   MeSH
• myši inbrední C57BL MeSH
• myši MeSH
• nádorové buněčné linie MeSH
• upregulace účinky léků   MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

N-Acetyl-D-glucosamine-coated polyamidoamine dendrimer (GlcNAc8) was shown previously to exhibit binding affinity to the rat recombinant NKR-P1 molecule (known in mice also as NK1.1) and to induce NK cell-mediated cytotoxicity. In this study, we investigated whether GlcNAc8 modulates antibody formation as activated NK cells were reported to participate in its regulation. C57BL/6 mice treated with GlcNAc8 and intact controls were immunized either with sheep red blood cells (SRBCs), 2,4-dinitrophenylated-lipopolysaccharide (DNP-LPS) or keyhole limpet hemocyanin (KLH) for evaluation of splenic antibody forming cell counts and serum immunoglobulin (Ig) levels. In vitro Ig formation was determined using supernatants of spleen mononuclear cells (SMCs) and CD49b or NK1.1-depleted SMC subpopulations. Serum antigen-specific IgG2a levels were also measured in DBA/2 and BALB/c mice (NK1.1-negative mouse strains on the basis of flow cytometric analysis) which possess different Nkr-p1c gene form than C57BL/6 ones. A significant increase in anti-SRBC IgG forming cells, serum levels of anti-KLH as well as anti-DNP IgG and IgG2a was observed after GlcNAc8 administration in C57BL/6 mice. IgM levels in supernatants of SMCs stimulated in vitro simultaneously with DNP-LPS and GlcNAc8 were significantly mounted compared with supernatants of SMCs primed with the antigen alone, but this enhancement was blocked after depletion of CD49b-positive or NK1.1-positive cells. In DBA/2 and BALB/c mice, GlcNAc8 influenced neither serum levels of anti-KLH nor anti-DNP IgG2a. These results indicate that GlcNAc8-induced upregulation of antibody formation is triggered by NK cell stimulation and depends on expressed NKR-P1 isoforms, particularly NKR-P1C.

• MeSH
• acetylglukosamin analogy a deriváty   farmakologie   chemie   MeSH
• antigeny Ly imunologie   metabolismus   MeSH
• B-lymfocyty imunologie   účinky léků   MeSH
• buňky NK imunologie   účinky léků   MeSH
• buňky produkující protilátky imunologie   účinky léků   MeSH
• dendrimery farmakologie   chemie   MeSH
• hemokyanin imunologie   MeSH
• imunoglobuliny krev   MeSH
• imunologické faktory farmakologie   chemie   MeSH
• krysa rodu rattus MeSH
• lektinové receptory NK-buněk - podrodina B imunologie   metabolismus   MeSH
• myši inbrední BALB C MeSH
• myši inbrední C57BL MeSH
• myši inbrední DBA MeSH
• myši MeSH
• NKT buňky imunologie   účinky léků   MeSH
• tvorba protilátek účinky léků   MeSH
• zvířata MeSH
• Check Tag
• krysa rodu rattus MeSH
• myši MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• práce podpořená grantem MeSH

Comblike glycodendrimers were prepared by the chemoselective ligation of cysteine-modified glycopeptides (1-7) with a 3-maleimidopropionate-modified linear synthetic carrier (8). Glycodendrimers bearing mono-, di-, or tri-Tn clusters (9-11) were tested as inhibitors using plant and mammalian lectins. In the former group, the Codium fragile lectin showed moderate discrimination among 9, 10, and 11. In the latter group, A and B isoforms of rat NKR-P1 lectin strongly discriminated between 9 and 10. 10 caused a 4-fold increase in killing of the NK resistant tumor cell lines at concentrations as low as 10(-8) M. Surprisingly, 11 interacted exclusively with the rat NKR-P1B isoform and inhibited efficiently natural killing in both rats and humans, even in the presence of the activating compounds 9 and 10. Dinitrophenol haptenization or influenza virus hemagglutinin T-cell epitope conjugation increased the immunogenicity of the parent compounds and resulted in the production of Tn specific antibodies.

• MeSH
• 2,4-dinitrofenol chemie   MeSH
• antigeny sacharidové asociované s nádorem chemie   imunologie   MeSH
• buňky NK imunologie   účinky léků   MeSH
• cystein chemie   MeSH
• cytotoxicita imunologická MeSH
• dendrimery farmakologie   chemická syntéza   chemie   MeSH
• epitopy MeSH
• financování organizované MeSH
• glykopeptidy chemie   MeSH
• hapteny MeSH
• hemaglutininové glykoproteiny viru chřipky chemie   MeSH
• imunoglobulin G biosyntéza   MeSH
• imunoglobulin M biosyntéza   MeSH
• Jurkat buňky MeSH
• krysa rodu rattus MeSH
• lektiny chemie   MeSH
• lidé MeSH
• myši inbrední BALB C MeSH
• myši MeSH
• protein - isoformy chemie   MeSH
• receptory imunologické chemie   MeSH
• rostlinné lektiny chemie   MeSH
• T-lymfocyty imunologie   MeSH
• tvorba protilátek MeSH
• vazba proteinů MeSH
• zvířata MeSH
• Check Tag
• krysa rodu rattus MeSH
• lidé MeSH
• myši MeSH
• ženské pohlaví MeSH
• zvířata MeSH