Protein Drug-Loaded Polymeric Nanoparticles

Considerable interest and research have focused on the administration of therapeutic proteins. For delivery of therapeutic proteins, bioavailability and stabilization of protein drugs to maintain therapeutically acceptable levels is an important challenge in clinical trials. To overcome these challenges, polymeric nanoparticles have become one of the best methods for protein delivery. In this review, we summarize the current available polymeric nanoparticles designed for protein delivery, current status, and advantages of protein delivery systems.

Dynamic changes and surveillance function of prion protein expression in gastric cancer drug resistance

AIM:To explore the dynamic changes of prion protein (PrPc) in the process of gastric cancer drug resistance and the role of PrPc expression in the prognosis of gastric cancer patients receiving chemotherapy.METHODS:A series of gastric cancer cell lines resistant to different concentrations of adriamycin was established,and the expression of PrPc,Bcl-2 and Bax was detected in these cells.Apoptosis was determined using Annexin V staining.Western blotting and immunohisto-chemistry were performed to detect the expression of PrPc in patients receiving chemotherapy and to explore the role of PrPc expression in predicting the chemosensitivity and the outcome of gastric cancer patients receiving chemotherapy.Follow-up was performed for 2 years.RESULTS:PrPc expression was increased with the increase in drug resistance.Bcl-2,together with PrPc,increased the level of anti-apoptosis of cancer cells.Increased PrPc expression predicted the enhanced level of anti-apoptosis and resistance to anticancer drugs.PrPc expression could be used as a marker for predicting the efficacy of chemotherapy and the prognosis of gastric cancer.Increased PrPc expression predicted both poor chemosensitivity and a low 2-year survival rate.Contrarily,low PrPc expression predicted favorable chemosensitivity and a relatively high 2-year survival rate.CONCLUSION:PrPc expression is associated with histological types and differentiation of gastric cancer cells;The PrPc expression level might be a valuable marker in predicting the efficacy of chemotherapy and the prognosis of gastric cancer patients receiving chemotherapy.

Relationship between Methylation Status of Multi-drug Resistance Protein(MRP) and Multi-drug Resistance in Lung Cancer Cell Lines

Objective： To study the relationship between the methylation status of multi-drug resistance protein （MRP） gene and the expression of its mRNA and protein in lung cancer cell lines. Methods： Human embryo lung cell line WI-38, lung adenocarcinoma cell line SPCA-1 and its drug-resistant cells induced by different concentrations of doxorubicin were treated with restriction endonuclease Eco47III. The methylation status of MRP was examined by PCR, and the expressions of its mRNA and protein were evaluated by in situ hybridization and immunohistochemistry. Results： MRP gene promoter region of WI-38 cells was in hypermethylation status, but the promoter region of MRP in SPCA-1 cells and their resistant derivatives induced by different concentrations of doxorubicin were in hypomethylation status. There were significant differences in the expression of MRP mRNA among WI-38 cell line, SPCA-1 cells and their drug-resistant derivatives induced by different concentration of doxorubicin. Consistently, MRP immunostaining presented similar significant differences. Conclusion： The promoter region of MRP in SPCA-1 lung adenocarcinoma cells was in hypomethylation status. The hypomethylation status of 5＇ regulatory region of MRP promoter is an important structural basis that can increase the activity of transcription and results in the development of drug resistance in lung cancer.

Synthesis and Characterization of Soy Protein Isolate/MMT Nanocomposite Film for the Control Release of the Drug Ofloxacin

Nanocomposites were prepared by blending soy protein isolate with different percentage of MMT by melt extrusion technique. The nanocomposites were characterized by using, XRD, TEM, SEM and TGA methods. The XRD studies indicated the absence of diffraction peaks for the bio-nanocomposites. From the TEM studies it was ascertained that the degree of exfoliation increased with increase in MMT content. The morphology of the nanocomposites was ascertained from the SEM studies. The degradation pattern of the nano-composites was evaluated from the TG analysis. The drug delivery system of the nanocmposites was investigated by blending the nanocomposites with ofloxacin at different pH media. The various kinetic parameters were evaluated and the mechanism of drug delivery has been postulated based on the kinetic data.

Interaction between human serum albumin and cholesterol-grafted polyglutamate as the potential carriers of protein drugs

Currently there is no successful platform technology for the sustained release of protein drugs.It seems inevitable to specifically develop new materials for such purpose, and hence the understanding of protein–material interactions is highly desirable. In this study, we synthesized cholesterol-grafted polyglutamate(PGA-g-Chol) as a hydrophobically-modified polypeptide, and thoroughly characterized its interaction with a model protein(human serum albumin) in the aqueous solution by using circular dichroism, fluorescence methods, and light scattering. With the protein concentration fixed at 5 μmol/L,adding PGA-g-Chol polymers into the solution resulted in continuous blue shift of the protein fluorescence(from 339 to 332 nm), until the polymer molar concentration reached the same value as the protein. In contrast, the un-modified polyglutamate polymers apparently neither affected the protein microenvironment nor formed aggregates. Based on the experimental data, we proposed a physical picture for such protein–polymer systems, where the polymer first bind with the protein in a 1:1 molar ratio via a fraction of their hydrophobic pendant cholesterol resides along the polymer chain. In this protein/polymer complex, there are excess unbound cholesterol residues. As the polymer concentration increases, the polymers form multi-polymer aggregates around 200 nm in diameter via the same hydrophobic cholesterol residues. The protein/polymer complex also participate in the aggregation via their excess cholesterol residues, and consequently the proteins are encapsulated into the nanoparticles. The encapsulation was also found to increase the thermal stability of the model protein.

Simple and Rapid Hollow Fiber Liquid Phase Microextraction Followed by High Performance Liquid Chromatography Method for Determination of Drug-protein Binding

A method was established using hollow fiber-liquid phase microextraction（HF-LPME） followed by high performance liquid chromatography（HPLC） to determine the concentration of the free（unbound） drug in the solution of the drug and protein. Measurements of drug-protein binding ratios and free drug concentrations were then analyzed with the Klotz equation to determine the equilibrium binding constant and number of binding sites for drug-protein interaction. The optimized method allows one to perform the efficient extraction and separation of free drug from protein-bound drug, protein, and other interfering substances. This approach was used to characterize the binding of the anticholinergic drugs atropine sulfate and scopolamine hydrobromide to proteins in human plasma and bovine serum albumin（BSA）. The results demonstrate the utility of HF-LPME method for measuring free drug concentrations in protein-drug mixtures and determining the protein binding parameters of a pharmacologically important class of drugs.

Computational Investigation of the Interaction of Anti-Influenza Drugs with CoVID-19 Protein

Coronavirus (CoVID-19) is a new outbreak of coronavirus disease which started in the Wuhan, China, the spread of this virus has now reached a global stage, urgent need is therefore needed to find new drug molecules which can either be used as a first aid intervention or slow down the multiplication rate of the virus within the system. In order to address this, this research looked into the existing antiviral drugs and screened them for their inhibitory properties towards the CoVID-19 protein. Recently, the crystal structure of the CoVID-19 (6LU7) protein has been established, this gives us the possible drug target site in CoVID-19. The binding affinity of the six compounds was screened using MOE (Molecular Operating Environment) software, four compounds (Zanamivir, Peramivir, Rimantidine, and Oseltamivir) out these six compounds have been approved by the Food Drug and Administration (FDA). The molecular docking calculation, Higher Occupied Molecular Orbital (HOMO) and Lowest Unoccupied Molecular Orbital (LUMO) calculation were used to hypothesise the bioactivity of the FDA approved drug against the CoVID-19 protein. The calculation showed that Pimodivir tops the list of the anti influenza drug which can be used as first aid treatment for patient. Apart from Pimodivir, Laninamivir Octanoate is also a very good drug which might be used to inhibit CoVID-19 protein. It was also discovered that based on binding property of Rimantadine, it might be suitable for Fragment Based Drug Design (FBDD) approach which might lead to the discovery of completely new drug entity. Stability of the new protein structure was studied using GROMACS molecular dynamic simulation software. The results showed that the stability of the protein structure was achieved over a range of time, this confirmed that 6LU7 crystal structure might be a suitable protein crystal structure suitable for the development of new drug towards the treatment of CoVID-19. Finally, based on the molecular docking result, Pimodivir and Laninamivir Octanoate might be useful in the treatment of infected patient.

主穹窿蛋白基因在高原地区癫痫儿童中的多态性分析

目的:探讨分析主穹窿蛋白(MVP)基因三个SNP位点rs4788187、rs3815824、rs3815823多态性与高原地区儿童癫痫的相关性。方法:选取2017年1月~2020年1月青海大学附属医院收治的80例癫痫患儿为研究对象,根据耐药性不同分为耐药组与非耐药组各40例,利用PCR-RELP方法检测rs4788187、rs3815824、rs3815823三个SNP位点的多态性分布,并进行统计分析。结果:两组年龄、性别、民族以及癫痫发作类型比较差异无统计学意义(P>0.05)。癫痫患儿rs4788187、rs3815824的等位基因C频率明显低于T,而rs3815823的等位基因T频率明显低于C;两组患儿SNP位点rs4788187、rs3815824、rs3815823的基因型频率和等位基因频率比较差异均无统计学意义(P>0.05)。结论:MVP基因的SNP位点(rs4788187、rs3815824、rs3815823)与癫痫耐药性不相关。

Predicted essential proteins of Plasmodium falciparum for potential drug targets

Objective:To identify novel drug targets for treatment of Plasmodium falciparum.Methods: Local BT.ASTP were used to find the proteins non-homologous to human essential proteins as novel drug targets.Functional domains of novel drug targets were identified by InterPro and Pfam.3D structures of potential drug targets were predicated by the SWISS-MODEL workspace. Ligands and ligand-binding sites of the proteins were searched by Ef-seek.Results:Three essential proteins were identified that might be considered as potential drug targets.AAN37254.1 belonged to 1-deoxy-D-xylulose 5-phosphate reductoisomerase,CAD50499.1 belonged to chorismale synthase,CAD51220.1 belonged to FAD binging 3 family,but the function of CAD51220.1 was unknown.The 3D structures,ligands and ligand-binding sites of AAM37254.1 and CAD50499.1 were successfully predicated.Conclusions:Two of these potential drug targets are key enzymes in 2-C-methyl-d-erythritol 4-phosphate pathway and shikimate pathway, which are absent in humans,so these two essential proteins are good potential drug targets.The function and 3D structures of CAD50499.1 is still unknown,it still need further study.

多药耐药相关蛋白转运体在药物性肝损伤中的作用研究进展

肝脏是人体新陈代谢最旺盛的器官,也是体内多种药物的解毒器官。当长期或过量使用药物时会增加药物性肝损伤(DILI)的风险。多药耐药相关蛋白(MRPs)是位于细胞膜上的功能蛋白,可转运多种药物,在DILI中发挥重要作用。MRPs功能的抑制、缺失是药物肝毒性产生的重要原因。本文对MRPs的结构、表达部位及功能进行归纳,并对MRPs与DILI的关系及其改善DILI的机制进行总结,期望更好地了解MRPs转运体与DILI的关系,为后续防治DILI提供参考。

蛋白质结构稳定性检测的实验课程设计

随着生物技术的突破和发展,多肽蛋白类药物的出现使得人类疾病的诊断和治疗发生了翻天覆地的变化。为了顺应多肽蛋白类药物的飞速发展,加强生命科学等专业本科生对此类药物制剂相关知识的普及已势在必行。此实验课程设计利用稳定性热检测法分析蛋白质结构的稳定性,为生命科学等专业本科生设计一堂深入了解蛋白质结构与功能关系的课程,以促进本科生拓宽学术视野,提升完成相关科学研究和适应医药产业发展新形势的能力。

间变性淋巴瘤激酶在治疗非小细胞肺癌中的作用及潜在医学应用

在非小细胞肺癌(non-small cell lung cancer,NSCLC)的治疗过程中,随着靶向表皮生长因子受体剂(epidermal growth factor receptor,EGFR)抑制剂的使用,不少患者出现了耐药,耐药的主要原因有EGFR位点的突变,旁路激活等原因。在旁路激活中主要有间变性淋巴瘤激酶(anaplastic lymphoma kinase,ALK)等通路的激活。最近有权威研究发现ALK与免疫治疗有密切的关系。该文从ALK的结构和生理功能,ALK的小分子抑制剂,ALK的生物学功能及其与NSCLC相关的应用现状进行综述,并展望了未来的发展方向,从而使ALK更好地应用于NSCLC的治疗。

Insights into the structural biology of G-protein coupled receptors impacts drug design for central nervous system neurodegenerative processes

In the last few years, there have been important new insights into the structural biology of G-protein coupled receptors. It is now known that allosteric binding sites are involved in the affinity and selec- tivity of ligands for G-protein coupled receptors, and that signaling by these receptors involves both G-protein dependent and independent pathways. The present review outlines the physiological and pharmacological implications of this perspective for the design of new drugs to treat disorders of the central nervous system. Specifically, new possibilities are explored in relation to allosteric and or- thosteric binding sites on dopamine receptors for the treatment of Parkinson＇s disease, and on muscarinic receptors for Alzheimer＇s disease. Future research can seek to identify ligands that can bind to more than one site on the same receptor, or simultaneously bind to two receptors and form a dimer. For example, the design of bivalent drugs that can reach homo/hetero-dimers of D2 dopa- mine receptor holds promise as a relevant therapeutic strategy for Parkinson＇s disease. Regarding the treatment of Alzheimer＇s disease, the design of dualsteric ligands for mono-oligomeric mus- carinic receptors could increase therapeutic effectiveness by generating potent compounds that could activate more than one signaling pathway.

N-钙黏蛋白与Wnt/β-catenin通路在骨髓间充质干细胞与白血病干细胞中的耐药机制研究

【目的】探讨N-钙黏蛋白(N-cadherin)与Wnt/β-catenin通路在骨髓间充质干细胞(MSC)与白血病干细胞(LSC)中的耐药机制研究。【方法】65例急性髓系白血病(AML)患者,完全缓解41例,未缓解24例,比较两组CD34+CD38-干细胞中N-cadherin的表达差异。根据不同MSC或去甲氧柔红霉素(IDA)处理下将CD34+CD38-Kg1α细胞分为6组,检测细胞增殖和凋亡情况及N-cadherin、β-catenin水平,检测细胞黏附及抗凋亡能力。【结果】未缓解患者的N-cadherin表达水平高于完全缓解者(P<0.01);LSC+IDA组(B组)细胞凋亡率显著高于LSC与MSC直接共培养+IDA组(D组)(P<0.05);在IDA浓度为100 nmol/L、200 nmol/L时,B组的细胞增殖抑制率显著高于D组(P<0.05);B组的集落形成率显著低于D组(P<0.05)。Western blot结果显示:N-cadherin及β-catenin在MSC直接接触培养条件下表达高于单独培养或分离培养;LSC在MSC共培养条件下,N-cadherin及β-catenin表达水平上升,伴随细胞核内β-catenin水平上升。【结论】骨髓微环境的MSC可以通过N-cadherin与LSC的黏附作用支持LSC的增殖能力,同时促进LSC Wnt/β-catenin通路的激活,对化疗药物产生耐药性。

人工智能辅助的蛋白质稳定性优化

蛋白质药物具有作用机制清晰、作用特异性强、不良反应少等优势,临床应用前景巨大。蛋白质的稳定性是蛋白质药物的一项非常重要的指标,对于其成药性、安全性和有效性都至关重要。近年来,人工智能辅助的蛋白质改造工程逐渐发展成为一种高效的蛋白质分子设计新策略,并被广泛应用于蛋白质稳定性预测、药物设计和抗体优化等方面。本文介绍主要的人工智能辅助的蛋白质稳定性优化方法,讨论不同种类优化方法的优劣及其在蛋白质药物设计和优化中的应用,探讨人工智能在蛋白质稳定性设计中的挑战和前景,以期为研究者们开发更稳定、更高效的蛋白质药物提供新的思路。

CYP3A4基因位点多态性对托法替布治疗类风湿关节炎临床疗效及药品不良反应的影响

目的探讨CYP3A4*4,CYP3A4*18,CYP3A4*1G基因位点多态性对托法替布治疗类风湿关节炎(RA)临床疗效及药品不良反应(ADR)的影响。方法选取医院风湿免疫科2020年2月至2022年8月收治的RA患者309例作为RA组,予枸橼酸托法替布片,每日2次,每次5 mg,共治疗6个月;选取同期的健康人群165例作为对照组。采用荧光聚合酶链反应(PCR)法检测CYP3A4*4,CYP3A4*18,CYP3A4*1G基因位点多态性;根据美国风湿病学学会(ACR)制订的ACR20标准评价托法替布的临床疗效,以是否符合ACR20标准,将RA患者分为改善组(181例)和未改善组(128例);统计治疗期间RA患者与托法替布相关的ADR,采用Karch和Lasagna评定法判定因果关系,以因果关系是否判定为肯定、很可能和可能,将RA患者分为ADR组(58例)和无ADR组(251例)。结果RA组和对照组患者的CYP3A4*4,CYP3A4*18,CYP3A4*1G基因位点多态性均无显著差异(P>0.05)。改善组和未改善组患者的疾病活动度差异显著(P<0.05),CYP3A4*4,CYP3A4*18,CYP3A4*1G基因位点多态性均无显著差异(P>0.05)。ADR累及系统为实验室检验异常、皮肤系统、消化系统、呼吸系统、血液系统,分别发生27例、11例、7例、5例、3例;ADR严重程度为轻度51例,中度7例。ADR组和无ADR组患者的CYP3A4*1G基因位点多态性差异显著(P<0.05),CYP3A4*4和CYP3A4*18基因位点多态性均无显著差异(P>0.05)。结论CYP3A4*1G基因位点多态性与托法替布治疗RA的ADR有相关性。使用托法替布时,应监测患者的CYP3A4*1G基因位点多态性,必要时调整剂量,保证用药安全。

PCI后损伤血管再内皮化的机制与治疗研究进展

晚期支架内血栓形成是经皮冠状动脉介入治疗(PCI)后主要的并发症,严重影响了冠状动脉粥样硬化性心脏病患者的预后,带来不可逆转的危害。PCI后损伤血管再内皮化可减少晚期支架内血栓的形成,再内皮化的机制主要包括内皮细胞黏附和增殖的调控、平滑肌细胞黏附和增殖的调控、血小板黏附聚集和活化以及纤维蛋白原的吸附。对冠状动脉粥样硬化性心脏病患者血管损伤再内皮化机制的研究,可为损伤后血管重塑提供新的思路和治疗靶点,有利于提升PCI患者的预后。现就PCI后损伤血管再内皮化的相关机制和治疗进行综述。

疱疹病毒利用宿主蛋白高效复制机理及其抑制剂研究进展

疱疹病毒的入侵、转录和蛋白合成等多个生命活动,都需要利用宿主细胞蛋白和微环境进行。该过程一般会导致细胞蛋白的合成和降解、修饰、细胞定位改变,以及与其他蛋白相互作用等一系列变化。本文从病毒感染引起的细胞凋亡和自噬体系、泛素-蛋白酶体系、核-质转运体系、生物合成体系和病毒诱导的先天免疫反应几个层面,综述病毒劫持或干扰细胞正常生命活动的研究进展,探讨病毒与宿主细胞的相互作用关系,并对目前抗疱疹病毒药物靶点和机制进行总结,以期为设计新型抗病毒制剂提供资料。

希特林蛋白缺乏症的营养及药物治疗

希特林蛋白缺乏症是由位于7q21.3染色体SLC25A13基因中的双等位基因致病变异引起的常染色体隐性遗传病,分为新生儿肝内胆汁淤积症(NICCD),希特林蛋白缺乏导致的发育不良和血脂异常(FTTDCD)及成人发病的Ⅱ型瓜氨酸血症(CTLN2)。饮食治疗是NICCD和FTTDCD的关键干预措施,过多、过量的碳水化合物摄入会加剧本病,而补充中链脂肪酸可以为肝细胞提供能量,促进脂肪生成。希特林蛋白缺乏症的根本治疗方案为肝移植,但由于供体稀缺和价格昂贵,营养干预和药物治疗仍然是临床治疗的主要手段。本文对希特林蛋白缺乏症的代谢途径、发病机制和临床表现进行简要概述,收集药物治疗和营养治疗方法并进行简要综述。