2440 J ournal of Medicinal Chemistry, 1996, Vol. 39, No. 13
Communications to the Editor
(5) Dolle, R. E.; Singh, J .; Rinker, J .; Hoyer, D.; Prasad, C. V. C.;
Helaszek, C. T.; Miller, R. E.; Ator, M. A. Aspartyl -((1-Phenyl-
3-(trifluoromethyl)pyrazol-5-yl)oxy)methyl Ketones as Interleu-
kin-1â Converting Enzyme Inhibitors. Significance of the P1 and
P3 Amido Nitrogens For Enzyme-Peptide Inhibitor Binding. J .
Med. Chem. 1994, 37, 3863 3865.
(6) Dolle, R. E.; Singh, J .; Whipple, D.; Osifo, I. K.; Speier, G.;
Graybill, T. L.; Gregory, J . S.; Harris, A. L.; Helaszek, C. T.;
Miller, R. E.; Ator, M. A. Aspartyl -((Diphenylphosphinyl)oxy)-
methyl Ketones as Novel Inhibitors of Interleukin-1â Converting
Enzyme. Utility of the Diphenylphosphinic Acid Leaving Group
For The Inhibiton of Cysteine Proteases. J . Med. Chem. 1995,
38, 220 222.
metic inhibitors of ICE have been described. The design
strategy we pursued sought to retain the P1 aspartic
acid residue and critical hydrogen-bonding functionality
(P1- and P3-NH) associated with peptides 1 3. Pyri-
midinone-based inhibitors 4 14 embody these design
elements (Figure 1). In addition, we have outlined a
convergent approach to the synthesis of mimetics 4 14.
This approach utilizes readily available aspartylamine
cassettes 19 21 in coupling with carboxylic acid part-
ners 15 18. As will be reported separately, the con-
vergent-based inhibitor synthesis permitted us to “mix
and match” 19 21 with virtually any carboxylic or
sulfonic acid (including other peptidomimetics), provid-
ing an expedient route to analog generation.
(7) (a) Wilson, K. P.; Black, J . F.; Thompson, J . A.; Kim, E. E.;
Griffith, J . P.; Navia, M. A.; Murcko, M. A.; Chambers, S. P.;
Aldape, R. A.; Raybuck, S. A.; Livingston, D. J . Structure and
Mechanism of Interleukin-1â Converting Enzyme. Nature 1994,
370, 270 252. (b) Walker, N. P. C.; Talanian, R. V.; Brady, K.
D.; Dang, L. C.; Bump, N. J .; Ferenz, C. R.; Franklin, S.; Ghayur,
T.; Hackett, M. C.; Hammill, L. D.; Herzog, L.; Hugunin, M.;
Houy, W.; Mankovich, J . A.; McGuiness, L.; Orlewicz, E.;
Paskind, M.; Pratt, C. A.; Reis, P.; Summani, A.; Terranova, M.;
Welch, J . P.; Xiong, L.; Moller, A.; Tracey, D. E.; Kamen, R.;
Wong, W. W. Crystal Structure of the Cysteine Protease Inter-
leukin-1â Converting Enzyme: A (p20/p10)2 Homodimer. Cell
1994, 78, 343 352.
(8) This ring system has been used as a mimetic for the P3 P2 (Val-
Pro) in the human leukocyte elastase inhibitor 4-((4-Cl-Ph)SO2-
NHCO)PhCO-Val-Pro-Val-COCF3. See: (a) Bernstein, P. R.;
Edwards, P. D.; Shaw, A.; Thomas, R. M.; Veale, C. A.; Warner,
P.; Wolanin, D. J . Pyrimidinyl Acetamides as Elastase Inhibitors.
European Patent Application, publication no. WO 9321210,
October 1993. (b) Bernstein, P. R.; Edwards, P. D.; Shaw, A.;
Shenvi, A. B.; Thomas, R. M.; Veale, C. A.; Warner, P.; Wolanin,
D. J . Alpha-Aminoboronic Acid Peptides and Their Use as
Elastase Inhibitors. PTC publication no. WO 9321214, October
1993. (c) Veale, C. A.; Steelman, G. B.; Chow, M. M. Efficient
Method for the Synthesis of 1,4-Disubstituted 5-Carbomethoxy-
pyrimidin-6-ones. J . Org. Chem. 1993, 58, 4490 4493. (d)
Brown, F. J .; Andisik, D. W.; Bernstein, P. R.; Bryant, C. B.;
Ceccarelli, C.; Damewood, J . R., J r.; Edwards, P. D.; Earley, R.
A.; Feeney, S.; Green, R. C.; Gomes, B.; Kosmider, B. J .; Krell,
R. D.; Shaw, A.; Steelman, G. B.; Thomas, R. M.; Vacek, E. P.;
Veale, C. A.; Tuthill, P. A.; Warner, P.; Williams, J . C.; Wolanin,
D. J .; Woolson, S. A. Design of Orally Active, Non-Peptidic
Inhibitors of Human Leukocyte Elastase. J . Med. Chem. 1994,
37, 1259 1261.
(9) For a related pyridone ring system having utility for elastase
inhibition, see: (a) Bernstein, P. R.; Shaw, A.; Thomas, R. M.;
Veale, C. A.; Warner, P.; Wolanin, D. J . Lactam Peptides Having
HLE Inhibiting Activity. European Patent Application, publica-
tion no. WO 9321209, October 1993. (b) Bernstein, P. R.; Andisik,
D.; Bradley, P. K.; Bryant, C. B.; Ceccarelli, C.; Damewood, J .
R., J r.; Earley, R.; Edwards, P. D.; Feeney, S.; Gomes, B. C.;
Kosmider, B. J .; Steelman, G. B.; Thomas, R. M.; Vacek, E. P.;
Veale, C. A.; Williams, J . C.; Wolanin, D. J .; Woolson, S. A. Non-
Peptidic Inhibitors of Human Leukocyte Elastase. 3. Design,
Synthesis, X-ray Crystallographic Analysis, and Structure-
Activity Relationships For a Series of Orally Active 3-Amino-6-
phenylpyridin-2-one Trifluoromethyl Ketones. J . Med. Chem.
1994, 37, 3313 3326 and references therein.
Refer en ces
(1) (a) Kostrua, M. J .; Tocci, M. J .; Limjuco, G.; Chin, J .; Cameron,
P.; Hillman, A. G.; Chartrain, N. A.; Schmidt, J . A. Identification
of a Monocyte Specific Pre-interleukin 1â Convertase Activity.
Proc. Natl. Acad. Sci. U.S.A. 1989, 86, 5227 5231. (b) Black,
R. A.; Kronheim, S. R.; Cantrell, M.; Deeley, M. C.; March, C.
J .; Prickett, K. S.; Wignall, J .; Conlon, P. J .; Cosman, D.; Hopp,
T. P.; Mochizuki, D. Y. Generation of Biologically Active Inter-
leukin-1â by Proteolytic Cleavage of the Inactive Precursor. J .
Biol. Chem. 1988, 263, 9437.
(2) (a) Sleath, P. R.; Hendrickson, R. C.; Kronheim, S. R.; March,
C. J .; Black, R. A. Substrate Specificity of the Protease That
Processes Human Interleukin-1â. J . Biol. Chem. 1990, 265,
14526. (b) Howard, A. D.; Kostura, M. J .; Thornberry, N.; Ding,
G. J . F.; Limjuco, G.; Weidner, J .; Salley, J . P.; Hogquist, K. A.;
Chaplin, D. D.; Mumford, R. A.; Schmidt, J . A.; Tocci, M. J . IL-
1-Converting Enzyme Requires Aspartic Acid Residues For
Processing of the IL-1â Precursor at Two Distinct Sites and Does
Not Cleave 31-kDa IL-1 . J . Immunol. 1991, 147, 2964. (c)
Thornberry, N. A.; Bull, H. G.; Calaycay, J . R.; Chapman, K. T.;
Howard, A. D.; Kostura, M. J .; Miller, D. K.; Molineaux, S. M.;
Weidner, J . R.; Aunins, J .; Elliston, K. O.; Ayala, J . M.; Casano,
F. J .; Chin, J .; Ding, G. J .-F.; Egger, L. A.; Gaffney, E. P.;
Limjuco, G.; Palyha, O. C.; Raju, S. M.; Rolando, A. M.; Salley,
J . P.; Yamin, T.-T.; Lee, T. D.; Shively, J . E.; MacCross, M.;
Mumford, R. A.; Schmidt, J . A.; Tocci, M. J . A Novel Het-
erodimeric Cysteine Protease is Required for Interleukin-1â
processing in Monocytes. Nature 1992, 356, 768. (d) Miller, D.
K.; Calaycay, J . R.; Chapman, K. T.; Howard, A. D.; Kostura,
M. J .; Molineaux, S. M.; Thornberry, N. A. The IL-1â Converting
Enzyme as a Therapeutic Target. In Immunosupressive and
Antiinflammatory Drugs; Allison, A. C., Lafferty, K. J ., Fliri, H.,
Eds.; Annals of the New York Academy of Sciences; New York
Academy of Sciences: New York, 1993; Vol. 696, pp 133 148.
(3) (a) Dinarello, C. A.; Wolff, S. M. The Role of Interleukin-1 in
Disease. N. Engl. J . Med. 1993, 328, 106. (b) Dinarello, C. A.
Interleukin-1 and Interleukin Antagonism. Blood 1991, 77, 1627.
(c) Dinarello, C. A.; Thompson, R. C. Blocking IL-1: Interleukin
1 Receptor Antagonist In Vivo and In Vitro Immunol. Today
1991, 12, 404 410.
(4) (a) Dolle, R. E.; Hoyer, D.; Prasad, C. V. C.; Schmidt, S. J .;
Helaszek, C. T.; Miller, R. E.; Ator, M. A. P1 Aspartate-Based
Peptide -((2,6-Dichlorobenzoyl)oxy)methyl Ketones as Potent
Time-Dependent Inhibitors of Interleukin-1â Converting En-
zyme. J . Med. Chem. 1994, 37, 563 564. (b) Assay: ICE was
partially purified from THP-1 cells using the DEAE-Sephacel
and Sephadex G-75 steps described by Black (Black, R. A.;
Kronheim, S. R.; Sleath, P. R. Activation of Interluekin-1â by a
Co-induced Protease. FEBS Lett. 1989, 247, 386). The ICE assay
contained 10 mM HEPES (pH 7.5), 25% glycerol, 1 mM dithiotre-
itol (DTT), and 10 µM Suc-Tyr-Val-Ala-Asp-AMC (BACHEM) in
a volume of 30 µL in a polystyrene 96-well microtiter plate.
Progress curves were obtained at 37 °C over 30 min. Kinetic data
were obtained on a Fluoroskan II fluorescence plate reader under
control of an Apple Macintosh computer running the DeltaSoft
data aquisition program (Biometallics, Inc.). Nonlinear progress
curves were analyzed as described by Tian and Tsou (Tian, W.-
X.; Tsou, C.-L. Determination of the Rate Constant of Enzyme
Modification by Measuring the Substrate Reaction in the Pres-
ence of the Modifier. Biochemistry 1982, 21, 1028 1032).
(10) Amine hydrochlorides 19 21 are stable in solid form to storage
at 0 °C for several months without decomposition.
(11) Knorr, R.; Trzeciak, A.; Bannwarth, W.; Gillessen, D. New
Coupling Reagents in Peptide Chemistry. Tetrahedron Lett.
1989, 30, 1927 1930.
(12) All new compounds gave physical and spectroscopic data con-
sistent with their structure.
(13) Wommack, J . B.; Pearson, D. E. Potential Antimalarials. IV.
Quinoline- , -dialkylmethanols. J . Med. Chem. 1970, 13, 383
386.
(14) As originally conceived, the pyrimidinyl acetamides are Val-Pro
mimetics for elastase (ref 8). Direct comparison of 5 (157 000
1
1
1
M
s
1) to Z-Val-Pro-Asp-CH2TPP (ref 5; 116 000 M
s )
shows these compounds to be essentially equipotent against
ICE.
J M9601516