胃舒散中碱式碳酸铋的鉴别

目的:建立胃舒散中主药碱式碳酸铋的化学反应鉴别方法。方法:将供试品进行有机炽灼破坏后,再根据《中国药典》碱式碳酸铋的碘化钾鉴别方法,选用稀硝酸为溶剂对铋盐进行定性分析。结果:供试品硝酸滤液均呈铋盐正反应,且可明显消除胃舒散中中药组分对碱式碳酸铋鉴别的干扰及假性反应,颜色鉴别直观。结论:本方法简便、稳定,可用于该制剂的质量控制。     

医院制剂强氯霜的鉴别方法的研究

目的:为制定强氯霜质量标准提供方法.方法:采用化学反应、薄层色谱法对强氯霜的鉴别实验进行方法学研究.结果:采用化学反应和薄层色谱法对强氯霜进行鉴别试验,阴性对照实验无干扰,专属性强.结论:本方法操作简单,重现性好,专属性强,可用于强氯霜的鉴别实验.     

糖尿病治疗新药——利拉鲁肽

目的:介绍治疗糖尿病新药利拉鲁肽。方法:根据文献,对利拉鲁肽的作用机制、药动学、临床疗效、不良反应等方面信息进行综述与评价。结果:利拉鲁肽通过胰高血糖素样肽-1受体调节体内血糖水平;皮下注射后体内半衰期达10h以上,与部分药物之间存在药动学相互作用;可显著降低糖化血红蛋白水平,具有降低体重和改善β细胞功能的作用;不良反应主要包括胃肠道反应和免疫原性反应等。结论:利拉鲁肽每日1次注射具有延缓糖尿病进展和减少心血管并发症发生的潜力。     

妇洁洗剂的制备及临床应用

对妇洁洗剂的制备与临床应用进行研究。方法 :以试剂法鉴别主药黄柏与蛇床子。结果 :鉴别呈正性反应。结论 :制备工艺简便 ,鉴别易行。临床总有效率为 96 %。     

兔抗乐果抗血清制备的初步实验研究

目的:对制备兔抗乐果抗血清进行初步研究。方法:采用生产乐果的中间体硫磷酯与1,4-二氨基丁烷反应合成乐果半抗原,利用碳二亚胺法,将半抗原与牛血清白蛋白(BSA)及卵清蛋白(OVA)偶联,分别制备免疫原和包被原。将合成的免疫原免疫新西兰白兔6次,制备抗乐果的抗血清。用间接ELISA检测抗血清的效价,用间接竞争ELISA检测与其他结构类似物的交叉反应率。结果:通过偶联,获得免疫原和包被原,半抗原与BSA和OVA的偶联比分别为13∶1和11∶1。免疫原免疫新西兰白兔获得的抗乐果抗血清效价分别为1∶256和1∶128;与氧化乐果的交叉反应率为23%,与其他所测结构类似的农药交叉反应率均小于2%。结论:通过免疫原免疫新西兰白兔,获得抗乐果的抗血清,但其抗血清效价较低,与制备检测试剂盒的要求尚存在较大的差距。     

壮药白楸的生药学及毒理学初步研究

目的 对大戟科植物白楸进行鉴别及急性毒性试验,确保用药安全.方法 采集药材标本并对其进行鉴别;白楸醇提取物以最大浓度及最大给药体积对实验组小鼠灌胃给药,观察14 d内不良反应情况.结果 化学反应结果显示白楸含有黄酮、皂苷等成分;急性毒性试验观察中未发现任何急性毒性反应.给药后14 d,空白组和药物组小鼠体质量增长率分别为73.5%和62.9% （P ＞0.05）.结论 该实验结果可为白楸临床用药指导和制定质量标准提供参考依据.     

桂灵丸鉴别方法的研究

目的研究桂灵丸的定性鉴别方法,制定其质量控制指标,以更好地控制其质量。方法采用显微鉴别、化学反应、薄层色谱方法鉴别桂灵丸处方中组分。结果①显微鉴别中将罂粟壳、五味子的显微特征列入控制指标,具有专属性;②采用化学反应对罂粟壳进行鉴别,结合其薄层色谱鉴别方法,更全面控制其质量;③薄层色谱鉴别对麻黄、罂粟壳、苦杏仁(炒)、五味子进行了研究,薄层色谱斑点清晰,阴性无干扰。结论文中的显微鉴别、化学反应和薄层色谱鉴别方法,可作为桂灵丸的定性控制指标。     

药物发现中的化学合成新策略

为了在短时间内发现性质优良的药物，药物化学家们需要对早期药物研发过程的效率和通量进行优化，包括利用新技术简化合成步骤及利用突破性反应来处理更为复杂的合成问题。本文主要讨论了部分实用的新技术，如聚合物辅助液相合成、微波辅助有机合成、流动化学等，并介绍了几种已被列入常规反应的突破性合成方法，如点击化学反应，多组分反应和关环易位反应。     

元胡止痛片快速显微、化学反应和荧光鉴别法

目的:建立元胡止痛片的快速显微、化学反应和荧光鉴别法。方法:根据法定的制备工艺,采用显微鉴别的方法,对不同厂家批号的元胡止痛片进行了比对试验。结果:通过显微成像系统准确地鉴别元胡止痛片的真伪;利用其各主要成分的化学和荧光鉴别也能鉴别其真伪。结论:适合于元胡止痛片的快速筛查。     

本刊郑重声明

目的建立医院制剂接骨胶囊的质量标准。方法采用化学反应方法和薄层色谱法，对处方中的自然铜、三七、红花进行定性鉴别。结果自然铜的化学反应呈正反应，在薄层色谱中均能检出接骨胶囊中的三七、红花两味药材的成分。结论建立的方法可靠、灵敏、重现性好，可用于接骨胶囊的质量控制。     

Peptimmunology: immunogenic peptides and sequence redundancy

Using short peptide fragments of proteins to elicit antibodies able to recognize the protein from which the peptide sequence was derived, is one of the main goals in immunotherapy today. Indeed, peptide-immunotherapy appears as an obliged way to obtain antibodies of predetermined specificity and exempt from the complications associated with whole cells/entire protein vaccines. However, effective peptide-immunotherapy remains an exciting theoretical speculation largely unrealized to date. The major obstacle in designing effective peptide vaccines is our incapacity to scientifically define peptide immunogenicity. This mini-review schematically describes: 1) the available methods to identify epitopic peptides; 2) the sequence redundancy concept as a possible basis for peptide immunogenicity.     

Removal of B cell epitopes as a practical approach for reducing the immunogenicity of foreign protein-based therapeutics

Immunogenicity of non-human proteins with useful therapeutic properties has prevented their development for use in the therapy of disease. However, this class of proteins could be very useful, if their immunogenicity could be markedly reduced so that many treatment cycles could be administered. One approach to reduce the immunogenicity of foreign proteins is to identify B cell epitopes on the protein and eliminate them by mutagenesis. In this article, theoretical aspects and experimental evidence for the feasibility of B cell epitope removal is reviewed. A special focus is given to our results with deimmunization of recombinant immunotoxins in which Fvs are fused to a 38 kDa portion of the bacterial protein, Pseudomonas exotoxin A (PE38). Immunotoxins targeting CD22 and CD25 have produced complete remissions in many patients with drug resistant Hairy Cell Leukemia and are being evaluated in other malignancies. Experimental data summarized in this review indicates that removal of B cell epitopes is a practical approach for making less immunogenic protein therapeutics from non-human functional proteins. This approach requires grouping of the epitopes to identify targets for deimmunization followed by quantitative analysis of the decrease in affinity produced by the mutations in B cell epitopes.     

The future of protein particle characterization and understanding its potential to diminish the immunogenicity of biopharmaceuticals: A shared perspective

Biopharmaceuticals represent an important and growing class of medicines. Immunogenic responses to biopharmaceuticals in patients can sometimes result in reduced safety and efficacy. Although multiple factors are known to influence immunogenicity, our understanding of the complex underlying mechanisms remains imperfect. In particular, the potential impact of protein aggregates (particulates) on immunogenicity is currently not well understood. This commentary discusses emerging technologies for particle assessment, what is known about the link between particulates and product safety and efficacy, and current regulatory guidances and perspectives. We consider approaches that in the future may permit specific particle attributes to be correlated with relative immunogenic risk, including the value of data derived from clinical and postmarketing surveillance. Finally, we identify some key remaining questions, which, when answered, may guide future strategies for decreasing the immunogenicity of biopharmaceuticals. © 2012 Wiley Periodicals, Inc. and the American Pharmacists Association J Pharm Sci 101:3580–3585, 2012     

聚乙二醇定点修饰蛋白质药物中巯基的研究进展

聚乙二醇(PEG)修饰是解决蛋白质药物溶解度低、稳定性差、半衰期短、具有免疫原性等问题的有效方法,通常在氨基上进行修饰,但在巯基上进行定点修饰更有利于获得结构明确、组成稳定的修饰产物。综述了聚乙二醇定点修饰蛋白质药物中巯基,包括在游离巯基、二硫键和引入的巯基上进行修饰的研究进展。     

生物技术药物免疫原性评价的技术发展概述

药物的免疫原性是指药物刺激机体形成特异抗体或致敏淋巴细胞的性质。评价非期望的免疫原性是生物技术药物临床前和临床评价的重要内容。免疫原性评价内容包括方法的开发、验证、药物诱导的抗体反应水平检测（滴度）、抗体的特性研究、抗体的中和活性测定、分析抗体产生对疗效、毒性和药动学的影响,并预测对人体潜在的免疫原性强弱。如何提高动物模型的预测性、优化检测技术、实现评价方法的规范化和标准化,是未来免疫原性评价亟需解决的问题。     

HAHA – nothing to laugh about. Measuring the immunogenicity (human anti-human antibody response) induced by humanized monoclonal antibodies applying ELISA and SPR technology

Immunogenicity induced by passively applied proteins is a serious issue because it is directly related to the patient's safety. The out-come of an immune reaction to a therapeutic protein can range from transient appearance of antibodies without any clinical significance to severe life threatening conditions. Within this article, enzyme-linked immunosorbent assay (ELISA) and surface plasmon resonance (SPR) methodology to measure immunogenicity are compared and the pros and cons are discussed.     

聚乙二醇修饰的牛血红蛋白血液代用品抗原性和免疫原性的研究

目的建立聚乙二醇修饰的牛血红蛋白血液代用品抗原性和免疫原性评价方法。方法用酶联方法测定供试品的抗原性残留量,用豚鼠过敏试验研究供试品的免疫原性,通过比较推算供试品抗原性残留量的安全性限值。结果供试品的抗原性残留量大于2 3%时,豚鼠会出现不同程度的过敏反应;供试品的抗原性残留量小于或等于2 3%时,豚鼠无过敏反应发生;供试品抗原性残留量的最高限值为2 3%。结论初步建立了牛血红蛋白血液代用品抗原性和免疫原性评价方法     

2012 AAPS National Biotech Conference Open Forum: A Perspective on the Current State of Immunogenicity Prediction and Risk Management

The immunogenicity profile of a biotherapeutic is determined by multiple product-, process- or manufacturing-, patient- and treatment-related factors and the bioanalytical methodology used to monitor for immunogenicity. This creates a complex situation that limits direct correlation of individual factors to observed immunogenicity rates. Therefore, mechanistic understanding of how these factors individually or in concert could influence the overall incidence and clinical risk of immunogenicity is crucial to provide the best benefit/risk profile for a given biotherapeutic in a given indication and to inform risk mitigation strategies. Advances in the field of immunogenicity have included development of best practices for monitoring anti-drug antibody development, categorization of risk factors contributing to immunogenicity, development of predictive tools, and development of effective strategies for risk management and mitigation. Thus, the opportunity to ask "where we are now and where we would like to go from here?" was the main driver for organizing an Open Forum on Improving Immunogenicity Risk Prediction and Management, conducted at the 2012 American Association of Pharmaceutical Scientists'' (AAPS) National Biotechnology Conference in San Diego. The main objectives of the Forum include the following: to understand the nature of immunogenicity risk factors, to identify analytical tools used and animal models and management strategies needed to improve their predictive value, and finally to identify collaboration opportunities to improve the reliability of risk prediction, mitigation, and management. This meeting report provides the Forum participant''s and author''s perspectives on the barriers to advancing this field and recommendations for overcoming these barriers through collaborative efforts.KEY WORDS: ADA, anti-drug antibodies, immunogenicity, neutralizing antibodies, prediction, predictive sciences, risk assesment, risk management, risk mitigation     

Evaluation of a 3D Human Artificial Lymph Node as Test Model for the Assessment of Immunogenicity of Protein Aggregates

The immunogenicity of protein aggregates has been investigated in numerous studies. Nevertheless, it is still unknown which kind of protein aggregates enhance immunogenicity the most. The ability of the currently used in vitro and in vivo systems regarding their predictability of immunogenicity in humans is often questionable, and results are partially contradictive. In this study, we used a 2D in vitro assay and a complex 3D human artificial lymph node model to predict the immunogenicity of protein aggregates of bevacizumab and adalimumab. The monoclonal antibodies were exposed to different stress conditions such as light, heat, and mechanical stress to trigger the formation of protein aggregates and particles, and samples were analyzed thoroughly. Cells and culture supernatants were harvested and analyzed for dendritic cell marker and cytokines. Our study in the artificial lymph node model revealed that bevacizumab after exposure to heat triggered a TH1- and proinflammatory immune response, whereas no trend of immune responses was seen for adalimumab after exposure to different stress conditions. The human artificial lymph node model represents a new test model for testing the immunogenicity of protein aggregates combining the relevance of a 3D human system with the rather easy handling of an in vitro setup.