长效化Exendin-4的研究进展

Exendin-4是一种新型GLP-1类似物,已被用于2型糖尿病的治疗。由于其优良的药学特性成为糖尿病治疗的亮点,而进一步开发设计长效Exendin-4药物成为近年2型糖尿病药物研究的热点。本文就长效Exendin-4药物的国内外研究进展、作用机制以及药效学性质做一综述。     

口服复方孕素1号片妇女血浆 FSH,LH,E_2-17β与 P(孕酮)的变化

本工作对9例正常育龄妇女与13例口服复方孕素1号片(一种新型的长效避孕药)妇女血浆中 FSH,LH,E_2-17β与孕酮进行了放射免疫测定。结果表明,药物使月经周期中的激素水平发生显著变化。同对照组正常值相比,服药后的激素变化可分三种类型。无排卵型(9例),排卵型(2例)与延迟排卵型(2例)。文中对各型激素变化特点作了描述。此外,根据本工作看来,作者以前观察到的孕素1号引起子宫内膜早熟,可能是药物的孕激素活性对内膜直接作用的结果。     

从类固醇到抗体更加激烈的新药开发竞争

市场对支气管哮喘和慢性阻塞性肺部疾病（COPD）治疗药的需求强烈。 全世界COPD患者仍任增加,新药开发竞争激烈展开。 从抗体药物到新型类固醇药物,候选新药多种多样。     

长效重组蛋白药物发展动态

部分重组蛋白药物存在半衰期短的缺陷,临床给药频率高,且大多为注射给药,严重影响患者使用依从性。长效重组蛋白药物是近年来生物技术药物发展的重要趋势之一。对蛋白分子进行改造或修饰,延长重组蛋白药物的半衰期,实现长效以减少给药频率主要通过4种方式:化学修饰、构建突变体、蛋白融合、糖基化修饰。针对上述4种长效化方式及已上市相关产品进行了综述。展望未来,紧跟国外先进技术和质量标准发展,进一步提高国产长效重组蛋白药物质量水平,推进国内相关产品标准升级,推动创新长效重组蛋白药物开发及专利布局是未来几年国内该领域的发展方向。     

长效GLP-1受体激动剂的研发进展

胰高血糖素样肽-1(glucagon-like Peptide-1,GLP-1)是机体在葡萄糖等刺激下释放的一类肠促胰岛素。GLP-1具有促进葡萄糖依赖性胰岛素分泌,促进胰岛β细胞增殖并抑制其凋亡、抑制摄食、减慢胃排空和减轻体重等作用。GLP-1在体内极不稳定,易被二肽基肽酶-IV(dipeptidyl peptidase-IV,DPP-IV)降解,半衰期短。从墨西哥巨蜥蜴唾液中分离得到GLP-1的天然类似物Exendin-4与其有着相似的作用,且不易被DPP-IV降解。以GLP-1及Exendin-4为基础,研发长效GLP-1受体激动剂,已成为研发新型糖尿病治疗药物的热点之一。本文就长效GLP-1受体激动剂的研发现状予以综述。     

新型长效重组促卵泡素的纯化和性质及药代动力学研究

促卵泡素(Follicle-stimulating hormone,FSH)用于治疗男女不孕不育症,由于FSH的半衰期较短,需要频繁注射给药,因此制备长效FSH制剂一直是该类药物的研发方向。首次将来自绒促性素(human chorionic gonadotropin,HCG)中的羧基端肽(carboxy-terminal peptide,CTP)和人免疫球蛋白G2(immunoglobulin G2,IgG2)Fc片段变体(vFc)与单链FSH有序连接,通过基因工程和哺乳动物细胞培养技术实现了其在中国仓鼠卵巢细胞中的稳定表达,然后利用Protein A柱层析手段进行纯化,制备了新型重组FSH-CTP-vFc融合蛋白,同时分析了不同细胞培养时间表达的重组融合蛋白的理化性质和体内外活性。动物试验结果表明,纯化的FSH-CTP-vFc融合蛋白具有显著的体内生物活性,药代动力学分析显示该重组融合蛋白的半衰期是重组FSH的近10倍。这种新型长效重组FSH-CTP-vFc融合蛋白在临床上可大大减少注射次数和患者痛苦,提高患者用药依从性。     

血友病B替代治疗药物研究现状及进展

血友病B是一种X染色体连锁的隐性遗传性出血性疾病,患者因体内缺乏凝血因子IX(FIX)而易发生出血事件,病情严重程度与FIX的缺乏程度相关,严重影响患者的寿命和生存质量。文中通过相关文献资料的查阅和整理,对血友病B替代治疗药物进行归纳和综述,重点阐述上市重组凝血因子IX制品和相关在研药物尤其是长效凝血因子IX的研究进展,为血友病B治疗药物的研发提供基础。     

β2受体激动剂：用于治疗慢性阻塞性肺病的支气管舒张剂

慢性阻塞性肺病(COPD) 是目前世界范围内发病率和病死率持续上升的常见慢性气道疾病。依据最新的2015 年版GOLD 指南, 对于稳定期COPD 的治疗目标是缓解症状和降低远期急性加重风险;支气管舒张剂可通过改变气道平滑肌张力,舒张支气管,改善气流 受限,缓解呼吸困难,提高患者生活质量,是治疗稳定期COPD 的一线基础药物。而β2 受体激动剂是临床上最常用的支气管舒张剂, 简介β2 受体激动剂的作用机制,主要对短效、长效和不同给药途径的β2 受体激动剂用于治疗COPD 的临床研究进展作一综述。     

长效重组药物研究进展

生物重组药物半衰期的长短显著影响药物在使用时的剂量以及治疗的效果,所以延长其在体内的生物半衰期,防止药物在体内迅速降解成为现在药物研究的重要方向之一.针对目前较为广泛使用的几种重组药物长效化方法.如定点突变、化学修饰、基因融合、药物剂型和给药方式的改变等,结合目前已经上市的和正在研发中的长效重组药物进行了综述.     

长效促胰岛素降糖酵母的构建及其对糖尿病模型小鼠的治疗效果

益生菌生物药物是指通过口服表达药用多肽(蛋白)的重组益生菌活细胞达到治疗疾病的新型口服给药系统。为了构建一种能有效防治2型糖尿病的酵母生物药物,文章首先构建了酿酒酵母(S.cerevisiae)整合型表达载体pNK1-PGK,并且通过绿色荧光蛋白(GFP)证明其表达功能正常,利用该载体将10×GLP-1 (Glucagon-like peptide-1)基因转化到酿酒酵母INVSc1中,通过营养缺陷型和Western blotting成功筛选出表达10×GLP-1的长效促胰岛素降糖酵母(Long-acting GLP-1 hypoglycemic yeast, LHY)。该酵母生长迅速,外源基因10×GLP-1表达稳定,表达量达到1.56 mg/g细胞湿重。通过链脲佐菌素和高脂高糖饮食联合诱导的方法构建了2型糖尿病小鼠模型,用LHY对其进行口服灌胃治疗,证明LHY具有较好疗效,明显降低血糖水平。     

信必可都保对稳定期非囊性纤维化支气管扩张的疗效研究

目的：观察信必可都保（布地奈德．福莫特罗）对稳定期非囊性纤维化支气管扩张患者的临床疗效。方法：将42例稳定期非囊性支扩患者随机分为对照组（20例,常规服用氨茶碱0．1g,tid）、观察组（22例,在常规基础上加用信必可都保吸入剂,160μg／4．5μg,tid）,疗程3个月,比较治疗前后两组肺功能指标（FEV1、FVC和PEF）72．SGRQ评分。结果：对照组试验前后肺功能、SGRQ评分无显著改变（P〉0．05）;观察组肺功能无显著改变（P〉0．05）,SGRQ评分显著降低,差异有统计学意义（P〈0．05）。结论：长期使用信必可都保吸入剂对稳定期非囊性纤维化支扩患者的生活质量有明显的改善作用,可有效控制气道炎症,但对改善肺功能无显著效果。     

Discovery of TD-4306, a long-acting β2-agonist for the treatment of asthma and COPD

A multivalent approach focused on amine-based secondary binding groups was applied to the discovery of long-acting inhaled β₂-agonists. Addition of amine moieties to the neutral secondary binding group of an existing β₂-agonist series was found to provide improved in vivo efficacy, but also led to the formation of biologically active aldehyde metabolites which were viewed as a risk for the development of these compounds. Structural simplification of the scaffold and blocking the site of metabolism to prevent aldehyde formation afforded a potent series of dibasic β₂-agonists with improved duration of action relative to their monobasic analogs. Additional optimization led to the discovery of 29 (TD-4306), a potent and selective β₂-agonist with potential for once-daily dosing.     

Multivalent design of long-acting β2-adrenoceptor agonists incorporating biarylamines

A series of potent β₂-adrenoceptor agonists incorporating a biarylamine secondary binding group was identified. The previously reported milveterol (5), identified by a multivalent approach and containing a typical β₂-agonist primary binding group linked via a phenethylamine linker to a hydrophilic secondary binding group, served as an initiation point. A more hydrophobic set of secondary binding groups was explored, prepared rapidly from a common intermediate by Buchwald–Hartwig amination. TD-5471 (25), a potent and selective full agonist of the human β₂-adrenoceptor, was identified as the most promising agent. It is potent, with slow onset in an in vitro guinea pig trachea model and shows a dose-dependent and long duration of action in an in vivo guinea pig model of bronchoprotection. TD-5471 is structurally differentiated from milveterol and its long duration of action is consistent with a correlation with hydrophobicity observed in other long-acting β₂-agonist discovery programs.     

Long-acting beta-agonists plus inhaled corticosteroids safety: a systematic review and meta-analysis of non-randomized studies

Background

Although several systematic reviews investigated the safety of long-acting beta–agonists (LABAs) in asthma, they mainly addressed randomized clinical trials while evidence from non-randomized studies has been mostly neglected. We aim to assess the risk of serious adverse events in adults and children with asthma treated with LABAs and Inhaled Corticosteroids (ICs), compared to patients treated only with ICs, from published non-randomized studies.

Methods

The protocol registration number was CRD42012003387 (http://www.crd.york.ac.uk/Prospero). Literature search for articles published since 1990 was performed in MEDLINE and EMBASE. Two authors selected studies independently for inclusion and extracted the data. A third reviewer resolved discrepancies. To assess the risk of serious adverse events, meta-analyses were performed calculating odds ratio summary estimators using random effect models when heterogeneity was found, and fixed effect models otherwise.

Results

Of 4,415 candidate articles, 1,759 abstracts were reviewed and 220 articles were fully read. Finally, 19 studies met the inclusion criteria. Most of them were retrospective observational cohorts. Sample sizes varied from 50 to 514,216. The meta-analyses performed (69,939-624,303 participants according to the outcome considered) showed that odds ratio of the LABAs and ICs combined treatment when compared with ICs alone was: 0.88 (95% CI 0.69-1.12) for asthma-related hospitalization; 0.75 (95% CI 0.66-0.84) for asthma-related emergency visits; 1.02 (95% CI 0.94-1.10) for systemic corticosteroids; and 0.95 (95% CI 0.9-1.0) for the combined outcome.

Conclusions

Evidence from observational studies shows that the combined treatment of LABAs and ICs is not associated with a higher risk of serious adverse events, compared to ICs alone. Major gaps identified were prospective design, paediatric population and inclusion of mortality as a primary outcome.     

山羊重组促卵泡素长效类似物基因在毕赤酵母中表达

为了获得长效FSH制剂, 利用重叠PCR技术将山羊FSH的a, b亚单位, 通过hCGb亚单位羧基末端延长肽(CTP)基因序列的连接, 构建成单链长效类似物基因FSHb-CTP-a。将其克隆至表达载体pPIC9K, 重组载体线性化处理后电转化至毕赤酵母GS115中, 经判型和G418筛选后获得高拷贝菌株His+Mut+。经甲醇诱导表达后, 进行SDS-PAGE和Western blot分析表明: 重组FSHb-CTP-a的转化子可以正确有效地表达目的蛋白, 分子量约为29 kD。放射免疫分析法(RIA)测定表达上清, 高拷贝转化子的平均表达量为91.849 mIU/mL, 显著高于低拷贝转化子的平均37.419 mIU/mL的表达量, 为FSH的结构研究和长效FSH制剂的生产奠定了基础。     

复方孕素1号(甲孕环酯)对猕猴月经与血浆孕酮的影响

本工作对14只猕猴的月经周期进行了观察。结果表明正常月经周期长短存在个体差异,范围为16—40天,不过多数周期(11/17,64.7%)为27—32天。经期大多不超过5天,经量约为5—6ml。于月经周期第10天给猴灌服一次复方孕素1号(10mg 孕素1号 0.05mg 炔雌醚/kg)后,月经周期延长或缩短,经量有所下降,但经期变化不大。放射免疫测定表明,猕猴正常月经周期的孕酮变化是有规则的:前半周期的孕酮含量一般低于1ng/ml,月经中期以后逐渐升高,于黄体中期形成高峰,最大值达8.7±(SD)4.4ng/ml,于周期末又下降至较低水平。于月经周期第10天给药后,7只猴11个周期中,3个周期的孕酮分泌无变化。另外8个周期中5个周期的孕酮分泌高峰受到抑制,3个周期的孕酮分泌峰延迟出现。这些结果表明药物的抑制排卵作用是不完全的,且存在着较大的个体差异。     

山羊重组促卵泡素长效类似物基因在毕赤酵母中表达

为了获得长效FSH制剂, 利用重叠PCR技术将山羊FSH的a, b亚单位, 通过hCGb亚单位羧基末端延长肽(CTP)基因序列的连接, 构建成单链长效类似物基因FSHb-CTP-a。将其克隆至表达载体pPIC9K, 重组载体线性化处理后电转化至毕赤酵母GS115中, 经判型和G418筛选后获得高拷贝菌株His+Mut+。经甲醇诱导表达后, 进行SDS-PAGE和Western blot分析表明: 重组FSHb-CTP-a的转化子可以正确有效地表达目的蛋白, 分子量约为29 kD。放射免疫分析法(RIA)测定表达上清, 高拷贝转化子的平均表达量为91.849 mIU/mL, 显著高于低拷贝转化子的平均37.419 mIU/mL的表达量, 为FSH的结构研究和长效FSH制剂的生产奠定了基础。