(N-acyl-N-alkyl)glycyl borolysine analogs: A new class of potent thrombin inhibitors

Bioorganic and Medicinal Chemistry Letters

Volumn 6, Issue 24, 1996, Pages 2913-2918

  Galemmo Jr , Robert A ^a   Fevig, John M ^a   Carini, David J ^a   Cacciola, Joseph ^a   Wells, Brian L ^a   Hillyer, Gregory L ^a   Buriak Jr , Joseph ^a   Rossi, Karen A ^a   Stouten, Pieter F W ^a   Alexander, Richard S ^a   Hilmer, Richard ^a   Bostrom, Lori ^a   Abelman, Matthew M ^a,b   Lee, Sheng Lian ^a   Weber, Patricia C ^a,c   Kettner, Charles A ^a   Knabb, Robert M ^a   Wexler, Ruth R ^a  

^a DUPONT COMPANY   (United States)

^b Dendreon   (United States)

^c MERCK RESEARCH LABORATORIES   (United States)

Author keywords

[No Author keywords available]

Indexed keywords

LYSINE DERIVATIVE; N (3 PHENYLPROPANOYL) N PHENETHYLGLYCYLBOROLYSINE; ORGANOBORON DERIVATIVE; PLASMIN; THROMBIN; THROMBIN INHIBITOR; TISSUE PLASMINOGEN ACTIVATOR; TRYPSIN; UNCLASSIFIED DRUG;

ARTICLE; CRYSTALLOGRAPHY; DRUG DESIGN; DRUG STRUCTURE; DRUG SYNTHESIS; ENZYME BINDING; HUMAN; IN VITRO STUDY;

EID: 0030591848     PISSN: 0960894X     EISSN: None     Source Type: Journal    
DOI: 10.1016/S0960-894X(96)00525-2     Document Type: Article

References (19)

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3. 3. Weber, P.C.; Lee, S-L.; Lewandowski, F.A.; Schadt, M.C.; Chang, C-H; Kettner, C. A. Biochemistry 1995, 34, 37540.
4. 4. Bode, W.; Stubbs, M. T. Actual. Chim. Ther. 1994, 21, 3.
5. 5. Kettner, C.; Knabb, R.; Fevig, J.; Hugli, T.; Lee, S.; Mantri, P.; Pangbum, M.; Reilly, T.; Stouten, P.; Thooten, M.; Weber, P.; Wexler, R. 212th American Chemical Society National Meeting, August 1996, Orlando, FL, MEDI 0112.
6. 6. Contemporaneous with our work a similar finding was published; see : Balasubramanian, N.; St. Laurent, D. R.; Federici, M. E.; Meanwell, N. A.; Wright, J. J.; Schumacher, W. A.; Seiler, S. M. J. Med. Chem. 1993, 36, 300.
7. 7. Fevig, J.M.; Abelman, M.M.; Brittelli, D.R.; Kettner, C.A.; Knabb, R.M.; Weber, P.C Bioorg. Med. Chem. Lett. 1996, 6, 295.
8. 8. Each ligand was modeled in various conformations and orientations into the thrombin active site and attached covalently to Ser195 with a tetrahedral, negatively charged carboxylate as a boronic acid surrogate. Energy minimizations with the Discover program and the CVFF force field (Molecular Simulations, Inc., Burlington, MA 01803-5297) were carried out for these different binding models keeping the protein rigid, but allowing the ligand to be completely flexible. The model with the lowest energy was assumed to represent the most probable binding mode.
9. 9. Wityak, J.; Earl, R.A.; Abelman, M.M.; Bethel, Y.B.; Fisher, B.; Kauffman, G.; Kettner, C. A.; Ma, P.; McMillan, J.L.; Mersinger, L.J.; Pesti, J.; Pierce, M.; Rankin, F.W.; Chorvat, R.J.; Confalone, P.N. J. Org. Chem. 1995, 60, 3717.
10. 2, 10% Pd-C); t-butyl N-cyclopropylglycinate for 4f was prepared according to Suh, J.T.; Youssefyeh, R. D.; Mann, W. S. J. Med. Chem. 1985, 28, 57;
11. 2O); all other glycine derivatives were commercially available. Satisfactory spectral data were obtained.
12. 11. Kettner, C.A. US Patent 5,384,410 1995 and ref 8.
13. i) assays were performed as described in ref 2.
14. (b) The thrombin time is defined as the concentration of inhibitor necessary to double the time required for clot formation induced by the addition of 4 NIH u/mL (final concentration) bovine thrombin; see: Knabb, R. M.; Kettner, C. A.; Timmermans, P. B. M. W. M.; Reilly, T. M. Thromb. Haemostasis 1992, 67, 56.
15. 50s were determined by incubating various concentrations of inhibitor with human α-thrombin for 10 min and measuring the rate of change in absorbance at 405 nm after adding chromogenic substrate S2238.
16. 13. Lim, M. S. L.; Johnston, E. R.; Kettner, C. A. J. Med. Chem. 1993, 36, 1831.
17. 14. Crystals of human thrombin-hirugen complex were prepared by the method described by Skrzypczak-Jankun, E., Carperos, V.E., Ravichandran, K.G., Tulinsky, A., Westbrook, M., and Marananore, J.M., J. Mol. Biol. 1991, 206, 755.
18. Crystals of the thrombin-hirugen complex were transferred to a well containing a sitting solution (0.58 M sodium phosphate buffer (pH 7.2), 0.05 mM sodium azide, and 33% w:v PEG 8000). These crystals were transferred to a solution containing the inhibitor. The inhibitor solution was prepared by first dissolving the inhibitor in DMSO followed by a 10-fold dilution of the inhibitor/DMSO solution into the sitting solution. Data were collecting on an R Axis image plate system mounted on a Rigaku RU300 rotating anode; crystals diffracted to 1.9 Å. The data were 88% complete with an R merge of 0.038. Data were xrefined using XPLOR (Brunger, A.T.: X-PLOR, Version 3.1. A System for X-ray Crystallography and NMR, Yale University Press, New Haven, CT) with a final R-value of 0.218. Clear electron densities were observed for all atoms of the inhibitor.
19. 15. Schacht, A. L.; Wiley, M. R.; Chirgadze, N.; Clawson, D.; Craft, T. J.; Coffman, W. J.; Jones, N. D.; Gifford-Moore, D.; Olkowski, J.; Shuman, R.T; Smith, G. F.; Weir, L. C. Bioorg. Med. Chem. Lett. 1995, 5, 2529.