High-throughput experimentation enables rapid reaction optimization by leveraging reaction miniaturization, automated processes, and advanced high-throughput analytical methods. To more effectively apply these principles to reactions mediated by photoredox catalysis, we developed the photoredox optimization (PRO) reactor. PRO is an automated platform that provides precise control over the delivered light irradiance to optically thin, temperature-controlled reaction volumes. Combined with high-intensity laser illumination, PRO facilitates accelerated photoredox reaction scouting using <10 mu L of reaction material. Crude products from PRO reactions are automatically transferred to microplates for analysis by infrared matrix-assisted laser desorption electrospray ionization mass spectrometry (IR-MALDESI-MS) which can quantify 384 reactions in under 6 minutes. Validation of the PRO reactor was achieved through a series of challenging decarboxylative cross-coupling reactions, which resulted in improved isolated yields up to 58%. PRO then enabled the design and execution of higher throughput 384-reaction HTE arrays which achieved improved yields for two previously unsuccessful photoredox cross-couplings, ultimately identifying novel reaction conditions outside the scope of our traditional 96-reaction arrays.

Over the last 5 years, IR-MALDESI-MS (Infrared Matrix-Assisted Laser Desorption Electrospray Ionization Mass Spectrometry) has been demonstrated for use in a range of high-throughput biochemical and cellular assays with remarkable sample acquisition rates up to 22 Hz for a single 384-well assay plate. With such high single plate acquisition rates, the rate limiting step becomes how fast subsequent plates can be presented to the MS for analysis. To make this transfer as fast as possible while maintaining safe operation in a laboratory environment, we developed a collaborative robotic plate transfer system (CRPTS) that combines a 6-axis robot with dual plate grippers, a 7th axis conveyor stage, and a 420-plate capacity sample loading window. As a demonstration of the throughput and flexibility of CRPTS, we performed a biochemical assay that monitored the oxidation of tris(2-carboxyethyl)phosphine (TCEP) to screen for nuisance compounds. Using continuous and step motion scan profiles, we analyzed 158,799 compounds contained in 448 assay plates over the course of 12.5 h (Z-Factor=0.87) and 17.5 h (Z-factor=0.99), respectively. Extrapolating these results enables the screening of a million compounds within 6-7 working days.

Intravenous (IV) administration of poorly water-soluble small molecule therapeutics can lead to precipitation during mixing with blood. This can limit characterization of pharmacological and safety endpoints in preclinical models. Most often, tests of kinetic and thermodynamic solubility are used to optimize the formulation for solubility prior to infusion in animals, but these do not capture the dynamic precipitation processes that take place during in-vivo administration. To better capture the fluid dynamic processes that occur during IV administration, we developed the Optical Spatial PREcipitation analYzer (OSPREY) as a method to quantify the amount and size of compound precipitates in whole blood using a flow-through system that mimics IV administration. Here, we describe the OSPREY device and its underlying imaging processing methods. We then validate the ability to accurately segment particles according to their size using monodisperse suspensions of microspheres (diameter 50 to 425 µm). Next, we use a tool compound, ABT-737, to study the effects of compound concentration, vessel flow rate, compound infusion rate and vessel diameter on precipitation. Finally, we use the physiological diameter and flow rate of rat femoral vein and dog saphenous vein to demonstrate the potential of OSPREY to model in-vivo precipitation in a controlled, dynamic in-vitro assay.

Infrared matrix-assisted laser desorption electrospray ionization (IR-MALDESI) mass spectrometry is an ambient-direct sampling method that is being developed for high-throughput, label-free, biochemical screening of large-scale compound libraries. Here, we report the development of an ultra-high-throughput continuous motion IR-MALDESI sampling approach capable of acquiring data at rates up to 22.7 samples per second in a 384-well microtiter plate. At top speed, less than 1% analyte carryover is observed from well-to-well, and signal intensity relative standard deviations (RSD) of 11.5% and 20.9% for 3 μM 1-hydroxymidazolam and 12 μM dextrorphan, respectively, are achieved. The ability to perform parallel kinetics studies on 384 samples with a ∼30 s time resolution using an isocitrate dehydrogenase 1 (IDH1) enzyme assay is shown. Finally, we demonstrate the repeatability and throughput of our approach by measuring 115200 samples from 300 microtiter plate reads consecutively over 5.54 h with RSDs under 8.14% for each freshly introduced plate. Taken together, these results demonstrate the use of IR-MALDESI at sample acquisition rates that surpass other currently reported direct sampling mass spectrometry approaches used for high-throughput compound screening.

Centralized high-throughput purification laboratories routinely produce large numbers of test tubes with fractions containing the purified compounds of interest interspersed with test tubes containing fractions collected from undesired peaks. Because the next step after purification entails the removal of the solvent in a centrifugal evaporator with multiple sample positions per rotor, select test tubes must be labeled prior to dry-down to track the identity of each compound. The diversity of test tube sizes and tray configurations from different chromatography system vendors complicates this labeling task. Therefore, the development of an automated tube labeler that can accommodate a multitude of test tube and tray sizes can reduce the chances of error as well as reduce the hands-on labor required to complete this tedious but essential task. Custom hardware and software have been implemented to inform and to enable the Pick-n-Place arm of a commercially available Tecan EVO robotic system to pick up and present select tubes, filled with purified chromatography fractions from a multitude of vendor trays, to a custom label application station integrated with a commercially available Zebra label printer. Particular challenges existed with accurately positioning tubes in Agilent G1364-84544 trays onto the deck of the instrument. The resulting instrument reduces hands-on time for labeling fractions by approximately 60%.

Automation of chemistry at a pharmaceutical company commonly entails bringing commercial solutions in-house, reproducing manual processes with a robot, or integrating multiple instruments to eliminate human intervention. A strategy of industrializing proven approaches, while financially justifiable, however, does not encourage innovation. On the other hand, trying to automate unproven or difficult processes may seem to be risky but can actually accelerate the adoption, modification, or rejection of novel technologies. Having chemists and engineers work together to develop automation that accelerates the development and evaluation of innovative concepts is one blueprint for delivering a competitive advantage to an organization.

Neural recording and stimulation with high spatial and temporal resolution are highly desirable in the study of neurocommunication and diseases. Planar multiple microelectrode arrays (MEA) or quasi-three-dimensional (3D) MEA with fixed height have been proposed by many researchers and become commercially available. In this paper, we present the design, fabrication, and test of a novel true 3D multiple electrode array for brain slice stimulation and recording. This MEA is composed of 105 microelectrodes with 50 μm diameter and 125 μm center-to-center spacing integrated in a 1.2 × 1.2 mm2 area. This "true" 3D MEA allows us to precisely position the individual electrodes by piezoelectric-based actuators to penetrate the inactive tissue layer and to approach the active neurons so as to optimize the recording and stimulation of electrical field potential. The capability to stimulate nerve fibers and record postsynaptic field potentials was demonstrated in an experiment using mouse brain hippocampus slice.

The efficient synthesis of cyclopropyl boronic esters in library format using a diazomethane flow reactor has been achieved. A pivotal component of the system is a fully automated tube-in-tube reactor allowing for safe handling of hazardous diazomethane on repeated small scale and for the generation of larger quantities of product. The setup enables the repeated execution of Pd-catalyzed cyclopropanation reactions without compromising its operation over time.

A flexible and integrated flow-chemistry–synthesis–purification compound-generation and sample-management platform has been developed to accelerate the production of small-molecule organic-compound drug candidates in pharmaceutical research. Central to the integrated system is a Mitsubishi robot, which hands off samples throughout the process to the next station, including synthesis and purification, sample dispensing for purity and quantification analysis, dry-down, and aliquot generation.

We report herein a high-throughput integrated synthesis—purification platform termed SWIFT (synthesis with integrated-flow technology) and processes that accelerate the rate at which validated small-molecule organic compounds are generated. A segmented-flow synthesizer was integrated to a preparative HPLC-MS, where each reaction product was purified immediately upon reaction completion. Further, automated structure-validation processes accelerate the rate at which drug discovery candidates are available for biological screening.

La presente invention concerne un procede rapide et efficace et un appareil pour detecter des effets electrophysiologiques, proarythmiques, contractiles, et autres de substances telles que des composes et des medicaments dans des preparations cardiaques cellulaires natives, les preparations representant une reponse pharmacologique cellulaire. Plus specifiquement, la presente invention concerne un procede pour (1) detecter rapidement et efficacement et verifier les effets de substances chimiques, composes et medicaments sur la repolarisation, la contractilite et l'excitabilite cardiaques en utilisant des techniques optiques et des protocoles de simulation personnalises, et (2) cribler rapidement et efficacement et selectionner des composes pour des effets electrophysiologiques et proarythmiques sur des myocytes cardiaques.

Experiments were performed to demonstrate the potential of counter-current chromatography (CCC) for the isolation of drugs and their metabolites from biological matrices relevant to the metabolism studies of pharmaceutical research. Examples of typical drugs are spiked into biological media ex vivo to provide test samples for analysis. A mass spectrometer hyphenated to a CCC allows for the detection of small molecule drugs within the matrix through selected ion monitoring, and fraction collection can provide material for further structural elucidation by NMR.

This manuscript details the construction of a fully automated flow hydrogenation apparatus for use in high-throughput organic synthesis. The instrument comprises of a Bohdan robot platform coupled with a ThalesNano H-cube hydrogenator and a series of solvent valves and pumping mechanisms. Using this instrument, we have been able to fully automate a number of key transformations that could not otherwise be conveniently undertaken in a high-throughput manner.

Experiments were performed to evaluate whether counter-current chromatography (CCC) could function as an alternative purification method to reversed-phase high-performance liquid chromatography (RP-HPLC) and normal-phase supercritical fluid chromatography (SFC). RP-HPLC and SFC are the routine methods currently used in our high-throughput purification (HTP) facility for the purification of high-throughput organic synthesis (HTOS) libraries and medicinal chemistry reaction mixtures. Pre-equilibration of the solvent mixture layers was not mandatory for effective chromatography when hexanes–ethyl acetate–methanol–water (HEMW) solvent mixtures were used. Key to the use of CCC for high-throughput applications is the ability to effectively select a solvent system appropriate to each library member. Pilot-scale CCC elution time was used to estimate a starting solvent ratio and RP-HPLC retention time was then used to adjust solvent ratios within a particular library. It was also found that dimethyl sulfoxide (DMSO) and DMSO–methanol were suitable as sample injection solvents when using the HEMW solvent systems.

The emerging field of biomedical microelectromechanical systems (bioMEMS) has the potential to revolutionize how drugs are discovered, how diseases are diagnosed, and how treatments are administered. Nevertheless, that potential is unlikely to translate into commercial success unless bioMEMS designers can prove that such novel functionality can be delivered each and every time without failure. The life-and-death ramifications of a system failure make absolute reliability a moral and regulatory requirement for many medical applications. This paper reveals the reliability hurdles that must be overcome when systems are used for pharmaceutical research, clinical diagnostics, or implantation into humans. Special attention is given to challenges that are likely to be exacerbated for miniaturized devices. Techniques useful for characterizing, minimizing, and monitoring for failures are also described. A strategy for addressing reliability problems is also presented. By refining bioMEMS for the research laboratory first and then applying the experience gained from that environment to clinical diagnostics applications, bioMEMS does not need to confront head-on the regulatory hurdles and established technologies that could otherwise impede its entry. Similarly, development of more robust, miniaturized clinical diagnostics systems can simplify the entry of implanted bioMEMS.

The modulation of fatty acid metabolism and especially the stimulation of fatty acid oxidation in liver or skeletal muscle are attractive therapeutic approaches for the treatment of obesity and the associated insulin resistance. However, current beta-oxidation assays are run in very low throughput, which represents an obstacle for drug discovery in this area. Here we describe results for a 48-well beta-oxidation assay using a new instrument design. A connecting chamber links two adjacent wells to form an experimental unit, in which one well contains the beta-oxidation reaction and the other captures CO(2). The experimental units are sealed from each other and from the outside to prevent release of radioactivity from the labeled substrate. CO(2) capture in this instrument is linear with time and over the relevant experimental range of substrate concentration. Cellular viability is maintained in the sealed environment, and cells show the expected responses to modulators of beta-oxidation, such as the AMP kinase activator 5-aminoimidazole carboxamide riboside. Data are presented for different lipid substrates and cell lines. The increased throughput of this procedure compared with previously described methods should facilitate the evaluation of compounds that modulate fatty acid metabolism.