Methoxy Ethyl Research Articles

Allele-Selective Thiomorpholino Antisense Oligonucleotides as a Therapeutic Approach for Fused-in-Sarcoma Amyotrophic Lateral Sclerosis.

Pathogenic variations in the fused in sarcoma (FUS) gene are associated with rare and aggressive forms of amyotrophic lateral sclerosis (ALS). As FUS-ALS is a dominant disease, a targeted, allele-selective approach to FUS knockdown is most suitable. Antisense oligonucleotides (AOs) are a promising therapeutic platform for treating such diseases. In this study, we have explored the potential for allele-selective knockdown of FUS. Gapmer-type AOs targeted to two common neutral polymorphisms in FUS were designed and evaluated in human fibroblasts. AOs had either methoxyethyl (MOE) or thiomorpholino (TMO) modifications. We found that the TMO modification improved allele selectivity and efficacy for the lead sequences when compared to the MOE counterparts. After TMO-modified gapmer knockdown of the target allele, up to 93% of FUS transcripts detected were from the non-target allele. Compared to MOE-modified AOs, the TMO-modified AOs also demonstrated reduced formation of structured nuclear inclusions and SFPQ aggregation that can be triggered by phosphorothioate-containing AOs. How overall length and gap length of the TMO-modified AOs affected allele selectivity, efficiency and off-target gene knockdown was also evaluated. We have shown that allele-selective knockdown of FUS may be a viable therapeutic strategy for treating FUS-ALS and demonstrated the benefits of the TMO modification for allele-selective applications.

Feasibility Evaluation and Numerical Simulation of Diesel–Diethyl Ether–2–Methoxy Ethyl Ether Blends Fueling in Stationary CI Engine

In many developed economies, the diesel engine is very important for transportation and farming. There is a lot of research being done in the field of renewable energy to replace conventional energy sources because of the major environmental issues and the rising expense of diesel. Diesel engine cannot be reinstate because of its efficient performance at higher power and reliability with alternative engines. Diesel engine emissions are very harmful to the atmosphere and human health. The main contaminants in diesel engines are smoke and NOx, which need to be effectively monitored. A numerous research is going on to diminish the emissions from CI engines by using some additives as well as the use of alternative fuels. This experimental inquiry was conducted to identify a suitable addition to lower exhaust emissions and improve CI engine performance. The trials used various mix ratios of pure diesel and blends of diesel and diethyl ether-2-methoxy ethyl ether. Mixing of 5% DEE and 5% MXEE with 90% diesel on volume basis (D90–DEE5–MXEE5) showed optimum results of emission and performance. Low exhaust emissions (HC 63.15%, CO 60.00%, and Smoke 18.29%) found to be substantial at peak loads and performance increase (decreasing in BSFC 5.00% and increasing in BTE 3.00%) were compared to mixture D90-DEE5-MXEE5 with diesel (at standard engine conditions). However, NOx rises (2.60%).

Oligonucleotides Containing C5-Propynyl Modified Arabinonucleic Acids: Synthesis, Biophysical and Antisense Properties.

The introduction of chemical modifications on the nucleic acid scaffold has allowed for the progress of antisense oligonucleotides (ASOs) in the clinic for the treatment of a variety of disorders. In contribution to the repertoire of gene-silencing nucleic acid modifications, herein we report the synthesis and incorporation of C5-propynyl arabinouridine (araUP ) and arabinocytidine (araCP ) into mixed-base ASOs containing a pyrimidine core. Substitution of the core with araUP and araCP resulted in stabilization of the duplex formed with RNA but not with DNA. Similar results were obtained with ASOs bearing phosphorothioate linkages or methoxyethyl (MOE) wings in a gapmer design. All modified ASOs were compatible with E. coli RNase H mediated degradation of target RNA. Substitution of DNA for araUP and araCP in the central portion of a gapmer with MOE wings demonstrated improved nuclease resistance. These results suggest C5-modified arabinonucleic acids may serve as a potential chemical modification for therapeutic ASOs.

Non-target impacts of pesticides on soil N transformations, abundances of nitrifying and denitrifying genes, and nitrous oxide emissions

Agriculture is the leading contributor to global nitrous oxide (N2O) emissions, mostly from soils. We examined the non-target impacts of four pesticides on N transformations, N cycling genes and N2O emissions from sugarcane-cropped soil. The pesticides, including a herbicide glyphosate (GLY), an insecticide imidacloprid (IMI), a fungicide methoxy ethyl mercuric chloride (MEMC) and a fumigant methyl isothiocyanate (MITC), were added to the soil and incubated in laboratory at 25 °C. The soil microcosms were maintained at two water contents, 55 % and 90 % water holding capacity (WHC), to simulate aerobic and partly anaerobic conditions, respectively. Half of the soil samples received an initial application of KNO3 and were then maintained at 90 % WHC for 38 d, whilst the other half received (NH4)2SO4 and were maintained at 55 % WHC for 28 d followed by 10 d at 90 % WHC to favour denitrification. Responses of individual functional genes involved in nitrification and denitrification to the pesticides and their relationships to N2O emissions varied with time and soil water. Overall, MITC had pronounced repressive effects on AOA and AOB amoA gene abundances and gross nitrification. Under 55 % WHC during the initial 28 d, N2O emissions were low for all treatments (≤62 μg N kg−1 soil). However, under 90 % WHC (either during the first 28 d or the increase in water content from 55 to 90 % WHC after 28 d) the cumulative N2O emissions increased markedly. Overall, under 90 % WHC the cumulative N2O emissions were 19 (control) to 79-fold (MITC) higher than under 55% WHC; with the highest emissions observed in the MITC treatment (3140 μg N kg−1 soil). This was associated with increases in gross nitrate consumption rates and abundances of denitrifying genes (nirK, nirS and qnorB). Therefore, to minimise N2O emissions, MITC should not be applied to field under wet conditions favouring denitrification.

Antisense Oligonucleotide-Related Macrovesicular Vacuolation of Hippocampal Neurons in Nonhuman Primates.

2'-methoxyethyl (MOE) antisense oligonucleotides (ASOs) tested in multidose intrathecal nonhuman primate (NHP) toxicity studies have consistently revealed the presence of single large vacuoles in pyramidal neurons of the hippocampus in the absence of any cellular response. Termed "macrovesicular," these vacuoles were characterized by immunohistochemistry and transmission electron microscopy which showed that these vacuoles are dilated lysosomes in neurons containing accumulated ASO. Additionally, two NHP studies were conducted to investigate the role of tissue fixation on their histogenesis. In Fixation Study 1, 6 doses of 5 mg 2'-MOE ASO with a full phosphorothioate backbone were administered by lumbar puncture over 5 weeks; in Fixation Study 2, 5 doses of 35 mg 2'-MOE ASO with a mixed phosphorothioate/phosphodiester backbone were administered over 12 weeks. At necropsy in each study, brain slices were either immersion fixed in neutral buffered 10% formalin or Carnoy's fixative; frozen at -80 °C; or perfusion fixed with modified Karnovsky's fixative. Fixed samples were processed to paraffin, sectioned, and stained with hematoxylin and eosin (H&E) and compared with H&E cryosections prepared from the frozen tissue of the same brain. The presence of vacuoles in fixed brain tissue but never in fresh frozen tissue showed that they arose during postmortem tissue fixation, and as such represent a processing artifact that is not relevant to human safety assessment of intrathecally administered 2'-MOE ASOs.

Anticorrosive behaviour of poly(vinyl carbazole-co-methoxy ethyl methacrylate) and its nanocomposites

A copolymer of vinyl carbazole with methoxy ethyl methacrylate [Poly (VK-co-MEM)] was prepared by free radical solution polymerization. The copolymer was reinforced with two different nanofillers namely CaCO3 and graphene by solution blending technique. The resultant polymer nanocomposites were characterized by Fourier Transform Infra-Red Spectroscopy (FTIR) and X-ray Diffraction (XRD) studies. The SEM micrographs of polymer and its nanocomposites were taken to observe the morphological changes. The IDT of the polymer and its nanocomposites were calculated using Thermogravimetric Analysis (TGA). The optical characteristic of the synthesized polymer and its nanocomposites were assessed using UV–Vis spectroscopy. The anticorrosive property of [Poly (VK-co-MEM)] and its nanocomposite coated on mild steel (MS) in 3.5% (w/v) NaCl were carried out using Potentiodynamic Polarization and Electrochemical Impedance Spectroscopy (EIS). The results show the prepared material has better protection against corrosion.

Low temperature ionothermal synthesis of TiO 2 nanomaterials for efficient photocatalytic H 2 production, dye degradation and photoluminescence studies

The photocatalytic hydrogen generation is a novel, eco-friendly and favourable method for production of green and clean energy using light energy. In this direction, we report low-temperature ionothermal method for the preparation of TiO2 nanoparticles (NPs) using methoxy ethyl methyl imidazolium tris (pentafluoroethyl) trifluoro phosphate (MOEMINtf2) as an ionic liquid (IL) at 120°C for 1 day. The synthesized nanomaterials were examined using different spectrochemical methods like UV-DRS, XRD, FT-IR, TEM, BET and TGA-DTA techniques. The mixed phase TiO2 is obtained with 81.7% of anatase and 18.3% of rutile phase by the XRD studies, and average crystallite size is found to be ∼7 nm. The stretching of Ti-O bond (∼555 cm−1) and few other bands related to ionic liquid were confirmed by FTIR spectrum. The band gap energy was observed to be ∼3.38 eV by UV-DRS analysis. TEM images reveal spherical shape with an average particles size of about 10 nm. Photocatalytic H2 generation was carried out using TiO2 NPs and observed the generation of 553 μmol h−1 g−1 via water splitting reaction. Furthermore, the prepared TiO2 NPs employed for the photocatalytic degradation of methylene blue dye (84.54%), and photoluminescence studies confirms the obtained material can be used in optoelectronic applications with green emission.

Increased Affinity of 2′‐O‐(2‐Methoxyethyl)‐Modified Oligonucleotides to RNA through Conjugation of Spermine at Cytidines

AbstractStructural modification at the 2′‐O‐position of riboses in oligonucleotide therapeutics is of critical importance for their use as drugs. To date, the methoxyethyl (MOE) substituent is the most important and features in dozens of antisense oligonucleotides that have been tested in clinical trials. Yet, the search for new improved modifications continues in a quest for increased oligonucleotide potency, improved transport in vivo and favorable metabolism. Recently, we described how the conjugation of spermine groups to pyrimidines in oligonucleotides vastly increases their affinity for complementary RNAs through accelerated binding kinetics. Here we describe how spermines can be linked to the exocyclic amino groups of cytidines in MOE‐oligonucleotides employing a straightforward ‘convertible nucleoside approach’ during solid phase synthesis. Singly‐ or doubly‐modified oligonucleotides show greatly enhanced affinity for complementary RNA, with potential for a new generation of MOE‐based oligonucleotide drugs.

Effect of DEE and MEA as additives on a CRDI diesel engine fueled with waste plastic oil blend

ABSTRACTThe unavoidable rise in plastic usage creates a dangerous threat to the environment due to its non-biodegradable nature. Disposal of waste plastics is carried out using various techniques. Among that pyrolysis process is effective one to convert waste plastics into oil. This plastic oil is used as an alternative fuel in internal combustion engines. In this present work, plastic oil is produced through thermal pyrolysis and catalytic pyrolysis processes using polyethylene plastic wastes (HDPE, LDPE) as feedstock. In the catalytic pyrolysis process, fly ash used as catalyst. Then, the effect of diethyl ether (DEE) and methoxy ethyl acetate (MEA) on the performance, combustion and emission characteristics of the CRDI diesel engine fueled with waste plastic oil blend is analyzed and compared with the diesel fuel operation. The experiments are carried out for six load conditions varying from 0% to 100% and without any engine modification. The experimental results showed that improvement in brake thermal efficiency is attained with test fuel D50W40MEA10 than neat diesel. It is observed from investigation that test fuel 50D40W10MEA has produced lower CO, HC and smoke emission than neat diesel. NOX emission with test fuel D50W40MEA10 is slightly higher than neat diesel, but lower than neat diesel with test fuel D50W40DEE10.

Intermolecular Interaction of Polymer Brushes Containing Phosphorylcholine and Inverse-Phosphorylcholine.

Choline phosphate (CP) is a phosphobetaine-type zwitterionic functional group, referred to as inverse phosphorylcholine (PC) due to the reverse orientation of a positively charged quaternary amine and anionic phosphate in contrast to PC lipids in nature. The A unique dipole paring between CP and PC groups has attracted much attention in the biointerface research field. Herein, to evaluate the molecular interaction between the CP and PC groups in water, force-distance curve measurements using scanning probe microscopy (SPM) with a PC-group-functionalized cantilever was carried out on the surface of polymer brushes bearing the CP groups. Three types of methacrylate monomers bearing CP with ethyl (Et), methoxyethyl (MOE), and isopropyl (iPr) phosphates were synthesized in 42-71% yields, and polymerized by surface-initiated atom transfer radical polymerization to form polymer brushes on silicon wafers. The surface free energy of CP-polymer brushes with Et, MOE, and iPr was estimated to be 64.0, 61.4, and 57.4 mN m-1, respectively, based on contact angle measurements. Force-distance curve measurements of polymer brushes having a CP group was conducted in water at 25 °C by SPM using a spherical probe produced by attaching a silica particle (SiP; d = 25 μm) covered with PC or CP groups to a tipless cantilever. Adhesion force larger than 14 nN was observed between the CP-polymer brushes and PC-SiP, whereas PC-polymer brushes revealed extremely low adhesion force of less than 0.6 nN with PC-SiP and propylsilane-modified SiP. The specific attractive molecular interaction between CP and PC groups was quantitatively evaluated.

Bioactive compounds from mangrove derived rare actinobacterium Saccharomonospora oceani VJDS-3.

A rare actinobacterium was isolated from Nizampatnam mangrove ecosystem of Andhra Pradesh, India, and was screened for its ability to produce bioactive compounds. The potential strain was identified as Saccharomonospora oceani VJDS-3 by polyphasic taxonomy. Purification of the biologically active compounds by column chromatography led to the isolation of three compounds, namely methoxy ethyl cinnamate (ethyl(E)-3-(4-methoxyphenyl)acrylate) (R1), 4-hydroxy methyl cinnamate (methyl(E)-3-(4-hydroxyphenyl)acrylate) (R2) and 4-methylbenzoic acid (R3). The structure of the compounds was elucidated on the basis of spectroscopic analysis including FTIR, EIMS, 1HNMR and 13CNMR spectroscopies. The antimicrobial activity of the bioactive compounds produced by the strain was tested against a panel of bacteria and fungi, and expressed in terms of minimum inhibitory concentration. Compound (R1) exhibited higher antimicrobial potential (50µg/ml) against Staphylococcus aureus, Bacillus megaterium and Candida albicans compared to R2 and R3. Antioxidant activity of compounds was determined by DPPH and ABTS radical scavenging activities. The results revealed that compound R3 effectively scavenged DPPH (73.08±1.29) and ABTS (99.74±0.00) radicals at a concentration of 25 and 50µg/ml, respectively. Antidiabetic and anti-obesity activities were evaluated by inhibitory potential of compounds against alpha-glucosidase, alpha-amylase and pancreatic lipase by spectrophotometric assays. Compound R1 showed effective inhibition against alpha-glucosidase (66.8±1.2) at 20µg/ml while moderate to weak activities were found against alpha-amylase and pancreatic lipase. To the best of our knowledge, this is the first report on the isolation of supra said compounds from the genus Saccharomonospora.

Synthesis, Self-Association, and Solubilizing Ability of an Amphiphilic Derivative of Poly(ethylene glycol) Methyl Ether

A new amphiphilic derivative–methoxy(polyethoxy)ethyl stearate has been synthesized with the goal of creating therefrom systems for delivery and enhancement of bioavailability of piperine. The aggregation and solubilizing properties of ω-methylpoly(oxyethylene) stearate have been studied by means of a set of physicochemical methods.

Oligonucleotide Therapies for the Lung: Ready to Return to the Clinic?

The most promising targets for treating lung diseases are often intracellular molecules recalcitrant to conventional pharmacotherapy. In this respect, antisense oligonucleotides (ASO) would be valuable tools to treat airways disease, as the plurality of their potential mechanisms of action is well-established, their efficiency has been extensively validated in-vitro, and their potential clinical value has been demonstrated with numerous regulatory approvals. Yet in spite of high confidence in the treatment rationale and mechanism of action, it appears that oligonucleotides delivered topically to the lung either rapidly access circulation via epithelial transcytosis or are removed by alveolar macrophages, exerting minimal if any action in the cytosol of cells relevant to lung disease. Moreover, use of cell penetrating peptides, liposomes or nanoparticle delivery systems has not so far been shown to eliminate circulatory clearance, may activate immune responses, or drive macrophage phenotypic changes that, together or in isolation, may present risks to patients.

Ionothermal synthesis of TiO2 nanoparticles for enhanced photocatalytic H2 generation

Photocatalytic hydrogen generation is one of the most promising solutions to convert light energy into green chemical energy. In the present work, methoxy ethyl methyl imidazolium methyl sulphonate ionic liquid is used for the synthesis of i-TiO2 nanoparticles via ionothermal method at 120 °C. The obtained products were characterized by various spectroscopic techniques like XRD, FTIR, Raman, UV–visible, DRS, TEM and TG-DSC analysis. XRD pattern confirmed the anatase phase with minor rutile phase having average crystallite size of 5 nm. From the FTIR spectrum, the band appeared at ∼547 cm−1 confirmed the Ti–O–Ti stretching and also few bands of ionic liquid. UV–vis spectrum clearly reveals the blue shift due to size effect of TiO2. The spherical surface structure and particle size (15–30 nm) have been studied in detail using TEM images. Finally, the practical applicability of the as synthesized i-TiO2 nanoparticles is shown by using it as a photocatalyst towards the generation of H2 through water splitting reaction and it is found to be 462 μmol h−1g−1.

Site-Specific Modification Using the 2'-Methoxyethyl Group Improves the Specificity and Activity of siRNAs.

Rapid progress has been made toward small interfering RNA (siRNA)-based therapy for human disorders, but rationally optimizing siRNAs for high specificity and potent silencing remains a challenge. In this study, we explored the effect of chemical modification at the cleavage site of siRNAs. We found that modifications at positions 9 and 10 markedly reduced the silencing potency of the unmodified strand of siRNAs but were well tolerated by the modified strand. Intriguingly, addition of the 2′-methoxyethyl (MOE) group at the cleavage site improved both the specificity and silencing activity of siRNAs by facilitating the oriented RNA-induced silencing complex (RISC) loading of the modified strand. Furthermore, we combined MOE modifications at positions 9 and 10 of one strand together with 2′-O-methylation (OMe) at position 14 of the other strand and found a synergistic effect that improved the specificity of siRNAs. The surprisingly beneficial effect of the combined modification was validated using siRNA-targeting endogenous gene intercellular adhesion molecule 1 (ICAM1). We found that the combined modifications eliminated its off-target effects. In conclusion, we established effective strategies to optimize siRNAs using site-specific MOE modifications. The findings may allow the creation of superior siRNAs for therapy in terms of activity and specificity.

SiRNA mediated inhibition of pancreatic tumor growth in vitro and in vivo

Rapid progress has been made toward small interfering RNA (siRNA)-based therapy for human disorders, but rationally optimizing siRNAs for high specificity and potent silencing remains a challenge. In this study, we explored the effect of chemical modification at the cleavage site of siRNAs. We found that modifications at positions 9 and 10 markedly reduced the silencing potency of the unmodified strand of siRNAs but were well tolerated by the modified strand. Intriguingly, addition of the 2′-methoxyethyl (MOE) group at the cleavage site improved both the specificity and silencing activity of siRNAs by facilitating the oriented RNA-induced silencing complex (RISC) loading of the modified strand. Furthermore, we combined MOE modifications at positions 9 and 10 of one strand together with 2′-O-methylation (OMe) at position 14 of the other strand and found a synergistic effect that improved the specificity of siRNAs. The surprisingly beneficial effect of the combined modification was validated using siRNA-targeting endogenous gene intercellular adhesion molecule 1 (ICAM1). We found that the combined modifications eliminated its off-target effects. In conclusion, we established effective strategies to optimize siRNAs using site-specific MOE modifications. The findings may allow the creation of superior siRNAs for therapy in terms of activity and specificity.

Synthesis of Methoxyethyl Nucleoside Dimer Phosphoramidites

Four types of methoxyethyl (MOE) nucleoside phosphoramidites, which are categorized as second‐generation building blocks of antisense oligonucleotide drugs, were synthesized. Also, three types of MOE nucleoside dimer phosphoramidites were synthesized to increase the efficiency and oligomer purity in solid phase synthesis. The block‐like dimer phosphoramidites can prevent or minimize the formation of the (N‐1) mer impurity, thereby affording the fabrication of pure oligonucleotides and reducing the synthesis time by performing coupling reactions in the order of 2 + 2 + 2.

Design, Synthesis and Molecular Docking Study of 6,7-Dioxo-4-aryl-aminocoumarin

Catechol-O-methyltransferase (COMT) inhibitors play an important role in the treatment of Parkinson's disease (PD). On the basis of the structure-activity relation (SAR) analysis of existing COMT inhibitors, entacapone and tolcapone, it has been derived that coumarin compounds containing catechol structures may have inhibitory bioactivities on COMT. Thus, the novel COMT inhibitors, 6,7-dioxo-4-arylaminocoumarin compounds, were designed, and their inhibitory bioactivities on COMT were investigated by theoretical calculation. The results show that ten 6,7-dioxo-4-aryl-aminocoumarin compounds exhibit good docking effect, especially 6,7-bis(2-methoxyethoxy)-4-phenylamino-2H-chromen-2-one (6b4) and 4-(3-ethynylphenyl)amino-6,7-bis(2-methoxyethoxy)-2H-chromen-2-one (6b5) which have the structure of catechol protected by methoxy ethyl group.

Mechanism of alternative complement pathway dysregulation by a phosphorothioate oligonucleotide in monkey and human serum.

The species sensitivity and mechanism of complement pathway activation by a phosphorothioate oligonucleotide were investigated in monkey and human serum. Increasing concentrations of a phosphorothioate oligonucleotide, ISIS 2302, were incubated in either monkey or human serum. Complement activation in monkey serum was selective for the alternative pathway and occurred at concentrations ≥ 50 μg/mL ISIS 2302. By comparison, complement activation in human serum was absent. A similar difference in sensitivity for activation was also observed for a representative 2'-methoxyethyl (MOE)-modified oligonucleotide. The absence of oligonucleotide-induced complement activation was also observed in dogs. Protein binding with ISIS 2302 and enzyme competition studies suggested that factor H was important in oligonucleotide-mediated complement activation process, and addition of factor H to serum effectively prevented the activation in monkey serum. Furthermore, based on the immunoassay for factor H, there was an apparent decrease in factor H concentration as the ISIS 2302 concentration increased. This result suggests that ISIS 2302 binds to factor H and interferes with the factor H antibody from the immunoassay. Factor H is a regulatory protein that limits alternative pathway activation. Disruption of factor H interaction with C3 convertase by oligonucleotide could promote activation in this pathway.

Virtual Screening of compounds to 1-deoxy-Dxylulose 5-phosphate reductoisomerase (DXR) from Plasmodium falciparum.

The 1-deoxy-D-xylulose 5-phosphate reductoisomerase (DXR) protein (Gen Bank ID AAN37254.1) from Plasmodium falciparum is a potential drug target. Therefore, it is of interest to screen DXR against a virtual library of compounds (at the ZINC database) for potential binders as possible inhibitors. This exercise helped to choose 10 top ranking molecules with ZINC00200163 [N-(2,2di methoxy ethyl)-6-methyl-2, 3, 4, 9-tetrahydro-1H-carbazol-1-amine] a having good fit (-6.43 KJ/mol binding energy) with the target protein. Thus, ZINC00200163 is identified as a potential molecule for further comprehensive characterization and in-depth analysis.