Pelitinib的合成

2-氨基-5-硝基苯酚经乙酰化、醚化及还原反应得3-乙氧基-4-乙酰胺基苯胺,与2-氰基-3-乙氧基丙烯酸乙酯反应后闭环,经三氯氧磷氯代后与3-氯-4-氟苯胺反应得3-氰基-4-(3-氯-4-氟苯胺基)-6-乙酰胺基-7-乙氧基喹啉,去乙酰保护基后与4-N,N-二甲胺基巴豆酰氯经酰化反应制得抗肿瘤药pelitinib,总收率约5％.     

琥珀酸拉米地坦的合成工艺改进

本研究对琥珀酸拉米地坦(1)的合成工艺进行改进.氯化亚砜作为溶剂和试剂,与N-甲基-4-哌啶甲酸盐酸盐(2)反应后,再与二甲胺反应生成N,N,N'-三甲基哌啶-4-甲酰胺(4);4与2,6-二溴吡啶经格氏反应制备2-溴-6-(1-甲基哌啶-4-基羰基)吡啶(5);5无需纯化在氯化铵/氢氧化钠体系中反应得到2-氨基-6-...     

4-(3-氯-4-氟苯胺基)-7-甲氧基-6-[3-(4-吗啉基)丙氧基]喹唑啉的合成

目的设计合成4-(3-氯-4-氟苯胺基)-7-甲氧基-6-[3-(4-吗啉基)丙氧基]喹唑啉(ZD1839).方法以3-羟基-4-甲氧基苯甲酸甲酯作为起始原料,经7步反应得到目标化合物.结果与结论目标化合物的结构经IR、1H-NMR和MS确证.本合成反应条件温和,原料易得,操作简便,反应步骤比文献方法(9步)减少两步,反应总收率为34.81%.     

吲哚酮的Mannich反应及其羟甲基化副产物的结构确证

在考察吗吲酮合成路线中Mannich反应的反应条件时，采用改变反应溶剂的方法，希望通过提高反应温度来提高收率。结果发现未进行Mannich反应，而进行羟甲基化副反应；产物经IR，^1H-NMR,13^C-NMR,MS确证为N-羟甲基-3-乙基-1，5，6，7-四氢-2-甲基-4H-吲哚-4-酮。因此3-乙基-1，5，6，7-四氢-2-甲基-4H-吲哚-4-酮（I）与多聚甲醛、吗啉盐酸盐进行Mannich反应时，不能采用冰乙酸作溶剂。     

厄贝沙坦的合成

环戊酮与氰化钠反应制得1-氨基环戊腈，成盐、水解成酰胺后与戊酰氯反应制得2-丁基-1，3-二氮杂螺[4-4]壬-1-烯-4-酮，相转移催化条件下与4’-溴甲基-2-氰基联苯反应制得2-丁基-3-[（2’-氰基联苯-4-基）甲基]-1，3-二氮杂螺[4-4]壬-1-烯-4-酮，最后与叠氮化钠反应制得厄贝沙坦，总收率约39％。     

盐酸雷洛昔芬的合成改进

目的:优化盐酸雷洛昔芬的合成路线.方法:以3-甲氧基苯硫酚和4-甲氧基-α-溴代苯乙酮为起始原料,经取代反应,环合反应得到6-甲氧基-2-(4-乙酰氧基苯基)苯并[b]噻吩,再与4-[2-(1-哌啶基)乙氧基]苯甲酰氯盐酸盐发生Friedel-Crafts反应,然后发生脱甲基反应,最后经成盐反应,共5步主要反应制得目标产物.结果:目标化合物结构经红外光谱、核磁共振氢谱及质谱确证.结论:本方法反应条件温和,操作简便,并且提高了产率.     

盐酸洛哌丁胺的合成

苯乙腈经溴化、傅-克反应得到二苯基乙腈(2)。2经与环氧乙烷反应,以36%溴化氢冰醋酸溶液开环得4-溴-2,2-二苯基丁酸(4)。后者的酰氯与二甲胺反应,续与4-对氯苯基-4-羟基哌啶(7)缩合以及成盐,得盐酸洛哌丁胺。收率58%(以7计)。     

(2R,3S)-2-[(R)-1-[3,5-二(三氟甲基)苯基]乙氧基]-3-(4-氟苯基)吗啉盐酸盐的合成

对甲氧基苯甲醛(3)和2-氨基乙醇进行还原胺化反应得2-(4-甲氧基苄胺基)乙醇(4),4和乙醛酸经成环反应得2-羟基-4-对甲氧基苄基吗啉-3-酮(5),5和三氟乙酐反应得6后与(R)-1-[3,5-二(三氟甲基)苯基]乙醇(7)缩合,再经结晶诱导不对称转化、格氏反应、氢化脱保护及成盐反应制得阿瑞吡坦关键中间体(2R,3S)-2-[(R)-1-[3,5-二(三氟甲基)苯基]乙氧基]-3-(4-氟苯基)吗啉盐酸盐,总收率约18%(以3计)。     

2-(3-氰基-4-羟基)苯基-4-甲基-5-噻唑甲酸乙酯的合成

目的优化非布司他关键中间体2-(3-氰基-4-羟基)苯基-4-甲基-5-噻唑甲酸乙酯(4)的合成方法。方法采用"一勺烩"方法,以4-羟基苯甲腈为起始原料,首先与硫氢化钠和无水氯化镁在N,N-二甲基甲酰胺中反应,所得中间体不经分离,直接加入2-氯乙酰乙酸乙酯进行环合反应,得到2-(4-羟基)苯基-4-甲基-5-噻唑甲酸乙酯(2);然后通过六亚甲基四胺/三氟乙酸进行Duff反应,得到2-(3-甲酰基-4-羟基)苯基-4-甲基-5-噻唑甲酸乙酯(3);再经盐酸羟胺/甲酸/甲酸钠体系脱水得到目标化合物。结果经四步反应合成非布司他关键中间体4,总收率为22.6%,其结构经核磁共振氢谱、质谱确证。结论改进后的工艺终产品无需柱色谱纯化,适合工业化生产。     

新型抗球虫药 ponazuril 的合成工艺改进

目的合成新型抗球虫药ponazuril。方法以4-三氟甲巯基苯酚和2-甲基-4-硝基氯苯为起始原料,与水合肼、固体光气、甲胺气反应制备中间体1-甲基-3-[3-甲基-4-(4-三氟甲巯基苯氧基)苯基]脲(7),再经与固体光气、氨气反应制备中间体3-甲基-5-[3-甲基-4-(4-三氟甲巯基苯氧基)苯基]缩二脲(8),最后经环合、氧化得到目标化合物。结果与结论经过7步反应制得目标化合物,总收率为50.7%,其结构经1H-NMR、MS确证,该法反应条件温和,操作方便,有利于工业化生产。     

Potential of neurotoxicity after a single oral dose of 4-bromo-, 4-chloro-, 4-fluoro- or 4-iodoaniline in rats

The potential for neurotoxicity after a single oral dose of four halogenated aniline derivatives--4-bromoaniline (4-BA), 4-chloroaniline (4-CA), 4- fluoroaniline (4-FA) and 4-iodoaniline (4-IA)--was given to rats was investigated at or near the lethal dosage level. Hindlimb paralysis was found in the 4-BA, 4-CA and 4-FA groups on clinical observation, with the maximum incidence of 100% in the 4-BA and 4-FA groups and 66.7% in the 4-CA group. Detailed clinical observations with functional tests identified the following effects: reduced response of hindlimb extensor thrust, gait abnormality in the open field and decreased grip strength in the fore- or hindlimbs in the 4-BA, 4-CA and 4-FA groups; decreased number of supported rearing episodes in the open field in the 4-BA and 4-CA groups; abnormal landing in the aerial righting reflex in the 4-BA and 4-FA groups; and prolonged surface righting reflex in the 4-BA group. Spongy change in the white matter of the spinal cord and brainstem and nerve fibre degeneration in the peripheral nerves were found in all haloaniline-treated groups. The central and peripheral nervous systems were most severely affected in the 4-BA group and the lesions in the 4-IA group were limited in grade. This study demonstrates that a bolus dose of 4-haloanilines to rats induces a neurotoxicity similar in character to that evoked by the parent aniline. The decreasing order of neurotoxic potential appears to be 4-BA > 4-FA > or = 4-CA > 4-IA when comparing at or near the lethal dosage level.     

SALL4 基因在肝内胆管癌中通过抑制KLF4 基因表达促进EMT 改变

目的探究SALL4基因与KLF基因在肝内胆管癌中的相互作用与影响。方法采用生物信息学方法预测SALL4与KLF4基因启动子之间的作用位点,并通过荧光素酶报告系统进行验证;RNA干扰技术对SALL4基因进行敲除,Western blotting检测KLF4蛋白的表达;免疫组化法检测临床样本中SALL4与KLF4蛋白的表达,并结合TCGA数据库进行验证。结果KLF4启动子上存在SALL4结合的模序,且二者的表达呈负相关。结论在肝内胆管癌中,SALL4能够结合KLF4启动子区域,抑制KLF4基因的表达。在肝内胆管癌细胞株中敲低SALL4基因,可使E-cadherin表达显著提高,提示SALL4能够促进上皮间质转化(EMT)的改变。     

Fenretinide metabolism in humans and mice: utilizing pharmacological modulation of its metabolic pathway to increase systemic exposure

BACKGROUND AND PURPOSE

High plasma levels of fenretinide [N-(4-hydroxyphenyl)retinamide (4-HPR)] were associated with improved outcome in a phase II clinical trial. Low bioavailability of 4-HPR has been limiting its therapeutic applications. This study characterized metabolism of 4-HPR in humans and mice, and to explore the effects of ketoconazole, an inhibitor of CYP3A4, as a modulator to increase 4-HPR plasma levels in mice and to increase the low bioavailability of 4-HPR.

EXPERIMENTAL APPROACH

4-HPR metabolites were identified by mass spectrometric analysis and levels of 4-HPR and its metabolites [N-(4-methoxyphenyl)retinamide (4-MPR) and 4-oxo-N-(4-hydroxyphenyl)retinamide (4-oxo-4-HPR)] were quantified by high-performance liquid chromatography (HPLC). Kinetic analysis of enzyme activities and the effects of enzyme inhibitors were performed in pooled human and pooled mouse liver microsomes, and in human cytochrome P450 (CYP) 3A4 isoenzyme microsomes. In vivo metabolism of 4-HPR was inhibited in mice.

KEY RESULTS

Six 4-HPR metabolites were identified in the plasma of patients and mice. 4-HPR was oxidized to 4-oxo-4-HPR, at least in part via human CYP3A4. The CYP3A4 inhibitor ketoconazole significantly reduced 4-oxo-4-HPR formation in both human and mouse liver microsomes. In two strains of mice, co-administration of ketoconazole with 4-HPR in vivo significantly increased 4-HPR plasma concentrations by > twofold over 4-HPR alone and also increased 4-oxo-4-HPR levels.

CONCLUSIONS AND IMPLICATIONS

Mice may serve as an in vivo model of human 4-HPR pharmacokinetics. In vivo data suggest that the co-administration of ketoconazole at normal clinical doses with 4-HPR may increase systemic exposure to 4-HPR in humans.     

Synthetic approaches to peptide analogues containing 4, 4-difluoro-L-proline and 4-keto-L-proline

An improved synthesis of 4, 4-difluoro-L-proline is described. A key step in this synthesis involves the fluorination of Z-4-keto-L-proline benzyl ester, using diethylaminosulfur trifluoride (DAST), to give the corresponding Z-4, 4-difluoro-L-proline benzyl ester. Preparation of dipeptide derivatives containing 4, 4-difluoro-L-proline has been accomplished by initial synthesis of the corresponding 4-keto-L-proline dipeptide derivatives, which were then fluorinated with DAST. A one-step synthesis of Boc-4-keto-L-proline from Boc-4-hydroxy-L-proline by chromium trioxide oxidation is reported. These synthetic procedures should facilitate the preparation of a variety of peptide analogues containing 4, 4-difluoro-L-proline and 4-keto-L-proline.     

Transfection of HepG2 cells with hGSTA4 provides protection against 4-hydroxynonenal-mediated oxidative injury

4-Hydroxynonenal (4-HNE) is a mutagenic ,β-unsaturated aldehyde produced during oxidative injury that is conjugated by several glutathione S-transferase (GST) isoforms. The alpha class human GSTA4-4 enzyme (hGSTA4-4) has a particularly high catalytic efficiency toward 4-HNE conjugation. However, hGST4-4 expression is low in most human cells and there are other aldehyde metabolizing enzymes that detoxify 4-HNE. In the current study, we determined the effect of over-expression of hGSTA4 mRNA on the sensitivity of HepG2 cells to 4-HNE injury. HepG2 cells transfected with an hGSTA4 vector construct exhibited high steady-state hGSTA4 mRNA, high GST–4-HNE catalytic activities, but lower basal glutathione (GSH) concentrations relative to insert-free vector (control) cells. Exposure to 4-HNE elicited an increase in GSH concentrations in the control and hGSTA4 cells, although the dose-response of GSH induction differed among the two cell types. Specifically, hGSTA4 cells had significantly higher GSH concentrations when exposed to 5–15 μM 4-HNE, but not at 20 μM 4-HNE, suggesting extensive GSH utilization at high concentrations of 4-HNE. The hGSTA4 cells exhibited a significant growth advantage relative to control cells in the absence of 4-HNE, and a trend towards increased growth at low dose exposures to 4-HNE. However, the hGSTA4 cells did not exhibit a growth advantage relative to control cells at higher 4-HNE exposures associated with increased GSH utilization. As expected, the hGSTA4 cells showed resistance to 4-HNE stimulated lipid peroxidation at all 4-HNE doses. In summary, our data indicates that over-expression of hGSTA4 at levels conferring high GST–4-HNE conjugating activity confers a partial growth advantage to HepG2 cells and protects against 4-HNE oxidative injury. However, the loss of proliferative capacity of hGSTA4 cells challenged with levels of 4-HNE associated with severe oxidative stress indicates a role of other aldehyde metabolizing enzymes, and/or GSH–electrophile transporter proteins, in providing full cellular protection against 4-HNE toxicity.     

An optimized derivative of an endogenous CXCR4 antagonist prevents atopic dermatitis and airway inflammation

Aberrant CXCR4/CXCL12 signaling is involved in many pathophysiological processes such as cancer and inflammatory diseases. A natural fragment of serum albumin, named EPI-X4, has previously been identified as endogenous peptide antagonist and inverse agonist of CXCR4 and is a promising compound for the development of improved analogues for the therapy of CXCR4-associated diseases. To generate optimized EPI-X4 derivatives we here performed molecular docking analysis to identify key interaction motifs of EPI-X4/CXCR4. Subsequent rational drug design allowed to increase the anti-CXCR4 activity of EPI-X4. The EPI-X4 derivative JM#21 bound CXCR4 and suppressed CXCR4-tropic HIV-1 infection more efficiently than the clinically approved small molecule CXCR4 antagonist AMD3100. EPI-X4 JM#21 did not exert toxic effects in zebrafish embryos and suppressed allergen-induced infiltration of eosinophils and other immune cells into the airways of animals in an asthma mouse model. Moreover, topical administration of the optimized EPI-X4 derivative efficiently prevented inflammation of the skin in a mouse model of atopic dermatitis. Thus, rationally designed EPI-X4 JM#21 is a novel potent antagonist of CXCR4 and the first CXCR4 inhibitor with therapeutic efficacy in atopic dermatitis. Further clinical development of this new class of CXCR4 antagonists for the therapy of atopic dermatitis, asthma and other CXCR4-associated diseases is highly warranted.     

Pharmacological profile of leukotrienes E4, N-acetyl E4 and of four of their novel omega- and beta-oxidative metabolites in airways of guinea-pig and man in vitro.

1, The biological effects of metabolites of leukotriene E4 (LTE4) i.e. N-acetyl LTE4 (N-AcLTE4), 20-COOH-LTE4, 20-COOH-N-AcLTE4, as well as 18-COOH-19,20-dinor-LTE4 (dinor-LTE4) and 16-COOH-17,18,19,20-tetranor-14,15-dihydro-LTE4 (tetranor-LTE4) were investigated on superfused strips of guinea-pig trachea (GPT) and lung parenchyma (GPP) in vitro. 2. The actions of LTE4 were studied in isolated, superfused strips of human lung parenchyma (HP) and bronchus (HBr), in comparison with LTD4 and histamine. Effects of N-AcLTE4, the 20-carboxy metabolites, dinor-LTE4 and tetranor-LTE4 were also investigated in HBr. 3. N-AcLTE4 (0.1-10 nmol) induced dose-related contractions of GPT and was approximately 100 times less active than LTD4 (3-100 pmol). 4. In GPP, N-AcLTE4 (0.01-3 nmol) was equiactive with LTE4 (0.01-1 nmol) and approximately one order of magnitude less active than LTD4 (1-300 pmol). Contractions caused by N-AcLTE4 and LTE4 were very similar and approximately twice as sustained as those due to LTD4. 5. LTE4 (0.1-30 nmol) contracted strips of HP and HBr and was about 2-3 orders of magnitude less active than LTD4. As in GPP, the effect of LTE4 was more protracted than that of LTD4. Actions of N-AcLTE4 were similar to those of LTE4 in HBr. 6. 20-carboxy-LTE4, 20-carboxy-N-AcLTE4, dinor-LTE4 and tetranor-LTE4, all at 0.3-30 nmol, were inactive in GPT, GPP and HBr.(ABSTRACT TRUNCATED AT 250 WORDS)     

抗髓过氧化物酶抗体阳性韦格纳肉芽肿病肺损害的临床分析

目的提高对抗髓过氧化物酶（MPO）抗体阳性的韦格纳肉芽肿（WG）病患者的肺损害的认识。方法自2002年3月至2008年1月,对确诊的8例有肺损害的韦格纳肉芽肿病患者的临床表现、实验室检查、影像学特点进行回顾性分析并结合文献进行复习。结果8例患者中,男4例、女4例,平均49.83岁。抗MPO抗体阳性4例,抗蛋白酶3（PR3）抗体阳性4例。抗MPO抗体阳性的WG患者,临床表现为发热4例,咳嗽、咳痰4例、咯血2例、呼吸困难1例、哮喘样发作1例;4例均累及上呼吸道和肾脏;胸部CT显示双肺单发或多发结节影2例、肿块影1例,双肺弥漫斑片影或磨玻璃影4例,肺实变影1例,结节性空洞1例及支气管扩张2例。误诊为肺癌、肺结核、军团菌肺炎、真菌性肺炎者各1例。结论在国人的WG患者中,抗MPO抗体阳性可能并不少见。对临床表现为发热、肺部阴影伴抗MPO抗体阳性的患者应考虑WG的可能。     

Comparative toxicity of 4-chlorobiphenyl and its metabolite 4-chloro-4'-biphenylol in isolated rat liver mitochondria

4-Chlorobiphenyl (4-CB) is converted by the microsomal cytochrome P-450 system to its hydroxylated metabolite 4-chloro-4'-biphenylol (4'-OH-4-CB). A study of the effects of 4-CB and 4'-OH-4-CB on the energy-linked functions of rat liver mitochondria was carried out. 4'-OH-4-CB was more effective than 4-CB in causing the inhibition of state 3 respiration of mitochondria with both succinate and glutamate/malate. As a substrate specificity, with glutamate/malate the inhibition by each compound (ID50, 30 microM for 4'-OH-4-CB, 76 microM for 4.CB) was more significant than that with succinate (ID50, 200 microM for 4'-OH-4-CB, never reached 50% for 4-CB). From the effects on DNP-stimulated respiration, it was indicated that the electron transport from both glutamate/malate and succinate to oxygen was more sensitively inhibited by 4'-OH-4-CB than by 4-CB, with the same substrate specificity as for state 3 respiration (i.e. the inhibition by both compounds was greater with glutamate/malate than with succinate). Since there existed a good coincidence in the inhibition between state 3 and DNP-stimulated respiration with both substrates, the inhibition of state 3 respiration by both compounds was due to the inhibition of the electron transport. With succinate, the uncoupling of oxidative phosphorylation by both compounds was observed, the extent of which was greater with 4'-OH-4-CB than with 4-CB, although the uncoupling by higher concentrations of 4'-OH-4-CB was masked because of the increased inhibition in respiration. With glutamate/malate, the uncoupling action of 4-CB was largely, while that of 4'-OH-4-CB was completely, masked by progressive respiratory inhibition. 4'-OH-4-CB was more effective than 4-CB in causing stimulation of latent ATPase in mitochondria. These results indicate that both 4-CB and 4'-OH-4-CB impair mitochondrial energy-transducing functions, but 4'-OH-4-CB is more effective than 4-CB in damaging these functions. Thus, the product of the metabolism is more biologically active than the parent compound. The impairment of energy-linked mitochondrial reactions by the metabolite as well as of the parent compound may be an important factor in the toxicity of 4-CB.     

4-酰硫基-4-脱氧-4-去甲表鬼臼毒素衍生物的合成和抗肿瘤活性

为研究4’-去甲表鬼臼毒素的C₄位不同原子及不同取代基类型同活性之间的关系,寻找活性高、毒性低的抗肿瘤新药,合成了11个4-酰硫基-4-脱氧-4’-去甲表鬼臼毒素,衍生物并进行了抗肿瘤活性试验。4-巯基-4-脱氧-4’-去甲表鬼臼毒素和不同的羧酸在二乙氧基磷酰氰酯存在下得相应的硫酯。该类化合物在L1210白血病细胞和KB细胞的体外抑制试验中普遍表现出抑制活性。化合物10的活性与依托泊甙相当。其余活性比依托泊甙差。与相应的C₄位酰氨基的4’-去甲表鬼臼毒素衍生物相比,活性明显弱。提示氮取代的衍生物活性更好。