《中国药物化学杂志》2007年第17卷第1期～第6期(总75～80期)总目次

研究论文2,2-二甲基-1,3-二氧戊环-4,5-二羧酸衍生物的设计、合成及其神经营养活性………………………任,聂爱华,肖军海,等(1)(1)含哌嗪环的三唑醇类化合物的合成及体外抗真菌活性………………………………………………何秋琴,刘超美,李科,等(1)(8)β-榄香烯醇酯类化合物的合成及抗癌活性研究…………………………………………………………张兴忠,徐莉英,陶淑娟,等(1)(13)5,7-二取代吡唑并[1,5-a]嘧啶类化合物的合成及其抗肿瘤活性……………………………………李冶,刘亚婧,袁孝烨,等(1)(18)卟啉树状大分子化合物的合成及其抗活性氧作用……     

人参酸性多糖体外抑制牙龈卟啉单胞菌粘附红细胞的作用

牙龈卟啉单胞菌Porphyromonas gingivalis为牙周炎的主要致病菌。以往研究表明,人参和菌陈蒿叶中的酸性多糖明显抑制幽门螺杆菌（HP）对宿主细胞的粘附。作者研究了人参和菌陈蒿酸性多糖抗牙龈卟啉单胞菌粘附红细胞的作用。     

丙-谷二肽树状大分子的合成

目的:合成以丙-谷二肽为构筑单元的新型结构多肽树状大分子。方法:以廉价易得的谷氨酸和丙氨酸为起始原料,经端基保护、脱保护、缩合等步骤得到一个有效的合成路线。结果:前三代大分子化合物总收率为42.8%,产物结构通过IR,1H-NMR,13C-NMR,MS等方法确证。结论:制备了丙-谷二肽树状分子的三代化合物。     

乙烯基烷氧乙基次卟啉Ⅳ衍生物的合成及其光敏活性

以原卟啉Ⅳ二甲酯为起始原料，合成并分离得到３－（１－羟乙基）－８－乙烯基次卟啉Ⅳ（Ⅱ１）和３－乙烯基－８－（１－羟乙基）次卟啉Ⅳ（Ⅱ２）二个异构体，两者分别经酰化后与醇（酚）进行烷氧裂解反应合成了８个全新的卟啉衍生物，在Ｄ２Ｏ中对ＮＡＤＰＨ光氧化作用的酶化效应进行实验表明：所有目标化合物均具有较强的光敏活性，明显高于参比药物ＨｐＤ，这类化合物为研究开发光敏剂新药提供了思考。     

双硫仑抗肿瘤作用及其机制研究进展

戒酒药双硫仑已有70多年临床应用的历史,其安全性得到了广泛验证.目前,大量细胞和动物实验研究结果表明,双硫仑可抑制多种人恶性肿瘤细胞增殖、迁移和侵袭,诱导肿瘤细胞死亡.二价铜离子可增强双硫仑的抗肿瘤效应.双硫仑的代谢产物与二价铜离子螯合,从多方面影响肿瘤细胞生物学功能,如使细胞内氧化还原反应失衡,活性氧水平上升,引起内...     

水溶性金属卟啉的合成、表征及其清除有毒活性氧的研究

目的合成并结构表征了4种水溶性金属卟啉配合物［5，10，15，20-四［4-(4′-吡啶-1)丁氧基苯基］金属卟啉溴化盐［锌卟啉(I)、铜卟啉(II)、锰卟啉(III)和钴卟啉(IV)］，作为抗活性氧(O^-₂，H₂O₂，HO·)模拟酶。方法 核黄素-蛋氨酸光照测其清除O^-₂作用，H₂O₂氧化Vit C法测其催化H₂O₂的分解，Fenton-苯甲酸钠荧光法测其对HO·清除作用，小鼠肝匀浆法测其抗脂质过氧化作用。结果1.0×10^-5~1.0×10^-6 mol·L^-1均具有良好的清除O^-₂作用；1.5×10^-6~1.0×10^-6 mol·L^-1均具有分解H₂O₂作用；2.0×10^-8~1.0×10^-8 mol·L^-1均具有清除HO·的功能；1.0×10^-7 mol·L^-1均可使脂质过氧化产物明显减少。结论合成4种水溶性金属卟啉配合物可作为抗多种活性氧(O^-₂，H₂O₂，HO·)模拟酶。     

重组抗人表皮生长因子受体2单克隆抗体的电荷异质体分析

目的  探讨重组抗人表皮生长因子受体2（human epidermal growth factor receptor 2,Her2）单克隆抗体（单抗）的酸碱异质体对其生物活性和亲和力的影响。方法  分别用过氧化氢、肽-N-糖苷酶F、羧肽酶B对抗Her2单抗进行氧化、脱糖、酶切处理。用阳离子交换高效液相色谱法和毛细管等电聚焦电泳分析抗Her2单抗的电荷异质体,使用微量差示扫描荧光法分析抗Her2单抗的热稳定性,用细胞增殖抑制法和表面等离激元共振技术分析抗Her2单抗的亲和力。结果  脱糖处理抗Her2单抗的酸性电荷异质体增加了5.58%,与免疫球蛋白G Fc受体1（high affinity immunoglobulin gamma Fc receptor Ⅰ,FcGR1A）的亲和力明显减弱,亲和力常数为9.032×10^-8 mol/L。氧化和酶切处理抗Her2单抗的热稳定性降低,去折叠温度分别为63.6和59.5 ℃,均低于未处理抗Her2单抗（66.7 ℃）。结论  3种处理均会使抗Her2单抗产生电荷异质性,脱糖处理抗Her2单抗与FcGR1A的亲和力减弱,氧化和酶切处理抗Her2单抗的热稳定性降低。     

四吡咯金属络合物的合成及其对再生障碍性贫血的实验性疗效研究

为了寻找再生障碍性贫血的治疗药物，设计合成了四吡咯化合物３（或８）－（１－乙氧基乙基）－８（或３）－羟乙基次卟啉ＩＸ（１）和３（或８）－（１－乙氧基乙基）－８（或３）－乙烯基次卟啉ＩＸ（２）的锰（Ⅲ）、（Ⅲ）、钴（Ⅲ）、镍（Ⅱ）、铜（Ⅱ）、锌（Ⅱ）金属络合物，在辐射致小鼠ＡＡ模型上（ｌｄ）和（ｌｅ）能明显延长ＡＡ小鼠的平均存活时间，增加２１ｄ后小鼠的存活个数，进一步考察（ｌｅ）对ＡＡ小鼠造血系统的     

CMIA法对HBV感染患者血清学标志物诊断准确率的影响

目的 探究化学发光微粒子免疫分析法（CMIA）在乙型肝炎病毒（HBV）感染患者中的应用,分析其对血清学标志物诊断准确率的影响。方法 选取340例疑似HBV感染患者,以实时荧光定量PCR检测结果为金标准,采用酶联免疫吸附法（ELISA）、CMIA法检测HBV感染血清学标志物[乙型肝炎表面抗体（抗-HBs）、乙型肝炎e抗原（HBeAg）、乙型肝炎表面抗原（HBsAg）、乙型肝炎核心抗体（抗-HBc）和乙型肝炎e抗体（抗-HBe）],观察对比两种检测方法血清学标志物检出率,比较两种检测方法的变异系数（CV）,分析CMIA法对HBV感染诊断准确率的影响。结果 340例疑似HBV感染患者中检出211例乙型肝炎,CMIA法检出220例,ELISA法检出174例,CMIA法检测HBV感染的准确率、灵敏度及阴性预测值高于ELISA法检测（P<0.05）;CMIA法检测HBV感染血清HBsAg、抗-HBs、抗-HBc、抗-HBe、HBeAg的CV值低于ELISA法检测（P<0.05）;CMIA法检测HBV感染血清HBsAg、抗-HBs、抗-HBc、抗-HBe、HBeAg的阳性率与ELISA法...     

2,7,12,18-四甲基-13,17-二(3-羟基丙基)卟啉的合成及其光敏化活性

由氯化高铁次卟啉经脱铁、酯化、还原合成了光敏剂２，７，１２，１８－四甲基－１３，１７－二（３－羟基丙基）卟啉，并测定其光敏活性     

Flavin-dependent antioxidant properties of a new series of meso-N,N'-dialkyl-imidazolium substituted manganese(III) porphyrins

A number of synthetic manganese complexes exhibit both in vitro and in vivo catalytic antioxidant activities. This study reports that the antioxidant potencies of a new series of meso-N,N'-dialkyl-imidazolium substituted manganese(III) porphyrins are dependent, in part, on their ability to redox cycle with endogenous flavin-dependent oxidoreductases. Inhibition of lipid peroxidation activities of these novel cationic porphyrins was compared using rat brain homogenate as a source of lipids and endogenous oxidoreductases. Iron and ascorbate was used as initiators of lipid peroxidation, and two indices of lipid peroxidation (thiobarbituric acid reactive species (TBARS) and F(2)-isoprostanes) were determined. All meso-N,N'-dialkyl-imidazolium substituted porphyrins tested were potent inhibitors of lipid peroxidation with IC(50) ranging from 0.1 to 34 microM with a metal-dependent potency of Mn(III)>Co(III)>Zn(II). A flavin-dependent oxidoreductase antioxidant process was supported by the ability of the diphenyleneiodonium chloride (DPI, a flavoenzyme inhibitor) to decrease the potency of Mn-porphyrins in the lipid peroxidation model and that Mn-porphyrins stimulate NADPH oxidation in rat brain homogenates. These data suggest that metalloporphyrins may have differential antioxidant effects in tissues due to the presence or absence of flavin-dependent oxidoreductases.     

Study of anaesthetic agents for their ability to elicit porphyrin biosynthesis in chick embryo liver

Effects of some anaesthetic drugs on the activity of delta-aminolevulinate synthetase and on the formation of porphyrins and cytochrome P-450 were studied in 18-day-old chick embryo livers in ovo. The drugs were either tested alone or with a small dose of 1,4-dihydro-3,5-dicarbethoxycollidine, which reproduces in the embryo liver a partial block in the heme biosynthesis pathway similar to that found in cells of human patients with porphyrias. Two series of local anaesthetics were tested: procaine and its derivatives (proxymetacaine, oxybuprocaine, butacaine and tetracaine) had no (or very slight) porphyrogenic effects. In contrast, lidocaine and its derivatives (bupivacaine, mepivacaine, etidocaine, pyrrocaine and prilocaine) were found to induce delta-aminolevulinate synthetase and to cause accumulation of porphyrins and cytochrome P-450. Some other drugs used in anaesthesiology were tested: fentanyl, morphine, sodium oxybate, pancuronium, pethidine and phenoperidine were found to be non-porphyrogenic; alcuronium was a slight inducer. It is suggested that the inducing drugs should be avoided in patients with hepatic porphyrias.     

Quality of potent Mn porphyrin-based SOD mimics and peroxynitrite scavengers for pre-clinical mechanistic/therapeutic purposes

Cationic Mn porphyrins are among the most potent SOD mimics and peroxynitrite scavengers. They have been widely and successfully used in different models of oxidative stress and are either progressing towards or are in phase I of clinical trials. The most frequently used compounds are Mn(III) meso-tetrakis(N-ethylpyridinium-2-yl)porphyrin (MnTE-2-PyP(5+) or AEOL10113), its methyl analogue (MnTM-2-PyP(5+) or AEOL10112), and Mn(III) meso-tetrakis(4-benzoic acid)porphyrin (MnTBAP). A great discrepancy between the in vivo data obtained with Calbiochem preparations and those of authentic MnTE-2-PyP(5+) and MnTM-2-PyP(5+) samples were recently observed. Surprisingly, the commercial samples were invariably of poor identity and consisted of mixtures of nearly equal contributions of non-alkylated, mono-, di-, tri- and tetraalkylated porphyrins, lacking thus the major structural entity that determines their antioxidant potency, i.e., the four positively charged orthoN-alkylpyridyl groups that afford thermodynamic tuning of the active site and electrostatic guidance of anionic superoxide and peroxynitrite species toward the metal center. The MnTE-2-PyP(5+) and MnTM-2-PyP(5+) compounds were not even the major species in the commercial samples sold as "MnTE-2-PyP" and "MnTM-2-PyP", respectively. While we have already reported the insufficient impurity of the MnTBAP samples from Alexis and other suppliers, in one more recent lot the situation is dramatic, as 25% of the sample was not MnTBAP, but metal-free ligand, H(2)TBAP. The (unintentional) use of the Mn porphyrins of low quality compromises therapeutic and/or mechanistic conclusions. Simple techniques, which include thin-layer chromatography, electrospray-mass spectrometry, UV-vis spectroscopy, and electrochemistry described here could be used routinely to check the overall quality of Mn porphyrins in order to avoid misleading conclusions and waste of valuable resources (animals, compounds, time, manpower).     

The potential of Zn(II) N-alkylpyridylporphyrins for anticancer therapy

Reactive oxygen species (ROS) are considered to be a main cause for cancer development, but they can also be used for cancer eradication. Because of this dual nature of ROS action, both antioxidant and prooxidant therapeutic agents have been developed and some have shown clinical promise. Selective uptake of porphyrins by malignant cells has for a long time been used for tumor imaging and for targeted delivery of ROS. Redox-active Mn porphyrins can act both as antioxidants and as prooxidants, and may thus be used in anticancer therapy. Porphyrins, which chelate redox inactive metals, for example Zn, demonstrate photo-sensitizing activity and thus can produce singlet oxygen and other reactive oxygen species in cancer cells on irradiation with visible light. Here we review the properties of Zn(II) N-alkylpyridylporphyrin-based photosensitizers, and their ability to damage selected cellular targets.     

Design of the complex between manganese porphyrins and catalase-poly(ethylene glycol) conjugates for a new antioxidant

A new design of antioxidant, the complex between manganese (Mn) porphyrins and catalase-poly(ethylene glycol) (PEG) conjugates, is reported. Gel filtration chromatography and a Langmuir-type adsorption isotherm proved that the catalase-PEG conjugate formed the complex with the Mn-porphyrin. The resulting complex exhibited significant superoxide dismutase (SOD) and catalase activity. These results suggest that the Mn-porphyrin/catalase-PEG complex with dual enzymatic activity, i.e., SOD and catalase, is promising for a new class of antioxidants.     

Gastrointestinal blood loss after non-steroidal anti-inflammatory drugs. Measurement by selective determination of faecal porphyrins.

1. A method for the detection of gastrointestinal blood loss based upon the selective measurement of faecal porphyrins was tested in two studies in healthy volunteers. 2. In the first study subjects (n = 6) received intragastric autologous blood (saline, 2 and 6 ml as a single dose) resulting in a dose dependent increase in faecal porphyrins. 3. In a subsequent placebo controlled cross over study in 12 subjects acetylsalicylic acid (ASA), nabumetone (a new NSAID) or placebo were administered for 5 days with a washout period of 9 days. They were no dietary restrictions. 4. All faeces were collected during the treatment period and both the full faecal homogenate and a random faecal sample were analyzed for deutero- and pemptoporphyrin content by h.p.l.c. Additionally a benzidine reaction was performed. 5. There was a highly significant correlation (r = 0.95) between the values obtained from random samples and the full homogenate. ASA increased the faecal porphyrin excretion (P less than 0.001) compared with placebo in contrast to nabumetone. Complaints of dyspepsia were most common after ASA. 6. Measurement of faecal porphyrins is useful for monitoring NSAID induced upper gastrointestinal blood loss and lacks some of the practical constraints of other methods.     

Nitric oxide, peroxynitrite and lipoxygenase in atherogenesis: mechanistic insights

Nitric oxide (*NO) is a free radical species that diffuses and concentrates in the hydrophobic core of low-density lipoprotein (LDL) to serve as a potent antioxidant. Peroxynitrite, the product of the diffusion-limited reaction between *NO and superoxide anion, as well as lipoxygenase, represent relevant mediators of oxidative modifications in LDL. The focus of this review is the analysis of interactions between *NO, peroxynitrite and lipoxygenase during LDL oxidation, which are relevant in the development of the early steps as well as progression of atherosclerosis. The role of CO2 to redirect peroxynitrite reactivity in LDL, as well as the lipophilic antioxidant sparing actions of *NO, ascorbate and CO2 is also analyzed. In this context, the effects of novel potential pharmacological strategies against atherosclerosis such as Mn(III)porphyrins will be discussed.     

The effect of some flavonoids on non-enzymatic lipid oxidation and enzymatic oxidation of arachidonic acid

Twenty flavonoids isolated from plants or transformed into methyl or acetyl derivatives were tested with regard to their influence on cyclooxygenase from the ram seminal vesicle microsomes and lipoxygenase from soya beans. Moreover, their antioxidant properties were evaluated by estimating the amount of the malonylaldehyde formed from arachidonic acid. Only rhamnetin and myricetin inhibited the soybean lipoxygenase. Most of the tested flavonoids stimulated cyclooxygenase at a high (100 microM) substrate concentration, myricetin being the most potent. Rhamnetin was the strongest antioxidant, while myricetin was about ten times weaker. Structural requirements for the cyclooxygenase stimulation, lipoxygenase inhibition and antioxidant properties were different in the case of the twenty tested flavonoids.     

Effect of antiepileptic drugs on the urinary excretion of porphyrins in non-porphyric subjects

The action of some anticonvulsant drugs as the causal agents of attacks of acute porphyria has been widely documented in the literature. However, little attention has been paid to the effect of these drugs on the urinary excretion of porphyrins in non-porphyric subjects. In a sample of 82 epileptic patients treated with phenobarbital (n = 54), phenytoin (n = 64), carbamazepine (n = 33), and valproate (n = 8), the daily doses were expressed according to a drug score that would reflect the capacity of these drugs as enzymatic inducers when administered in polytherapy. A significantly increased urinary excretion of D-glucaric acid (DGA) and porphyrins was found in this group of patients (P<0.001), with coproporphyrin being the major fraction in all cases (>60%). Urinary DGA had a highly significant correlation with the drug score (r = 0.783, P<0.001); however, no significant correlations were found between the urinary porphyrins and DGA (r = 0.005) or the drug score (r = 0.053). Neither was any significant relationship found between the urinary porphyrins and the serum activity of 5'-nucleotidase (r = 0.066) or the presence of a cholestasis objectivized through the presence of the isoform of gamma-glutamyltransferase with beta-globulins electrophoretic mobility. However, in a group of 10 patients a significant correlation was found between the urinary excretion of porphyrins and beta-N-acetylhexosaminidase (r = 0.790, P<0.01). Therefore, it does not appear that the liver enzyme induction, or even a subclinical cholestasis, produced by the antiepileptic drugs administered to these patients may serve to explain the increase in the urinary excretion of porphyrins. A possible renal origin is proposed for the increase of urinary porphyrins in these cases.     

In vitro effects of trichothecenes on human dendritic cells.

The aim of this work was to study the in vitro effects of trichothecenes on human dendritic cells. Trichothecenes are mycotoxins produced by fungi such as Fusarium, Myrothecium, and Stachybotrys. Two aspects have been explored in this work: the cytotoxicity of trichothecenes on immature dendritic cells to determine IC 50 (inhibition concentration), and the effects of trichothecenes on dendritic cell maturation process. Two mycotoxins (T-2 and DON) known to be immunotoxic have been tested on a model of monocyte-derived dendritic cells culture. Cytotoxic effects of T-2 toxin and DON on immature dendritic cells showed that DON is less potent than T-2 toxin. The exposure to trichothecenes during dendritic cell maturation upon addition of LPS or TNF-alpha markedly inhibited the up-regulation of maturation markers such as CD-86, HLA-DR and CCR7. Features of LPS or TNF-alpha -mediated maturation of dendritic cells, such as IL-10 and IL-12 secretions and endocytosis, were also impaired in response to trichothecenes treatment. These results suggest trichothecenes have adverse effects on dendritic cells and dendritic cell maturation process.