Methyl Group Research Articles

Development of LAT1-Selective Nuclear Medicine Therapeutics Using Astatine-211

We investigated nuclear medicine therapeutics targeting the L-type amino acid transporter 1 (LAT1). We previously reported that a nuclear medicine therapeutic drug using astatine 211 (211At), an alpha-emitting nuclide that can be produced in an accelerator and targets LAT1 as a molecular target, is effective. The seed compound was 3-[211At] Astato-α-methyl-L-tyrosine (211At-AAMT-OH-L). We used a unique labeling method. By changing the OH group of phenol to a methyl group, retention was successfully increased. It was also found that the amount of the L-isomer taken up by the D-isomer and L-isomer was clearly higher, and the L-isomer was superior as a therapeutic drug. Compounds in which the methyl group was replaced with an ethyl or propyl group were also examined, but their retention did not increase significantly. In fact, we observed increased non-specific accumulation and dynamics, suggesting that labeling may be off. In addition, 211At-AAMT-O-Me-L, which has a simple structure, was clearly superior in terms of uptake speed for several candidate compounds. As a result, we were able to develop a compound that can be easily labeled, has high specific radioactivity, is stable, and has a strong therapeutic effect.

Cooperative role of LSD1 and CHD7 in regulating differentiation of mouse embryonic stem cells

AbstractLysine-specific histone demethylase 1 (LSD1) is a histone demethylase that plays a critical role in epigenetic regulation by removing the methyl group from mono- and di-methylated lysine 4 on histone H3 (H3K4me1/2), acting as a repressor of gene expression. Recently, catalytically independent functions of LSD1, serving as a scaffold for assembling chromatin-regulator and transcription factor complexes, have been identified. Herein, we show for the first time that LSD1 interacts with chromodomain-helicase-DNA-binding protein 7 (CHD7) in mouse embryonic stem cells (ESCs). To further investigate the CHD7–LSD1 crosstalk, we engineered Chd7 and Chd7/Lsd1 knockout (KO) mouse ESCs. We show that CHD7 is dispensable for ESC self-renewal and survival, while Chd7 KO ESCs can differentiate towards embryoid bodies (EBs) with defective expression of ectodermal markers. Intriguingly, Chd7/Lsd1 double KO mouse ESCs exhibit proliferation defects similar to Lsd1 KO ESCs and have lost the capacity to differentiate properly. Furthermore, the increased co-occupancy of H3K4me1 and CHD7 on chromatin following Lsd1 deletion suggests that LSD1 is required for facilitating the proper binding of CHD7 to chromatin and regulating differentiation. Collectively, our results suggest that LSD1 and CHD7 work in concert to modulate gene expression and influence proper cell fate determination.

S-adenosyl-L-methionine is the unexpected methyl donor for the methylation of mercury by the membrane-associated HgcAB complex

Mercury (Hg) is a heavy metal that exhibits high biological toxicity. Monomethylmercury and dimethylmercury are neurotoxins and a significant environmental concern as they bioaccumulate and biomagnify within the aquatic food web. Microbial Hg methylation involves two proteins, HgcA and HgcB. Here, we show that HgcA and HgcB can be heterologously coexpressed, and the HgcAB complex can be purified. We demonstrated that HgcA is a membrane-associated cobalamin-dependent methyltransferase and HgcB is a ferredoxin-like protein containing two [4Fe-4S] clusters. Further, spectroscopic and kinetic results demonstrate that S-adenosyl-L-methionine (SAM) donates the methyl group to Hg in a two-step reaction involving a methylcob(III)alamin intermediate including Co-thiolate ligation from a conserved Cys residue. Our findings uncover a biological role for SAM in microbial Hg methylation.

Enzyme stabilisation due to incorporation of a fluorinated non-natural amino acid at the protein surface.

We have previously engineered E. coli transketolase (TK) enzyme variants that accept new substrates such as aliphatic or aromatic aldehydes, and also with improved thermal stability. Irreversible aggregation is the primary mechanism of deactivation for TK in the buffers used for biocatalysis, and so we were interested in determining the extent to which this remains true in more complex media, crude cell lysates or even in vivo. Such understanding would better guide future protein engineering efforts. NMR offers a potential approach to probe protein structure changes, aggregation, and diffusion, and19F-labelled amino acids are a useful NMR probe for complex systems with little or no background signal from the rest of the protein or their environment. Here we labelled E. coli TK with two different19F probes, trifluoromethyl-L-phenylalanine (tfm-Phe), and 4-fluoro phenylalanine (4F-Phe), through site specific non-natural amino acid incorporation. We targeted them to residue K316, a highly solvent exposed site located at the furthest point from the enzyme active sites. Characterisation of the19F-labelled TK variants revealed surprising effects of these mutations on stability, and to some extent on activity. While variant TK-tfm-Phe led to a 7.5°C increase in the thermal transition midpoint (Tm) for denaturation, the TK-4F-Phe variant largely abolished the aggregation of the enzyme when incubated at 50°C19. F-NMR revealed different behaviours in response to temperature increases for the two TK variants, displaying opposite temperature gradient chemical shifts, and diverging motion regimes, suggesting that the mutations affected differently both the local environment at this site, and its temperature-induced dynamics. A similar incubation of TK at 40-55°C is also known to induce higher cofactor-binding affinities, leading to an apparent heat activation under low cofactor concentration conditions. We have hypothesised previously that a heat-inducible conformational change in TK leads to this effect1. H-NMR revealed a temperature-dependent re-structuring of methyl groups, also at 30-50°C, which may be linked to the heat activation. While our kinetic studies were not expected to observe the heat activation event due to the high cofactor concentrations used, this was not the case for TK-4F-Phe, which did appear to heat activate slightly at 45°C. This implied that the mutations at K316 could influence cofactor-binding, despite their location at 47 Å from either active site. Such long-distance effects of mutations are not unprecedented, and indeed we have previously shown how distant mutations can influence active-site loop stability and function in TK, mediated via dynamically coupled networks of residues. Molecular dynamics simulations of the two19F containing variants similarly revealed networks of residues that could couple the changes in dynamics at residue K316, through to changes in active site dynamics. These results independently highlight the sensitivity of active-site function to distant mutations coupled through correlated dynamic networks of residues. They also highlight the potential influence of surface-incorporated probes on protein stability and function, and the need to characterise them well prior to further studies.

Comprehensive Study on Cell Components in High‐Voltage Pouch Cells with Lithium Perchlorate: Decomposition, Transesterification, Chlorination, Deposition, and Self‐Discharge

AbstractBattery development has traditionally focused on high energy and long lifetime cells, but there is now a shift towards their sustainability and safety. One example of this trend is the search for fluorine‐free conductive salts. The overwhelming majority of lithium‐ion conductive salts contain fluorine, which is critical regarding their environmental impact, sustainability, and toxicology. In this study, we perform a comprehensive investigation of the performance and aging mechanisms of cell components with LiClO4 as conductive salt in high‐voltage NMC622‖Graphite pouch cells. The cells containing LiClO4 show poorer electrochemical performance compared to their LiPF6 equivalents. However, to the best of our knowledge, a mechanistic understanding of the effect of LiClO4 on the aging of electrode and electrolyte components for high‐voltage cells is largely missing. Developing such an understanding will pave the way toward designing alternative salts to LiPF6, ultimately leading to fluorine‐free and more sustainable battery cells. Our results show, that the chlorination of ethyl methyl carbonate at both methyl and ethyl groups and the formation of large (Liw)AlxOyClz composite deposits on the cathode surface result from perchlorate degradation at the cathode. This leads to increased cell resistance, reduced capacity retention, and accelerated degradation of the LiClO4‐containing electrolytes.

Vinylene‐Linking of Polycyclic Aromatic Hydrocarbons to π‐Extended Two‐Dimensional Covalent Organic Framework Photocatalyst for H2O2 Synthesis

AbstractPolycyclic aromatic hydrocarbons (PAHs) hold the predominant role either as individual molecules or building blocks in the field of organic semiconductors or nanocarbons. Connecting PAHs via sp2‐carbon bridges to form high‐crystalline π‐extended structures is highly desired not only for enlarging the regimes of two‐dimensional materials but also for achieving exceptional properties/functions. In this work, we developed 5,10‐dimethyl‐4,9‐diazapyrene as a key monomer, whose two methyl groups at the positions adjacent to nitrogen atoms, can helpfully increase the solubility, and serve as the active connection sites. In the presence of organic acids, this monomer enables smoothly conducting Knoevenagel condensation to form two vinylene‐linked PAH‐cored COFs, which show high‐crystalline honeycomb structures with large surface areas up to 1238 m2 g−1. Owing to the direct connection mode of PAH building blocks with vinylene, the as‐prepared COFs possess spatially extended π‐conjugation and substantial semiconducting properties. Consequently, their visible‐light photocatalysis with exceptional activity and durability was manifested to generate H2O2 up to 3820 μmol g−1 h−1 in pure water, and even 17080 μmol g−1 h−1 using benzyl alcohol as a hole sacrificial agent.

A Genetically Engineered Reporter System Designed for 2H-MRI Allows Quantitative In Vivo Mapping of Transgene Expression.

The ability to obtain quantitative spatial information on subcellular processes of deep tissues in vivo has been a long-standing challenge for molecular magnetic resonance imaging (MRI) approaches. This challenge remains even more so for quantifying readouts of genetically engineered MRI reporters. Here, we set to overcome this challenge with a molecular system designed to obtain quantitative 2H-MRI maps of a gene reporter. To this end, we synthesized deuterated thymidine, d3-thy, with three magnetically equivalent deuterons at its methyl group (-CD3), showing a singlet peak with a characteristic 2H-NMR frequency (δ = 1.7 ppm). The upfield 3.0 ppm offset from the chemical shift of the HDO signal (δ = 4.7 ppm) allows for spectrally resolving the two 2H NMR signals and quantifying the concentration of d3-thy based on the known concentration of a tissue's HDO. Following systemic administration of d3-thy, its accumulation as d3-thy monophosphate in cells expressing the human thymidine kinase 1 (hTK1) transgene was mapped with 2H-MRI. The data obtained in vivo show the ability to use the d3-thy/hTK1 pair as a reporter probe/reporter gene system to quantitatively map transgene expression with MRI. Relying on a structurally unmodified reporter probe (d3-thy) to image the expression of unmutated human protein (hTK1) shows the potential of molecular imaging with 2H-MRI to monitor gene reporters and other relevant biological targets.

Modifications of Prenyl Side Chains in Natural Product Biosynthesis

AbstractIn recent years, there has been a growing interest in understanding the enzymatic machinery responsible for the modifications of prenyl side chains and elucidating their roles in natural product biosynthesis. This interest stems from the pivotal role such modifications play in shaping the structural and functional diversity of natural products, as well as from their potential applications to synthetic biology and drug discovery. In addition to contributing to the diversity and complexity of natural products, unique modifications of prenyl side chains are represented by several novel biosynthetic mechanisms. Representative unique examples of epoxidation, dehydrogenation, oxidation of methyl groups to carboxyl groups, unusual C−C bond cleavage and oxidative cyclization are summarized and discussed. By revealing the intriguing chemistry and enzymology behind these transformations, this comprehensive and comparative review will guide future efforts in the discovery, characterization and application of modifications of prenyl side chains in natural product biosynthesis.

Pop the bubbles and let the current flow: mechanochemistry of micron and nano-sized openings in dielectric layers

Thin or ultra-thin dielectric layers have been widely used in various applications such as capacitors, piezo-electrics, and solar cells. This study explains the mechanism and chemistry of creating nano- and micron-sized openings in atomic-layer-deposited aluminum oxide-based dielectric layers using the alkali metal salt selenization technique. The necessary components for this mechanism are excess methyl groups present in the dielectric layer, supply of selenium and alkali metals, and a minimum annealing temperature. It is shown and explained that to create openings in the dielectric layer, heavier alkali halide metal salts require less energy, or - in other words - a lower annealing temperature. The overall hypothesis is explained via a thermodynamic approach with supportive thermochemical reactions. Thus, an easy way to engineer the dielectric layer to form openings at low temperatures is presented, beneficial for various applications like photovoltaics, optoelectronics, or micro-electro-mechanical systems (MEMS).

Synthesis of (2,3-dihydrobenzofuran-3-yl)acetic acids from salicylic aldehydes: Trimethylsilyl as traceless activating group

Salicylic aldehydes were readily alkylated with (chloromethyl)trimethylsilane to give the silylated ortho-methoxy benzaldehydes in 85–96 % yields. These aldehydes were converted into arylidenemalonates in excellent yields. O-(Trimethylsilyl)methyl group was applied as the synthetic equivalent of aryloxymethyl anion in the nucleophilic cyclization at the alkene moiety promoted by cesium fluoride in DMF at 140 °C. In this way, (2,3-dihydrobenzofuran-3-yl)acetic acids and the corresponding esters were synthesized from salicylic aldehydes in 41–70 % overall yields.

Structural and Photophysical Differences in Crystalline Trigonal Planar Copper Iodide Complexes with 1,2-Bis(methylpyridin-2-yl)disilane Ligands.

We synthesized trigonal planar Cu(I) iodide complexes with 1,2-bis(methylpyridin-2-yl)disilane ligands L1-L4 and investigated how the substitution position of the methyl group on the pyridine ring in σ-π conjugation affects their structure and physical properties. The structures were characterized by NMR, elemental analysis, and single-crystal X-ray diffraction. In the crystalline state, the methylpyridyl groups of CuIL1-CuIL3 were coordinated with Cu(I) in an anticlinal conformation relative to the Si-Si σ bond, whereas those of CuIL4 were coordinated with Cu(I) in a synperiplanar conformation relative to the Si-Si σ bond. The conformational difference in the crystalline state was influenced by the N-Cu-N bite angle and the emission wavelength. CuIL1-CuIL3 exhibited blue-green emission (λem: 476-494 nm), and CuIL4 exhibited green-yellow emission (λem: 512 nm) with high emission quantum yields (Φ: 0.59-0.86) in the crystalline state at 293 K. These Cu(I) complexes exhibited thermally activated delayed fluorescence from the S1 state at 293 K and phosphorescence from the T1 state at 77 K in the crystalline state. The optical properties in the crystalline state were discussed by DFT and TD-DFT calculations. These complexes also displayed aggregation-induced emission in THF-water solution (fw > 80%), although they did not show emission in dehydrated THF.

Disappearance of Odd-Even Effects at the Substrate Interface of n-Alkanes.

The physical and chemical properties of organic compounds having alkyl chains are frequently influenced by the parity of the chain length, which is known as the odd-even effect. Understanding the molecular origin of this phenomenon is particularly important for designing materials used in organic thin-film devices. In this work, we focus on thin films of n-alkanes as the simplest model to study the odd-even effect at the substrate interface and analyze the aggregation structure using p-polarized multiple-angle incidence resolution spectrometry in combination with grazing incidence X-ray diffraction. The spectroscopic analysis shows a pronounced odd-even alternation of the molecular tilt angles in the multilayer films. In addition, high-resolution Brewster-angle transmission spectroscopy reveals that the conformation of the methyl group highly depends on whether the carbon number is even or odd. In contrast to the multilayer films, the odd-even effects do not appear in the monolayer films. We demonstrate that, in other words, the interlayer interactions of the molecules are responsible for the odd-even effects. This study also highlights the first identification of the monolayer phase of n-alkanes by using grazing incidence X-ray diffraction in combination with high-resolution infrared spectroscopy. These results not only reveal the molecular origin for the odd-even effect of n-alkanes but also provide analytical techniques for discussing the monolayer structure of various alkylated compounds on a functional group basis.

Epigenetic Effects of Natural Products in Inflammatory Diseases: Recent Findings.

Inflammation is an essential step for the etiology of multiple diseases. Clinically, due to the limitations of current drugs for the treatment of inflammatory diseases, such as serious side effects and expensive costs, it is urgent to explore novel mechanisms and medicines. Natural products have received extensive attention recently because of their multi-component and multi-target characteristics. Epigenetic modifications are crucial pathophysiological targets for developing innovative therapies for pharmacological interventions. Investigations examining how natural products improving inflammation through epigenetic modifications are emerging. This review state that natural products relieve inflammation via regulating the gene transcription levels through chromosome structure regulated by histone acetylation levels and the addition or deletion of methyl groups on DNA duplex. They could also exert anti-inflammatory effects by modulating the proteins in typical inflammatory signaling pathways by ubiquitin-related degradation and the effect of glycolysis derived free glycosyls. Studies on epigenetic modifications have the potential to facilitate the development of natural products as therapeutic agents. Future research directed at better understanding of how natural products modulate inflammatory processes through less studied epigenetic modifications including neddylation, SUMOylation, palmitoylation and lactylation, may provide new implications. Meanwhile, higher quality preclinical studies and more powerful clinical evidence are still needed to firmly establish the clinical efficacy of the natural products. Trial Registration: ClinicalTrials.gov Identifier: NCT01764204; ClinicalTrials.gov Identifier: NCT05845931; ClinicalTrials.gov Identifier: NCT04657926; ClinicalTrials.gov Identifier: NCT02330276.

The ‘photosynthetic C1 pathway’ links carbon assimilation and growth in California poplar

Although primarily studied in relation to photorespiration, serine metabolism in chloroplasts may play a key role in plant CO2 fertilization responses by linking CO2 assimilation with growth. Here, we show that the phosphorylated serine pathway is part of a ‘photosynthetic C1 pathway’ and demonstrate its high activity in foliage of a C3 tree where it rapidly integrates photosynthesis and C1 metabolism contributing to new biomass via methyl transfer reactions, imparting a large natural 13C-depleted signature. Using 13CO2-labelling, we show that leaf serine, the S-methyl group of leaf methionine, pectin methyl esters, and the associated methanol released during cell wall expansion during growth, are directly produced from photosynthetically-linked C1 metabolism, within minutes of light exposure. We speculate that the photosynthetic C1 pathway is highly conserved across the photosynthetic tree of life, is responsible for synthesis of the greenhouse gas methane, and may have evolved with oxygenic photosynthesis by providing a mechanism of directly linking carbon and ammonia assimilation with growth. Although the rise in atmospheric CO2 inhibits major metabolic pathways like photorespiration, our results suggest that the photosynthetic C1 pathway may accelerate and represents a missing link between enhanced photosynthesis and plant growth rates during CO2 fertilization under a changing climate.

The contribution of methyl groups to electron spin decoherence of nitroxides in glassy matrices.

Long electron spin coherence lifetimes are crucial for high sensitivity and resolution in many pulse electron paramagnetic resonance (EPR) experiments aimed at measuring hyperfine and dipolar couplings, as well as in potential quantum sensing applications of molecular spin qubits. In immobilized systems, methyl groups contribute significantly to electron spin decoherence as a result of methyl torsional quantum tunneling. We examine the electron spin decoherence dynamics of the nitroxide radical 2,2,6,6-tetramethylpiperidin-1-oxyl (TEMPO) in both a methyl-free solvent and a methyl-containing solvent at cryogenic temperature. We model nitroxide and solvent methyl effects on decoherence using cluster correlation expansion (CCE) simulations extended to include methyl tunneling and compare the calculations to experimental data. We show that by using the methyl tunneling frequency as a fit parameter, experimental Hahn echo decays can be reproduced fairly well, allowing structural properties to be investigated in silico. In addition, we examine the Hahn echo of a hypothetical system with an unpaired electron and a single methyl to determine the effect of geometric configuration on methyl-driven electron spin decoherence. The simulations show that a methyl group contributes the most to electron spin decoherence if it is located between 2.5 and 6-7Å from the electron spin, with its orientation being of secondary importance.

The role of choline in the physiology of reproductivity

Choline is an essential nutrient, but intake among pregnant women is insufficient – only 7% of them consume adequate amounts of choline. The discrepancy between choline requirements and actual intake raises questions about the need for choline supplementation and its potential benefits for pregnant women. The diet of the expectant mother, rich in choline, in the preconception period ensures folliculogenesis and better quality of oocytes, changes the foetal epigenome, counteracting genomic defects during intrauterine development and reducing the susceptibility to metabolic changes in adulthood. Choline is a source of methyl groups and its consumption reduces the risk of NTDs by 7 times, so a promising trend in nutritional support of pregnancy and prevention of NTDs is the intake of folates and choline in combination with other micronutrients. Choline promotes the growth, proliferation and functioning of placental cells, transport of macronutrients, and has an anti-inflammatory effect. The risk of low foetal weight is reduced by 69% when choline is added to the mother’s regular diet. A significant connection has been shown between high levels of choline in the mother and a 2-fold reduction in the risk of preeclampsia, premature birth and a significant effect on the neurodevelopment and mental health of the child – improving neurocognitive functions and reducing the risk of autism and attention deficit hyperactivity disorder. A choline supplement at a dose of 130 mg/day as part of the vitamin-mineral complex Femibion 1 is sufficient to meet the needs for this nutrient during the preconception period and in the first trimester of pregnancy. The need for choline supplementation in the pregravid period and the first trimester of pregnancy is because on folliculogenesis and oocyte protection, epigenetic programming, the effectiveness of preventing congenital malformations of the foetus, participation in metabolic and physiological processes in the placenta, reducing the risk of placenta-associated complications, influence on the development of nervous tissue and brain of the foetus and newborn.

Exploring the Relationship Between Electrical Characteristics and Changes in Chemical Composition and Structure of OSG Low-K Films Under Thermal Annealing

The influence of annealing temperature on the chemical, structural, and electrophysical properties of porous OSG low-k films containing terminal methyl groups was investigated. The films were deposited via spin coating, followed by drying at 200 °C and annealing at temperatures ranging from 350 °C to 900 °C. In the temperature range of 350–450 °C, thermal degradation of surfactants occurs along with the formation of a silicon-oxygen framework, which is accompanied by an increase in pore radius from 1.2 nm to 1.5 nm. At 600–700 °C, complete destruction of methyl groups occurs, leading to the development of micropores. FTIR spectroscopy reveals that after annealing at 700 °C, the concentration of silanol groups and water reaches its maximum. By 900 °C, open porosity is no longer observed, and the film resembles dense SiO2. JV measurements show that the film annealed at 450 °C exhibits minimal leakage currents, approximately 5 × 10−11 A/cm2 at 700 kV/cm. This can be attributed to the near-complete removal of surfactant residues and non-condensed silanols, along with non-critical thermal degradation of methyl groups. Leakage current models obtained at various annealing temperatures suggest that the predominant charge carrier transfer mechanism is Poole–Frenkel emission.

Hole-Transporting Materials Based on a Fluorene Unit for Efficient Optoelectronic Devices

Solution-processable hole-transporting materials (HTMs) that form highly soluble films and thermally stable amorphous states are essential for advancing optoelectronic devices. However, the currently commercialized HTM, N,N-bis(3-methylphenyl)-N,N0-bis(phenyl)benzidine (TPD), exhibits poor solubility and limited carrier transport when spin-coated into thin films. Herein, to address these issues, a fluorenyl group was ingeniously incorporated into a series of molecules structurally similar to TPD. The resulting compounds, namely, 2,7-di-(N,N-diphenylamino)-9,9-dimethyl-9H-fluorene (DDF), 2,7-di-p-tolyl-(N,N-diphenylamino)-9,9-dimethyl-9H-fluorene (2M-DDF), and 2,7-di-tetra-p-tolyl-(N,N-diphenylamino)-9,9-dimethyl-9H-fluorene (4M-DDF), offered tunable energy levels, carrier transport, crystallinity, and steric configuration via adjustment of the number of terminal methyl groups. Owing to its satisfactory performance, 2M-DDF can serve as an effective alternative to TPD in OLED devices as well as a guest molecule in host–guest systems for long-afterglow materials. Devices incorporating 2M-DDF as the HTM, with an Alq3 emitter, achieved a maximum CE of 4.78 cd/A and a maximum L (Lmax) of 21,412 cd m−2, with a turn-on voltage (Von) of 3.8 V. The luminous efficiency of 2M-DDF was approximately five times that of TPD (4106 cd m−2). Furthermore, when 2M-DDF and TPD were utilized as guest molecules in afterglow materials, the afterglow duration of 2M-DDF (10 s) was 2.5 times that of TPD (4 s). This study provides a theoretical basis for the development of high-performance HTMs and long-afterglow materials, establishing a framework for the application of fluorene-based compounds in emerging fields such as long-afterglow materials.

Supercomputer-Based Virtual Screening for Deoxyribonucleic Acid Methyltransferase 1 Inhibitors as Novel Anticancer Agents

Targeting epigenetics is a new strategy to treat cancer and develop novel epigenetic drugs with anti-tumor activity. DNA methyltransferases transfer the methyl group from S-adenosyl-L-methionine (SAM) to the cytosine residue in a CpG island, leading to the transcription silencing of the gene. Hypermethylation can frequently be observed in several tumor types. Hence, the inhibition of DNMT1 has become a novel approach to cure cancer. In this study, virtual screening and molecular docking were performed for more than 11,000 ligands from the ZINC15 database to discover new hypomethylation agents. Four candidate compounds were further tested for their effects on DNMT1 in silico and in vitro. Compounds 2 and 4 showed the best DNMT1 inhibitory activity, but only compound 4 was able to inhibit the growth of several cancer cell lines. The hypomethylation of the luciferase gene by compound 4 was verified by a CMV- luciferase assay using KG-1 cells. Additionally, compound 4 suppressed cell migration in a dose- and time-dependent manner in the wound healing assay. Moreover, cell cycle analyses demonstrated that compound 4 arrested CCRF-CEM cells and MDA-MB-468 cells in the G0/G1 phase. Also, compound 4 significantly induced early and late apoptosis in a dose-dependent manner. In conclusion, we introduce compound 4 as a novel DNMT1 inhibitor with anticancer activity.

Systematic study on the hydrogen abstraction reactions from oxygenated compounds by H and HO2

AbstractTo extend the rule‐based approach for hydrogen abstraction reactions from oxygenated compounds, a systematic investigation was performed to examine the reactivity of gas‐phase hydrogen abstraction reactions from alkyl groups (methyl and ethyl groups) bound to oxygen atoms in five types of oxygenated compounds (alcohols, ethers, formate esters, acetate esters, and carbonate esters) by H atoms and HO2 radicals comprehensively considering rotational conformers. Quantum chemical calculations were conducted at the CBS‐QB3 level for stationary points. Rate constants were determined employing conventional transition state theory (TST). For hydrogen abstraction reactions by H, the rotational conformer distribution partition function was employed to approximate partition functions, owing to the similarity in vibrational energy‐level structures among conformers. In hydrogen abstraction reactions by HO2, the vibrational structures of transition‐state (TS) conformers varied significantly due to the hydrogen bonding, leading to an inappropriate evaluation of rate constants when using the lowest‐energy conformer as a representative. Therefore, the rate constants were calculated by the multi‐structural TST. It was revealed that the differences in functional groups containing O atoms mainly affect the bond dissociation energies of the C–H bonds and the activation energies of hydrogen abstraction reactions only when the C atoms are adjacent to the O atoms. Additionally, it was found that hydrogen bonds formed in the TSs show minor effect on rate parameters for the overall rate constants, apart from the reduction of the pre‐exponential factors for the H‐abstraction reactions from the methylene position of ethyl groups. The comparison with the rate constants from previous studies showed reasonable results, indicating that the rate constants in this study, which thoroughly consider rotational conformers, can be the current best estimates.