Aminopyrrolidineamide inhibitors of site-1 protease

Bioorganic and Medicinal Chemistry Letters

Volumn 17, Issue 16, 2007, Pages 4411-4414

  Hay, Bruce A ^a   Abrams, Barbara ^a   Zumbrunn, Allice Y ^a   Valentine, James J ^a   Warren, Laurie C ^a   Petras, Stephen F ^a   Shelly, Lorraine D ^a   Xia, Angela ^a   Varghese, Alison H ^a   Hawkins, Julie L ^a   Van Camp, Jennifer A ^b   Robbins, Michael D ^a   Landschulz, Katherine ^a   Harwood Jr , H James ^a  

^a PFIZER GLOBAL RESEARCH AND DEVELOPMENT   (United States)

^b PFIZER INC   (United States)

Author keywords

Metabolic syndrome; Site 1 protease; SREBP; Sterol regulatory element binding protein

Indexed keywords

AMINE; AMINOPYRROLIDINEAMIDE; PF 429242; PROTEINASE INHIBITOR; PYRROLIDINE DERIVATIVE; SERINE PROTEINASE; SITE 1 PROTEASE; SITE 1 PROTEASE INHIBITOR; STEROL REGULATORY ELEMENT BINDING PROTEIN; UNCLASSIFIED DRUG;

ARTICLE; CHOLESTEROL METABOLISM; DRUG CLEARANCE; DRUG STRUCTURE; DRUG SYNTHESIS; DYSLIPIDEMIA; ENZYME INHIBITION; FATTY ACID METABOLISM; HIGH THROUGHPUT SCREENING; HUMAN; METABOLIC SYNDROME X; NONHUMAN; SUBSTITUTION REACTION;

ACTIVE TRANSPORT, CELL NUCLEUS; ANIMALS; ANTILIPEMIC AGENTS; COMBINATORIAL CHEMISTRY TECHNIQUES; GENE EXPRESSION REGULATION; HUMANS; INHIBITORY CONCENTRATION 50; MALE; MICE; MOLECULAR STRUCTURE; PROPROTEIN CONVERTASES; PYRROLES; SERINE ENDOPEPTIDASES; STEROL REGULATORY ELEMENT BINDING PROTEINS;

EID: 34447322283     PISSN: 0960894X     EISSN: None     Source Type: Journal    
DOI: 10.1016/j.bmcl.2007.06.031     Document Type: Article

References (13)

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8. The enzymatic activity and inhibition of S1P were measured fluorometrically using the MCA-conjugated peptidyl substrate, Ac-VFRSLK-MCA, essentially as described in Cheng, D.; Espenshade, P. J.; Slaughter, C. A.; Jaen, J. C.; Brown, M. S.; Goldstein, J. L. J. Biol. Chem. 1999, 274, 22805, with the following modifications: The reaction was conducted in 96-well plate format with an assay volume of 40 μL, the fluorogenic peptide substrate concentration was 20 μM, the purified His-tagged human S1P enzyme (secreted into serum-free pH 8.0 medium from stably transfected CHO-K1 cells and purified by nickel column affinity chromatography) concentration was 25 μg/mL (1 μg/well), the reaction was conducted for 4 h at 37 °C, compounds dissolved in DMSO were added such that the final DMSO concentration in the assay was 2.5%. Under these conditions, the assay exhibited a S/N ratio of 8 and a coefficient of variation of 15%. Confirmed S1P inhibitors did not exhibit fluorescence at either 360 or 460 nm nor did they quench fluorescence from a control well-containing MCA.
9. Enantiomers were separated and analyzed on preparative and analytical chiralpak AD columns, respectively, using a heptane/ethanol gradient with 0.2% DEA as mobile phase.
10. Proteolytic processing and nuclear translocation of SREBP were assessed as described by Cheng et al. (1999) op cit (Ref. 8).
11. HMG-CoA synthase, fatty acid synthase, and LDL receptor gene expression in cultured cells and in hepatic tissue samples was measured as outlined in. Hawkins J.L., Gafvels M., Olin M., Lund E.G., Andersson U., Schuster G., Bjorkhem I., Russell D.W., and Eggertsen G. J. Clin. Invest. 110 (2002) 1191
12. Cholesterol and fatty acid synthesis in cultured cells and in experimental animals were measured as outlined in. Harwood Jr. H.J., Petras S.F., Shelly L.D., Zaccaro L.M., Perry D.A., Makowski M.R., Hargrove D.M., Martin K.A., Tracey W.R., Chapman J.G., Magee W.P., Dalvie D.K., Soliman V.F., Martin W.H., Mularski C.J., and Eisenbeis S.A. J. Biol. Chem. 278 (2003) 37099
13. LDL receptor mediated LDL internalization was measured in cultured cells as outlined in:. Harwood Jr. H.J., and Pellarin L.D. Biochem. J 323 (1997) 649 using DiI-LDL as the probe