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Development and Application of an Automated Solution Stability Assay for Drug Discovery

P.O. Box CN 8000, Princeton, NJ 08543-8000 dil{at}wyeth.com

Edward H. Kerns

Hong Chen

Susan L. Petusky

Screening of solution stability provides an early alert on potential liabilities of drug candidates so that strategies can be developed to overcome the challenges. Afully automated solution stability assay has been developed to accelerate traditionalmanual operation. The assay uses the advanced capabilities of a high-performance liquid chromatography instrument that is present in many pharmaceutical research laboratories. The samples are prepared automatically by a temperature-controlled autosampler. The samples are delivered to the stability matrices, mixed, incubated, and injected at selected time points during the reaction time course. This automated process occurs without operator intervention, thus allowing 96 experiments to be run with0.5hof a scientist's time compared to 8 h for the same studywhenperformedmanually. Automationnotonly eliminates themanual operation but also improves accuracy and throughput. The assay protocol has been optimized to achieve homogenous mixing and eliminate carryover. The assay is robust, flexible, and high throughput. It can be used to study stability for a large number of samples undermultiple incubation conditions and has awide range of applications in drug discovery and development, such as screening compound stability in biological assaymedia, obtaining a stability-pH profile, surveying compound stability in physiological fluids, and performing development forced degradation and excipient compatibility.

Key Words: stability • automation • high throughput • stability-pH profile • hydrolysis • chemical stability • HPLC • LC-MS • kinetics

References

• Kerns EH, Di L: Pharmaceutical profiling in drug discovery. Drug Disc Today 2003;8:316-323.[CrossRef][Medline] [Order article via Infotrieve]
• Kerns EH, Di L: Accelerated stability profiling in drug discovery. Log P 2004 Proceeding 2004.
• Kibbey CE, Poole SK, Robinson E, Jackson JD, Durham D: An integrated process for measuring the physicochemical properties of drug candidates in a preclinical discovery environment. J Pharm Sci2001;90:1164-1175.[Medline] [Order article via Infotrieve]
• Shah KP, Zhou J, Lee R, Schowen RL, Elsbernd R, Ault JM, et al: Automated analytical systems for drug development studies. I—asystem for the determination of drug stability. J Pharm Biomed Anal1994;12:993-1001.[Medline] [Order article via Infotrieve]
• von Rauch M, Schlenk M, Gust R: Effects of C2-alkylation, N-alkylation, and N,N'-dialkylation on the stability and estrogen receptor interaction of (4R,5S)/ (4S,5R)-4,5-Bis(4-hydroxyphenyl)-2-imidazolines. J Med Chem 2004;47:915-927.[Medline] [Order article via Infotrieve]
• Magnin DR, Robl JA, Sulsky RB, Augeri DJ, Huang Y, Simpkins LM, et al: Synthesis of novel potent dipeptidyl peptidase IV inhibitors with enhanced chemical stability: interplay between the N-terminal amino acid alkyl side chain and the cyclopropyl group of -aminoacyl-L-cis-4,5-methanoprolinenitrile-based inhibitors. J Med Chem2004;47:2587-2598.[Medline] [Order article via Infotrieve]
• Chang CWT, Hui Y, Elchert B, Wang J, Li J, Rai R: Pyranmycins, a novel class of aminoglycosides with improved acid stability: the SAR of Dpyranoses on ring III of pyranmycin. Org Lett2002;4:4603-4606.[Medline] [Order article via Infotrieve]
• Regueiro-Ren A, Leavitt K, Kim SH, Hofle G, Kiffe M, Gougoutas JZ, et al: SAR and pH stability of cyano-substituted epothilones. Org Lett 2002;4:3815-3818.[CrossRef][Web of Science][Medline] [Order article via Infotrieve]
• Posner GH, Paik IH, Sur S, McRiner AJ, Borstnik K, Xie S, et al: Orally active, antimalarial, anticancer, artemisinin-derived trioxane dimers with high stability and efficacy. J Med Chem2003;46:1060-1065.[CrossRef][Medline] [Order article via Infotrieve]
• Chong Y, Gumina G, Mathew JS, Schinazi RF, Chu CK: L-2',3'-didehydro-2',3'-dideoxy-3'-fluoronucleosides: synthesis, anti-HIV activity, chemical and enzymatic stability, and mechanism of resistance. J Med Chem 2003;46:3245-3256.[CrossRef][Medline] [Order article via Infotrieve]
• Guilford WJ, Bauman JG, Skuballa W, Bauer S, Wei GP, Davey D, et al: Novel 3-oxa lipoxin A4 analogueswith enhanced chemical andmetabolic stability have anti-inflammatory activity in vivo. J Med Chem2004;47:2157-2165.[CrossRef][Web of Science][Medline] [Order article via Infotrieve]
• Jung M, Lee K, Kendrick H, Robinson BL, Croft SL: Synthesis, stability, and antimalarial activity of new hydrolytically stable and water-soluble (+)deoxoartelinic acid. J Med Chem2002;45:4940-4944.[Medline] [Order article via Infotrieve]
• Akers MJ: Excipient-drug interactions in parenteral formulations. J Pharm Sci2002;91:2283-2300.[CrossRef][Medline] [Order article via Infotrieve]
• Jonkman-DeVries JD, Doppenberg WG, Henrar REC, Bult A, Beijnen JH: Systematic study on the chemical stability of the prodrug antitumor agent carzelesin (U-80,244). J Pharm Sci1996;85:1227-1233.[Medline] [Order article via Infotrieve]
• Bonina FP, Arenare L, Palagiano F, Saija A, Nava F, Trombetta D, et al: Synthesis, stability, and pharmacological evaluation of nipecotic acid prodrug. J Pharm Sci1999;88:561-567.[Medline] [Order article via Infotrieve]
• Vabeno J, Nielsen CU, Ingebrigtsen T, Lejon T, Steffansen B, Luthman K: Dipeptidomimetic ketomethylene isosteres as pro-moieties for drug transport via the human intestinal di-/tripeptide transporter hPEPT1: design, synthesis, stability, and biological investigations. J Med Chem2004;47:4755-4765.[Medline] [Order article via Infotrieve]
• Di L, Kerns EH, Hong Y, Kleintop TA, McConnell OJ, Huryn DM: Optimization of a higher throughput microsomal stability screening assay for profiling drug discovery candidates. J Biomol Screen2003;8:453-462.[Abstract/Free Full Text]
• Di L, Kerns EH, Gao N, Li SQ, Huang Y, Bourassa JL, et al: Experimental design on single-time-point high-throughput microsomal stability assay. J Pharm Sci2004;93:1537-1544.[CrossRef][Web of Science][Medline] [Order article via Infotrieve]
• Di L, Kerns EH, Hong Y, Chen H: Development and application of high throughput plasma stability assay for drug discovery. Int J Pharm2005;297:110-119.[Medline] [Order article via Infotrieve]
• Clapton SD, Anand BS, Mitra AK: Effect of mono-and di-acylation on the ocular disposition of ganciclovir: physicochemical properties, ocular bioreversion, and antiviral activity of short chain ester prodrugs. J Pharm Sci2002;91:660-668.[CrossRef][Medline] [Order article via Infotrieve]
• Patel P, Osechinskiy S, Koehler J, Zhang L, Vajjhala S, Philips C, et al: Micro parallel liquid chromatography for high-throughput compound purity analysis and early ADMET profiling. J Assoc Lab Automation2004;9:185-191.
• Kyranos JN, Cai H, Wei D, Goetzinger WK: High-throughput highperformance liquid chromatography/mass spectrometry for modern drug discovery. Curr Opin Biotechnol2001;12:105-111.[CrossRef][Medline] [Order article via Infotrieve]
• Kyranos JN, Lee H, Goetzinger WK, Li LYT: One-minute full-gradient HPLC/UV/ELSD/MS analysis to support high-throughput parallel synthesis. J Comb Chem2004;6:796-804.[Medline] [Order article via Infotrieve]
• Lai F, Khojasteh-Bakht SC: Automated online liquid chromatographic/mass spectrometric metabolic study for prodrug stability. J Chromatogr B Analyt Technol Biol2005;814:225-232.
• Kerns EH, Kleintop T, Little D, Tobien T, Mallis L, Di L, et al: Integrated high capacity solid phase extraction-MS/MS system for pharmaceutical profiling in drug discovery. J Pharm Biomed Anal2004;34:1-9.[Medline] [Order article via Infotrieve]
• Lim HK, Chan KW, Sisenwine S, Scatina JA: Simultaneous screen for microsomal stability and metabolite profile by direct injection turbulentlaminar flow LC-LC and automated tandem mass spectrometry. Anal Chem2001;73:2140-2146.[Medline] [Order article via Infotrieve]
• Zeng H, Wu JT, Unger SE: The investigation and the use of high flowcolumnswitching LC/MS/MS as a high-throughput approach for direct plasma sample analysis of single and multiple components in pharmacokinetic studies. J Pharm Biomed Anal2002;27:967-982.[CrossRef][Medline] [Order article via Infotrieve]
• Peng SX, Strojnowski MJ, Bornes DM: Direct determination of stability of protease inhibitors in plasma by HPLC with automated column-switching. J Pharm Biomed Anal1999;19:343-349.[Medline] [Order article via Infotrieve]
• Xu R, Nemes C, Jenkins KM, Rourick RA, Kassel DB, Liu CZC: Application of parallel liquid chromatography/mass spectrometry for high throughput microsomal stability screening of compound libraries. J Am Soc Mass Spectrom2002;13:155-165.[Medline] [Order article via Infotrieve]
• U.S. Pharmacopeia: The National Formulary (USP-NF). Rockville, MD: United States Pharmacopeial Convention, 2004.
• Song Y, Schowen RL, Borchardt RT, Topp EM: Effect of "pH" on the rate of asparagine deamidation in polymeric formulations: "pH"-rate profile. J Pharm Sci2001;90:141-156.[Medline] [Order article via Infotrieve]
• Fubara JO, Notari RE: Influence of pH, temperature and buffers on cefepime degradation kinetics and stability predictions in aqueous solutions. J Pharm Sci1998;87:1572-1576.[CrossRef][Web of Science][Medline] [Order article via Infotrieve]
• Zhou M, Notari RE: Influence of pH, temperature, and buffers on the kinetics of ceftazidime degradation in aqueous solutions. J Pharm Sci1995;84:534-538.[CrossRef][Medline] [Order article via Infotrieve]
• Muangsiri W, Kirsch LE: The kinetics of the alkaline degradation of daptomycin. J Pharm Sci2001;90:1066-1075.[Medline] [Order article via Infotrieve]

This version was published on February 1, 2006

Journal of Biomolecular Screening, Vol. 11, No. 1, 40-47 (2006)
DOI: 10.1177/1087057105281363


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