Clive Green

AZD5438, a potent oral inhibitor of cyclin-dependent kinases 1, 2, and 9, leads to pharmacodynamic changes and potent antitumor effects in human tumor xenografts

Deregulation of the cell cycle has long been recognized as an essential driver of tumorigenesis, and agents that selectively target key cell cycle components continue to hold promise as potential therapeutics. We have developed AZD5438, a 4-(1-isopropyl-2-methylimidazol-5-yl)-2-(4-methylsulphonylanilino) pyrimidine, as a potent inhibitor of cyclin-dependent kinase (cdk) 1, 2, and 9 (IC50, 16, 6, and 20 nmol/L, respectively). In vitro, AZD5438 showed significant antiproliferative activity in human tumor cell lines (IC50 range, 0.2–1.7 μmol/L), causing inhibition of the phosphorylation of cdk substrates pRb, nucleolin, protein phosphatase 1a, and RNA polymerase II COOH-terminal domain and blocking cell cycling at G2-M, S, and G1 phases. In vivo, when orally administered at either 50 mg/kg twice daily or 75 mg/kg once daily, AZD5438 inhibited human tumor xenograft growth (maximum percentage tumor growth inhibition, range, 38–153; P < 0.05). In vivo, AZD5438 reduced the proportion of actively cycling cells. Further pharmacodynamic analysis of AZD5438-treated SW620 xenografts showed that efficacious doses of AZD5438 (>40% tumor growth inhibition) maintained suppression of biomarkers, such as phospho-pRbSer249/Thr252, for up to 16 hours following a single oral dose. A comparison of different schedules indicated that chronic daily oral dosing provided optimal cover to ensure antitumor efficacy. These data indicate that broad cdk inhibition may provide an effective method to impair the dysregulated cell cycle that drives tumorigenesis and AZD5438 has the pharmacologic profile that provides an ideal probe to test this premise. [Mol Cancer Ther 2009;8(7):1856–66]

Identification, optimisation and in vivo evaluation of oxadiazole DGAT-1 inhibitors for the treatment of obesity and diabetes

A novel series of DGAT-1 inhibitors was discovered from an oxadiazole amide high throughput screening (HTS) hit. Optimisation of potency and ligand lipophilicity efficiency (LLE) resulted in a carboxylic acid containing clinical candidate 53 (AZD3988), which demonstrated excellent DGAT-1 potency (0.6 nM), good pharmacokinetics and pre-clinical in vivo efficacy that could be rationalised through a PK/PD relationship.

Imidazoles: Sar and Development of a Potent Class of Cyclin-Dependent Kinase Inhibitors.

An imidazole series of cyclin-dependent kinase (CDK) inhibitors has been developed. Protein inhibitor structure determination has provided an understanding of the emerging structure activity trends for the imidazole series. The introduction of a methyl sulfone at the aniline terminus led to a more orally bioavailable CDK inhibitor that was progressed into clinical development.

Imidazole pyrimidine amides as potent, orally bioavailable cyclin-dependent kinase inhibitors

The development of a novel series of imidazole pyrimidine amides as cyclin-dependent kinase (CDK) inhibitors is described. The series was found to have much improved CDK2 inhibition and potent in vitro anti-proliferative effects against cancer cell lines. Control of overall lipophilicity was important to achieve good in vitro potency along with acceptable physiochemical properties and margins against inhibition of both CYP isoforms and the hERG potassium ion channel. A compound with an attractive overall balance of properties was profiled in vivo and possessed suitable physiochemical and pharmacokinetic profiles for oral dosing.

Imidazole Piperazines: Sar and Development of a Potent Class of Cyclin-Dependent Kinase Inhibitors with a Novel Binding Mode.

A piperazine series of cyclin-dependent kinase (CDK) inhibitors have been identified. The compounds exhibit excellent physiochemical properties and a novel binding mode, whereby a bridging interaction via a water molecule with Asp 86 of CDK2, leads to selectivity for the CDK family of enzymes over other kinases. Piperazines 2e and 2i were subsequently shown to inhibit tumour growth when dosed orally in a nude mouse xenograft study. Additional chemical series that exploit this unexpected interaction with Asp 86 are also described.

Identification, optimization, and pharmacology of acylurea GHS-R1a inverse agonists.

Ghrelin plays a major physiological role in the control of food intake, and inverse agonists of the ghrelin receptor (GHS-R1a) are widely considered to offer utility as antiobesity agents by lowering the set-point for hunger between meals. We identified an acylurea series of ghrelin modulators from high throughput screening and optimized binding affinity through structure-activity relationship studies. Furthermore, we identified specific substructural changes, which switched partial agonist activity to inverse agonist activity, and optimized physicochemical and DMPK properties to afford the non-CNS penetrant inverse agonist 22 (AZ-GHS-22) and the CNS penetrant inverse agonist 38 (AZ-GHS-38). Free feeding efficacy experiments showed that CNS exposure was necessary to obtain reduced food intake in mice, and it was demonstrated using GHS-R1a null and wild-type mice that this effect operates through a mechanism involving GHS-R1a.

Why a Special Issue on Acoustic Liquid Handling

Certain topics stir excitement in the hearts and minds of scientists. Synthesizing new genes with never before seen capabilities, discovering cures for Ebola and other devastating diseases, devising new breakthrough drug therapy for cancer sufferers, expanding the understanding of protein structure via crystallography—those are topics that thrill and motivate. By comparison, liquid handling may appear prosaic. Whether it is a hose, a pipette, an eye-dropper, pistonor peristaltic-driven systems, or an ink-jet printer, there is a “been there, done that” attitude. It seems as though everything that could be said about a liquid handler has already been said many times over. So why, then, are we devoting an entire issue of the Journal of Laboratory Automation to one specific type of liquid handling—acoustic droplet ejection (ADE)? Precisely because ADE, a relative newcomer to liquid handling, enables new, far-reaching science. Experiments that were impossible for practical reasons become simple and easy. The ability to use sound waves to move liquids in precise nanoliter increments has wide-sweeping effects on driving exciting advances in science, including those topics mentioned above. ADE fundamentally changes what can be done, and it has facilitated breakthroughs in new areas of innovative research. ADE is not another evolutionary step in the lab. It is correctly described as truly revolutionary. The importance of control and precision with increasingly lower transfer volumes has become elevated as detection technologies have significantly improved, making miniaturization to highdensity formats not only possible but also desirable for both throughput and financial reasons. Already broadly employed in drug discovery, ADE is now becoming an essential part of many labs. Synthetic biology? Cures for emerging diseases? New therapies for cancer? All of these have exploited the advantages of ADE. Kanigowska et al. use ADE to generate new genes at radically lower costs. Their process would have been financially unfeasible were it not for ADE. This reduction in expenses echoes the drug discovery efforts reported by Wingfield et al. Rasmussen et al. use assay-ready plates made via ADE to test drugs against Ebola and other BSL-4 pathogens and open up faster, simpler ways to collaborate to end these scourges. Advances in individualized medicine that identify exactly what drugs or combinations of drugs are the best anticancer therapy for any specific individual are covered by Blom et al. and Kulesskiy et al. The miniaturization and speed provided by ADE move individualized systems medicine from theory to reality. Ericson et al. and Wu et al. use ADE to expand protein crystallography via miniaturization and crystallization with multiple ligands. Of great interest to attendees of SLAS2015, the SLAS International Conference and Exhibition in Washington, D.C., as well as to all people interested in faster high-throughput assays is the article describing the first ever acoustic interface to a mass spectrometer. In “Novel Acoustic Loading of a Mass Spectrometer: Toward Next-Generation High-Throughput MS Screening,” Sinclair et al. elaborate upon the 2015 SLAS Innovation Award–winning presentation on the coupling of mass spectrometry with ADE to analyze as many as three assays per second. In a related article, Chin et al. show how they combine ADE with matrix-assisted laser desorption/ ionization mass spectrometry (MALDI MS). As mentioned, ADE is used extensively in drug discovery. In this issue, Naylor et al. expand the use of ADE in compound management to show how it preserves the potency of peptides. Edwards et al. describe ADE-enabled high-throughput epigenetic methyltransferase assays. Acoustic droplet ejection is fundamentally different from earlier technologies. It uses focused sound to propel droplets of liquids upward from a source location to a destination. The roots of ADE go back a century, when Wood and Loomis used acoustic energy to propel droplets of liquid from a vessel. With modern ADE, a low level of sound energy is focused at the meniscus, causing a mound to form on the surface of the liquid. When a small additional amount of energy is put into the liquid, droplets are propelled upward to an inverted target or microtiter plate with unparalleled precision and accuracy. Most important for achieving high scientific impact, ADE is a completely touchless transfer method. Traditionally, the term noncontact transfer has meant that the device transferring 619712 JLAXXX10.1177/2211068215619712Journal of Laboratory AutomationOlechno et al. research-article2015

Consolidating a Distributed Compound Management Capability into a Single Installation: The Application of Overall Equipment Effectiveness to Determine Capacity Utilization

Compound management (CM) is a critical discipline enabling hit discovery through the production of assay-ready compound plates for screening. CM in pharma requires significant investments in manpower, capital equipment, repairs and maintenance, and information technology. These investments are at risk from external factors, for example, new technology rendering existing equipment obsolete and strategic site closures. At AstraZeneca, we faced the challenge of evaluating the number of CM sites required to support hit discovery in response to site closures and pressure on our operating budget. We reasoned that overall equipment effectiveness, a tool used extensively in the manufacturing sector, could determine the equipment capacity and appropriate number of sites. We identified automation downtime as the critical component governing capacity, and a connection between automation downtime and the availability of skilled staff. We demonstrated that sufficient production capacity existed in two sites to meet hit discovery demand without the requirement for an additional investment of

Alkynes to (E)-enolates using tandem catalysis: stereoselective anti-aldol and syn-[3,3]-rearrangement reactions

7 million in new facilities. In addition, we developed an automated capacity model that incorporated an extended working-day pattern as a solution for reducing automation downtime. The application of this solution enabled the transition to a single site, with an annual cost saving of

IMIDAZO [1,2-A] PYRIDINE HAVING ANTI-CELL-PROLIFERATION ACTIVITY

2.1 million.