RAPID AND ROBUST EARLY AD ME SOLUTI FOR PHARMACEUTICAL RESEARCH AND DEVELOPMENT

AGILENT EARLY ADME SOLUTIONS REALIZE EFFECTIVE PHARMACEUTICALS Risk Mitigation Through Early ADME Studies The discovery and development of novel therapeutics is highly complex, with companies constantly adjusting the balance between efcacy, safety, speed, and cost. Rising regulatory pressures and increasing efforts to reduce costly late-stage attrition puts a greater emphasis on obtaining robust ADME data as early as possible for drug candidates.1 By eliminating compounds with toxic liabilities during discovery, instead of during clinical development, more resources can be focused on compounds...

Analyses of samples from in vivo ADME studies, including bile, urine, feces, and plasma In vitro ADME assays, including metabolic stability, CYP inhibition/induction, permeability, and PgP inhibition High-throughput ADME assays Qualitative and quantitative ADME analyses Drug metabolite characterization, quantitation, and mixture analysis Metabolite purication Drug metabolite characterization Agilent Solutions ADME Workow ADME Applications Condently increase your odds of success with Agilent— Solutions for early ADME studies Agilent RapidFire/MS System p. 4 Agilent Bravo Automated Liquid Handling...

ACCELERATE ANALYSIS Unparalleled Throughput with RapidFire/MS Quickly advancing the safest drug candidates involves making go/no-go decisions on a large number of compounds based on early ADME data. This requires performing ADME studies on very large numbers of compounds in the discovery setting at the throughputs necessary to provide data quickly, without compromising on the quality of the information generated. The Agilent RapidFire/MS platform provides the unmatched throughput at the plate-reader speeds required to do just that. Each sample is processed in 6–10 seconds and a single 384-well...

Greater versatility For highly exible ADME studies, the RapidFire system is optimized for integration with Agilent Triple Quadrupole MS, TOF/MS, and Q-TOF/MS instruments. With TOF or Q-TOF/MS, quantitative data can be obtained in the same experiment that provides qualitative identication of the major metabolites, further increasing efciency (see p. 6). Figure 4. Data from eight different CYP enzyme/substrate pairs using traditional LC/MS and RapidFire/MS technology. The dotted line represents a line of unity.5 (See Agilent application note 5990-9184EN.) CLint RapidFire/MS (μL/min/mg) Uncompromising...

STREAMLINE WORKFLOWS Simultaneous Qualitative and Quantitative Measurements As the pressure to increase productivity continues to challenge the pharmaceutical industry, ADME scientists have been looking for ways to conduct experiments faster and more efciently. The Agilent 6500 series of Q-TOF LC/MS instruments offers the sensitivity, mass accuracy, and resolution required to enable simultaneous qualitative and quantitative data collection, greatly increasing efciency (Figure 6). High sensitivity and mass accuracy for increased productivity While Agilent’s 6400 Series Triple Quadrupole instruments...

MS/MS spectrum Figure 8. MS and MS/MS spectra of a buspirone monohydroxy metabolite from a 10-minute incubation sample (A). The sub-ppm mass accuracy of the precursor and fragment ions, along with the excellent isotopic fidelity (overall score > 99 %), provided high-confidence metabolite identification and structure elucidation Excellent separation and broad metabolite coverage was achieved in three and a half minutes (B). (See Agilent To achieve more efficient data acquisition, the Agilent 6550 Q-TOF provides a number of features that simplify and streamline experimental setup. With Q-TOF technology,...

MAXIMIZE UNDERSTANDING NMR Analysis for Detailed Structure Elucidation NMR is the gold-standard, unmatched analytical technique for structure verication and identication of partial or complete unknowns. Thus, it is the method of choice for determining structures of metabolites directly from biological matrices or, more routinely, following purication. Agilent offers a wide range of NMR systems for the in-depth structure analysis of candidate compounds and their metabolites, including high-eld NMR systems, advanced probe technologies, automated sample loading, and our powerful VnmrJ 3 software...

Figure 10. Agilent cold probes deliver data with higher signal-to-noise using limited sample in less time. This HSQC spectrum, collected on a 10 µg sample of retrorsine, represents routine analysis with a 600 MHz 3mm MicroSample Cold Probe. Sophisticated experiments, easy implementation The sample-centric design of VnmrJ 3 software makes it easy to set up experiments, collect data, and analyze results. With a large variety of built-in pulse sequences and experimental protocols, all organized for convenient setup and use, sophisticated experiments are easy to implement. While used most often for...

SIMPLIFY PURIFICATION Achieve the Highest Recovery and Purity High recovery rates and purity levels are key issues for the isolation and purication of valuable drug metabolites. Agilent offers robust preparative LC solutions for the purication of nanogram to gram sample quantities. The Agilent 1260 Innity Series Analytical-scale Purication System (Figure 12) handles ow rates from 100 μL/min to 10 mL/min at pressures up to 600 bar, making it the system of choice for metabolite purication in the nanogram to low milligram range, on columns with internal diameters between 2.1 and 9.4 mm. Additionally,...

Agilent fraction collector Conventional o t fraction collector action collec Fraction trigger from detector Lost proportion of fraction Spoilt proportion of fraction Delay volume (peak dispersion) Figure 13. Agilent fraction collectors are designed to have the lowest delay volumes to avoid peak dispersion and carry-over between fractions, ensuring the highest recovery and purity for your fractions, especially at low ow rates. (See Agilent publication 5990-8840EN) Inject Delay Calibration Sample (no column) mAU 1400 1000 600 200 Diverter Valve Fraction Delay Sensor Mass-triggered metabolite purication...