Aminooxy analogues of spermine and their monoacetyl derivatives

Convenient methods of synthesis of 1-aminooxy-3,8-diaza-11-aminoundecane, its earlier unknown N1-and N1 -acetyl derivatives, and also 1,10-bis(aminooxy)-3,8-diazadecane are suggested. It is shown a possibility to selectively delete the acid-labile ethoxyethylidene protection of aminooxy group by hydrosulfates in the presence of N-tert-butyloxycarbonyl group.     

Aminooxy analogues of spermine and their monoacetyl derivatives

Convenient methods of synthesis of 1-aminooxy-3,8-diaza-11-aminoundecane, its earlier unknown N ¹-and N ¹¹-acetyl derivatives, and also 1,10-bis(aminooxy)-3,8-diazadecane are suggested. It is shown that it is possible to selectively delete the acid-labile ethoxyethylidene protection of aminooxy group by hydrosulfates in the presence of N-tert-butyloxycarbonyl group.     

Aminooxy pluronics: synthesis and preparation of glycosaminoglycan adducts

Novel biomaterials have been prepared in which glycosaminoglycans (GAGs) are chemically modified to create amphiphilic multiblock copolymers that are able to adhere to hydrophobic surfaces and can self-assemble into cross-linker-free hydrogels. First, the triblock poly(ethylene oxide)-polypropylene oxide copolymers (Pluronics) were converted into the previously unknown aminooxy (AO) derivatives. Both mono-AO and bis-AO Pluronics (AOPs) were synthesized and fully characterized in order to prepare tetrablock and pentablock copolymers, respectively. Second, the AOPs were coupled to the uronic acid carboxylates of heparin (HP) and hyaluronic acid (HA) using carbodiimide chemistry in order to give the previously undescribed amidooxy GAG derivatives. The coupling chemistry was confirmed using a newly prepared fluorescent AO reagent. Third, AOP-heparin and AOP-fluorescently labeled heparin were shown to adsorb efficiently to polystyrene surfaces, as determined by IL-8 based ELISA and fluorescence measurements, respectively. Fourth, AOP-linked fluorescently labeled HA was shown to adsorb efficiently to plastic surfaces. Finally, three different AOPs were evaluated for self-assembling hydrogel formation by AOP-HA pentablock polymers. In short, AOP-GAG adducts are semisynthetic amphiphilic biomacromolecules that offer a range of valuable practical opportunities for surface modification, preparation of cross-linker-free hydrogels, and formation of self-assembling mimics of the extracellular matrix.     

Impact of adsorbents selection on capture efficiency of cell culture derived human influenza viruses

The study aims on affinity matrix selection for a cell culture derived influenza virus capture step in downstream processing. Euonymus europaeus lectin (EEL) was used as an affinity ligand. Human influenza A/Puerto Rico/8/34 (H1N1) virus produced in MDCK cells was chosen as a model strain. The chromatographic separation characteristics of reinforced cellulose membranes and different matrices such as agarose, cellulose, polymer and glass particles with immobilized EEL have been determined. Results obtained were compared to affinity matrices, which are currently used in large-scale vaccine manufacturing. Mass balances for the viral membrane protein hemagglutinin showed that EEL affinity chromatography results in higher recoveries than conventional processes using Cellufine sulphate and heparinized agarose. The most efficient media, a polymer and a cellulose membrane, have been further characterized by protein and host cell DNA measurements. Separations based on the polymer matrix and the cellulose membrane removed contaminating DNA to 0.2 and 1%, respectively. Total protein contents were decreased to 50 and 31%, respectively. The EEL-membrane showed the highest influenza virus binding capacity. These characteristics demonstrate that EEL affinity chromatography is a promising candidate for capturing influenza viruses from MDCK cell culture broths in addition to currently applied chromatographic media.     

Aminooxy analogues of spermidine as inhibitors of spermine synthase and substrates of hepatic polyamine acetylating activity

Aminooxy analogues of spermidine, 1-aminooxy-3-N-[3-aminopropyl]- aminopropane (AP-APA) and N-[2-aminooxyethyl]-1,4-diaminobutane (AOE-PU), were tested as substrates or inhibitors of the enzymes involved in methionine and polyamine metabolism. Both compounds were good competitive inhibitors and poor substrates of spermine synthase, good substrates of cytosolic polyamine acetyltransferase, inactivators of S-adenosylmethionine decarboxylase and inhibitors of ornithine decarboxylase. AP-APA and AOE-PU showed K1-values of 1.5 and 186 microM as inhibitors of purified spermine synthase, and Km-values of 1.4 and 2.1 mM as substrates of the crude hepatic polyamine acetyltransferase activity. AP-APA was more potent than AOE-PU in crude enzyme preparations. Neither drug had any significant effect at 1 mM concentration on the activities of spermidine synthase, methionine adenosyltransferase, S-adenosylhomocysteine hydrolase, and methylthioadenosine phosphorylase. The results suggest that compounds of this type are valuable tools in unraveling the physiology of polyamines.     

In-Situ recovery of ethanol from fermentation broth by hydrophobic adsorbents

The in-situ recovery of ethanol from fermentation by different hydrophobic adsorbents has been studied. Liquid phase experiments have demonstrated that pentasil zeolites have a high selectivity for ethanol in the low concentration range. It is shown that the contact of the fermentation broth with the zeolite NaZSM-5 avoids the product inhibition. On the other hand, the ethanol production rate dramatically changes in dependence on the glucose-to-adsorbent ratio.     

(Aminooxy)acetic acid inhibits petunia growth and gibberellin- and cytokinin-stimulated growth in bioassays

(Aminooxy)acetic acid (AOA) was applied to greenhouse-grown petunias and was used in bioassays for three plant growth hormones so that its growth regulator properties could be studied. In greenhouse studies foliar sprays of 4.8–12 mm AOA inhibited vegetative growth of petunia seedlings (Petunia xhybrida Vilm. White Flash). When gibberellin A ₃ (GA₃) was applied to shoot tips previously treated with AOA, plant growth was stimulated, but there was no AOA x GA₃ interaction. Some changes in petunia leaf morphology induced by AOA were reversed by GA₃. AOA inhibited elongation of corn coleoptile segments (Zea mays L. B73 x Mol7) whether or not 10 m indole-3-acetic acid (IAA) was present, but there was no AOA x IAA interaction. AOA reduced lettuce hypocotyl (Lactuca sativa L. Grand Rapids) elongation induced by GA₃ and radish cotyledon (Raphanus sativus L. Champion) expansion induced by benzyladenine (BA). We propose that AOA interferes with postsynthetic metabolism of plant hormones during cell elongation induced by GA₃ and cell expansion induced by BA.Abbreviations AOA (aminooxy)acetic acid - GA₃ gibberellin A₃ - IAA indole-3-acetic acid - BA benzyladenine     

Evaluation of adsorbents for separation and purification of paclitaxel from plant cell cultures

In this study, we evaluated the efficiency of different adsorbents for the removal of plant-derived impurities during the pre-purification of paclitaxel from plant cell cultures. Using the synthetic adsorbents sylopute and active clay and their major components SiO₂ and MgO, we performed adsorbent treatment and analyzed the paclitaxel precipitates recovered from hexane precipitation. When SiO₂ was used, the highest purity (～58.1%) and yield (～91.5%) of paclitaxel were obtained. We also determined differences in the effectiveness of the adsorbent treatment according to changes in the surface area, pore volume and pore diameter of SiO₂. Adsorbent treatment was more effective when pore diameter was larger (silica I [2.19 nm] < silica II [4.92 nm] < silica III [9.07 nm]). The highest purity (～74.3%) and yield (～92.9%) of paclitaxel were obtained when silica III was used in the adsorbent treatment. Pore diameter had a greater effect on the removal of plant-derived impurities during the pre-purification of paclitaxel compared with surface area and pore volume. This result could be confirmed by HPLC analysis of the absorbent after treatment and TGA of the organic substances that were bonded to the adsorbent.     

Aminooxy functionalized oligonucleotides: preparation, on-support derivatization, and postsynthetic attachment to polymer support.

Three novel phosphoramidites, each bearing a phthaloyl-protected aminooxy tail, were prepared and applied in automated oligonucleotide synthesis. After chain assembly, the phthaloyl protection was removed with hydrazinium acetate. Normal succinyl linker turned to be stable under these conditions, and hence the support-bound oligonucleotide could be converted to a pyrene oxime conjugates by reacting with pyrene carbaldehyde or cis-retinal. Standard ammonolytic deprotection then released the deprotected conjugate in solution. Alternatively, the crude aminooxy-tethered oligonucleotide was immobilized to microscopic polymer particles by reacting the aminooxy function with the particle-bound aldehyde or epoxide groups. These immobilized oligonuceotides were shown to serve properly as probes in a mixed phase hybridization assay.     

Removal of organic pollutants from aqueous solutions by adsorbents prepared from an agroalimentary by-product

Two series of crosslinked starch polymers were tested for their ability to adsorb organic pollutants in aqueous solutions. The polymers were prepared by a crosslinking reaction of starch-enriched flour using epichlorohydrin as the crosslinking agent, without and in the presence of NH(4)OH. These polymers were used as sorbent materials for the removal of phenolic derivatives from wastewater. The influence of several parameters (kinetics, pH and polymer structure) on the sorption capacity was evaluated using the batch and the open column methods. Results of adsorption experiments showed that the starch-based materials exhibited high sorption capacities toward phenolic derivatives. The study of the kinetics of pollutant uptake revealed that the adsorbents presented a relatively fast rate of adsorption. The experimental data were examined using the Langmuir and Freundlich models and it was found that the Freundlich model appeared to fit the isotherm data better than the Langmuir model.     

Diterpenes derived from clerodanes from Pulicaria angustifolia

The investigation of Pulicaria angustifolia afforded, in addition to known triterpenes and caryophyllene epoxide, four methyl esters of diterpenes, all derived from clerodane. Three of them are derivatives of seco-nidoresedic acid. The structures were elucidated by spectroscopic methods.     

Amino acids derived from ornithine

Summary The use of copper complexes in the protection of lysine and tyrosine side chains has been extensively described. Based on this work and in order to produre a peptide library consisting of unnatural amino acids, three amino acids were prepared by acylation/sulfonation of the -amino function of ornithine and they were incorporated in a random sequence. Depending upon the acidity of the remaining proton on the -nitrogen, an intramolecular cyclization competed with the amide formation using standard coupling conditions (DIC/HOBt/DIEA). This side reaction could be suppressed by simple removal of the base from the condensation mixture. The obtained peptides were purified on a PLRP-s^® column and their identity was confirmed by mass spectrometry.Abbreviations Fmoc 9-fluorenylmethyloxycarbonyl - Inp isonipecotic acid - Thi -(2-thienylcarbonyl)ornithine - Tos -tosylornithine - Qui -(8-quinolinesulfonyl)ornithine - Ala -alanine - SPPS solid phase peptide synthesis     

Angiogenesis inhibitors derived from thalidomide

5-Hydroxy-2-(2,6-diisopropylphenyl)-1H-isoindole-1,3-dione (5HPP-33: 10), which was obtained during our previous structural development studies on thalidomide, was revealed to possess potent anti-angiogenic activity in a human umbilical vein endothelial cell (HUVEC) assay. Thalidomide (1) and its metabolite, 5-hydroxythalidomide (5-HT: 2), which possesses a hydroxyl group at the position corresponding to that of 5HPP-33, as well as IMiDs (immunomodulatory derivatives of thalidomide: 3 and 5), also showed weak or moderate activity in the same assay.     

Development of low-concentration mercury adsorbents from biohydrogen-generation agricultural residues using sulfur impregnation

Mercury adsorbents were derived from waste biohydrogen-generation barley husk and rice husk via carbonization, steam activation, and sulfur impregnation at 300-650 °C. The samples derived from agricultural residues showed a greater Hg⁰ adsorption than that of a coal-based activated carbon, confirming the feasibility of resource recovery of these agricultural residuals for low-concentration gaseous Hg adsorption. Sulfur impregnation reduced both the surface area and pore volume of the samples, with lower temperature causing a greater decrease. Elevating the impregnation temperature increased the organic sulfur contents, suggesting that in addition to elemental sulfur, organic sulfur may also act as active sites to improve Hg⁰ adsorption. Oxygen and sulfur functional groups accompanying the microporous structures may account for the enhancing Hg⁰ adsorption of the raw and sulfur-treated samples, respectively. The pseudo-second-order model can best describe the chemisorption characteristics, implying that Hg⁰ adsorption on the samples was in a bimolecular reaction form.     

Exosomes derived from dendritic cells

Dendritic cells (DC) are potent antigen presenting cells and the only ones capable of inducing primary cytotoxic immune responses both in vivo and vitro. DCs secrete a 60-80 nm membrane vesicle population of endocytic origin, called exosomes. The protein composition of exosomes was analyzed using a systematic proteomic approach. Besides MHC and costimulatory molecules, exosomes bear several adhesion proteins, probably involved in their specific targeting. Exosomes also accumulate several cytosolic factors, most likely involved in exoxome's biogenesis in late endosomes. Like DCs, exosomes induce potent anti tumor immune responses in vivo. Indeed, a single injection of DC-derived exosomes sensitized with tumor peptides induced the eradication of established mouse tumors. Tumor-specific cytotoxic T lymphocytes were found in the spleen of exosome treated mice, and depletion of CD8+ T cells in vivo inhibited the anti tumor effect of exosomes. These results strongly support the implementation of human DC-derived exosomes for cancer immunotherapy.     

Novel macrocycles derived from nucleosides

A novel nucleoside-derived macrocycle formed from three thymidine 5'-carboxylic acid monomer units is described.