Exploring Rigid and Flexible Scaffolds to Develop Potent Glucuronic Acid Glycodendrimers for Dengue Virus Inhibition.

Multivalent glycodendrimers are valuable tools for studying carbohydrate-protein interactions, and their scaffolds represent important components to increase specificity and affinity. Previous work by our group described the preparation of a tetravalent glucuronic acid rigid dendron that binds with good affinity to the dengue virus envelope protein (KD = 22 µM). Herein, the chemical synthesis and binding analysis of three new sets of rigid, semirigid, and flexible glucuronic acid-based dendrimers bearing different levels of multivalency and their interactions with the dengue virus envelope protein are described. The different oligoalkynyl scaffolds were coupled to glucuronic acid azides by a copper-catalyzed azide-alkyne cycloaddition reaction through optimized synthetic strategies to afford the desired glycodendrimers with good yields. Surface plasmon resonance studies have demonstrated that glycodendrimers 12b and 12c, with flexible scaffolds, give the best binding interactions with the dengue virus envelope protein (12b: KD = 0.487 µM and 12c: KD = 0.624 µM). Their binding constant values were 45 and 35 times higher than the one obtained in previous studies with a rigid tetravalent glucuronic acid dendron (KD = 22 µM), respectively. Molecular modeling studies were carried out in order to understand the difference in behavior observed for 12b and 12c. This work reports an efficient glycodendrimer chemical synthesis process that provides an appropriate scaffold that offers an easy and versatile strategy to find new active compounds against the dengue virus.

Evaluation of Anti-Neuroinflammatory Activity of Isatin Derivatives in Activated Microglia.

Neuroinflammation plays a crucial role in the progression of Alzheimer's disease and other neurodegenerative disorders. Overactivated microglia cause neurotoxicity and prolong the inflammatory response in many neuropathologies. In this study, we have synthesised a series of isatin derivatives to evaluate their anti-neuroinflammatory potential using lipopolysaccharide activated microglia as a cell model. We explored four different substitutions of the isatin moiety by testing their anti-neuroinflammatory activity on BV2 microglia cells. Based on the low cytotoxicity and the activity in reducing the release of nitric oxide, pro-inflammatory interleukin 6 and tumour necrosis factor α by microglial cells, the N1-alkylated compound 10 and the chlorinated 20 showed the best results at 25 µM. Taken together, the data suggest that 10 and 20 are promising lead compounds for developing new neuroprotective agents.

Efficient multi-click approach to well-defined two-faced octasilsesquioxanes: the first perfect Janus nanocube.

The preparation of the first structurally well-defined Janus nanocube showing two chemically distinct opposed faces is described. The synthetic approach is based on a highly efficient and symmetry-controlled CuAAC functionalization of an octa-azido cubic silsesquioxane with a conformationally constrained tetra-alkyne with an appropriate spatial orientation of the triple bonds.

Nucleophilic addition of amines to ruthenium carbenes: ortho-(alkynyloxy)benzylamine cyclizations towards 1,3-benzoxazines.

A new ruthenium-catalyzed cyclization of ortho-(alkynyloxy)benzylamines to dihydro-1,3-benzoxazines is reported. The cyclization is thought to take place via the vinyl ruthenium carbene intermediates which are easily formed from [Cp*RuCl(cod)] and N2 CHSiMe3 . The mild reaction conditions and the efficiency of the procedure allow the easy preparation of a broad range of new 2-vinyl-2-substituted 1,3-benzoxazine derivatives. Rearrangement of an internal C(sp) in the starting material into a tetrasubstituted C(sp(3) ) atom in the final 1,3-benzoxazine is highly remarkable.

Nonafluorobutanesulfonyl azide as a shelf-stable highly reactive oxidant for the copper-catalyzed synthesis of 1,3-diynes from terminal alkynes.

Nonafluorobutanesulfonyl azide is a highly efficient reagent for the copper-catalyzed coupling of terminal alkynes to give symmetrical and unsymmetrical 1,3-diynes in good to excellent yields and with good functional group compatibility. The reaction is extremely fast (<10 min), even at low temperature (−78 °C), and requires substoichiometric amounts of a simple copper(I) or copper(II) salt (2­5 mol %) and an organic base (0.6 mol %). A possible mechanistic pathway is briefly discussed on the basis of model DFT theoretical calculations. The quantitative assessment of the safety of use and shelf stability of nonafluorobutanesulfonyl azide has confirmed that this reagent is a superior and safe alternative to other electrophilic azide reagents in use today.

Rhodium-catalyzed intermolecular C-H amination of simple hydrocarbons using the shelf-stable nonafluorobutanesulfonyl azide.

A new procedure has been developed for the direct intermolecular C-H amination of simple hydrocarbons using shelf-stable nonafluorobutanesulfonyl azide in the presence of a dirhodium(II) tetracarboxylate catalyst under mild reaction conditions. Some mechanistic details are briefly discussed on the basis of control experiments.

Application of the lithiation-borylation reaction to the preparation of enantioenriched allylic boron reagents and subsequent in situ conversion into 1,2,4-trisubstituted homoallylic alcohols with complete control over all elements of stereochemistry.

The reactions of Hoppe's lithiated carbamates with vinylboranes and boronic esters give allylic boranes/boronic esters, and subsequent addition of aldehydes provides a new route to enantioenriched homoallylic alcohols with high enantiomeric ratios and diastereomeric ratios. Specifically, reactions of sparteine-complexed lithiated carbamates with trans-alkenyl-9-BBN derivatives followed by addition of aldehydes gave (Z)-anti-homoallylic alcohols in greater than 95:5 er and 99:1 dr. However, in the special case of the methyl-substituted lithiated carbamate, diamine-free conditions were required to achieve high selectivity. Reactions of sparteine-complexed lithiated carbamates with (Z)-alkenyl pinacol boronic esters and (E)-alkenyl neopentyl boronic esters gave (E)-syn- and (E)-anti-homoallylic alcohols, respectively, in greater than 96:4 er and 98:2 dr. In these reactions, a Lewis acid (MgBr(2) or BF(3) x OEt(2)) was required to promote both the 1,2-metalate rearrangement and the addition of the intermediate allylic boronic ester to the aldehyde. This methodology provides a general route to each of the three classes of homoallylic alcohols with high selectivity.

P-TEFb is critical for the maturation of RNA polymerase II into productive elongation in vivo.

Positive transcription elongation factor b (P-TEFb) is the major metazoan RNA polymerase II (Pol II) carboxyl-terminal domain (CTD) Ser2 kinase, and its activity is believed to promote productive elongation and coupled RNA processing. Here, we demonstrate that P-TEFb is critical for the transition of Pol II into a mature transcription elongation complex in vivo. Within 3 min following P-TEFb inhibition, most polymerases were restricted to within 150 bp of the transcription initiation site of the active Drosophila melanogaster Hsp70 gene, and live-cell imaging demonstrated that these polymerases were stably associated. Polymerases already productively elongating at the time of P-TEFb inhibition, however, proceeded with elongation in the absence of active P-TEFb and cleared from the Hsp70 gene. Strikingly, all transcription factors tested (P-TEFb, Spt5, Spt6, and TFIIS) and RNA-processing factor CstF50 exited the body of the gene with kinetics indistinguishable from that of Pol II. An analysis of the phosphorylation state of Pol II upon the inhibition of P-TEFb also revealed no detectable CTD Ser2 phosphatase activity upstream of the Hsp70 polyadenylation site. In the continued presence of P-TEFb inhibitor, Pol II levels across the gene eventually recovered.

[Parental smoking and lung function in healthy children and adolescents]. / Tabaquismo parental y función pulmonar en niños y adolescentes sanos.

OBJECTIVE: The debate continues on the effect of passive smoking on nonsmokers. The effect of parental smoking on the lung function of children varies considerably according to geographic area, source of passive smoking, and sex. The objective of this study was to evaluate the effect of parental smoking on the lung function of children. POPULATION AND METHODS: A cross-sectional study was performed on a sample of the population of healthy children and adolescents between 6 and 18 years of age in Galicia. Subjects were selected by means of 2-stage cluster sampling grouped by sex and age. RESULTS: Approximately 56% of the children were exposed to the tobacco smoke of one of their parents. Children whose fathers were smokers presented a 40% higher risk of reduced forced expiratory flow at 75% of forced vital capacity (FEF75%) and a 30% higher risk of reduced FEF25%-75%. Children whose mothers were smokers presented a 30% higher risk of reduced forced expiratory volume in the first second and a 40% higher risk of reduced FEF50%. There was a 60% increase in risk of reduced FEF75%. The fact that both parents smoked did not appear to increase the risk of reduced lung function. CONCLUSIONS: Parental smoking has a considerable effect on the lung function of children and adolescents. Smoking by either the mother or the father has a decisive influence. The fact that this effect is independent of the growth of the child and that the obstructive effect is located principally in the distal airways appears to confirm the hypothesis that this effect is produced after birth.

Tabaquismo parental y función pulmonar en niños y adolescentes sanos / Parental smoking and lung function in healthy children and adolescents

Objetivo: Continúa la controversia sobre el efecto del tabaquismo pasivo en los no fumadores. El efecto del tabaquismo de los padres sobre la función pulmonar de los hijos presenta gran variabilidad entre diferentes zonas geográficas, fuente del tabaquismo pasivo y sexos. El objetivo del presente estudio ha sido valorar el efecto del tabaquismo de los padres sobre la función pulmonar de sus hijos. Población y métodos: Hemos llevado a cabo un estudio transversal en una muestra de la población de niños y adolescentes sanos de 6 a 18 años de Galicia, seleccionada mediante un muestreo bietápico en racimos y estratificada por sexo y edad. Resultados: Alrededor del 56% de los niños estaban expuestos al humo del tabaco de alguno de sus padres. Los niños de padres fumadores presentaban un 40% más de riesgo de reducción del flujo espiratorio forzado al 75% de la capacidad vital forzada (FEF75%), y un 30% de reducción del FEF25-75%. Los niños cuyas madres eran fumadoras tenían un 30% más de riesgo de reducción del volumen espiratorio forzado en el primer segundo, y un 40% de reducción del FEF50%. El incremento de riesgo de reducción del FEF75% fue del 60%. El hecho de que fumaran ambos progenitores no pareció incrementar el riesgo de función pulmonar reducida. Conclusiones: El tabaquismo parental tiene un importante efecto sobre la función pulmonar de niños y adolescentes. Tanto el tabaquismo materno como el paterno influyen decisivamente. El hecho de que este efecto sea independiente del crecimiento del niño y que el efecto obstructivo se localice fundamentalmente en la vía aérea distal parece confirmar la hipótesis de que este efecto se produce después del nacimiento

Objective: The debate continues on the effect of passive smoking on nonsmokers. The effect of parental smoking on the lung function of children varies considerably according to geographic area, source of passive smoking, and sex. The objective of this study was to evaluate the effect of parental smoking on the lung function of children. Population and methods: A cross-sectional study was performed on a sample of the population of healthy children and adolescents between 6 and 18 years of age in Galicia. Subjects were selected by means of 2-stage cluster sampling grouped by sex and age. Results: Approximately 56% of the children were exposed to the tobacco smoke of one of their parents. Children whose fathers were smokers presented a 40% higher risk of reduced forced expiratory flow at 75% of forced vital capacity (FEF75%) and a 30% higher risk of reduced FEF25%-75%. Children whose mothers were smokers presented a 30% higher risk of reduced forced expiratory volume in the first second and a 40% higher risk of reduced FEF50%. There was a 60% increase in risk of reduced FEF75%. The fact that both parents smoked did not appear to increase the risk of reduced lung function. Conclusions: Parental smoking has a considerable effect on the lung function of children and adolescents. Smoking by either the mother or the father has a decisive influence. The fact that this effect is independent of the growth of the child and that the obstructive effect is located principally in the distal airways appears to confirm the hypothesis that this effect is produced after birth

Flavopiridol administered using a pharmacologically derived schedule is associated with marked clinical efficacy in refractory, genetically high-risk chronic lymphocytic leukemia.

Despite promising preclinical studies with the cyclin-dependent kinase inhibitor flavopiridol in chronic lymphocytic leukemia (CLL) and other diseases, previous clinical trials with this agent have been disappointing. The discovery of differential protein binding of flavopiridol in human and bovine serum contributed to an effective pharmacokinetic-derived schedule of administration of this agent. On the basis of pharmacokinetic modeling using our in vitro results and data from a previous trial, we initiated a phase 1 study using a 30-minute loading dose followed by 4 hours of infusion administered weekly for 4 of 6 weeks in patients with refractory CLL. A group of 42 patients were enrolled on 3 cohorts (cohort 1, 30 mg/m2 loading dose followed by 30 mg/m2 4-hour infusion; cohort 2, 40 mg/m2 loading dose followed by 40 mg/m2 4-hour infusion; and cohort 3, cohort 1 dose for treatments 1 to 4, then a 30 mg/m2 loading dose followed by a 50 mg/m2 4-hour infusion). The dose-limiting toxicity using this novel schedule was hyperacute tumor lysis syndrome. Aggressive prophylaxis and exclusion of patients with leukocyte counts greater than 200x10(9)/L have made this drug safe to administer at the cohort 3 dose. Of the 42 patients treated, 19 (45%) achieved a partial response with a median response duration that exceeds 12 months. Responses were noted in patients with genetically high-risk disease, including 5 (42%) of 12 patients with del(17p13.1) and 13 (72%) of 18 patients with del(11q22.3). Flavopiridol administered using this novel schedule has significant clinical activity in refractory CLL. Patients with bulky disease and high-risk genetic features have achieved durable responses, thereby justifying further study of flavopiridol in CLL and other diseases.

Synthesis of enantiopure (alphaS,betaS)- or (alphaR,betaS)-beta-amino alcohols by complete regioselective opening of aminoepoxides by organolithium reagents LiAlH4 or LiAlD4.

The reaction of chiral (2R,1'S)- or (2S,1'S)-2-(1-aminoalkyl)epoxides, 1 or 2 with a variety of organolithium compounds to obtain the corresponding (alphaS,betaS)- or (alphaR,betaS)- beta-amino alcohols in enantiopure form is reported. In both cases, the opening of the oxirane ring at C-3 proceeded with total regioselectivity. Moreover, the ring opening of aminoepoxides 1 or 2 by hydride (utilizing LiAlH4) to obtain the corresponding (2S,3S)- or (2R,3S)-3-aminoalkan-2-ols is also described. The reaction of 1 or 2 with LiAlD4 in place of LiAlH4 gave the corresponding (2S,3S)- or (2R,3S)-3-amino-1-deuterioalkan-2-ols.

Synthesis of enantiopure allylamines by reductive alkylation of amino epoxides with organolithium reagents.

[reaction: see text] Transformation of enantiopure (2R,1'S)-2-(1-aminoalkyl)epoxides 1 into the corresponding allylamines 2 is described. The opening of the epoxide ring with different organolithium compounds takes place with total selectivity and in high yields.

Total selective synthesis of enantiopure O1-acyl-3-aminoalkane-1,2-diols by ring opening of aminoepoxides with carboxylic acids.

[reaction: see text] Synthesis of (2R,3S)- or (2S,3S)-O1-acyl-3-aminoalkane-1,2-diols by ring opening of enantiopure (2R,1'S)- or (2S,1'S)-2-(1-aminoalkyl)epoxides 1 or 2, with carboxylic acids in the presence of BF3 x Et2O and chlorotrimethylsilane, is described. The conversion takes place with total selectivity and in good yield. In addition, (2R,3S)-O,O-diacyl-3-aminoalkane-1,2-diols 3 were also prepared from reaction of (2R,1'S)-2-(1-aminoalkyl)epoxides 1 with carboxylic acids under the same reaction conditions and without chlorotrimethylsilane. Mechanisms to explain both transformations are proposed.

Selective ring-opening of nonactivated amino aziridines by thiols and unusual nucleophilic substitution of a dibenzylamino group.

[Reaction: see text]. The reaction of chiral 2-(1-aminoalkyl)aziridines 1 with different thiols, in the presence of BF3*Et2O, is reported. The obtained products were dependent on the structure of the starting amino aziridines 1. Thus, enantiopure (2S,3S)-2-(alkylthio)alkane-1,3-diamines 2 were obtained from aziridines with C-2 substituents with lower steric congestion and partially racemized (2S,3S)-2,3-bis(alkylthio)alkan-1-amines 3 (ee = 56-66%) from aziridines with larger C-2 subtituents. In both cases, the opening of the nonactivated aziridine ring at C-2 took place with retention of configuration and proceeded with regio- and stereoselectivity at C-2. In the synthesis of 3, 2 equiv of thiol reacts with 1 and the opening of aziridine ring at C-2 was followed by an unusual displacement of the dibenzylamino group by a second equivalent of thiol. The regiochemistry and relative configuration of compounds 3 was established by single-crystal X-ray analysis. A mechanism is proposed to explain the results obtained.

Regioselective ring opening of amino epoxides with nitriles: an easy synthesis of (2R,3S)- and (2S,3S)-1,3-diaminoalkan-2-ols with differently protected amine functions.

[reaction: see text] Transformation of enantiopure (2R,1'S)- or (2S,1'S)-2-(1-aminoalkyl)epoxides 1 or 2 into the corresponding (2R,3S)- and (2S,3S)-1,3-diaminoalkan-2-ols 3 or 4 is described. The opening of the epoxide ring with different nitriles (Ritter reaction) takes place with total selectivity and in high yields in the presence of BF3.Et2O. Interestingly, the two amine groups are differently protected. A mechanism to explain this transformation is proposed.

Stereoselective functionalization of 2-(1-aminoalkyl)aziridines via lithiation of aziridine-borane complexes.

Highly selective functionalization of the aziridine ring of (2S,1'S)-2-(1'-aminoalkyl)aziridines 1, through successive formation of aziridine-borane complexes, lithiation, treatment with a variety of electrophiles and final decomplexation is described. The influence of the structure of the starting complexes 2 and of the electrophiles in the stereoselectivity of this process has been studied. Finally, successive double lithiation-electrophile reactions were carried out affording enantiopure 1,2,3,3-tetrasubstituted aziridine-borane complexes with high selectivity.

Totally selective ring-opening of amino epoxides with ketones: a general entry to enantiopure (2R,3S)- and (2S,3S)-3-aminoalkano-1,2-diols.

[Reaction: see text] Transformation of enantiopure diastereoisomers (2R,1'S)- and (2S,1'S)-2-(1-aminoalkyl)epoxides into the corresponding 4-(1-aminoalkyl)-1,3-dioxolanes is achieved by reaction with different ketones in the presence of BF3.Et2O. The conversion takes place in very high yields, total selectivity, and without epimerization. A mechanism to explain this transformation is proposed. The obtained 1,3-dioxolanes can be deprotected, and (2R,3S)- and (2S,3S)-3-aminoalkano-1,2-diols were isolated.

Synthesis of enantiopure imidazolines through a Ritter reaction of 2-(1-aminoalkyl)aziridines with nitriles.

The Ritter reaction of enantiopure 2-(1-aminoalkyl)aziridines 1 with different nitriles afford enantiopure tetrasubstituted imidazolines 2. The opening of the aziridine ring takes place with total regio- and stereoselectivity. A mechanism to explain the described addition reaction is proposed. [reaction: see text]