The role of the automation development group in analytical research and development at Dupont Merck

Laboratory robotics has been firmly established in many non-QC laboratories as a valuable tool for automating pharmaceutical dosage form analysis. Often a single project or product line is used to justify an initial robot purchase thus introducing robotics to the laboratory for the first time. However, to gain widespread acceptance within the laboratory and to justify further investment in robotics, existing robots must be used to develop analyses for existing manual methods as well as new projects beyond the scope off the original purchase justification. The Automation Development Group in Analytical Research and Development is a team of analysts primarily devoted to developing new methods and adapting existing methods for the robot. This team approach developed the expertise and synergy necessary to significantly expand the contribution of robotics to automation in the authors'' laboratory.     

Recent advances in microbial production of phenolic compounds

Phenolic compounds (PCs) are a group of compounds with various applications in nutraceutical, pharmaceutical and cosmetic industries. Their supply by plant extraction and chemical synthesis is often limited by low yield and high cost. Microbial production represents as a promising alternative for efficient and sustainable production of PCs. In this review, we summarize recent advances in this field, which include enzyme mining and engineering to construct artificial pathways, balance of enzyme expression to improve pathway efficiency, coculture engineering to alleviate metabolic burden and side-reactions, and the use of genetic circuits for dynamic regulation and high throughput screening. Finally, current challenges and future perspectives for efficient production of PCs are also discussed.     

Pluripotent Stem Cell-Derived Hepatocytes Phenotypic Screening Reveals Small Molecules Targeting the CDK2/4-C/EBPα/DGAT2 Pathway Preventing ER-Stress Induced Lipid Accumulation

Non-alcoholic fatty liver disease (NAFLD) has a large impact on global health. At the onset of disease, NAFLD is characterized by hepatic steatosis defined by the accumulation of triglycerides stored as lipid droplets. Developing therapeutics against NAFLD and progression to non-alcoholic steatohepatitis (NASH) remains a high priority in the medical and scientific community. Drug discovery programs to identify potential therapeutic compounds have supported high throughput/high-content screening of in vitro human-relevant models of NAFLD to accelerate development of efficacious anti-steatotic medicines. Human induced pluripotent stem cell (hiPSC) technology is a powerful platform for disease modeling and therapeutic assessment for cell-based therapy and personalized medicine. In this study, we applied AstraZeneca’s chemogenomic library, hiPSC technology and multiplexed high content screening to identify compounds that significantly reduced intracellular neutral lipid content. Among 13,000 compounds screened, we identified hits that protect against hiPSC-derived hepatic endoplasmic reticulum stress-induced steatosis by a mechanism of action including inhibition of the cyclin D3-cyclin-dependent kinase 2-4 (CDK2-4)/CCAAT-enhancer-binding proteins (C/EBPα)/diacylglycerol acyltransferase 2 (DGAT2) pathway, followed by alteration of the expression of downstream genes related to NAFLD. These findings demonstrate that our phenotypic platform provides a reliable approach in drug discovery, to identify novel drugs for treatment of fatty liver disease as well as to elucidate their underlying mechanisms.     

利用EROD生物测试法快速筛选焚烧废气中二恶英类化合物

二恶英类化合物的检测是垃圾焚烧废气检测中的重要项目,以HRGC-HRMS高分辨气质法为主的仪器分析方法的通量低且检测成本高,不利于对大量样品进行快速的检测。本文结合一套新型样品前处理柱,开发了EROD生物测试法用于垃圾焚烧炉烟道废气样品中二恶英类化合物的检测,该方法具有通量大、检测成本低等优点。在检测过程中,同时比对了EROD生物测试法和HRGC-HRMS法两种方法,发现两者具有强相关性(R2=0.95)。这一结果表明,结合新型前处理柱的EROD生物测试法能够得到与高分辨气质仪器分析方法具有相近TEQ值的检测结果,适合于垃圾焚烧废气样品中的二恶英类化合物的快速筛查检测。     

Inhibition of Eimeria tenella CDK‐Related Kinase 2: From Target Identification to Lead Compounds

Apicomplexan parasites encompass several human‐ and animal‐pathogenic protozoans such as Plasmodium falciparum, Toxoplasma gondii, and Eimeria tenella. E. tenella causes coccidiosis, a disease that afflicts chickens, leading to tremendous economic losses to the global poultry industry. The considerable increase in drug resistance makes it necessary to develop new therapeutic strategies against this parasite. Cyclin‐dependent kinases (CDKs) are key molecules in cell‐cycle regulation and are therefore prominent target proteins in parasitic diseases. Bioinformatics analysis revealed four potential CDK‐like proteins, of which one—E. tenella CDK‐related kinase 2 (EtCRK2)—has already been characterized by gene cloning and expression. 1 By using the CDK‐specific inhibitor flavopiridol in EtCRK2 enzyme assays and schizont maturation assays (SMA), we could chemically validate CDK‐like proteins as potential drug targets. An X‐ray crystal structure of human CDK2 (HsCDK2) served as a template to build protein models of EtCRK2 by comparative homology modeling. Structural differences in the ATP binding site between EtCRK2 and HsCDK2, as well as chicken CDK3, were addressed for the optimization of selective ATP‐competitive inhibitors. Virtual screening and “wet‐bench” high‐throughput screening campaigns on large compound libraries resulted in an initial set of hit compounds. These compounds were further analyzed and characterized, leading to a set of four promising lead compounds for development as EtCRK2 inhibitors.     

Managing laboratory automation in a changing pharmaceutical industry

The health care reform movement in the USA and increased requirements by regulatory agencies continue to have a major impact on the pharmaceutical industry and the laboratory. Laboratory management is expected to improve effciency by providing more analytical results at a lower cost, increasing customer service, reducing cycle time, while ensuring accurate results and more effective use of their staff. To achieve these expectations, many laboratories are using robotics and automated work stations. Establishing automated systems presents many challenges for laboratory management, including project and hardware selection, budget justification, implementation, validation, training, and support. To address these management challenges, the rationale for project selection and implementation, the obstacles encountered, project outcome, and learning points for several automated systems recently implemented in the Quality Control Laboratories at Eli Lilly are presented.     

Recent advances in flexible sensors for wearable and implantable devices

Flexible devices are emerging as important applications for future display, robotics, in vitro diagnostics, advanced therapies, and energy harvesting. In this review, we provide an overview of recent achievements in flexible mechanical and electrical sensing devices, focusing on the properties and functions of polymeric layers. In the order of historical development, sensing platforms are classified into four types: electronic skins for robotics and medical applications, wearable devices for in vitro diagnostics, implantable devices for human organs or tissues for surgical applications, and advanced sensing devices with additional features such as transparency, self‐power, and self‐healing. In all of these examples, a polymer layer is used as a versatile component including a flexible structural support and a functional material to generate, transmit, and process mechanical and electrical inputs in various ways. We briefly discuss some outlooks and future challenges toward the next steps for flexible devices. © 2013 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 130: 1429–1441, 2013     

Systematic Extraction of Structure–Activity Relationship Information from Biological Screening Data

A data mining approach is introduced that automatically extracts SAR information from high‐throughput screening data sets and that helps to select active compounds for chemical exploration and hit‐to‐lead projects. SAR pathways are systematically identified consisting of sequences of similar active compounds with gradual increases in potency. Fully enumerated SAR pathway sets are subjected to pathway scoring, filtering, and mining, and pathways with the most significant SAR information content are prioritized. High‐scoring SAR pathways often reveal activity cliffs contained in screening data. Subsets of SAR pathways are analyzed in SAR trees that make it possible to identify microenvironments of significant SAR discontinuity from which hits are preferentially selected. SAR trees of alternative pathways leading to activity cliffs identify key compounds and help to develop chemically intuitive SAR hypotheses.     

A NIR luminescent copolymer based on platinum porphyrin as high permeable dissolved oxygen sensor for microbioreactors

Microbioreactors with multioptical sensors have become increasingly important because of their small working volumes, high degree of parallelization and the available robotics. A novel hydrophobic luminescent copolymer P(Pt‐TPP‐TFEMA) along with reference P(Pt‐TPP‐EMA) containing the pendant group of 5,10,15,20‐tetraphenylporphyrin (TPP) moiety as low‐cost dissolved oxygen (DO) chemosensor film for high‐throughput microbioreactors is designed. Its sensor film exhibits fast response to DO with good stability and fatigue resistance, being capable of applying as a low‐cost DO indicator for high‐throughput bioprocess measurement. Results show that the quenching response of DO increases with the enhancement in the copolymeric hydrophobicity using the presented hybrid fluorinated ethyl methacrylate. Furthermore, the long emission band at 650 nm of chromophore TPP with large Stoke's shift about 250 nm brings several advantages, such as low scattering, deep penetration, and minimal interferences of absorption and fluorescence from the fermentation system, which shows high‐promising application in bioprocess monitoring. © 2013 American Institute of Chemical Engineers AIChE J, 59: 2743–2752, 2013     

客户管理系统开发与应用

本软件开发的主要任务是完成与客户相关的资料的系统化、自动化和规范化。系统为满足使用者的功能需求,要求用面向对象(OPP)的程序设计、开放式关系数据库连接(ODBC)及对象嵌入与链接等技术,按照软件工程原理设计系统,面向对象软件开发模式,将软件分解成模块。本系统具有完善的基础维护信息和客户信息维护功能,并增置了金牌客户服务模块,满足了所有客户日常管理的需求。     

Prospects and Challenges of Developing Plant-Derived Snake Antivenin Natural Products: A Focus on West Africa

Snakebite envenomation (SBE) is an important public health issue that is now receiving renewed attention following its reclassification as a Neglected Tropical Disease (NTD). Most incidences occur in rural areas of resource-limited countries, as such, timely and appropriate medical care for SBE is often inaccessible. The administration of anti-snake venom serum (ASV) is the only effective definitive treatment of SBE, but treatment failure to available ASVs is not uncommon. Emerging evidence highlights the potential of small-molecule compounds as inhibitors against toxins of snake venom. This presents an encouraging prospect to develop an alternative therapeutic option for the treatment SBE, that may be amenable for use at the point of care in resource-constraint settings. In view of the pivotal role of natural products in modern drug discovery programmes, there is considerable interest in ethno-pharmacological mining of medicinal plants and plant-derived medicinal compounds toward developing novel snake venom-neutralising therapeutics. In this review, we compile a collection of medicinal plants used in the treatment of SBE in West Africa and highlight their promise as potential botanical drugs or as sources of novel small-molecule compounds for the treatment of SBE. The challenges that must be surmounted to bring this to fruition including the need for (sub) regional collaboration have been discussed.     

Rationalization of Activity Cliffs of a Sulfonamide Inhibitor of DNA Methyltransferases with Induced-Fit Docking

Inhibitors of human DNA methyltransferases (DNMT) are of increasing interest to develop novel epi-drugs for the treatment of cancer and other diseases. As the number of compounds with reported DNMT inhibition is increasing, molecular docking is shedding light to elucidate their mechanism of action and further interpret structure–activity relationships. Herein, we present a structure-based rationalization of the activity of SW155246, a distinct sulfonamide compound recently reported as an inhibitor of human DNMT1 obtained from high-throughput screening. We used flexible and induce-fit docking to develop a binding model of SW155246 with a crystallographic structure of human DNMT1. Results were in excellent agreement with experimental information providing a three-dimensional structural interpretation of ‘activity cliffs’, e.g., analogues of SW155246 with a high structural similarity to the sulfonamide compound, but with no activity in the enzymatic assay.     

Modeling promiscuity based on in vitro safety pharmacology profiling data

This study describes a method for mining and modeling binding data obtained from a large panel of targets (in vitro safety pharmacology) to distinguish differences between promiscuous and selective compounds. Two naïve Bayes models for promiscuity and selectivity were generated and validated on a test set as well as publicly available drug databases. The model shows a higher score (lower promiscuity) for marketed drugs than for compounds in early development or compounds that failed during clinical development. Such models can be used in triaging high‐throughput screening data or for lead optimization.     

明日仓储今日观

由于有了电子商务、订单完成与产品质量日益变得同样重要，你的仓库是否也能胜任呢？挑选灵活性高、适应性强的设施、你就会永远能够及时地为你的客护提供所需的服务，他们也将会继续成为你忠实的客户。     

基于服务利润链视角的营销服务文化建设——以中国石化化工销售有限公司为例

中国石化化工销售有限公司应用服务利润链理论研究，连续4年开展的“服务年”系列活动，提炼了营销服务文化理念，传播服务理念，得到员工的认同，并转化为服务行为。实践证明，利润、员工、企业和客户确实存在着直接的联系，内部员工是基础，通过员工服务来提高企业和客户的满意度、忠诚度，最终促进企业自身持续发展。     

Perspectives on weatherability testing of automotive coatings

Automotive coatings continue to evolve rapidly in order to reduce the environmental impact of coating operations and to meet rising customer expectations for appearance and performance. The anticipated changes in chemistry, together with improved appearance retention and the need to respond rapidly to customer desires, have placed new challenges on weathering protocols. This paper discusses these issues and describes several methodologies that are likely to be part of the next generation of accelerated testing protocols. Of critical importance is the need to understand and minimize where possible sources of variability in testing that can lead to uncertain results and increased risk of failure in service. Research Laboratory MD-3182, P.O.Box 2053, Dearborn, MI 41821.     

Fragment-Based Screening by Protein Crystallography: Successes and Pitfalls

Fragment-based drug discovery (FBDD) concerns the screening of low-molecular weight compounds against macromolecular targets of clinical relevance. These compounds act as starting points for the development of drugs. FBDD has evolved and grown in popularity over the past 15 years. In this paper, the rationale and technology behind the use of X-ray crystallography in fragment based screening (FBS) will be described, including fragment library design and use of synchrotron radiation and robotics for high-throughput X-ray data collection. Some recent uses of crystallography in FBS will be described in detail, including interrogation of the drug targets β-secretase, phenylethanolamine N-methyltransferase, phosphodiesterase 4A and Hsp90. These examples provide illustrations of projects where crystallography is straightforward or difficult, and where other screening methods can help overcome the limitations of crystallography necessitated by diffraction quality.     

Inhibitors of the salicylate synthase (MbtI) from Mycobacterium tuberculosis discovered by high-throughput screening

A simple steady‐state kinetic high‐throughput assay was developed for the salicylate synthase MbtI from Mycobacterium tuberculosis, which catalyzes the first committed step of mycobactin biosynthesis. The mycobactins are small‐molecule iron chelators produced by M. tuberculosis, and their biosynthesis has been identified as a promising target for the development of new antitubercular agents. The assay was miniaturized to a 384‐well plate format and high‐throughput screening was performed at the National Screening Laboratory for the Regional Centers of Excellence in Biodefense and Emerging Infectious Diseases (NSRB). Three classes of compounds were identified comprising the benzisothiazolones (class I), diarylsulfones (class II), and benzimidazole‐2‐thiones (class III). Each of these compound series was further pursued to investigate their biochemical mechanism and structure–activity relationships. Benzimidazole‐2‐thione 4 emerged as the most promising inhibitor owing to its potent reversible inhibition.     

Unlocking the potential of asymmetric hydrogenation at Merck

This Account outlines the efforts of Merck scientists toward implementing asymmetric hydrogenation as a core competency within Merck Research Laboratories. Several key factors are discussed including (i) a focus on efficient chemical synthesis, (ii) implementation of high throughput screening (HTS) techniques, (iii) demonstration of robustness on scale, and (iv) diligence to ensure freedom of operation and catalyst supply for manufacturing. Several examples of the development of efficient asymmetric hydrogenation processes are described.     

High‐Throughput Screening Technology for Automotive Applications

The development of advanced emission control systems to meet the strict regulations requires efficient and flexible material screening capabilities. Here, a high throughput test unit is described. Two case studies demonstrate the rapid screening of relevant parameter spaces and material functionalities which can be used in product development. One involves steady‐state testing of hydrocarbon oxidation in Diesel aftertreatment systems, while the other shows the evaluation of oxygen storage capacity in the optimization of three‐way catalysts for gasoline engines.