Protein kinase inhibitors: insights into drug design from structure

Protein kinases are targets for treatment of a number of diseases. This review focuses on kinase inhibitors that are in the clinic or in clinical trials and for which structural information is available. Structures have informed drug design and have illuminated the mechanism of inhibition. We review progress with the receptor tyrosine kinases (growth factor receptors EGFR, VEGFR, and FGFR) and nonreceptor tyrosine kinases (Bcr-Abl), where advances have been made with cancer therapeutic agents such as Herceptin and Gleevec. Among the serine-threonine kinases, p38, Rho-kinase, cyclin-dependent kinases, and Chk1 have been targeted with productive results for inflammation and cancer. Structures have provided insights into targeting the inactive or active form of the kinase, for targeting the global constellation of residues at the ATP site or less conserved additional pockets or single residues, and into targeting noncatalytic domains.     

Rational design of peptide-based HIV proteinase inhibitors

A series of peptide derivatives based on the transition-state mimetic concept has been designed that inhibit the proteinase from the human immunodeficiency virus (HIV). The more active compounds inhibit both HIV-1 and HIV-2 proteinases in the nanomolar range with little effect at 10 micromolar against the structurally related human aspartic proteinases. Proteolytic cleavage of the HIV-1 gag polyprotein (p55) to the viral structural protein p24 was inhibited in chronically infected CEM cells. Antiviral activity was observed in the nanomolar range (with one compound active below 10 nanomolar) in three different cell systems, as assessed by p24 antigen and syncytium formation. Cytotoxicity was not detected at 10 and 5 micromolar in C8166 and JM cells, respectively, indicating a high therapeutic index for this new class of HIV proteinase inhibitors.     

体外Src蛋白酪氨酸激酶抑制剂筛选模型的建立

在体外构建一个以Src蛋白为靶位点的蛋白酪氨酸激酶抑制剂快速筛选模型,为筛选Src蛋白酪氨酸激酶抑制剂奠定基础.基因工程表达GST-v-Src蛋白,收集包涵体蛋白,并经变性复性处理.以生物素化的聚Glu∶Tyr(4∶1)为激酶反应底物、以辣根过氧化物酶(horseradish peroxidase,HRP)标记的磷酸酪氨酸特异性单克隆抗体(HRP-PY20)检测磷酸酪氨酸残基的酶联免疫吸附实验(enzyme-linked immunosorbent assay,ELISA)测定获得蛋白的激酶活性,并进行抑制剂筛选.该模型具有靶向性强、快速、简便可行、高通量的特点.用该方法对6种治疗肿瘤配方中常用的中草药进行筛选,筛选出3种中草药水煎剂富含Src蛋白酪氨酸激酶抑制剂成分.     

Blocking of EGF-dependent cell proliferation by EGF receptor kinase inhibitors

A systematic series of low molecular weight protein tyrosine kinase inhibitors were synthesized; they had progressively increasing affinity over a 2500-fold range toward the substrate site of epidermal growth factor (EGF) receptor kinase domain. These compounds inhibited EGF receptor kinase activity up to three orders of magnitude more than they inhibited insulin receptor kinase, and they also effectively inhibited the EGF-dependent autophosphorylation of the receptor. The most potent compounds effectively inhibited the EGF-dependent proliferation of A431/clone 15 cells with little or no effect on the EGF-independent proliferation of these cells. The potential use of tyrosine protein kinase inhibitors as antiproliferative agents is demonstrated.     

Structural mechanism for STI-571 inhibition of abelson tyrosine kinase

The inadvertent activation of the Abelson tyrosine kinase (Abl) causes chronic myelogenous leukemia (CML). A small-molecule inhibitor of Abl (STI-571) is effective in the treatment of CML. We report the crystal structure of the catalytic domain of Abl, complexed to a variant of STI-571. Critical to the binding of STI-571 is the adoption by the kinase of an inactive conformation, in which a centrally located "activation loop" is not phosphorylated. The conformation of this loop is distinct from that in active protein kinases, as well as in the inactive form of the closely related Src kinases. These results suggest that compounds that exploit the distinctive inactivation mechanisms of individual protein kinases can achieve both high affinity and high specificity.     

Biogenic methane, hydrogen escape, and the irreversible oxidation of early Earth

The low O2 content of the Archean atmosphere implies that methane should have been present at levels approximately 10(2) to 10(3) parts per million volume (ppmv) (compared with 1.7 ppmv today) given a plausible biogenic source. CH4 is favored as the greenhouse gas that countered the lower luminosity of the early Sun. But abundant CH4 implies that hydrogen escapes to space (upward arrow space) orders of magnitude faster than today. Such reductant loss oxidizes the Earth. Photosynthesis splits water into O2 and H, and methanogenesis transfers the H into CH4. Hydrogen escape after CH4 photolysis, therefore, causes a net gain of oxygen [CO2 + 2H2O --> CH4 + 2O2 --> CO2 + O2 + 4H(upward arrow space)]. Expected irreversible oxidation (approximately 10(12) to 10(13) moles oxygen per year) may help explain how Earth's surface environment became irreversibly oxidized.     

成品油管网调运决策平台的结构设计

为了满足复杂成品油管网调运决策和安全运行的需求,通过引入供应链管理理念,提出油品调运决策平台的结构设计.在提高管网内信息可视性的基础上,通过针对管网供应链模型的仿真、管网和中转库区调运计划的统筹制定,解决油品调运各链接问题,实现调运风险控制,确保调运顺畅和库存优化.阐明了成品油管网调运决策平台结构及平台中信息系统、决策仿真系统、计划编制系统、状态显示系统的功能和关联,为了解决油品调运各链接问题,实现调运风险控制,确保调运顺畅和库存优化.     

啤酒花自动采摘机的结构设计及机理分析

介绍了1种啤酒花自动采摘机的主要结构设计及工作机理.通过对啤酒花品种‘青岛大花’的力学特征分析,采用了捋花采摘的设计思想,机器摘花时作用于花瓣上的力较小,采摘动作柔和,使啤酒花采摘时的破碎率降低,损失率减小,并通过调整摘花轮之间的差速度提高啤酒花的摘净率.对于不同品种啤酒花,因其力学特性与‘青岛大花’不同,可通过调整摘花轮之间的差速系统达到对不同品种啤酒花的采摘要求.     

甘蔗ADP/ATP转运蛋白酶基因克隆及其序列的生物信息学分析

甘蔗(Saccharum officinarum L.)是C4光合途径作物,有着特殊的能量代谢系统,研究其能量代谢过程的ADP/ATP转运蛋白酶基因具有重要的意义.采用同源克隆原理结合PCR技术克隆甘蔗AAC转运蛋白酶基因,并应用生物信息学方法,对其进行多序列比对分析、物理性质分析、三级结构预测、亚细胞定位、跨膜区域预测和疏水性分析.结果表明,克隆获得了甘蔗AAC cDNA序列的全长为1170bp,包括一个可编码387个氨基酸的完整ORF.生物信息学分析显示,AAC家族基因的N端序列相对不保守,C端序列高度保守;其分子量是42.265 kD,理论等电点是9.79,预测其为稳定的蛋白;GRAVY值为-0.099;含有6个跨膜区域;其6个跨膜螺旋(TM1-TM6)跨越磷脂双分子层结构,盐桥可能是通过中间的3个连接螺旋α1～α3共同构成的;定位在细胞质的概率为60.9%.获得了甘蔗AAC完整ORF序列,并通过生物信息学预测该基因所编码的蛋白为一个稳定跨膜蛋白,且可能定位在细胞质,为甘蔗AAC基因的深入研究和应用奠定初步基础.     

可变径管道检测器皮碗结构设计及密封性能

可变径油气管道检测器是一种可以同时对一个发、收球站间距内的不同口径管道进行在役检测的管道检测设备,其优点是通过能力强、适用范围广、经济性好。通过对实现变径检测的可变径皮碗的结构设计及密封性能进行研究,提出一套适用于油气介质的40～48 in(1 in=25.4 mm)可变径皮碗结构。利用有限元仿真分析、可变径管道检测器牵拉、气体密封驱动测试方法,验证了可变径皮碗结构方案的合理性和可行性。通过现场工业应用,证明可变径皮碗结构设计及密封性能均满足设计指标和现场应用要求。打破了国外长期对可变径清管与检测技术的垄断,为中国自主研发可变径清管及检测装备早日实现系列化工业现场应用奠定了坚实的技术基础。(图7,表2,参20)     

南方冬种马铃薯分级机结构设计

针对南方冬种马铃薯的物理特性和分级要求,分析已有果蔬分级设备的优缺点,采用辊杆式分级方法,确定了分级机的整体结构,并用Solidworks绘出其结构图.对主要零部件进行了结构设计和参数计算,为样机制作提供了理论基础.     

南方冬种马铃薯分级机结构设计

针对南方冬种马铃薯的物理特性和分级要求,分析已有果蔬分级设备的优缺点,采用辊杆式分级方法,确定了分级机的整体结构,并用Solidworks绘出其结构图.对主要零部件进行了结构设计和参数计算,为样机制作提供了理论基础.     

Crystal growth inhibitors for the prevention of L-cystine kidney stones through molecular design

Crystallization of L-cystine is a critical step in the pathogenesis of cystine kidney stones. Treatments for this disease are somewhat effective but often lead to adverse side effects. Real-time in situ atomic force microscopy (AFM) reveals that L-cystine dimethylester (L-CDME) and L-cystine methylester (L-CME) dramatically reduce the growth velocity of the six symmetry-equivalent {100} steps because of specific binding at the crystal surface, which frustrates the attachment of L-cystine molecules. L-CDME and L-CME produce l-cystine crystals with different habits that reveal distinct binding modes at the crystal surfaces. The AFM observations are mirrored by reduced crystal yield and crystal size in the presence of L-CDME and L-CME, collectively suggesting a new pathway to the prevention of L-cystine stones by rational design of crystal growth inhibitors.     

六旋翼植保无人机飞行框架的结构设计与试验

为了达到无人机轻量化的要求，以碳纤维材质为机身主体，设计了一套六旋翼植保无人机的飞行框架。基于ANSYS对该框架关键结构部件的刚度和强度进行了静力学分析，根据分析结果，将单层中心固定板改为了双层结构并在机臂之间加入了加强筋。分析表明：优化设计后，无人机中心板形变量降低了104.36倍，最大应力降低了13.15倍，而质量仅增加了1.9%。最后通过实际飞行振动试验，基于LabVIEW对植保无人机机身3个不同位置竖直方向的振动信号进行小波变换降噪处理和时、频域计算评估，振动最大RMS值为1.69 g，小于飞行控制系统振动报警阈值2.5 g，表明其在稳定性和操控能力上具有良好的表现。     

气力式谷子精量排种器结构设计及性能试验

针对现有气力式排种器对谷子等小粒径种子难以实现精量播种问题,根据谷子形状等物理特性,通过优化排种器结构,设计一种舀勺-气吸组合式谷子精量排种器,研究排种器关键部件结构与参数对谷子精量排种性能的影响。应用JPS-12排种器性能检测试验台,以排种轴转速、舀种勺位置夹角和舀种勺圆心角为试验因素,以穴粒数合格指数、穴距合格指数、空穴指数及穴距变异系数为试验指标,进行三因素三水平正交试验,分析各试验因素对于性能指标影响的显著性。结果表明:排种器较优的工作参数为,舀种勺圆心角45°、舀种勺位置夹角-15°、排种轴转速10 r/min;优化定型后舀勺-气吸组合式谷子精量排种器综合性能试验指标为,穴粒数合格指数88.7%,穴距合格指数87.7%,空穴指数1.3%,穴距变异系数12.3%。该排种器作业性能满足小粒径作物精量播种和农艺要求。     

管径508 mm单节双封型内封堵器结构设计及封堵性能

随着油气管道管内高压封堵技术的发展,为了提高管内高压封堵器封堵的安全性和可靠性,基于对国外管内封堵器结构设计现状的充分调研,提出了一种适用于管径为508 mm油气管道的单节双封型内封堵器结构方案,并依据设计性能指标进行了封堵能力的理论分析计算。利用有限元仿真分析方法对设计的单节双封型管内封堵器的封堵能力进行仿真分析,仿真分析结果高于指标要求的理论计算值,说明设计的单节双封型管内封堵器封堵能力满足设计指标要求。该研究可为单节双封型管内封堵器结构设计提供参考,对中国管内高压封堵技术水平的提升及相关装备的高质量发展具有一定的推动作用。     

油气管道的锚固法兰结构设计与有限元分析

锚固法兰按照常规计算方法得到的各部位尺寸较为保守,重要部位的结构尺寸往往不合理,局部应力过大,缺少必要的分析优化过程.以某规格高压、大口径锚固法兰为例,采用理论计算与有限元分析方法相结合进行设计计算.结果表明:在法兰的圆角处和管颈处有最大的应力强度分布,其余位置的各应力分量均较小,强度校核满足要求,结构设计合理.因此,锚固法兰设计计算结合有限元分析方法,可确保设计质量,满足不同工程项目锚固的技术要求.