3-取代吲哚-2-酮类化合物的设计、合成及抗肿瘤活性研究

目的设计合成对微管蛋白和血管内皮细胞生长因子受体2(VEGFR-2)激酶具有双重抑制作用的3-取代吲哚-2-酮类化合物,考察其体外抑瘤活性。方法以取代的苯胺为起始原料,经缩合、环合、还原、取代等反应制得系列目标化合物,并考察该系列化合物对微管蛋白和肿瘤细胞的抑制活性。结果共合成了11个新的目标化合物。实验结果显示,化合物j9对微管蛋白和VEGFR-2激酶具有双重抑制活性。所有目标化合物对3种肿瘤细胞株均有中等强度的抑制活性。结论该类化合物是一类具有多靶点作用的抗肿瘤化合物。

植物性食品对尿激酶活性抑制的测定实验

目的 了解水果、蔬菜和茶叶等植物性食物对于促进细胞恶性转化的尿激酶活性抑制效果。方法 利用谷氨酸 甘氨酸 精氨酸肽为底物的尿激酶活性测定原理 ,分别用 2 5种水果、37种蔬菜的榨汁和 5 1种茶叶的水浸提液与尿激酶反应 ,再加入光活性尿激酶反应底物反应后用酶标仪测定剩余酶活性。结果 2 5种水果、37种蔬菜的测试样中有近一半的尿激酶活性抑制率为 2 0 %以上 ,其中 ,柠檬、猕猴桃、豌豆、菠菜、南瓜等的抑制率超过 80 % ,大蒜、萝卜、蜂斗菜、旱芹、李子、菠萝、葡萄等的抑制率为 5 0 %～ 79% ;5 1个茶叶样的平均抑制率为 83 2 % ,其中 2 8个茶叶样的抑制率 >90 % ,10个茶叶样为 80 %～ 89% ,10个茶样本为 70 %～ 79% ,绿茶类的平均抑制率比红茶类的抑制率高出10 %～ 15 % ;蔬菜的加工方式对抑制效果有影响 ,有刺激味的蔬菜以有机溶剂浸提效果好于水浸提效果 ,无刺激味的则以水浸提的效果为好 ;多数新鲜蔬菜经冷藏和热处理后的效果为减弱。结论 日常生活中的植物性食物有抑制尿激酶活性的作用。

新型查耳酮衍生物的合成及其初步抗蛋白酪氨酸激酶(PTKs)活性研究

目的设计合成一系列全新的查耳酮类衍生物,并初步测试其蛋白酪氨酸激酶(PTKs)抑制活性。方法以间二甲苯为原料,经硝化、还原、水解、甲氧甲基保护等反应得到中间体取代苯乙酮,该中间体再与取代苯甲醛发生羟醛缩合反应后脱保护基得到目标化合物。采用酶联免疫吸附法(ELISA),以金雀异黄素为阳性对照,对目标化合物进行体外PTKs抑制活性检测。结果与结论合成了10个查耳酮衍生物,其中9个是未见报道的新化合物,10个化合物的结构经核磁共振氢谱和质谱确证。化合物6a及新化合物6b、6c和6d对PTKs具有良好的抑制活性。

Effect of Lysophosphatidylcholine on ATP and ρ-Nitrophenyl Phosphate Hydrolysis by the Plasma Membrane H^+-ATPase from Soybean Hypocotyls

The stimulatory effect of lysophosphatidylcholine (lyso_PC) on ATP and ρ_nitrophenyl phosphate (PNPP) hydrolysis by the plasma membrane H +_ATPase from soybean (Glycine max (L.) Merr.) hypocotyls was studied. Results showed that lyso_PC stimulated the hydrolysis of ATP; ATP hydrolysis was enhanced dramatically when lyso_PC was within 0-0.03%, and increased slightly when lyso_PC was higher than 0.03%. At the concentration of 0.03%, lyso_PC stimulated ATP hydrolysis by 80.5%. Kinetics analysis showed that V max increased from 0.46 μmol P i·mg -1 protein·min -1 to 0.87 μmol P i·mg -1 protein·min -1 while K m increased from 0.88 mmol/L to 1.15 mmol/L under lyso_PC treatment. The optimum pH of ATP hydrolysis was shifted from 6.5 to 7.0 . Moreover, it was found lyso_PC enhanced the inhibition of ATP hydrolysis by hydroxylamine. In the presence of 200 mmol/L hydroxylamine, ATP hydrolysis was inhibited by 74.4%, while it was inhibited by 84.4% when treated with lyso_PC. However, PNPP hydrolysis and the inhibitory effect of vanadate were not affected by lyso_PC. The above results indicated that the kinase domain might be an action site or regulatory region of the C_terminal autoinhibitory domain in the plant plasma membrane H +_ATPase.

Effect of Calpain inhibitor I on glucocorticoid receptor-dependent degradation and its transactivation ability

Objective: To investigate the effect of Calpain inhibitor I on glucocorticoid receptor-dependent proteasomal degradation and its transcriptional activity. Methods: After Raw-264.7 cells were treated with Calpain inhibitor I, dexamethasone, or both for about 12 h, the change of glucocorticoid receptor was detected by western blot analysis. COS-7 cells were transfected with PRsh-GRα expression vector and glucocorticoid-responsive receptor pMAMneo-CAT, then the effect of Calpain inhibitor I on glucocorticoid receptor transcriptional activation ability was determined by CAT activity. Results: The glucocorticoid receptor levels decreased after RAW-264.7 cells were treated with dexamethasone for 12 hours, which effect can be inhibited by Calpain inhibitor I to some extent. CAT activity assay showed that Calpain inhibitor I enhance glucocorticoid receptor transcriptional activity. Conclusion: Calpain inhibitor I can inhibit the down-regulation of dexamethasone on glucocorticoid receptor, and enhances glucocorticoid receptor transactivation ability.

FR167653,a p38 mitogen-activated protein kinase inhibitor,aggravates experimental colitis in mice

AIM: To investigate the effects of FR167653 on the development of dextran sulfate sodium (DSS)-induced colitis in mice. METHODS: BALB/c mice were fed rodent chow containing 3.5% (wt/wt) DSS. The recipient mice underwent intra-peritoneal injection of vehicles or FR167653 (30 mg/kg per day). The mice were sacrificed on day 14, and the degree of colitis was assessed. Immunohistochemical analyses for CD4+ T cell and F4/80+ macrophage infiltration were also performed. Mucosal cytokine expression was analyzed by RT-PCR. RESULTS: The body weight loss was more apparent in the FR167653-treated DSS mice than in the vehicle- treated DSS mice. The colon length was shorter in the FR167653-treated DSS mice than in the vehicle-treated DSS mice. Disease activity index and histological colitis score were significantly higher in FR167653- than in vehicle-treated DSS animals. Microscopically, mucosal edema, cellular infiltration (CD4 T cells and F4/80 macrophages), and the disruption of the epithelium were much more severe in FR167653-treated mice than in controls. Mucosal mRNA expression for interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α) were found to be markedly reduced in FR167653-treated DSS mice. CONCLUSION: Treatment with FR167653 aggravated DSS colitis in mice. This effect was accompanied by a reduction of mucosal IL-1β and TNF-α expression, suggesting a role of p38 mitogen-activated protein kinase (MAPK)-mediated proinflammatory cytokine induction in host defense mechanisms.

Effects of p38 MAPK inhibitor on the rat pain behavior and proinflammatory cytokines in a metastatic bone cancer pain model

Objective： To observe the effects of p38 mitogen activated protein kinase （MAPK） inhibitor SB203580 by intrathecal injection on the pain behavior and the spinal proinflammatory cytokines in a rat model of bone cancer pain induced by breast cancer cells. Methods： Eleven rats were used to establish the models of bone cancer pain, six rats were treated by intrathecal SB203580 injection, and the other 5 were as the controls. The paw withdrawal latency （PWL）, histology and the spinal levels of IL-1β and TNF-α were detected. Results： All the 11 rats presented evident bone destruction and thermal hyperalgesia after intra-tibial injection of breast cancer cells. No effect of SB203580 on the bone destruction was observed. However, following intrathecal injection of SB203580, the left PWLs （12.12± 1.26 s at 16 days and 12.99 ± 1.65 s at 19 days） were significant higher than that of controls （9.05 ± 1.08 s at 16 days and 8.55 ± 1.60 s at 19 days）, P 〈 0.05. Meanwhile, inkathecal injection of SB203580 evidently reduced the levels of spinal IL-1β and TNF-α. Conclusion： Intrathecal injection of SB203580 in a rat model of bone cancer pain cannot prevent the tibial destruction but significantly depress the thermalgia sensitivity, which might result from inhibiting inkacellular p38 MAPK signaling transduction, and thereby reducing the release of the proinflammatory cytokines.

Prognostic factors to predict survival in non-small-celllung cancer with brain metastasis

Objective: The purpose of the study was to assess prognostic factors to predict overall survival(OS) and progression-free survival(PFS) in non-small-cell lung cancer(NSCLC) with brain metastasis(BM). Methods: From November 2011 to March 2013, the clinical data of 31 NSCLC cases with BM treated with multiple modalities including brain radiotherapy alone, systemic chemotherapy, whole brain radiotherapy(WBRT) combined with tyrosine kinase inhibitor(TKIs). The efficacy and adverse reaction were evaluated after treatment. Results: In terms of intracranial lesions, the objective response rate(ORR) and the disease control rate(DCR) were 22.6% and 90.3%, respectively. As for systemic disease, ORR and DCR were 32.3% and 93.5%, respectively. The median time to progression-free survival(PFS) was 298 days(95% CI: 258.624–337.376 days), whereas in the epidermal growth factor receptor(EGFR) mutation patients was 331 days. Patients who received EGFR-TKIs combined with brain radiation had better response rate(RR) than those only brain radiation. Univariate analysis showed that the EGFR-mutations could predictive factors for PFS, and not to other clinical pathological features. The most common toxicities were rash and diarrhea, but all were well-tolerated. Conclusion: EGFR-mutations is the independent prognostic factors affecting the survival rates of NSCLC patients with BM. Through the clinical observation, icotinib combined with WBRT may be effective on brain metastases in NSCLC patients, and toxicities are tolerable, which worth further study.