雷珠单抗的大肠埃希菌胞外分泌表达、纯化及结构表征

抗血管内皮细胞生长因子(VEGF)抗体可抑制新生血管的形成和生长,在年龄相关性黄斑变性眼科疾病的临床治疗中具有重要应用价值,特别是已上市的商品名为雷珠单抗的VEGF抗体Fab片段,在市场上取得巨大成功。本研究探索了雷珠单抗在大肠埃希菌中的胞外分泌表达技术,所表达的产品利用SDS-PAGE、Western Blot、肽谱图全序列分析和纳米差示扫描量热法(Nano DSC)等手段进行了初步的结构表征。结果显示,本研究成功实现了雷珠单抗的大肠埃希菌胞外分泌表达,经过进一步的Capto L亲和色谱获得了有正确氨基酸序列的Fab产品,经质谱分析具有正确的二硫键配对,且具有与标准品相似的热稳定性,为其进一步的工业生产奠定了基础。     

雷珠单抗治疗息肉状脉络膜血管病变的临床效果分析

目的探讨研究雷珠单抗治疗息肉状脉络膜血管病变(PCV)的临床疗效及预后。方法选取2014年~2016年我院收治的48例息肉状脉络膜血管病变患者,随机分为对照组24例与观察组24例,对照组采用常规治疗,观察组采用雷珠单抗治疗,对比两组疗效及预后。结果两组患者治疗后Log Mar视力明显低于治疗前,(P<0.05)。观察组治疗后1周、3月Log Mar视力评分明显低于对照组(P<0.05)。对比两组患者治疗前黄斑中心视网膜厚度(CRT),差异无统计学意义(P>0.05)。观察组患者治疗后1周、3个月CRT明显低于对照组(P<0.05)。结论对于息肉状脉络膜血管病变患者的治疗方式,采用雷珠单抗治疗,可有效提高其临床治疗效果,改善患者视力、减轻患者息肉病灶渗漏。     

玻璃体腔内注射雷珠单抗治疗早产儿视网膜病变的临床疗效及其对VEGF水平的影响

目的 探讨玻璃体腔内注射雷珠单抗治疗早产儿视网膜病变的临床疗效及其对血管内皮生长因子(VEGF)水平的影响.方法 选择南昌大学第二附属医院儿童眼科2018年4月—2020年3月收治的80例视网膜病变早产儿为研究对象,随机分为对照组与观察组,各40例.对照组采用激光治疗,观察组给予雷珠单抗玻璃体腔内注射治疗.2组均连续治...     

超声乳化术联合雷珠单抗对青光眼并白内障的效果及对角膜功能和相关蛋白的影响

目的 探讨超声乳化术联合雷珠单抗对青光眼并白内障的效果及对角膜功能和相关蛋白的影响。方法 将106例青光眼并白内障患者随机分为雷珠单抗组(超声乳化术联合雷珠单抗治疗)与对照组(超声乳化术治疗)各53例。检测患者角膜功能及相关蛋白表达情况。结果 术后3个月,两组最佳矫正视力、角膜内皮细胞密度、血清IP-10水平均明显高于术前1 d,差异有统计学意义(P<0.05),且雷珠单抗组更高,差异有统计学意义(P<0.05),两组血清VEGF水平明显低于术前1 d,差异有统计学意义(P<0.05),且雷珠单抗组更低,差异有统计学意义(P<0.05)。术后3个月,雷珠单抗组的并发症发生率低于对照组,差异有统计学意义(P<0.05)。结论 超声乳化术联合雷珠单抗在青光眼并白内障患者的应用能促进血清IP-10的表达,抑制VEGF的表达,还可提高角膜内皮细胞密度,促进改善患者的视力,降低并发症的发生。     

糖尿病黄斑水肿应用玻璃体内注射雷珠单抗、曲安奈德的疗效分析

目的 比较玻璃体内注射雷珠单抗、曲安奈德在糖尿病黄斑水肿中的临床疗效.方法 收集本院2014年10月至2015年10月入院的80例糖尿病黄斑水肿患者,随机分为两组,雷珠单抗组患者给予雷珠单抗治疗,曲安奈德组患者给予曲安奈德治疗,比较两组患者治疗前后最佳矫正视力、RNV渗透面积、黄斑区中心视网膜厚度、眼压与并发症等.结果 雷珠单抗组患者BCVA为(0.556±0.155),显著高于曲安奈德组的(0.409±0.143);RNV渗透面积为(5.405±3.274),显著低于曲安奈德组的(8.332±4.058);雷珠单抗组患者CMT为(289.231±64.040) μm,显著低于曲安奈德组的(370.127±88.172)μm;雷珠单抗组患者治疗后眼压为(15.157±2.351) mmHg,显著低于曲安奈德组(17.264±3.197)mmHg;两组比较,t=4.4897、3.550、4.695、3.358,差异有统计学意义(P＜0.01).结论 玻璃体内注射雷珠单抗在糖尿病黄斑水肿中的临床疗效更为显著,安全性较高,具有借鉴性.     

血管内皮生长因子抑制剂治疗视网膜血管瘤样增生药理及临床研究进展

视网膜血管瘤样增生(retinal angiomatous proliferation,RAP)是一种血管增生导致的眼科疾病,因血管生长因子及血管抑制因子失衡,促使微血管瘤形成,可严重影响患者视力。高表达的血管内皮生长因子(vascular endothelial growth factor,VEGF)可加快RAP发生发展,抗VEGF可加快瘤体萎缩而改善临床疗效。现阶段已知的抗VEGF药物包括雷珠单抗(ranibizumab)、贝伐珠单抗(bevacizumab)及阿柏西普(aflibercept)等。近年来,抗VEGF药物在RAP治疗中的作用备受关注,现对VEGF在RAP中的作用机制及抗VEGF治疗进展作一综述,旨在为临床提供参考。     

雷珠单抗治疗视网膜静脉阻塞合并黄斑水肿的临床观察

目的:观察并对比玻璃体腔内注射雷珠单抗治疗视网膜分支静脉阻塞(BRVO)合并黄斑水肿(ME)的疗效。方法:采用回顾性临床系列研究方法,将2015年6月—2018年6月确诊为BRVO合并ME的患者46例(46只眼)纳入研究。所有患眼给予玻璃体腔内注射雷珠单抗治疗。分别于治疗前、治疗后1个月、2个月、3个月及6个月后记录最佳矫正视力(BCVA),并行光相断层扫描(OCT)检查。观察分析治疗前后BCVA、黄斑中心凹厚度(CMT)以及患者局部及全身并发症情况。结果:玻璃体腔内注射雷珠单抗治疗视网膜分支静脉阻塞合并黄斑水肿前后在1个月、2个月、3个月及6个月各时间点的BCVA和CMT相比,差异均有统计学意义(P<0.01)。结论:玻璃体腔注射雷珠单抗可以有效提高BRVO合并ME患者的视力,减轻黄斑水肿,预防新生血管类疾病的发生,其疗效是肯定的。     

具有抗肿瘤血管生成作用的VEGFR2抑制剂的研究进展

肿瘤血管生成是肿瘤生长与转移的重要过程。血管内皮生长因子(vascular endothelial growth factor, VEGF)和它们的受体(VEGFR1, 2, 3)调控内皮细胞的增殖与迁移,在很多肿瘤内高表达。对VEGFR的抑制已成为许多癌症的有效疗法。VEGFR2在肿瘤血管生成中起重要作用,VEGFR2磷酸化是肿瘤血管生成的重要过程,抑制这个过程是利用VEGF信号通路治疗癌症的主要机理之一。使用抗体和小分子与VEGF结合或者干扰不同的VEGFR结构域可以阻断VEGF信号通路。许多小分子VEGFR2抑制剂得到开发,大部分为多靶点抑制剂,FDA已批准一系列VEGFR2抑制剂上市。本文对已上市及进入临床研究晚期的VEGFR2小分子抑制剂及其活性、临床应用进行了综述,并对它们的构效关系作出了简要分析。发挥多靶点抑制剂的优点,规避其带来的毒副作用,是有希望的研究方向。     

23G玻璃体切割术联合雷珠单抗治疗增生性糖尿病视网膜病变的临床观察

目的探讨23G玻璃体切割术联合雷珠单抗治疗增生性糖尿病视网膜病变的临床效果。方法选取2016年2月~2017年1月间我院收治的在23G玻璃体切割术前未进行雷珠单抗注射的增生性糖尿病视网膜病变患者40例(40眼)为非注射组,选取2017年2~10月我院收治的在23G玻璃体切割术前通过雷珠单抗注射的增生性糖尿病视网膜病变患者40例(40眼)为注射组,比较两组患者的临床治疗效果。结果注射组患者的手术时间、电凝使用次数和新生血管出血的次数、不良反应发生情况明显少于非注射组,两组相比差异有统计学意义(P0.05)。通过对患者进行术后6个月随访发现非注射组患者视力为0.4,注射组患者视力是0.6,两组相比存在明显差异性(P0.05)。结论用23G玻璃体切割术联合雷珠单抗,可使手术耗时降低、简化手术程序;降低手术中医源性视网膜裂孔以及大出血,并发症等。     

雷珠单抗联合玻璃体切割术治疗增生性糖尿病视网膜病变合并Ⅰ、Ⅱ期青光眼临床观察

目的：探讨雷珠单抗联合玻璃体切割术( PPV)对增生性糖尿病视网膜病变( PDR)合并Ⅰ、Ⅱ期新生血管性青光眼( NVG)的临床效果。方法回顾性分析2011年6月—2014年6月眼科确诊为PDR合并Ⅰ、Ⅱ期NVG患者60例(60只眼)的临床资料,根据术前有无注射雷珠单抗分为观察组34例(34只眼)和对照组26例(26只眼),均行23G PPV,观察组术前注射雷珠单抗,对照组术前未注射雷珠单抗。观察两组手术时间、新生血管平均出血次数、电凝使用次数、术中及术后6个月并发症发生率;术前、术后1个月测定最佳修正后视觉灵敏度( BCVA)、房水中血管内皮细胞生长因子( VEGF)、色素上皮衍生因子( PEDF)及术后1个月黄斑中心凹视网膜厚度( CRT)值。结果观察组手术时间短于对照组,新生血管出血次数、电凝使用次数少于对照组( P<0．05);观察组术前、术后房水VEGF、PEDF低于对照组,术后BCVA高于对照组(P<0．01);观察组术中医源性视网膜裂孔、术中大量出血及术后一过性高眼压、玻璃体出血发生率均低于对照组( P<0．05)。结论雷珠单抗联合PPV治疗PDR合并NVG,可降低房水VGEF、PEDF,提高BCVA水平,降低术中及术后并发症发生率。     

肝癌组织血管内皮生长因子与受体的表达及其临床意义

目的研究血管内皮生长因子(VEGF)及受体(VEGFR)在肝癌组织中的表达及其临床意义。方法采用免疫组化SP法检测76例肝癌切除标本中VEGF、VEGFR的表达,分析VEGF、VEGFR与肝癌临床发病特征、术后生存时间、术后复发的关系。结果本组76例1年后复发29例、2年后复发16例,累计1、2年平均无瘤生存率分别为61.85%±5.57%、40.97%±5.63%;76例肝癌手术标本中VEGF阳性表达61例,阳性率80.3%,VEGFR阳性表达67例,阳性率88.2%,并且统计发现VEGF、VEGFR的阳性表达和肿瘤的大小、数目、有无包膜、术前是否肝内转移都是影响肝癌术后复发与生存率的关键因素,而VEGF及VEGFR的阳性表达与肿瘤体积大小、有无包膜、术前是否转移更是密切相关;VEGF、VEGFR的表达二者之间也相互关联。结论 VEGF、VEGFR过度表达与肝癌术后复发及生存时间密切相关,联合分析二者的表达对肝癌手术治疗的预后判断有更大指导价值。     

润肺止嗽丸对慢性阻塞性肺疾病大鼠细胞凋亡的改善作用及其机制研究

目的研究中成药润肺止嗽丸对慢性阻塞性肺疾病（COPD）大鼠细胞凋亡的影响,并探讨其作用机制。方法 32只雄性Wistar大鼠随机分为对照组、模型组、润肺1月和3月组;使用烟熏联合气道滴入脂多糖法制备COPD大鼠模型;润肺1、3月组ig给予润肺止嗽丸0.9 g/kg,每天给药1次,分别连续给药1、3个月,对照组和模型组给予等体积的超纯水1个月;HE染色观察慢阻肺大鼠肺组织变化;TUNEL法检测凋亡细胞并计算凋亡指数（AI）;Elisa法检测肺泡灌洗液（BALF）中血管内皮生长因子（VEGF）水平;Western blotting法检测肺组织中VEGF和血管内皮生长因子受体2（VEGFR2）的蛋白表达。结果与对照组比较,模型组大鼠肺组织形态病变明显;AI显著升高;BALF中VEGF、肺组织中VEGF和VEGFR2水平均显著下降。与模型组比较,润肺1月和3月组肺组织形态明显好转;AI显著下降;BALF中VEGF、肺组织中VEGF和VEGFR2水平显著升高;且润肺3月组效果好于1月组。结论润肺止嗽丸可以改善慢阻肺细胞凋亡,机制可能与上调VEGF和VEGFR2相关。     

Emerging oral VEGF inhibitors for the treatment of renal cell carcinoma

Introduction: The incidence of renal cell carcinoma (RCC) has increased in recent years and, unfortunately, many patients initially present with metastatic disease. When surgery is not an option, treatment involves administration of targeted therapies. The vascular endothelial growth factor (VEGF) pathway has been identified as an important mediator for the development of RCC. Numerous agents target VEGF-mediated signaling, yet resistance and progressive disease still persists. Novel small molecule VEGF inhibitors with high affinity for the VEGF receptor (VEGFR) have been discovered and are currently under investigation for the management of RCC.

Areas covered: The VEGFR pathway, its aberrant signaling, and the agents under development that inhibit VEGFR signaling are discussed. The mechanism(s), pharmacokinetics, pharmacodynamics, efficacy, and toxicity of these investigational agents are also reviewed.

Expert opinion: Management of metastatic RCC involves combination immunotherapy or administration of oral VEGFR inhibitors and largely depends on risk stratification. Emerging and investigational oral VEGFR inhibitors, given as monotherapy or in combination with immunotherapy, could augment current treatment approaches and may mitigate toxicities associated with VEGFR inhibition.     

The clinical application of fruquintinib on colorectal cancer

ABSTRACT

Introduction: Anti-angiogenetic agents are currently the most commonly used drugs for the treatment of colorectal cancer (CRC) patients, including various inhibitors targeting the epidermal growth factor receptor (EGFR), vascular endothelial growth factor (VEGF), and VEGF receptors (VEGFRs). Fruquintinib (HMPL-013), a highly selective and long-term small-molecule inhibitor of VEGFR (VEGFR1, 2, and 3) was recently approved in China for CRC treatment. Clinical studies have shown it has many advantages, such as low off-target toxicity, good drug tolerance, and strong effect.

Areas covered: In the review, the molecular structure, mechanism of action, pharmacokinetics, clinical efficacy, and safety of fruquintinib are introduced in detail. The potential clinic application on non-small cell lung cancer (NSCLC) and gastric cancer is also discussed.

Expert commentary: Fruquintinib was approved for patients with metastatic colorectal cancer (RAS wild type) who have previously received fluorouracil, oxaliplatin, and irinotecan-based chemotherapy and who have received or are not suitable for anti- VEGF therapy and anti- EGFR therapy. As a novel, therapeutic approach to CRC, Fruquintinib could be used as a third-line drug for the treatment of CRC patients. Due to drug resistance during the long-term therapy, the combination of fruquintinib with other targeted therapy drugs may be an effective option for CRC treatment.     

血清中VEGF，VEGFR1与VEGFR2在肺癌中的表达及其相互关系

目的本研究为探讨肺癌患者血清及组织中VEGF,VEGFR1,VEGFR2的表达在肺癌发生发展中的变化,为肺癌的诊断、发病机制提供生物学指标。方法选择山西省肿瘤医院胸外科2008年1月到2008年10月收治的初治原发性肺癌患者65例作为病例组,术后均经组织病理学证实。按照频数匹配的方法选择本院健康体检中20例正常人作为健康对照组。采集外周静脉血,用EHSA法检测病例组外周血中VEGF,VEGFR1,VEGFR2含量,并与健康对照组比较。采用SPSS12．0软件包进行统计学分析VEGF,VEGFR1和VEGFR2与患者病理类型、临床分期、性别、年龄之间的关联性及其三者之间的相关性。结果①肺癌患者血清VEGF、VEGFR：表达与健康对照组比较差异有显著性（均P〈0．01）;VEG—FR,表达在两组之间比较差异无显著性（P〉0．05）;②血清VEGF、VEGFR1、VEGFR2表达在性别,年龄及临床分期之间比较差异均无显著性（均P〉0．05）。结论①肺癌血清中VEGF,VEGFR2的表达在腺癌,鳞癌及小细胞癌患者中明显高于正常对照,而VEGFR1的表达与对照组比较无显著性差异;②患者肺癌血清中VEGF,VEGFR1,VEGFR2的表达在性别,年龄,临床分期中差异均无显著性。     

柠檬苦素靶向VEGF/VEGFR2通路对肺癌A549细胞的增殖和血管生成的影响

目的:探讨柠檬苦素靶向血管内皮生长因子(VEGF)/血管内皮细胞生长因子受体2(VEGFR2)通路对肺癌A549细胞的增殖和血管生成的影响。方法:肺癌A549细胞分为空白对照组、氟尿嘧啶组(500μg·ml^-1)、柠檬苦素低、高剂量组(100,1000μg·ml^-1);MTT法测定各组细胞增殖水平,甲醇固定吉姆沙染色测定单克隆形成数目,BD Matrigel基质测定血管形成水平,transwell板测定相对迁移率,Axv-FiTC试剂盒测定细胞凋亡水平,RT-PCR法及酶联免疫吸附法测定VEGF、VEGFR2 mRNA及MEK1/2、ERK1/2蛋白表达。结果:柠檬苦素低、高剂量组和氟尿嘧啶组A值、存活率、克隆形成数目、相对总血管长度、相对总血管数目、相对迁移率、VEGF、VEGFR2 mRNA及MEK1/2、ERK1/2蛋白水平低于空白对照组,凋亡率高于空白对照组(P<0.05),且剂量-效应关系明显(P<0.05);柠檬苦素低剂量组A值、存活率、克隆形成数目、相对总血管长度、相对总血管数目、相对迁移率、VEGF、VEGFR2 mRNA及MEK1/2、ERK1/2蛋白水平高于氟尿嘧啶组,凋亡率低于氟尿嘧啶组(P<0.05);柠檬苦素高剂量组上述指标与氟尿嘧啶组比较差异无统计学意义(P>0.05)。结论:柠檬苦素能抑制肺癌A549细胞增殖、血管生成、侵袭、诱导肺癌A549细胞凋亡;其机制可能与柠檬苦素抑制VEGF/VEGFR2诱导的MEK1/2、ERK1/2磷酸化有关。     

Systemic safety of anti-VEGF drugs: a commentary

Introduction: VEGF is a mediator of angiogenesis. Thus, concerns have been expressed following the use of VEGF inhibitors for the treatment of neovascular age-related macular degeneration (nAMD). Ranibizumab, and more recently aflibercept, are VEGF inhibitors licensed for the treatment of nAMD. Bevacizumab is also used but unlicensed for this application.

Areas covered: A non-systematic review of nAMD trials was undertaken to investigate four outcomes: all-cause mortality, all systemic serious adverse events (SSAEs), arteriothrombotic events (ATEs) and gastrointestinal (GI) complications. Differences in event rates with injections of ranibizumab compared to bevacizumab, aflibercept, photodynamic therapy (PDT) and sham were explored and quantified using fixed-effect meta-analyses.

Expert opinion: Anti-VEGF agents can influence vascular health; however, the data suggest no difference in the risk of an ATE or death between anti-VEGF agents. Clinical trials are limited in their size and eligibility criteria and databases of patients treated in routine practice should also be scrutinized.     

血管内皮生长因子及其受体在抗肿瘤治疗应用的研究进展

肿瘤的生长依赖肿瘤新生血管的形成。血管内皮生长因子（VEGF）是一类能促进血管生成的细胞因子，能诱导细胞的有丝分裂和调节内皮细胞的通透性。VEGF及其受体介导的肿瘤血管新生在肿瘤的生长和转移中具有重要的作用。本文综述了VEGF及其受体的分类、作用机制及在抗肿瘤治疗中的应用。     

The antitumor effect of tanshinone IIA on anti-proliferation and decreasing VEGF/VEGFR2 expression on the human non-small cell lung cancer A549 cell line

The effects of tanshinone IIA on the proliferation of the human non-small cell lung cancer cell line A549 and its possible mechanism on the VEGF/VEGFR signal pathway were investigated. The exploration of the interaction between tanshinone IIA and its target proteins provides a feasible platform for studying the anticancer mechanism of active components of herbs. The CCK-8 assay was used to evaluate the proliferative activity of A549 cells treated with tanshinone IIA (2.5−80 μmol/L) for 24, 48 and 72 h, respectively. Flow cytometry was used for the detection of cell apoptosis and cell cycle perturbation. VEGF and VEGFR2 expression were studied by Western blotting. The binding mode of tanshinone IIA within the crystal structure of the VEGFR2 protein was evaluated with molecular docking analysis by use of the CDOCKER algorithm in Discovery Studio 2.1. The CCK-8 results showed that tanshinone IIA can significantly inhibit A549 cell proliferation in a dose- and time-dependent manner. Flow cytometry results showed that the apoptosis rate of tested group was higher than the vehicle control, and tanshinone IIA-treated cells accumulated at the S phase, which was higher than the vehicle control. Furthermore, the expression of VEGF and VEGFR2 was decreased in Western blot. Finally, molecular docking analysis revealed that tanshinone IIA could be stably docked into the kinase domain of VEGFR2 protein with its unique modes to form H-bonds with Cys917 and π–π stacking interactions with Val848. In conclusion, tanshinone IIA may suppress A549 proliferation, induce apoptosis and cell cycle arrest at the S phase. This drug may suppress angiogenesis by targeting the protein kinase domains of VEGF/VEGFR2.Abbreviations: ADM, adriamycin; CAM, chorioallantoic membrane; CCK-8, cell counting kit-8; DMSO, dimethylsulfoxide; EPCs, endothelial progenitor cells; FBS, fetal bovine serum; FCM, flow cytometry; HRP, horseradish peroxidase; IC₅₀, 50% inhibitory concentration; MD, molecular dynamics; mOS, median overall survival; NS, normal saline; NSCLC, non-small cell lung cancer; PI, propidium iodide; PKB/AKT, protein kinase B; RMSD, root-mean-square deviation; Tan IIA, tanshinone IIA; tRR, tumor response rate; vdW, van der Waals force; VEGF, vascular endothelial growth factorKEY WORDS: Non-small cell lung cancer, Tanshinone IIA, VEGF/VEGFR signal pathway, Molecular docking     

靶向血管内皮生长因子及其受体的抗肿瘤药物研究进展

血管生成对肿瘤的生长和转移起着关键作用,血管内皮生长因子（VEGF）及其受体信号通路是调节肿瘤新生血管生成的重要途径,因此,近年来以VEGF及其受体为作用靶标的抗肿瘤血管生成治疗已经成为研究热点,目前已有多种药物上市或处于临床试验阶段。本文主要综述了VEGF及其受体在肿瘤血管生成调节机制中的作用,同时着重介绍靶向VEGF及其受体的抗肿瘤药物的新近研究进展、临床应用及存在的问题。