Quinazoline-2,4(1H,3H)-diones inhibit the growth of multiple human tumor cell lines

Quinazoline-2,4( $1H,3H$ )-diones exhibit a wealth of biological activities including antitumor proliferation. We established an improved method for the synthesis of quinazoline-2,4( $1H,3H$ )-dione derivatives with three points of molecular diversity. Data indicate that compounds 60 (average $\text{ logGI}_{50} \!=\! -6.1$ ), 65 (average $\text{ logGI}_{50} \!=\! -6.13$ ), 69 (average $\text{ logGI}_{50} \!=\! -6.44$ ), 72 (average $\text{ logGI}_{50} \!=\! -6.39$ ), and 86 (average $\text{ logGI}_{50} = -6.45$ ) significantly inhibited the in vitro growth of 60 human tumor cell lines tested. Structure–activity relationship analyses indicate that chlorophenethylureido is the necessary substituent at the $\text{ D}_{3}$ diversity point (7-position of quinazoline-2,4( $1H,3H$ )-dione), in particular, $o$ -chlorophenethylurea (69) achieved optimal activity. $o$ - or $m$ -Chlorophenethyl substitutions (69 and 72) at the $\text{ D}_{2}$ diversity point (3-position of quinazo line-2,4( $1H,3H$ )-dione) gave the most potent compounds. Methoxyl and 4-methylpiperazin-1-yl substitution at the $\text{ D}_{1}$ diversity point (6-position of quinazoline-2,4( $1H,3H$ )-dione skeleton) may yield better activity than other groups. The quinazoline-2,4( $1H,3H$ )-dione scaffold can be effectively replaced by 2 $H$ -benzo[b][1,4]thiazin-3(4 $H$ )-one.     

新疆肉苁蓉多糖清除活性氧自由基

研究了肉苁蓉多糖体外抗氧化活性。以抗坏血酸为对照,以类Fenton反应产生羟自由基,多巴胺自氧化反应产生超氧阴离子自由基,次氯酸钠与过氧化氢反应产生单线态氧、二苯代苦味肼基自由基为实验模型,采用紫外-可见分光光度法和化学发光分析法,首次测定新疆肉苁蓉多糖对超氧阴离子自由基、羟自由基、DPPH.自由基（二苯代苦味肼基自由基）、单线态氧的清除作用。结果表明：新疆肉苁蓉多糖对上述自由基均有明显清除作用,且清除作用与多糖浓度呈正相关性。     

Synthesis and Antitumor,Antioxidant Effects Studies of N-Ethylpiperazine Substitute Thiourea Ligands and Their Copper(II) Complexes

A series of N-ethylpiperazine substitute thioureas [C₆N₂H₁₃NHCSNHR], where R = -C₃H₅ (L ₁ ), -C₁₀H₇ (L ₂ ), and -C₇H₇ (L ₃ ), and their copper (II) complexes have been synthesized. These ligands and complexes have been characterized by elemental analyses, IR, ¹H and ¹³C-NMR spectra, UV-Vis, magnetic susceptibility, thermogravimetrical analyses, and MALDI-TOF MS. In vitro antitumor activity of ligands and their complexes has been screened toward several tumor cell lines. The effects on these complexes of the growth of L1210 and MCF7 were studied comparatively with that of free ligands. Antioxidant and radical scavenging activities of synthesized compounds were determined by various in vitro assays including 1,1-diphenyl-2-picryl-hydrazyl free radicals (DPPH), 2,2′-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid radicals (ABTS+), and ferrous ion (Fe²⁺) chelating activities. Moreover, these activities were compared to synthetic and standard antioxidant trolox. The results showed that the synthesized compounds had effective antioxidant power.     

甘青铁线莲花不同溶剂提取物的抗氧化性比较

采用邻二氮菲-Fe²⁺氧化法,邻苯三酚自氧化法以及卵黄脂蛋白不饱和脂肪酸（PUFA）过氧化体系,对甘青铁线莲花乙醚提取物、甲醇提取物、乙醇提取物和槲皮素的抗氧化活性进行测定和比较。结果表明,甘青铁线莲花不同溶剂提取物对不同的自由基清除作用差异较大。与槲皮素相比较,各提取物对超氧阴离子自由基（·O₂^-）清除的强弱为:槲皮素>乙醚提取物>乙醇提取物>甲醇提取物。对于羟基自由基（·OH）的清除能力乙醚提取物优于其他2种提取物,但低于槲皮素,对抑制卵黄脂蛋白脂质过氧化（LPO）作用的大小依次为乙醇提取物>槲皮素>乙醚提取物>甲醇提取物。因此,不同溶剂的提取物均具有一定的抗氧化性,其中乙醚提取物对几种自由基的清除作用相对较好,三种提取物对脂质过氧化物作用抑制最强,可以作为潜在的天然抗氧化剂开发利用。     

Mechanisms of the Reactions of Radicals with Ozone

The available experimental data (rate constants and activation energies) for the reactions of the hydrogen atom and the R^?, RO^?, RO ₂ ^? , HO^?, and HO ₂ ^? radicals with ozone are analyzed using the method of intersecting parabolas (potential curves). The conclusion is drawn that the primary event in the reactions of H^?, R^?, HO^?, and RO^? with ozone is the addition of the radical to the ozone molecule with the subsequent fast decomposition of the labile polyoxide radical formed. The classical potential barrier for this addition reaction is close to that for the addition of radicals to molecules with multiple bonds. Peroxy radicals react with ozone by the associative decomposition mechanism, RO ₂ ^? + O₃ → RO^? + 2O₂. The ozone molecule reacts with the HO ₂ ^? radical by abstracting its hydrogen atom, O₃ + HO ₂ ^? → HO ₃ ^? + O₂. The experimental data were used to determine the parameters required to calculate the activation energies for the reactions under study.     

Spectroscopic Study of 2-(2-pyridyliminomethyl)phenol as a Novel Fluorescent Probe for Superoxide Anion Radicals and Superoxide Dismutase Activity

A novel spectrofluorometric method, using 2-(2-pyridyliminomethyl)phenol as a fluorescent probe, was developed for the determination of superoxide anion radical (O₂ ^•−) and superoxide dismutase activity (SOD). The new fluorescent probe was synthesized and characterized with elemental analysis and IR spectra. It was oxidized by O₂ ^•− to form a less fluorescence product. Based on this reaction, a spectrofluorometric method was proposed and successfully used to determine superoxide anion radicals and SOD activity. The effects of interferences were studied. The reaction was simple, precise and sensitive. It was applied to determine SOD activity in garlic, papaya and spinach successfully.     

电子顺磁共振法(EPR)对银杏叶提取物及其卷烟捕获自由基能力研究

自由基存在于人体和卷烟烟气中,会引起人体病理反应和氧化损伤,种类和形态多样,如：O-·₂,HO·,R·,RO·,ROO·。电子顺磁共振法(EPR)是检测和定量分析自由基的重要方法之一。银杏叶提取物(EGB)是由黄酮、内酯、原花青素类成分组成的植物药,具有较好清除自由基和抗氧化能力。本文以槲皮素、芦丁、原花青素等化合物为对照,运用EPR法结合自旋捕集技术研究EGB和EGB卷烟对DPPH,HO·,O2-·、烟气气相自由基、烟气粒相自由基的抑制水平。结果表明,EGB能够有效抑制化学/生物系统和卷烟烟气产生的自由基。尽管抑制自由基整体水平比黄酮单体和原花青素弱,但EGB提取方法简单容易得到,可作为天然抗氧化剂和自由基清除剂用于食品和烟草等行业。EPR法灵敏度高,稳定性好,是用于自由基研究的可靠方法。捕集剂DEPMPO捕集O2-·更加高效,PBN捕集烟气气相自由基比DMPO更有优势。该研究讨论了EGB清除自由基的机理,丰富的酚羟基结构是其抗氧化作用的主要原因。     

臭牡丹黄酮类化合物提取及其抗氧化作用

研究臭牡丹中黄酮类化合物的最佳提取工艺条件及其抗氧化活性.以提取时间、料液比、提取温度、乙醇体积分数为主要影响因素,以黄酮类化合物提取率为考察指标,确定最佳提取工艺条件,并通过对亚硝酸盐、超氧阴离子自由基、羟自由基的清除效果及对猪油的抗氧化研究其抗氧化活性.结果表明,臭牡丹中黄酮类化合物的最佳提取工艺条件为提取时间2.0h、料液比1∶40（g/mL）、提取温度70℃、乙醇体积分数70％的条件下提取效果最好,臭牡丹中黄酮类化合物在各抗氧化体系中均表现出较强的抗氧化活性,且其作用具有剂量效应关系,其抗氧化活性均强于维生素C.     

The interaction between hydrogen peroxide and metal oxides: EPR investigations

The interaction of hydrogen peroxide with various metal oxides has been followed by EPR in the aim of investigating the paramagnetic species formed along the complex processes of H₂O₂ decomposition and interaction with the oxide. Data are reported about three systems (MgO, ZrO₂ and Me²⁺/Y zeolite) which are representative of three classes of oxides (insulators, semiconductors and aluminosilicate molecular sieves). Several radical species have been put into evidence in the various cases. The superoxide O ₂ ^? ion has been observed in all cases while in the case of MgO also the OH radical and the O^? radical ion have been identified. In the case of Zn²⁺/Y zeolite, beside the superoxide radical anion, a new signal is observed which shows up upon treating the sample at 640 K and which is preliminarily assigned to a species in the triplet state in the zeolitic cavity.     

The role of guaiacyl moiety in free radical scavenging by 3,5-dihydroxy-4-methoxybenzyl alcohol: thermodynamics of 3H+/3e− mechanisms

ABSTRACT

Participation of guaiacyl moiety of 3,5-dihydroxy-4-methoxybenzyl alcohol (DHMBA) in inactivation of free radicals was investigated using the DFT method. The thermodynamics of triple (3H⁺/3e^?) free radical scavenging mechanisms was investigated. The Gibbs free energies of reactions of inactivation of selected 10 free radicals indicate DHMBA as a potent scavenger. Obtained results allow us to suggest that the contribution of guaiacyl moiety to antioxidant activity of phenolic compounds should be taken into account, what has been scarcely considered until now.     

大叶白麻根中化学成分的预实验及其抗氧化能力探究

为给大叶白麻的合理利用提供理论依据,对大叶白麻根中可能的化学成分进行了预实验。初步探索证明,大叶白麻根中含有酚类、糖、多糖、苷类、有机酸、内酯类,香豆素、甾醇、三萜等化学成分。还对大叶白麻的根提取物运用AB-8型大空树脂进行了纯化富集,并对其中两个主要馏分采用DPPH.和O-2.自由基体系和亚硝酸盐体系进行了抗氧化性能研究。结果表明这两个馏分对DPPH.和亚硝酸盐具有较好的清除作用,且清除作用随着二者的用量的增加而增强。另外,这两个馏分对O-2.自由基体系也有较好的清除作用,且清除作用略强于维生素C。     

High-frequency EPR studies on cofactor radicals in photosystem I

Electron paramagnetic resonance (EPR) spectroscopy at W-band (94 GHz) is used to resolve theg-tensors of the radical ions of the primary chlorophyll donor P ₇₀₀ ^+? and the quinone acceptor A ₁ ^?? in photosystem I. The obtainedg-tensor principal values are compared with those of the isolated pigment radicals in organic solvents and the shifts are related to an impact of the protein environment. P ₇₀₀ ^+? has been investigated in photosystem I single crystals at 94 GHz. W-band EPR applied to the photoinduced radical pair P ₇₀₀ ^+? A ₁ ^?? is used to correctly assign theg-tensor axes of P ₇₀₀ ^+? to the molecular structure of the primary donor. Density functional theory calculations on a model of A ₁ ^?? in its binding pocket derived from the recent crystal structure of photosystem I were utilized to correlate experimental magnetic resonance parameters with structural elements of the protein.     

Free-radical scavenging and mechanism study of flavonoids extracted from the radix ofScutellaria baicalensis Georgi

Free-radical scavenging activities of baicalein, baicalin, wogonin and wogonoside, the four major flavonoids in a traditional Chinese herb medicine, the radix ofScutellaria baicalensis Georgi, were examined by electron paramagnetic resonance (EPR). The results showed that baicalein and baicalin scavenged hydroxyl radical, superoxide anion, 2,2-diphenyl-1-picrylhydrazyl radical and alkyl radical in a dose-dependent manner, while wogonin and wogonoside showed subtle or no effect on these radicals. Baicalein was the most effective free-radical scavenger among the four tested compounds. When 10 mmol/l tested flavonoids dissolved in alkaline solution, only baicalein and baicalin form stable free radicals which can be detected by EPR technique, it was found that the signal came from theiro-di-hydroxyl structure in ring A. To our knowledge, it was the first time demonstrated that flavonoids witho-di-hydroxyl structure in ring A could also form stable semiquinone free radicals. These results demonstrated that flavonoids in radix ofScutellaria baicalensis witho-di-hydroxyl group in ring A such as baicalein and baicalin are good free-radical scavengers and might contribute to some of their pharmaceutical effects.     

Structural analysis of three-spin systems of photosystem II by PELDOR

The pulsed electron electron double resonance (PELDOR) pulse sequence is applied to a three-spin system consisting of three radicals (Y _D ^· , Y _Z ^· and Q _A ^? ) generated in spinach PS II. The distance between Y_Z and Q_A has been determined to be 3.4 nm with the previously derived distances of the other radical pairs, 2.9 nm for Y _D ^· -Y _Z ^· and 3.9 nm for Y _D ^· -Q _A ^? . This distance has been derived from the Y _Z ^· -Q _A ^? radical pair trapped in Y_D-less mutants ofChlamydomonas reinhardtii. Furthermore the method was applied to the Y _D ^· -Q _A ^? -Chl _Z ⁺ system to find the unknown distance between Q_A and Chl_Z. The derived distance was 3.4 nm. A triangular configuration was found in the membrane system that gives the relative positions of the electron transfer components.     

掌裂草葡萄提取物的抗氧化性

通过测定掌裂草葡萄提取物对羟自由基(.OH)的清除率、对超氧阴离子(O2-.)的抑制率,研究其抗氧化活性。结果显示:提取物对羟自由基(·OH)、超氧阴离子(O2-·)有较强的清除抑制作用,并随着提取物添加量的增加,抗氧化作用以及对羟自由基及超氧阴离子的清除抑制作用也随之增强。     

Abilities of Antioxidants to Eliminate the Peroxyl Radical Derived from 2,2′-Azobis(2-Amidinopropane) Dihydrochloride (AAPH)

Electron spin resonance (ESR) of 5-(2,2-dimethyl-1,3-propoxy cyclophosphoryl)-5-methyl-1-pyrroline N-oxide (CYPMPO) spin adducts of free radicals derived from the UV irradiation of 2,2′-azobis(2-amidinopropane) dihydrochloride (AAPH) under rapid scanning condition was examined. The ESR signal obtained was the superposition of two spectra, the spin adduct of the alkoxyl radical [CYPMPO-ORa, Ra = C(CH₃)₂-C(⁺NH₂Cl^?)NH₂] and the peroxyl radical (CYPMPO-OORa). The decay rate of CYPMPO-OORa was 8,400 times that of CYPMPO-ORa. The order of the k _SB/k _ST values (the slope of the Stern–Volmer’s plot) for the peroxyl radical (RaOO·) was l-ascorbic acid > caffeic acid > rutin ~ Trolox ~ (+)-catechin ~ glutathione (reduced), which was almost the same order as that for the alkoxyl radical (RaO·). Though the k _SB/k _ST value of each antioxidant for the peroxyl radical was about half of that for the alkoxyl radical, the ratios of the values of antioxidants to that of trolox (the relative ORAC values) were almost the same between the peroxyl and alkoxyl radicals. The relative oxygen radical absorbance capacity (ORAC) values of polyphenols estimated by the ORAC-ESR assay using both peroxyl and alkoxyl radicals were smaller than those by the ORAC-FL assay.     

Pulse ENDOR studies on the radical pair P 700 +· A 1 ·− and the photoaccumulated quinone acceptor A 1 ·− of photosystem I

The secondary acceptor A₁ of the electron transport chain(s) of photosystem (PS) I is a phylloquinone (vitamin K₁, VK₁). Pulse electron paramagnetic resonance and electron nuclear double resonance (ENDOR) experiments at X-band frequencies were performed on the photoaccumulated acceptor radical A ₁ ^·? and the radical pair state P ₇₀₀ ^·+ A ₁ ^·? in PS I ofThermosynechococcus elongatus. The data obtained were compared with data from the respective radical anion of VK₁ in organic solvents. The unusualg tensor magnitude of A ₁ ^·? is explained by the hydrophobic binding pocket of this radical. The hyperfine couplings and the spin (and charge) density distribution is very different for A ₁ ^.? in PS I and VK ₁ ^·? in frozen alcoholic solution. This is attributed to a rather strong one-sided hydrogen bond to A ₁ ^·? . The presence of a hydrogen bond to A ₁ ^·? has only a minor effect ong. The hyperfine coupling constants of A ₁ ^·? determined from the radical pair spectra deviate only slightly from those derived from photoaccumulated A ₁ ^·? in PS I treated with dithionite at high pH. ENDOR resonances of the proton in a H bond were detected and an estimate of the strength and geometry of this bond to A ₁ ^·? was obtained. The significance of the hydrogen bond and other (hydrophobic) interactions of A₁ with the surrounding are briefly discussed.     

Electron spin polarization in consecutive spin-correlated radical Pairs: Application to short-lived and long-lived precursors in type 1 photosynthetic reaction centres

An analytical treatment of the spin dynamics in sequential photoinduced correlated coupled radical pairs is presented and applied to the spectra of the states P⁺A ₁ ^? and P⁺F _x ^? in type 1 photo-synthetic reaction centres. Expressions for the spin polarized spectra are derived for the specific limiting cases of a very short-lived and very long-lived primary radical pair which correspond to the situation found in heliobacteria and photosystem I (PSI), respectively. The inhomogeneous line-broadening due to the unresolved hyperfine couplings is taken explicitly into account. It is shown that the density matrix of the secondary pair ρ₂ can be written as the sum of two terms corresponding to (i) the part which is independent of the spin dynamics in the precursor, (ii) the additional spin polarization which is generated during the lifetime of the precursor and transferred to the secondary pair. The latter term contains two contributions which arise from the difference of the Zeeman interactions of the radicals in the primary pair and from the inhomogeneous line broadening. The predicted polarization patterns are compared to those established for chemically induced dynamic electron polarization (CIDEP) when uncoupled radicals are generated from a radical pair precursor. The expressions are then used to simulate the experimental spectra of the consecutive pairs P⁺A ₁ ^? and P⁺F _x ^? in PSI using parameters derived entirely from independent experimental data. Excellent agreement with the experimental results is obtained. The spectra of P⁺F _x ^? in heliobacteria at X- and K-band are also simulated and it is shown that the observed polarization patterns can be reproduced assuming direct electron transfer from A₀ to F_x with a time constant ofτ = 600 ps.     

Proton ENDOR of dimethylnitramine free radicals

Electron nuclear double resonance (ENDOR) spectra of free radicals produced by ultraviolet (UV) photolysis of polycrystalline and single crystal dimethylnitramine (or N-methyl-N-nitromethanamine) [DMN; (CH₃)₂NNO₂ were studied atca. ?30°C. Results suggest that multiple radical species are formed during UV photolysis of DMN, perdeutero-DMN-d₆, and¹⁵N-labeled DMN. Proton ENDOR spectra are consistent with assignment of a cation radical (CH₃)₂NNO ₂ ⁺ as the major DMN radical species. Proton hyperfine coupling anisotropy, which is observed from the ENDOR spectra, is attributed to inequivalence of the two DMN methyl groups.     

Electronic properties of charged fullerenes characterized by EPR and Vis-NIR spectroscopy

EPR and Vis-NIR absorption spectra have both been measured for clarification of contradictory statements about the para-and diamagnetic states of fullerenes. Thereby identification of one sharp EPR signal in solution at room temperature to C ₆₀ ^? (g=2.000±4.0001; ΔB=0.07±0.01 mT) could be made upon using different fullerene sources (TechnoCarbo, Hoechst) and methods of anion generation (chemically, electrochemically, and photochemically). This fact is also supported by the similar observation for a monosubstituted derivative (g=1.9999; ΔB=0.10 mT), in which a small broadening of this sharp signal is found originating from additional¹H hyperfine interactions. Furthermore theg-values of the radical anions of C₆₀ increase with charge (g(C ₆₀ ^? <g(C ₆₀ ^2? ) < <g(C ₆₀ ^3? ) <g(C ₆₀ ^5? )) indicating largest contributions from spin-orbit coupling for the monoanion. No diamagnetic states for the dianions of [60]- and [70]- fullerenes could be found so far but biradical species with largest zero field splittingsD=2.7 mT (C ₆₀ ^2? ), andD=3.1 mT (C ₇₀ ^2? ), respectively. The cation formation of C₆₀ (g=2.0023-2.0029; ΔB=0.15-0.20 mT) with antimony pentachloride was controlled by mass spectrometry. Stable cations were found only in methylenechloride. In other solvents like toluene addition reactions seem to occur.