TRPM离子通道与人类疾病

TRPM蛋白家族是一类表达于多种哺乳动物细胞中广泛存在的离子通道。近年来发现它们在维持某些特定生理功能中起关 键作用且与人类疾病密切相关。研究显示氧化应激可使TRPM离子通道功能异常导致疾病发生、发展。TRPM亚家族的三个成 员，TRPM2，TRPM4 和TRPM7 均受氧化应激的调控，其功能改变、增加或缺失与炎症及免疫系统的激活、神经退行性疾病和神经 系统疾病、心血管疾病、癌症及糖尿病，代谢紊乱和骨疾病等疾病紧密联系。本文就近年来氧化应激调控的TRPM离子通道与人 类疾病的关系做简要综述。此外，文章也将探讨它们作为药物设计靶点和工具的应用前景。     

Aggregation prone regions in human proteome: Insights from large‐scale data analyses

Protein aggregation leads to several burdensome human maladies, but a molecular level understanding of how human proteome has tackled the threat of aggregation is currently lacking. In this work, we survey the human proteome for incidence of aggregation prone regions (APRs), by using sequences of experimentally validated amyloid‐fibril forming peptides and via computational predictions. While approximately 30 human proteins are currently known to be amyloidogenic, we found that 260 proteins (～1% of human proteome) contain at least one experimentally validated amyloid‐fibril forming segment. Computer predictions suggest that more than 80% of the human proteins contain at least one potential APR and approximately two‐thirds (65%) contain two or more APRs; spanning 3–5% of their sequences. Sequence randomizations show that this apparently high incidence of APRs has been actually significantly reduced by unique amino acid composition and sequence patterning of human proteins. The human proteome has utilized a wide repertoire of sequence‐structural optimization strategies, most of them already known, to minimize deleterious consequences due to the presence of APRs while simultaneously taking advantage of their order promoting properties. This survey also found that APRs tend to be located near the active and ligand binding sites in human proteins, but not near the post translational modification sites. The APRs in human proteins are also preferentially found at heterotypic interfaces rather than homotypic ones. Interestingly, this survey reveals that APRs play multiple, often opposing, roles in the human protein sequence‐structure‐function relationships. Insights gained from this work have several interesting implications towards novel drug discovery and development. Proteins 2017; 85:1099–1118. © 2017 Wiley Periodicals, Inc.     

2-Acylamino-5-nitro-1,3-thiazoles: Preparation and in vitro bioevaluation against four neglected protozoan parasites

The 2-acylamino-5-nitro-1,3-thiazole derivatives (1–14) were prepared using a one step reaction. All compounds were tested in vitro against four neglected protozoan parasites (Giardia intestinalis, Trichomonas vaginalis, Leishmania amazonensis and Trypanosoma cruzi). Acetamide (9), valeroylamide (10), benzamide (12), methylcarbamate (13) and ethyloxamate (14) derivatives were the most active compounds against G. intestinalis and T. vaginalis, showing nanomolar inhibition. Compound 13 (IC₅₀ = 10 nM), was 536-times more active than metronidazole, and 121-fold more effective than nitazoxanide against G. intestinalis. Compound 14 was 29-times more active than metronidazole and 6.5-fold more potent than nitazoxanide against T. vaginalis. Ureic derivatives 2, 3 and 5 showed moderate activity against L. amazonensis. None of them were active against T. cruzi. Ligand efficiency indexes analysis revealed higher intrinsic quality of the most active 2-acylamino derivatives than nitazoxanide and metronidazole. In silico toxicity profile was also computed for the most active compounds. A very low in vitro mammalian cytotoxicity was obtained for 13 and 14, showing selectivity indexes (SI) of 246,300 and 141,500, respectively. Nitazoxanide showed an excellent leishmanicidal and trypanocidal effect, repurposing this drug as potential new antikinetoplastid parasite compound     

Probing binding mechanism of interleukin-6 and olokizumab: in silico design of potential lead antibodies for autoimmune and inflammatory diseases

Computer-aided antibody engineering has been successful in the design of new biologics for disease diagnosis and therapeutic interventions. Interleukin-6 (IL-6), a well-recognized drug target for various autoimmune and inflammatory diseases such as rheumatoid arthritis, multiple sclerosis, and psoriasis, was investigated in silico to design potential lead antibodies. Here, crystal structure of IL-6 along with monoclonal antibody olokizumab was explored to predict antigen–antibody (Ag???Ab)-interacting residues using DiscoTope, Paratome, and PyMOL. Tyr56, Tyr103 in heavy chain and Gly30, Ile31 in light chain of olokizumab were mutated with residues Ser, Thr, Tyr, Trp, and Phe. A set of 899 mutant macromolecules were designed, and binding affinity of these macromolecules to IL-6 was evaluated through Ag???Ab docking (ZDOCK, ClusPro, and Rosetta server), binding free-energy calculations using Molecular Mechanics/Poisson Boltzman Surface Area (MM/PBSA) method, and interaction energy estimation. In comparison to olokizumab, eight newly designed theoretical antibodies demonstrated better result in all assessments. Therefore, these newly designed macromolecules were proposed as potential lead antibodies to serve as a therapeutics option for IL-6-mediated diseases.     

Role of IL-6 in Asthma and Other Inflammatory Pulmonary Diseases

The incidence and severity of chronic lung diseases is growing and affects between 100 and 150 million people worldwide and is associated with a significant rate of mortality. Unfortunately, the initial cause that triggers most chronic lung diseases remains unknown and current available therapies only ameliorate, but do not cure the disease. Thus, there is a need for identification of new targets and development of novel therapies especially for those most severely affected. IL-6, like other inflammatory cytokines, has been shown to be elevated in different lung diseases, but it was considered a byproduct of ongoing inflammation in the lung. However, recent studies support a dissociation of IL-6 from inflammation in the lung and suggest that this cytokine plays an active role in pathogenesis of asthma and, in all likelihood, COPD. IL-6 may therefore be a germane target for treatment of these and other chronic lung disease. Here, we provide an overview of the studies in mouse models and human patients that provide support for the involvement of IL-6 in lung diseases.     

Host identification in unfed ticks from stable isotope compositions (δ13C and δ15N)

Determination of the ratios of natural stable isotopes (¹³C/¹²C and ¹⁵N/¹⁴N) in unfed Ixodes ricinus nymphs and adults, which, in their previous stage, fed on captive wild rodents (Apodemus sylvaticus and Myodes glareolus), wild birds (Parus major and Cyanistes caeruleus) or domestic ruminants (Ovis aries and Bos taurus), demonstrated that it is possible to identify each host category with confidence. First, the tick–blood spacing, which is the difference between values obtained from ticks and the blood of hosts that they had fed on in the previous stage, was consistent (152 spacings investigated from 15 host individuals in total). Second, potential confounding factors (tick age and sex) did not affect the discriminatory power of the isotope patterns, nor did different rearing conditions (room temperature vs. 4 °C) or the duration of development (maximum of 430 days). The findings that the tick–blood isotope spacings, across a diverse range of hosts, were similar and predictable, and that confounders had little or no effect on this, strongly support the usage of the isotope approach. Because each of the host categories has a different role in the population dynamics of I. ricinus and in tick‐borne pathogen ecology, the method described here has great potential for the clarification of tick and tick‐borne pathogen ecology in the field.     

纳米孔测序技术在病毒性传染病检测及研究中的应用

快速、准确鉴定出病原体是临床感染性疾病诊断和传染病预防控制的基础。高通量测序基因检测技术突破了传统检测手段的时效性、灵敏度等的局限，为病原体检测和研究提供了便捷、高效的途径。本综述以高通量测序技术发展过程为基础，回顾纳米孔三代测序技术，及其在病毒性传染病检测鉴定及研究中的应用，并对该技术的应用前景及可能存在的问题进行阐述，期望它能在病毒性传染病的防控方面发挥更大的作用。     

计算机辅助设计可溶性BLyS受体, eBCMA-Fc 融合蛋白及其在大肠杆菌中的表达

B 细胞成熟抗原 (BCMA)是 B淋巴细胞刺激因子(BLyS)的受体之一．它的胞外区与人IgG1 Fc的融合蛋白eBCMA-Fc，又称为诱饵受体，具有拮抗BLyS的活性．为了设计新的拮抗肽，基于BCMA和Fc的晶体结构，通过计算机图形学技术、分子模拟方法，建立了eBCMA-Fc融合蛋白的三维理论结构．利用均方根位移（root mean square distance, RMSD）对eBCMA-Fc融合蛋白与单体eBCMA、Fc构象差异进行分析．融合蛋白eBCMA-Fc中的eBCMA段与单体eBCMA的主链碳原子间RMSD值为0.036 nm，Fc段与单体Fc的主链碳原子间RMSD值为0.064 nm．结果表明，对比单体，融合蛋白eBCMA-Fc并未因eBCMA与Fc直接连接而发生构象的变化．分子对接方法显示,融合蛋白eBCMA-Fc中的BCMA与BLyS作用，而Fc扮演着稳定BCMA构象的支架作用．为进一步验证上述理论分析，构建eBCMA-Fc融合基因，并将载有eBCMA-Fc融合基因的原核表达质粒转化BL21 (DE3)菌、在细菌中表达．目的蛋白经蛋白A亲和柱纯化大约为36 kD，与理论预测值34 kD相近．免疫印迹表明抗人IgG抗体能够识别eBCMA-Fc融合蛋白．ELISA证实,eBCMA-Fc融合蛋白能够结合BLyS．随着eBCMA-Fc融合蛋白增加，结合BLyS的融合蛋白也相应增加．而对照人IgG,即使在高浓度条件下，也不结合BLyS．此外，eBCMA-Fc 融合蛋白能够抑制BLyS对B细胞肿瘤Daudi细胞的作用．这些研究为下一步设计和筛选BLyS拮抗肽提供了实验基础．     

Symmetrical aryl linked bis-iminothiazolidinones as new chemical entities for the inhibition of monoamine oxidases: Synthesis,in vitro biological evaluation and molecular modelling analysis

The multifactorial nature of Parkinson’s disease necessitates the development of new chemical entities with inherent ability to address key pathogenic processes. To this end, two series of new symmetrical 1,2- and 1,4-bis(2-aroyl/alkoylimino-5-(2-methoxy-2-oxoethylidene)-4-oxo-thiazolidin-3-yl)benzene derivatives (3a–g and 5a–e) were synthesized in good yields by the cyclization of 1,2- and 1,4-bis(N′-substituted thioureido)benzene intermediates with dimethyl acetylenedicarboxylate (DMAD) in methanol at ambient temperature. The bis-iminothiazolidinone compounds were investigated in vitro for their inhibition of monoamine oxidase (MAO-A & MAO-B) enzymes with the aim to identify new and distinct pharmacophores for the treatment of neurodegenerative disorders like Parkinson’s disease. Most of the designed compounds exhibited good inhibitory efficacy against monoamine oxidases. Compound 5a was identified as the most potent inhibitor of MAO-A depicting an IC₅₀ value of 0.001 μM, a 4-fold stronger inhibitory strength compared to standard inhibitor (clorgyline: IC₅₀ = 0.0045 μM). Molecular docking studies provided insights into enzyme-inhibitor interactions and a rationale for the observed inhibition towards monoamine oxidases.