Nucleobase-Functionalized 7-Deazaisoguanine and 7-Deazapurin-2,6-diamine Nucleosides: Halogenation, Cross-Coupling, and Cycloaddition.

The functionalization in position-7 of 7-deazaisoguanine and 7-deazapurin-2,6-diamine ribo- and 2'-deoxyribonucleosides by halogen atoms (chloro, bromo, iodo), and clickable alkynyl and vinyl side chains for copper-catalyzed and copper-free cycloadditions is described. Problems arising during the synthesis of the 7-iodinated isoguanine ribo- and 2'-deoxyribonucleosides were solved by the action of acetone. The impact of side chains and halogen atoms on the pKa values and hydrophobicity of nucleosides was investigated. Halogenated substituents increase the lipophilic character of nucleosides in the order Cl < Br < I and decrease the pK values of protonation. Photophysical properties (fluorescence, solvatochromism, and quantum yields) of azide-alkyne click adducts bearing pyrene as sensor groups were determined. Pyrene fluorescence was solvent-dependent and changed according to the linker lengths. Excimer emission was observed in dioxane for the long linker adduct. Bioorthogonal inverse-electron-demanding Diels-Alder cycloadditions (iEDDA) were conducted on the electron-rich vinyl groups of 7-deazaisoguanine and 7-deazapurin-2,6-diamine nucleosides as dienophiles and 3,6-dipyridyl-1,2,4,5-tetrazine as diene. The initially formed complex reaction mixture of isomers could be easily oxidized with iodine in tetrahydrofuran (THF)/pyridine leading to single aromatic tetrazine adducts within a short time and in excellent yields.

The synthesis and bioactivity of pyrrolo[2,3-d]pyrimidine derivatives as tyrosine kinase inhibitors for NSCLC cells with EGFR mutations.

EGFR mutations are an ongoing challenge in the treatment of NSCLC, and demand continuous updating of EGFR TKI drug candidates. Pyrrolopyrimidines are one group of versatile scaffolds suitable for tailored drug development. However not many precedents of this type of pharmacophore have been investigated in the realm of third generation of covalent EGFR-TKIs. Herein, a series of pyrrolo[2,3-d]pyrimidine derivatives able to block mutant EGFR activity in a covalent manner were synthesized, through optimized Buchwald-Hartwig C-N cross coupling reactions. Their preliminary bioactivity and corresponding inhibitory mechanistic pathways were investigated at molecular and cellular levels. Several compounds exhibited increased biological activity and enhanced selectivity compared to the control compound. Notably, compound 12i selectively inhibits HCC827 cells harboring the EGFR activating mutation with up to 493-fold increased efficacy compared to in normal HBE cells. Augmented selectivity was also confirmed by kinase enzymatic assay, with the test compound selectively inhibiting the T790 M activating mutant EGFRs (IC50 values of 0.21 nM) with up to 104-fold potency compared to the wild-type EGFR (IC50 values of 22 nM). Theoretical simulations provide structural evidence of selective kinase inhibitory activity. Thus, this series of pyrrolo[2,3-d]pyrimidine derivatives could serve as a starting point for the development of new EGFR-TKIs.

Acyclic Janus-AT Nucleoside Host Channels Precisely Lock Water into Single-File Wires with Local Rotational Flexibility.

Confining polar water molecules to particular geometries demands sophisticated intermolecular interactions, and not many small synthetic molecules have accomplished such a task. Herein, regioisomeric acyclic Janus-AT nucleosides (1 and 2), with a self-complementary fused genetic alphabet and conformationally flexible side chains, have been selectively synthesized. 1 and 2 adopt disparate base-pair motifs from the π-π stacked hydrophobic base moieties and distinct hydrogen bond (HB) interconnections from the hydrophilic sugar residues, which in turn lead to divergent, intricate intermolecular interaction networks with different capacities to confine water molecules. Under the precise control of the host framework of the N8 -regioisomer, separate ordered single-file water wires can be locked through special three-HB clamps into unique inter- and intra-wire geometrical alignments. Localized dynamic synchronized rotations within the fixed framework coordinated by both the host hydroxy groups and guest water molecules were observed in a temperature-induced reversible single-crystal-to-single-crystal transition (SCSCT).

Effects of glutathione on the survival, growth performance and non-specific immunity of white shrimps (Litopenaeus vannamei).

Reduced glutathione (GSH) is the most abundant non-enzymatic antioxidant present in mammalian cells and the main intracellular defence mechanism against oxidative stress. This study investigated the effects of GSH on survival rate, mean body gain weight, feed efficiency (FE), phenoloxidase (PO) activity, superoxide dismutase (SOD) activity, acid phosphatase (ACP), alkaline phosphatase (AKP) activity, GSH peroxidase (GPx) and susceptibility to Vibrio alginolyticus when Pacific white shrimps (Litopenaeus vannamei) were fed with GSH-containing diets. GSH was added to diets at 0.10, 0.20 and 0.30 g/kg during the eight-week breeding experiment. Oral administration of GSH had significantly increased mean body weight gain, FE, PO activity, SOD activity, ACP activity, AKP activity, GPx activity and susceptibility to V. alginolyticus compared with those of the control group (p < .05). Results indicate that GSH exerts both growth-promoting and immunostimulatory effects on Pacific white shrimps (L. vannamei).

High-yield production of canine parvovirus virus-like particles in a baculovirus expression system.

An optimized VP2 gene from the current prevalent CPV strain (new CPV-2a) in China was expressed in a baculovirus expression system. It was found that the VP2 proteins assembled into virus-like particles (VLPs) with antigenic properties similar to those of natural CPV and with an especially high hemagglutination (HA) titer (1:2(20)). Dogs intramuscularly or orally immunized with VLPs produced antibodies against CPV with >1:80 hemagglutination inhibition (HI) units for at least 3 months. The CPV VLPs could be considered for use as a vaccine against CPV or as a platform for research on chimeric VLP vaccines against other diseases.

Preparation of the oligosaccharides derived from Flammulina velutipes and their antioxidant activities.

The oligosaccharides were prepared from Flammulina velutipes by hydrolysis of F. velutipes polysaccharides with hydrogen peroxide (H2O2). The yields of F. velutipes derived oligosaccharides (FVOs) were monitored during the hydrolysis process. FVOs yields were affected by three factors, i.e. reaction temperature, H2O2 concentration, and time, which were optimized by using an orthogonal design experiments as follows: reaction temperature 70°C, H2O2 concentration 3%, and reaction time 6h. Under these optimum conditions, the maximal yield of the oligosaccharides reached 17.10%, which was higher than that of hot water extraction method. The oligosaccharides were partially characterized by Fourier transform infrared spectrum, monosaccharide composition, and antioxidant activity. The results indicate that the oligosaccharides derived from F. velutipes showed strong hydroxyl radical activity and reducing capacity at the concentration of 100 µg/mL.

Effect of Tween 80 on the production of curdlan by Alcaligenes faecalis ATCC 31749.

This study aims to investigate the effects of Tween 80 on curdlan production, cell growth, and glucosyltransferase activity. The addition of Tween 80 to the culture medium increased curdlan production. However, curdlan production did not increase further when excessive Tween 80 (>0.3% Tween 80) was added to the culture medium. The addition of Tween 80 to the culture medium did not affect cell growth. The glucosyltransferase activity involved in the curdlan synthesis increased with the increase of Tween 80 concentration. The glucosyltransferase activity did not increase further when excessive Tween 80 (>0.3% Tween 80) was added to the culture medium. Maximum curdlan was observed at day 5 and then levelled off. The biomass continued to increase until the end of the experimental period (6d). Maximum glucosyltransferase activity was also observed at day 5 and decreased thereafter. The results indicate that the enhanced curdlan production by Tween 80 is highly correlated with glucosyltransferase activity.

Preparation of water soluble chitosan by hydrolysis using hydrogen peroxide.

Chitosan is not soluble in water, which limits its wide application particularly in the medicine and food industry. In the present study, water soluble chitosan (WSC) was prepared by hydrolyzing chitosan using hydrogen peroxide under the catalysis of phosphotungstic acid in homogeneous phase. Factors affecting hydrolysis were investigated and the optimal hydrolysis conditions were determined. The WSC structure was characterized by Fourier transform infrared spectroscopy. The resulting products were composed of chitooligosaccharides of DP 2-9. The WSC content of the product and the yield were 94.7% and 92.3% (w/w), respectively. The results indicate that WSC can be effectively prepared by hydrolysis of chitosan using hydrogen peroxide under the catalysis of phosphotungstic acid.

Hypolipidemic activity of the chitooligosaccharides from Clanis bilineata (Lepidoptera), an edible insect.

Clanis bilineata (CB) is an edible insect, and CB larvae (CBL) are widely consumed in China, yielding a large number of CBL skin (CBLS). In this study, chitooligosaccharides (COS) from CBLS were prepared and their hypolipidemic activity was determined in rats. Thirty-six male Sprague-Dawley rats in 3 groups were fed diets containing either high fat (HF) or COS, or a normal fat (NF) for 6 weeks. Compared with HF group, COS group showed decreased body weight gain, plasma triacylglycerol (TG), total cholesterol (TC), and plasma low-density lipoprotein cholesterol (LDL-C) level. Fecal fat and cholesterol of the COS group were higher than those of NF and HF groups. The results indicated that COS could be a suitable alterative hypolipidemic source for humans.

Nutritional evaluation of protein from Clanis bilineata (Lepidoptera), an edible insect.

BACKGROUND: This study was carried out to determine the nutritive quality of protein from Clanis bilineata (CB), an edible insect. RESULTS: The protein content of dried CB was 685.3 g kg(-1) and its essential amino acid content was 528.4 g kg(-1), both of which are comparable to those of milk and eggs. Three groups of rats were fed diets containing either casein or CB protein (CBP) as protein source or a protein-free diet for 10 days. The nutritional parameters measured for the group fed the diet containing CBP showed a positive nitrogen balance of 1.37, a net protein retention of 2.9 and a true digestibility of 95.8%, comparable to those measured for the group fed the casein diet. CONCLUSION: The results suggest that CBP may be a suitable alternative dietary protein source for humans.