Fluorinated N,N′-diarylureas as AMPK activators

Adenosine monophosphate-activated kinase (AMPK) plays a central role in regulating energy homeostasis in eukaryotic cells. AMPK also regulates lipid synthesis by inhibiting acetyl-CoA carboxylase (ACC) and regulates mTOR signaling by activating TSC2. Due to its important roles in cell metabolism, AMPK is an attractive target for metabolic diseases, such as type II diabetes and obesity. AMPK activators, such as metformin, that are used for diabetes treatment are also effective anticancer agents. However, the efficacies of many known AMPK activators are relatively low. For example, metformin activates AMPK at millimolar levels. In this study, we identified a novel family of AMPK activators, namely fluorinated N,N′-diarylureas, that activate AMPK at 1–3 μM concentrations. These novel agents strongly inhibit the proliferation of colon cancer cells. We studied the potential mechanisms of these agents, performed a structure–activity relationship (SAR) study and identified several fluorinated N,N′-diarylureas as potent AMPK activators.     

N的影响

植物和土壤中的¹⁵N自然丰度值(δ¹⁵N)是评价生态系统N循环的一个重要指标, 而放牧是草原生态系统的主要土地利用方式, 对草原生态系统的N循环过程的改变起着重要作用。该研究测定了内蒙古锡林河流域放牧和围封条件下草原群落主要优势植物和土壤的δ¹⁵N值, 探讨放牧对草原N循环的影响。研究中所测定的8种植物叶片δ¹⁵N变化很大(–4.04‰–4.34‰), 但与植物功能型有一定的相关性。放牧显著降低了大针茅(Stipa grandis)、杂类草和小半灌木木地肤(Kochia prostrata)的δ¹⁵N值。具有潜在共生固氮能力的豆科植物δ¹⁵N偏低负值(–4.04‰ – –1.90‰), 但在放牧和围封条件下无显著差异; 而被认为具有联合固氮能力的羊草(Leymus chinensis), 放牧后δ¹⁵N显著增加, 一定程度上表明了豆科植物和羊草生物固氮能力的存在。所有植物中, 除无菌根侵染的木地肤外, 其他有丛枝菌根真菌侵染记录的物种δ¹⁵N值较低, 通常接近0或为负值, 说明在N限制的内蒙古草原, 菌根转运N可能也是一种重要的N源途径。放牧显著降低了0–20 cm土壤δ¹⁵N值, 这也与过去的研究结果不同。δ¹⁵N的测定为生态系统提供了一个整合时空N循环过程的综合指标, 反映出放牧改变了草原生态系统的N循环。     

Stereoselective and driving-force-dependent photoinduced electron-transfer reactions of zinc myoglobin with optically active N,N′-dimethylcinchoninium and N,N′-dimethylcinchonidinium ions

In order to understand the detailed mechanism of the stereoselective photoinduced electron-transfer (ET) reactions of zinc-substituted myoglobin (ZnMb) with optically active molecules by flash photolysis, we designed and prepared new optically active agents, such as N,N′-dimethylcinchoninium diiodide ([MCN]I₂) and N,N′-dimethylcinchonidinium diiodide ([MCD]I₂). The photoexcited triplet state of ZnMb, ³(ZnMb)*, was successfully quenched by [MCN]²⁺ and [MCD]²⁺ ions to form the radical pair of ZnMb cation (ZnMb^·+) and reduced [MCN]^·+ and [MCD]^·+, followed by a thermal back ET reaction to the ground state. The rate constants (k _q) for the ET quenching at 25 °C were obtained as k _q(MCN)=(1.9±0.1)×10⁶ M⁻¹ s⁻¹ and k _q(MCD)=(3.0±0.2)×10⁶ M⁻¹ s⁻¹, respectively. The ratio of k _q(MCD)/k _q(MCN)=1.6 indicates that the [MCD]²⁺ preferentially quenches ³(ZnMb)*. The second-order rate constants (k _b) for the thermal back ET reaction from [MCN]^·+ and [MCD]^·+ to ZnMb^·+ at 25 °C were k _b(MCN)=(0.79±0.04)×10⁸ M⁻¹ s⁻¹ and k _b(MCD)=(1.0±0.1)×10⁸ M⁻¹ s⁻¹, respectively, and the selectivity was k _q(MCD)/k _q(MCN)=1.3. Both quenching and thermal back ET reactions are controlled by the ET step. In the quenching reaction, the energy differences of ΔΔH ^≠(MCD–MCN) and ΔΔS ^≠(MCD–MCN) at 25 °C were obtained as −1.1 and 0 kJ mol⁻¹, respectively. On the other hand, ΔΔH ^≠(MCD–MCN)=11±2 kJ mol⁻¹ and TΔΔS ^≠(MCD–MCN)=−10±2 kJ mol⁻¹ were given in the thermal back ET reaction. The highest stereoselectivity of 1.7 for [MCD]^·+ found at low temperature (10 °C) was due to the ΔΔS ^≠ value obtained in the thermal back ET reaction. Electronic Supplementary Material Supplementary material is available for this article at and is accessible for authorized users.     

The Structure of “N1, N6-Carbonyladenosine”

Abstract

Re-interpretation of the available data led to structural assignment of the title N¹, N⁶carbonyladenosine (1b) as N⁶,N⁶-carbonyldiadenosine (4b).     

应用N端Cys-free断裂蛋白质内含子标记蛋白质N端

蛋白质的特异位点修饰可以帮助了解蛋白质的结构与功能.但是,现有的蛋白质特异位点标记方法种类有限,而且存在局限性,所以有必要开发新的蛋白质特异位点标记方法．以谷胱甘肽-S-转移酶(GST)为研究对象,借助蛋白质反式剪接技术,建立了利用新型断裂蛋白质内含子对蛋白质进行N 端标记的新方法.在这个方法中,通过简单的重组表达、标记和纯化得到带有荧光基团的小肽,经过蛋白质反式剪接,荧光基团被标记到蛋白质的N 端. 初步研究结果显示,标记效率可达到12 %.     

Antimicrobial efficacy of some N,N’-Dialkyl-N,N’-dimethyl-1, 6-hexanediamine dioxides

A total of 17 N,N'-dialkyl-N,N'-dimethyl-1,6-hexanediamine dioxides were tested for activity against three microorganisms. A relationship was found between the length of the alkyl substituent and antimicrobial activity.     

蓝藻抗病毒蛋白 N

蓝藻抗病毒蛋白N(cyanovirin-N)是Boyd等1997年在蓝藻中发现的一种抗病毒蛋白,其抗病毒机制是与病毒表面衣壳蛋白上的甘露寡糖结合,阻止病毒与宿主细胞表面的受体结合。由于近年不断发现它对各种严重的RNA逆转录病毒的抗病毒活性而倍受关注。     

人禽流感H7N9、H5N1、H10N8临床特征

人禽流感(human-avian influenza)是一种由禽流感病毒中某些亚型感染人所引起的急性呼吸道传染病,目前能够感染人的禽流感病毒主要有H5、H7、H9和H10亚型。人感染禽流感病毒A(H5N1)、A(H7N9)、A(H10N8)与重症季节性流感临床表现相似,主要表现为重症肺炎,病死率高。人禽流感与重症季节性流感和重症新甲型H1N1流感患者有相同的危险因素,如高龄、合并基础疾病等,但人禽流感临床表现更重。     

Nα-formyl-norleucyl-leucyl-phenylalanyl chloromethylketone

Summary N-formyl-norleucyl-leucyl-phenylalanine-chloromethyl ketone is chemotactic for, and induces lysosomal enzyme release from rabbit peritoneal neutrophils over essentially the same range of concentrations as does the free acid form of the same peptide (Na-formyl-norleucyl-leucyl-phenylalanine-OH). The chloromethyl ketone derivative does however differ from the free acid in respect to its ability to interact with the neutrophil and cause deactivation or desensitization to cytochalasin B. Neutrophils preincubated in the cold with the chloromethyl ketone followed by washing have cytochalasin B sensitivity conferred upon them, as measured by the release of lysosomal enzymes. The degree of release induced by this pre-treatment appears to be related to the initial responsiveness of the cells. This is in contrast to the free acid where no cytochalasin B sensitivity in conferred under any circumstances. Thus, the chloromethyl ketone, unlike the free acid, appears to irreversibly activate the cell. Desensitization to the late addition of cytochalasin B is also significantly retarded when the chloromethyl ketone derivative is compared to the free acid form of the peptide. These studies suggest that the chloromethyl ketone derivative of the peptide may covalently interact with the neutrophil receptor.     

利用N糖苷酶F对单克隆抗体N糖酶解条件的优化

为了优化利用N糖苷酶F（PNGase F）酶解单克隆抗体中N糖的方法，应用本公司生产的单抗对PNGase F酶的酶解条件进行优化，包括缓冲液pH、酶种类、仪器、酶解程度、变性缓冲液及酶加入量等，总结酶解条件对N糖谱结果的影响。结果显示，置换缓冲液至1×PBS中可以避免某些单抗在低pH酶解时G0F转化为G0F-GN并可改善峰型；快速PNGase F和加入变性缓冲液能有效提高酶解效率；UPLC和1.7 μm粒径色谱柱能提高分离度；不完全酶解影响N糖含量结果。研究结果表明，采用优化的酶解条件可快速、有效的酶解单抗上的N糖，使N糖谱结果准确可靠，为细胞株筛选和单抗药物质量控制提供有效手段。     

Removal of239Pu from mice with 3,4,3 LICAM(C) or N,N′, N″, N‴-tetra-(2,3-dihydroxybenzoyl)-spermine

Summary Male mice SAS/4 were injected i.v. with²³⁹Pu citr(IV) 0.27 µCikg^–1–9.99 kBqkg^–1. After 1 h 30 µmol kg^–1 of 3,4,3 LICAM(C), N, N, N, N-tetra-(2,3-dihydroxybenzoyl)-spermine or Na₃CaDTPA as a reference compound was given intraperitoneally. After 4 days the animals were sacrified and the Pu content in livers, kidneys, femurs and carcasses was determined by the liquid scintillation method. It was found that, as compared with the control, 3,4,3 LICAM(C) removed 83% of the Pu activity deposited in the liver, 71% of that in the femur and 79% of the Pu in the whole body. The Pu content in the kidneys exceeded the control value by about 50%. Na₃CaDTPA removed 96, 86, 40 and 72% of plutonium from the liver, kidneys, femurs and carcasses respectively.Tetra-DHB-spermine caused the excretion of 50, 57 and 39% of Pu from liver, bone and whole body respectively. The retention of Pu in the kidneys was increased to 400% of the control value.     

Polarisationsoptischer Nachweis von Neurosekret mit N,N′-Diäthylpseudoisozyaninchlorid

Zusammenfassung An neurosekretorischen Zellen des Nucleus praeopticus der Bachforelle wurde nach oxydativer Vorbehandlung die Anlagerung von N,N-Diäthylpseudoisozyaninchlorid vergleichend fluoreszenz-und polarisationsmikroskopisch untersucht. Dicht angeordnete dissoziierte Sulfonsäuregruppen binden die Farbstoffassoziate elektrostatisch. Eine orientierte Einlagerung der reversiblen Polymerisate des Pseudoisozyanins in die neurosekretorischen Zellen ist polarisationsoptisch nachweisbar. Die polarisationsmikroskopischen Erscheinungen entsprechen den fluoreszenzoptischen. Gleichartige Anwendung von Akridinderivaten bei niedrigem pH-Wert und histochemische Kontrollversuche unterstützen die angegebene Erklärung der Befunde. Auf die Möglichkeit der polarisationsoptisch-quantitativen Untersuchung des Neurosekrets wird hingewiesen.

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Summary A comparative study (fluorescence and polarization microscopy) was carried out in the neurosecretory cells of the nucleus praeopticus in trouts. The object of the investigation was to study the adsorption of N,N-diethyl pseudoisocyanine chloride following oxidation. Densely arranged dissociated sulphonic acid groups seem to bind the dye electro-staticly. An orientated inclusion of the reversible polymers of the pseudoisocyanine into the neurosesecretory cells is demonstrable by polarization microscopy. The polarization optical image corresponds to the fluorescence microscopic image. The results of a similar investigation using acridine derivatives at a low pH and histochemical controls seem to support the results of the findings. The possibility of quantitative polarization optical studies of the neurosecretion is discussed.

     

Nδ-benzoyl-l-ornithine and Nδ-benzoyl-l-γ-hydroxyornithine from Vicia pseudo-orobus

Two new non-protein amino acids, N^δ-benzoyl-l-ornithine and N^δ-benzoyl-l-γ-hydroxyornithine have been characterized from the seeds of Vicia pseudo-orobus.     

Soybean yield response to residual NO3−N and applied N

Summary During 1976 through 1978, 10N treatments (combinations of N application times and rates) were used in a corn study. Those treatments created different levels of soil NO ₃ ^– –N content that were well-suited to a study of the influence of residual NO ₃ ^– –N and applied N on soybean yield. In April 1979 we applied ammonium nitrate at rates of 0, 75, or 150 kg N/ha to three subplots formed from each of the whole plots (previous N treatment plots). With N fertilization in 1979, seed yield increased where the residual NO ₃ ^– –N amount was less than 190 kg/ha but decreased where the residual amount was greater than 190 kg/ha. As the NO ₃ ^– –N content in the soil increased by 1 kg/ha, the soybean yield increase due to N fertilization in 1979 decreased by approximately 4 kg/ha.Contribution no. 82-368-J, Dep. of Agronomy, Kansas Agric. Exp. Stn., Manhattan, KS 66506, USA     

土壤动物与N素循环及对N沉降的响应

以主要的生态过程之一——N循环为对象,论述了土壤动物不仅对凋落物的分解有重要影响,而且在N素矿化和植物对N的吸收过程中也起着重要作用。同时,日益严重的全球变化问题之一——N沉降对土壤动物的多样性及其在生态系统中的功能构成了极大的威胁。另还对土壤动物与N循环研究的方法、土壤动物在N循环过程中的作用机制、热带地区的需求及N沉降下土壤动物的响应作了探讨,并提出,开展大尺度的专类研究及长期定位研究成为下一步研究的需要。     

N,N,N′,N′-tetramethyl-p-phenylenediamine initiates the appearance of a well-resolved I peak in the kinetics of chlorophyll fluorescence rise in isolated thylakoids

Addition of N,N,N′,N′-tetramethyl-p-phenylendiamine (TMPD) to thylakoid membranes isolated from pea leaves initiates the appearance of peak I in the polyphasic rise of chlorophyll (Chl) fluorescence observed during strong illumination, making it similar to that observed in leaves or intact chloroplasts. This effect depends on TMPD concentration and incubation period of isolated thylakoids with TMPD. The resolution of I-peak in the presence of weak concentrations of TMPD which reduced the overlap between I- and P-peaks, resulted from a decreased reduction of both fast and slow plastoquinone (PQ) pools of the granal and stromal thylakoids, respectively, as TMPD effectively accepts electrons from reduced PQ. High concentrations of TMPD markedly decreased the J–I–P phase of fluorescence rise and greatly retarded the I–P step rise. Accumulation of oxidized TMPD in the thylakoid lumen accelerated the re-oxidation of the acceptor side of Photosystem II (PSII) as illustrated by a two-fold increase in the magnitude of the fast component and complete suppression of the middle component of the variable Chl fluorescence (F_v) decay in the dark. Evidently, exogenous addition of high concentrations of TMPD prevented the light-induced reduction of the slow PQ pool.     

Antibiotic N", N""-Dibenzyleremomycin with a Reduced 1,2-Peptide Bond

Eremomycin derivatives with benzylated amino groups of both residues of eremosamine and with (R) or (S)-2-amino-4-methylpentyl substituted for N-methyl-D-Leu, the first amino acid residue of its heptapeptide, were synthesized in order to study the role of the peptide bond between the first and second amino acid residues of the heptapeptide moiety of the antibiotic in its interaction with the precursors of the bacterial cell wall peptidoglycan and the exhibition of its antibacterial activity. Comparison of the antibacterial activities of N",N"-dibenzyleremomycin, de-(N-methyl-D-Leu)-N",N"-dibenzyleremomycin, and its N-(2-amino-4-methylpentyl)-derivative (1,2-deoxo-N",N"-dibenzyleremomycin) demonstrated that cleavage or replacement of the first amino acid residue by the corresponding aminoalkyl residue results in a decrease in its antibacterial activity towards both vancomycin-sensitive and vancomycin-resistant strains of microorganisms.     

《柳叶刀》：H7N9死亡风险低于H5N1高于H1N1

中国科学家6月24日在线发表于《柳叶刀》上的研究论文称,H7N9禽流感患者死亡风险远低于人感染H5N1禽流感。但高于季节性流感和甲型H1N1流感。