Identified using recombinant NL4.3 computer virus (T124A IN mutant). Table 4 Orotic acid (6-Carboxyuracil) In Vitro ADME Profile of Compound 26 Open in a separate window EC50 range,a nM11C27serum shift (50% HS), fold-changeb2.1HLM/RLM ( em t /em 1/2),?min210/ 300Caco-2 ( em P /em app), ?106?cm/s14CyP450 inh. mg/mL 0.85 Open in a separate window aDetermined with HxB2 virus (A124/T125 IN variant), NL4.3 computer virus (T124/T125), or recombinant NL4.3 computer virus (A124/T125, A124/A125, N124/T125, or N124/A125 IN variants) while previously described. bDetermined by measurement of EC50 ideals 50% human being serum. cFor the amorphous powder. In conclusion, we have used an assay based on the 3 control activity of HIV-1 IN to display the Boehringer Ingelheim compound collection and determine hit compound 1.23?29 Hit-to-lead and lead optimization effort founded the importance of the C3 and C4 substituents to binding to the KIAA1575 CCD of IN, which translated into excellent antiviral potency against a number of viruses with different em aa /em 124/ em aa /em 125 variants of IN. We also founded the importance of the C7 position within the serum shifted potency. Balancing good potency with superb metabolic stability was accomplished through the intro of a quinoline substituent in the C4 position. Combination of these findings ultimately led to the finding of compound 26 (BI 224436), the 1st NCINI to advance into a phase Ia medical trial. Acknowledgments We gratefully acknowledge the contribution of the following colleagues: Christine Martens for development of the LTR DNA 3-processing assay that was utilized for the HTS, Kevork Mekhssian for development of the displacement assay, Patrick Salois for IC50 determinations, Cline Plouffe for em K /em d-app determinations, Elizabeth Wardrop and Sonia Tremblay for EC50 determinations, Hugo Poirier for Caco-2 permeability data, Josie DeMarte for microsomal stability data, and Laibin Luo, Danhui Sun, and Eduard Bugan for logD and solubility determinations. Glossary ABBREVIATIONScARTcombination antiretroviral therapyHTShigh-throughput screenLTRlong terminal repeatHLMhuman liver microsomesRLMrat liver microsomes Supporting Info Available Synthetic techniques for preparation of NCINIs and characterization of important compounds. This material is available free of charge via the Internet at http://pubs.acs.org. Author Present Orotic acid (6-Carboxyuracil) Address ? Boehringer Ingelheim Pharmaceuticals, Inc., 900 Ridgebury Road, Ridgefield, Connecticut 06877, United States. Author Present Address ? Division of Chemistry and Biochemistry, Concordia University Orotic acid (6-Carboxyuracil) or college, 7141 Sherbrooke Street Western, Montreal, QC H4B 1R6, Canada. Author Present Address Bristol-Myers Squibb, Virology, 5 Study Parkway, Wallingford, Connecticut 06492, United States. Author Present Address Division of Chemistry, McGill University or college, Orotic acid (6-Carboxyuracil) 801 Sherbrooke Street Western, Montreal, QC H3A 0B8, Canada. Author Present Address EMD Serono, 45 Middlesex Turnpike, Billerica, Massachusetts 01821, United States. Author Present Address # Toronto Study Chemicals, 2 Brisband Road, North York, ON M3J 2J8, Canada. Author Present Address Woman Davis Institute, Jewish General Hospital, 3755 C?te Ste-Catherine Road, Montreal, QC H3T 1E2, Canada. Notes The authors declare no competing financial interest. Supplementary Material ml500002n_si_001.pdf(234K, pdf).
Identified using recombinant NL4