U.S. patent application number 10/418012 was filed with the patent office on 2003-11-13 for matrix metalloprotease inhibitors.
Invention is credited to Bender, Steven Lee, Castelhano, Arlindo Lucas, Deal, Judith Gail, Horne, Stephen, Liak, Teng J., Yuan, Zhengyu.
Application Number | 20030212067 10/418012 |
Document ID | / |
Family ID | 23344942 |
Filed Date | 2003-11-13 |
United States Patent
Application |
20030212067 |
Kind Code |
A1 |
Castelhano, Arlindo Lucas ;
et al. |
November 13, 2003 |
Matrix metalloprotease inhibitors
Abstract
Compounds of the formula (I) 1 and their pharmaceutically
acceptable salts inhibit matrix metalloproteases, such as
stromelysin, gelatinase, matrilysin and collagenase, and are useful
in the treatment of mammals having disease-states alleviated by the
inhibition of such matrix metalloproteases.
Inventors: |
Castelhano, Arlindo Lucas;
(Pleasanton, CA) ; Bender, Steven Lee; (Oceanside,
CA) ; Deal, Judith Gail; (Temecula, CA) ;
Horne, Stephen; (Burlington, CA) ; Liak, Teng J.;
(Mississauga, CA) ; Yuan, Zhengyu; (Santa Clara,
CA) |
Correspondence
Address: |
HELLER EHRMAN WHITE & MCAULIFFE LLP
275 MIDDLEFIELD ROAD
MENLO PARK
CA
94025-3506
US
|
Family ID: |
23344942 |
Appl. No.: |
10/418012 |
Filed: |
April 16, 2003 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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10418012 |
Apr 16, 2003 |
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09468762 |
Dec 21, 1999 |
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6579890 |
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09468762 |
Dec 21, 1999 |
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08343158 |
Nov 22, 1994 |
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6037472 |
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Current U.S.
Class: |
514/237.8 ;
514/114; 514/603; 544/162; 558/190; 564/86 |
Current CPC
Class: |
C07C 323/60 20130101;
C07F 9/3241 20130101; C07C 237/22 20130101; A61P 3/08 20180101;
C07C 327/12 20130101; C07F 9/65583 20130101; C07D 263/26 20130101;
C07C 323/59 20130101; A61P 27/02 20180101; C07C 279/14 20130101;
C07C 311/46 20130101; C07C 327/32 20130101; C07C 317/40 20130101;
C07C 255/57 20130101; C07C 2601/14 20170501; C07F 9/306 20130101;
C07D 295/13 20130101; A61P 35/00 20180101; A61P 3/10 20180101; C07C
237/42 20130101; A61P 29/00 20180101; C07F 9/58 20130101; C07C
259/06 20130101; C07C 2603/18 20170501; A61P 43/00 20180101; C07D
213/75 20130101; C07F 9/301 20130101; C07F 9/3211 20130101; A61P
1/02 20180101; C07F 9/60 20130101; C07C 255/60 20130101; C07C
323/63 20130101; C07F 9/4816 20130101 |
Class at
Publication: |
514/237.8 ;
514/114; 514/603; 544/162; 558/190; 564/86 |
International
Class: |
A61K 031/66; A61K
031/537; A61K 031/18 |
Claims
What is claimed is:
1. A compound of the formula 44wherein: R.sup.1 is mercapto,
acetylthio, carboxy, hydroxycarbamoyl, N-hydroxyformamide,
alkoxycarbonyl, aryloxycarbonyl, aralkoxycarbonyl,
benzyloxycarbamoyl or a group of the formula 45 where R.sup.6 is
aryl or heteroaryl; R.sup.2 is alkyl, cycloalkyl, aryl,
heterocycloalkyl, or heteroaryl; R.sup.3 is alkyl, cycloalkyl,
aralkyl, or heteroaralkyl; R.sup.7 is aryl, heteroaryl or
heterocycloalkyl; X is a group of the formula
--(CH.sub.2).sub.m--Y--(CH.- sub.2).sub.n--, where: Y is O, S, or a
single bond, m is an integer from 0 to 4, n is an integer from 0 to
4, and m+n is an integer from 0 to 4; p is an integer from 0 to 4,
provided that R.sup.2--X is biphenylalkyl when p is not 0; or a
pharmaceutically acceptable salt thereof.
2. The compound or salt of claim 1 wherein: R.sup.1 is carboxy;
R.sup.3 is cyclohexyl; R.sup.7 is optionally substituted phenyl or
N-morpholino; X is propanyl; and p is 2 or 3.
3. The compound or salt of claim 2 wherein R.sup.7 is
4-(aminosulfonyl)phenyl or N-morpholino.
4. The compound or salt of claim 1 wherein: R.sup.2 is alkyl,
optionally substituted phenyl, or a group of the formula: 46where:
A is CH.sub.2, O, NH, S, CH.sub.2--CH.sub.2, or NH--CH.sub.2;
R.sup.10 is H, alkyl, alkoxy, alkylamino or acylamide; and R.sup.11
is H or halo; R.sup.7 is 4-pyridyl or optionally substituted
phenyl; and p is 0.
5. The compound or salt of claim 4 wherein: R.sup.1 is carboxy,
hydroxycarbamoyl, or N-hydroxyformamide; R.sup.2 is phenyl,
biphenyl, 4-(pyridyl)phenyl, or 2-methylpropyl; R.sup.3 is t-butyl,
4-aminobutyl, dimethylaminobutyl, 4-(N,N'diethylguanido)butyl,
propyl, 2-methylpropyl, 1-hydroxyisopropyl, 1-hydroxyethyl, or
cyclohexyl; and X is a single bond, ethylene or propanyl.
6. The compound or salt of claim 4 wherein R.sup.2 is biphenyl.
7. The compound or salt of claim 6 wherein: R.sup.1 is carboxy,
N-hydroxyformamide, or hydroxycarbamoyl; R.sup.3 is alkyl; and
R.sup.7 is 4-pyridyl.
8. The compound or salt of claim 7 wherein R.sup.3 is t-butyl and X
is propanyl.
9. The compound or salt of claim 8 wherein R.sup.1 is carboxy.
10. The compound or salt of claim 8 wherein R.sup.1 is
N-hydroxyformamide.
11. The compound or salt of claim 8 wherein R.sup.1 is
hydroxycarbamoyl.
12. The compound or salt of claim 4 wherein: R.sup.2 is a group of
the formula: 47where: A is CH.sub.2; R.sup.10 is H or acylamide;
and R.sup.11 is H; R.sup.7 is optionally substituted phenyl; and X
is propanyl.
13. The compound or salt of claim 12 wherein: R.sup.1 is carboxy,
hydroxycarbamoyl, or N-hydroxyformamide; R.sup.1 is alkyl; and
R.sup.7 is alkoxycarbonylphenyl.
14. The compound or salt of claim 13 wherein: R.sup.1 is carboxy;
R.sup.3 is 2-methylpropyl; and R.sup.7 is
4-(methoxycarbonyl)-phenyl.
15. The compound or salt of claim 6 wherein R.sup.7 is optionally
substituted phenyl.
16. The compound or salt of claim 15 wherein R.sup.1 is carboxy;
R.sup.3 is alkyl or cycloalkyl; and X is propanyl.
17. The compound or salt of claim 16 wherein R.sup.3 is cyclohexyl;
and R.sup.7 is 4-(hydroxyethylaminosulfonyl)phenyl or
4-(dimethylaminoethyl-a- minosulfonyl)phenyl.
18. The compound or salt of claim 16 wherein R.sup.3 is
4-(amino)butyl or 4-(isopropylamino) butyl; and R.sup.7 is
4-(ethoxycarbonyl) phenyl.
19. The compound or salt of claim 16 wherein R.sup.3 is
1-hydroxyisopropyl; and R.sup.7 is phenyl.
20. The compound or salt of claim 16 wherein R.sup.3 is
tert-butyl.
21. The compound or salt of claim 20 wherein R.sup.7 is
4-(N-morpholinopropylaminosulfonyl)phenyl.
22. The compound or salt of claim 20 wherein R.sup.7 is
4-(methylaminosulfonyl)phenyl.
23. The compound or salt of claim 20 wherein R.sup.7 is
4-(hydroxyethylaminosulfonyl)phenyl.
24. The compound or salt of claim 20 wherein R.sup.7 is
4-(methylsulfinyl)phenyl.
25. The compound or salt of claim 4 wherein: R.sup.1 is carboxy;
R.sup.2 is phenyl; R.sup.3 is alkyl or cycloalkyl; and R.sup.7 is
optionally substituted phenyl.
26. The compound or salt of claim 25 wherein: R.sup.3 is
4-(amino)butyl; R.sup.7 is 4-(ethoxycarbonyl)phenyl; and X is
propanyl.
27. The compound or salt of claim 25 wherein: R.sup.3 is
(N,N'-diethylguanido)N-butyl; R.sup.7 is 4-(ethoxycarbonyl)phenyl;
and X is propylene.
28. The compound or salt of claim 25 wherein: R.sup.3 is
cyclohexyl; R.sup.7 is
4-(N",N"-dimethylaminoethylaminosulfonyl)phenyl; and X is
ethylene.
29. The compound or salt of claim 4 wherein: R.sup.1 is mercapto,
carboxy, hydroxycarbamoyl, or N-hydroxyformamide; R.sup.2 is
2-methylpropyl; R.sup.3 is alkyl, cycloalkyl, or heteroaralkyl;
R.sup.7 is optionally substituted phenyl; and X is a single
bond.
30. The compound or salt of claim 29 wherein: R.sup.1 is
hydroxycarbamoyl; R.sup.3 is propyl, 2-methylpropyl, cyclohexyl or
3-methylindolyl; and R.sup.7 is 4-(methoxy)phenyl,
4-(carboxy)phenyl, 4-(methoxycarbonyl)pheny- l or
4-(dimethylaminoethylcarbamoyl)phenyl.
31. The compound or salt of claim 30 wherein R.sup.7 is
4-(dimethylaminoethylcarbamoyl)phenyl.
32. The compound or salt of claim 30 wherein R.sup.7 is
4-(methoxycarbonyl)phenyl.
33. The compound or salt of claim 29 wherein: R.sup.1 is
N-hydroxyformamide; R.sup.3 is 2-methylpropyl; and R.sup.7 is
4-(methoxycarbonyl)phenyl.
34. The compound or salt of claim 29 wherein: R.sup.1 is carboxy;
R.sup.3 is cyclohexyl or 2-methylpropyl; and R.sup.7 is
4-(methoxycarbonyl)phenyl- .
35. The compound or salt of claim 29 wherein: R.sup.1 is mercapto;
R.sup.3 is 2-methylpropyl; and R.sup.7 is
4-(methoxycarbonyl)phenyl.
36. The compound or salt of claim 4 wherein: R.sup.1 is carboxy;
R.sup.2 is 4-(2-hydroxyethyl)phenyl, 4-(2-hydroxypropyl)-phenyl,
4-(2-hydroxybutyl)phenyl, 4-pyridylphenyl, biphenyl,
4'-(aminoethoxy)biphenyl, 4'-(cyano)biphenyl, or
4'-(hydroxy)biphenyl; R.sup.3 is 2-methylpropyl; R.sup.7 is
4-(methoxycarbonyl)phenyl; and X is propanyl.
37. The compound or salt of claim 36 wherein R.sup.2 is
4-(pyridyl)phenyl.
38. A pharmaceutical composition comprising a compound or salt of
claim 1 and a pharmaceutically acceptable excipient.
39. A method of inhibiting matrix metalloproteinase activity in a
mammal comprising administering to a mammal in need thereof a
therapeutically effective amount of a compound or salt of claim
1.
40. A method of treating matrix metalloproteinase-mediated
disorders selected from the group consisting of arthritis, abnormal
wound healing, tissue ulceration, bone resorption disease,
diabetes, tumor invasion, tumor metastasis and periodontal disease
comprising administering to a mammal in need thereof a
therapeutically effective amount of a compound or salt of claim
1.
41. A compound of the formula: 48wherein: R.sup.2 is alkyl, aryl or
heteroaryl; and X is a group of the formula
--(CH.sub.2).sub.m--Y--(CH.su- b.2).sub.n, where: Y is O, S, or a
single bond, m is an integer from 0 to 4, n is an integer from 0 to
4, and m+n is an integer from 0 to 4; or R.sup.2 and X together are
lower alkenyl.
42. A process for making a compound of the formula: 49wherein
R.sup.2 is aryl or heteroaryl, said process comprising
hydrogenating a compound of the formula: 50in the presence of a
palladium/carbon catalyst.
43. The process of claim 42 wherein said compound of the formula:
51is made by contacting
N-(2R-(t-butoxycarbonyl)-methyl-4-pentenoyl)-4S-phenyl-
methyl-2-oxazolidinone with an R.sup.2-halide in the presence of a
base and a palladium catalyst.
44. A process for making a compound of the formula: 52wherein
R.sup.2 is aryl or heteroaryl, said process comprising contacting a
compound of the formula: 53with sodium hexamethyldisilazide and
t-butylbromoacetate.
45. A compound of the formula: 54wherein R.sup.2 is aryl or
heteroaryl.
46. A process for making a compound of the formula: 55wherein
R.sup.2 is aryl or heteroaryl, said process comprising: (a)
contacting a compound of the formula: 56where R.sup.2 is hydrogen,
aryl or heteroaryl, with an excess of mesyl chloride in pyridine
followed by refluxing under basic conditions, and (b) where R.sup.2
is hydrogen in step (a), reacting the product of step (a) with an
aryl halide or a heteroaryl halide in the presence of a base and a
palladium catalyst.
47. A process for the preparation of a compound of formula (I)
57wherein, R.sup.1 is mercapto, acetylthio, carboxy,
benzyloxycarbamoyl, hydroxycarbamoyl, N-hydroxyformamide,
alkoxycarbonyl, aralkoxycarbonyl, benzyloxycarbamoyl, or a
thiomethylphosphinoyl group of the formula 58where R.sup.6 is aryl
or heteroaryl; R.sup.2 is alkyl, aryl, or heteroaryl; R.sup.3 is
alkyl, cycloalkyl, aralkyl or heteroaralkyl; X is a group of the
formula --(CH.sub.2).sub.m--Y--(CH).sub.n, where: Y is O, S, or a
single bond, m is an integer from 0 to 4, n is an integer from 0 to
4, and m+n is an integer from 0 to 4; R.sup.7 is aryl, heteroaryl
or heterocycloalkyl; and p is an integer from 0 to 4; said process
comprising: (A) contacting a compound of formula (D) 59with a
compound of formula (P) 60where R.sup.1 is alkoxycarbonyl,
aralkoxycarbonyl, aryl- or heteroaryl-thiomethylphosphinoyl, or
acetylthio; in the presence of a base and an amide coupling reagent
to give the corresponding compound of formula (I); or (B)
catalytically hydrogenating the corresponding compound where X and
R.sup.2 together are optionally aryl- or heteroaryl-substituted
alkenyl; or (C) treating a compound of formula (I), where R.sup.1
is alkoxycarbonyl or aralkoxycarbonyl, under mild acidic conditions
to give the corresponding compound of formula (I) where R.sup.1 is
carboxy; or (D) contacting a compound of formula (I), where R.sup.1
is carboxy, with O-benzylhydroxylamine to give the corresponding
compound of formula (I) where R.sup.1 is benzyloxycarbamoyl; or (E)
catalytically hydrogenating a compound of formula (I), where
R.sup.1 is benzyloxycarbamoyl, to give the corresponding compound
of formula (I) where R.sup.1 is hydroxycarbamoyl; or (F) contacting
a compound of formula (I), where R.sup.1 is carboxy, with
hydroxylamine to give the corresponding compound of formula (I)
where R.sup.1 is hydroxycarbamoyl; or (G) catalytically
hydrogenating a compound of the formula 61where BnO is benzyloxy,
to give the corresponding compound of formula (I) where R.sup.1 is
N-hydroxyformamide; or (H) treating a compound of formula (I),
wherein R.sup.1 is acetylthio, with ammonium hydroxide in a protic
solvent to give the corresponding compound of formula (I) where
R.sup.1 is mercapto.
Description
CROSS-REFERENCE TO RELATED APPLICATIONS
[0001] This application is a continuation-in-part of Ser. No.
08/147,811, filed Nov. 4, 1993, incorporated herein by
reference.
FIELD OF THE INVENTION
[0002] The present invention is directed to compounds and their
pharmaceutically acceptable salts, which inhibit matrix
metalloproteases, and are therefore useful in the treatment of
mammals having disease-states alleviated by the inhibition of such
matrix metalloproteases.
BACKGROUND OF THE INVENTION
[0003] Matrix metalloproteases ("MMPs)" are a family of proteases
(enzymes) involved in the degradation and remodeling of connective
tissues. Members of this family of endopeptidase enzymes are
present in various cell types that reside in or are associated with
connective tissue, such as fibroblasts, monocytes, macrophages,
endothelial cells, and invasive or metastatic tumor cells. MMP
expression is stimulated by growth factors and cytokines in the
local tissue environment, where these enzymes act to specifically
degrade protein components of the extracellular matrix, such as
collagen, proteoglycans (protein core), fibronectin and laminin.
These ubiquitous extracellular matrix components are present in the
linings of joints, interstitial connective tissues, basement
membranes and cartilage. Excessive degradation of extracellular
matrix by MMPs is implicated in the pathogenesis of many diseases,
including rheumatoid arthritis, osteoarthritis, periodontal
disease, aberrant angiogenesis, tumor invasion and metastasis,
corneal ulceration, and in complications of diabetes. MMP
inhibition is, therefore, recognized as a good target for
therapeutic intervention.
[0004] The MMPs share a number of properties, including zinc and
calcium dependence, secretion as zymogens, and 40-50% amino acid
sequence homology. The MMP family includes collagenases,
stromelysins, gelatinases, and matrilysin, as discussed in greater
detail below.
[0005] Interstitial collagenases catalyze the initial and
rate-limiting cleavage of native collagen types I, II, III and X.
Collagen, the major structural protein of mammals, is an essential
component of the matrix of many tissues, for example, cartilage,
bone, tendon and skin. Interstitial collagenases are very specific
matrix metalloproteases which cleave collagen to give two fragments
which spontaneously denature at physiological temperatures and
therefore become susceptible to cleavage by less specific enzymes.
Cleavage by the collagenase results in the loss of structural
integrity of the target tissue, essentially an irreversible
process.
[0006] The gelatinases include two distinct, but highly related,
enzymes: a 72-kD enzyme secreted by fibroblasts and a wide variety
of other cell types, and a 92-kD enzyme released by mononuclear
phagocytes, neutrophils, corneal epithelial cells, tumor cells,
cytotrophoblasts and keratinocytes. These gelatinases have been
shown to degrade gelatins (denatured collagens), collagen types IV
(basement membrane) and V, fibronectin and insoluble elastin.
[0007] The stromelysins (1 and 2) have been shown to cleave a broad
range of matrix substrates, including laminin, fibronectin,
proteoglycans, and collagen types IV and IX in their non-helical
domains.
[0008] Matrilysin (putative metalloprotease or PUMP) is a recently
described member of the matrix metalloprotease family. Matrilysin
has been shown to degrade a wide range of matrix substrates
including proteoglycans, gelatins, fibronectin, elastin, and
laminin. Its expression has been documented in mononuclear
phagocytes, rat uterine explants and sporadically in tumors.
[0009] Inhibitors of MMPs provide useful treatments for diseases
associated with the excessive degradation of extracellular matrix,
such as arthritic diseases (rheumatoid arthritis and
osteoarthritis), bone resorptive diseases (such as osteoporosis),
the enhanced collagen destruction associated with diabetes,
periodontal disease, corneal ulceration, ulceration of the skin,
tumor invasion and metastasis, and aberrant angiogenesis.
[0010] The design and uses of MMP inhibitors is described, for
example, in J. Enzyme Inhibition (1987), Vol. 2, pp. 1-22; Drug
News & Prospectives (1990), Vol. 3, No. 8, pp. 453-458;
Arthritis and Rheumatism (1993), Vol. 36, No. 2, pp. 181-189;
Arthritis and Rheumatism (1991), Vol. 34, No. 9, pp. 1073-1075;
Seminars in Arthritis and Rheumatism (1990), Vol. 19, No. 4,
Supplement 1 (February), pp. 16-20; Drugs of the Future (1990),
Vol. 15, No. 5, pp. 495-508; and J. Enzyme Inhibition (1987), Vol.
2, pp. 1-22. MMP inhibitors are also the subject of various patents
and patent applications, for example, U.S. Pat. Nos. 5,189,178
(Galardy) and 5,183,900 (Galardy), European Published Patent
Applications 0 438 223 (Beecham) and 0 276 436 (F. Hoffmann-La
Roche), and Patent Cooperation Treaty International Applications
92/21360 (Merck), 92/06966 (Beecham) and 92/09563 (Glycomed).
SUMMARY OF THE INVENTION
[0011] The invention provides new compounds which are useful as
inhibitors of matrix metalloproteases and which are effective in
treating disease-states characterized by excessive activity of
matrix metalloproteases.
[0012] Accordingly, one aspect of the invention is directed to
compounds of formula (I): 2
[0013] wherein:
[0014] R.sup.1 is mercapto, acetylthio, carboxy, hydroxycarbamoyl,
N-hydroxyformamide, alkoxycarbonyl, aryloxycarbonyl,
aralkoxycarbonyl, benzyloxycarbamoyl or a group of the formula
3
[0015] where R.sup.6 is aryl or heteroaryl;
[0016] R.sup.2 is alkyl, cycloalkyl, aryl, heterocycloalkyl, or
heteroaryl;
[0017] R.sup.3 is alkyl, cycloalkyl, aralkyl, or heteroaralkyl;
[0018] R.sup.7 is aryl, heteroaryl or heterocycloalkyl;
[0019] X is a group of the formula
--(CH.sub.2).sub.m--Y--(CH.sub.2).sub.n- --, where:
[0020] Y is O, S, or a single bond,
[0021] m is an integer from 0 to 4,
[0022] n is an integer from 0 to 4, and
[0023] m+n is an integer from 0 to 4;
[0024] p is an integer from 0 to 4, provided that R.sup.2--X is
biphenylalkyl when p is not 0;
[0025] and the pharmaceutically acceptable salts thereof.
[0026] Another aspect of the invention provides processes for
synthesizing the compounds and salts of formula (I).
[0027] In another aspect, the invention is directed to a sub-genus
of formula (I), i.e., the compounds of formula (II), as follows:
4
[0028] wherein:
[0029] R.sup.1 is mercapto, acetylthio, carboxy, hydroxycarbamoyl,
alkoxycarbonyl, aryloxycarbonyl, aralkoxycarbonyl,
benzyloxyaminocarbonyl or a group of the formula 5
[0030] where R.sup.6 is aryl or heteroaryl;
[0031] R.sup.2 is alkyl, aralkyl or cycloalkylalkyl;
[0032] R.sup.3 is cycloalkyl, alkyl (optionally substituted by
cycloalkyl, hydroxy, mercapto, alkylthio, aralkoxy, carboxy, amino,
alkylamino, guanidino, carbamoyl, pyridinyl or indolyl), or aralkyl
(optionally substituted by hydroxy, carboxy, alkyl or alkoxy);
[0033] R.sup.4 is nitro, amino, cyano, hydroxy, alkoxy, carboxy,
alkoxycarbonyl, alkylsulfonyl, haloalkyl, alkoxycarbonylalkyl,
tetrazolyl, carbamoyl (optionally substituted by alkyl or
dialkylaminoalkyl), or aminosulfonyl (optionally substituted by
alkyl); and
[0034] R.sup.5 is hydrogen, halo or hydroxy,
[0035] as a single stereoisomer or as a mixture thereof; and the
pharmaceutically acceptable salts thereof.
[0036] Another aspect of the invention is directed to compounds of
the formula 6
[0037] wherein:
[0038] R.sup.2 is alkyl, aryl or heteroaryl; and
[0039] X is a group of the formula
--(CH.sub.2).sub.m--Y--(CH).sub.n, where:
[0040] Y is O, S, or a single bond,
[0041] m is an integer from 0 to 4,
[0042] n is an integer from 0 to 4, and
[0043] m+n is an integer from 0 to 4;
[0044] or R.sup.2 and X together are lower alkenyl.
[0045] Another aspect of the invention is directed to processes for
synthesizing a compound of the formula 7
[0046] wherein R.sup.2 is aryl or heteroaryl, by
[0047] (a) hydrogenating a compound of the formula: 8
[0048] in the presence of a palladium/carbon catalyst; or
[0049] (b) contacting a compound of the formula 9
[0050] with sodium hexamethyldisilazide and
t-butylbromoacetate.
[0051] Other aspects of the invention are directed to compounds of
the formula 10
[0052] wherein R.sup.2 is aryl or heteroaryl, and a process for
synthesizing these compounds by
[0053] (a) contacting a compound of the formula: 11
[0054] where R.sup.2 is hydrogen, aryl or heteroaryl, with an
excess of mesyl chloride in pyridine followed by refluxing under
basic conditions, and
[0055] (b) where R.sup.2 is hydrogen in step (a), reacting the
product of step (a) with an aryl halide or a heteroaryl halide in
the presence of a base and a palladium catalyst.
[0056] Another aspect of the invention is directed to methods of
inhibiting matrix metalloprotease activity in a mammal, which
methods comprise administering to the mammal in need thereof a
therapeutically effective amount of a compound of formula (I) as
defined above, as a single stereoisomer, or as a mixture thereof,
or a pharmaceutically acceptable salt thereof.
[0057] Another aspect of the invention is directed to a
pharmaceutical composition useful in inhibiting matrix
metalloprotease activity in a mammal, which composition comprises a
therapeutically effective amount of a compound of formula (I) as
defined above, as a single stereoisomer or as a mixture thereof; or
a pharmaceutically acceptable salt thereof; and a pharmaceutically
acceptable excipient.
DETAILED DESCRIPTION OF THE INVENTION
[0058] Definitions
[0059] As used in the specification and appended claims, unless
specified to the contrary, the following terms have the meaning
indicated:
[0060] "BOC" refers to t-butoxycarbonyl.
[0061] "CBZ" refers to benzyloxycarbonyl (carbobenzyloxy).
[0062] "DCC" refers to N,N-dicyclohexylcarbodiimide.
[0063] "DMAP" refers to N,N-dimethylaminopyridine.
[0064] "DMF" refers to N,N-dimethylformamide.
[0065] "EDCI" refers to
N-ethyl-N'-(3-dimethylaminopropyl)-carbodiimide.
[0066] "HOBT" refers to 1-hydroxybenzotriazole.
[0067] "Hydroxy" refers to the radical --OH.
[0068] "Amino" refers to the radical --N.
[0069] "Acetylthio" refers to the radical --SC(O)CH3.
[0070] "Halo" refers to bromo, chloro or fluoro.
[0071] "Carbamoyl" refers to the radical --C(O)NH,.
[0072] "Carboxy" refers to the radical --C(O)OH.
[0073] "Hydroxyamino" refers to the radical --NHOH.
[0074] "Hydroxycarbamoyl" refers to the radical --C(O)NHOH.
[0075] "N-Hydroxyformamide" refers to the radical --N(OH)C(O)H
[0076] "Benzyloxycarbamoyl" refers to
--C(O)N(H)OCH.sub.2C.sub.6H.sub.5.
[0077] "Acylamide" refers to --NHC(O)R.sub.a where R.sub.a is
alkyl.
[0078] "Mercapto" refers to the radical --SH.
[0079] "Alkyl" refers to a straight or branched chain monovalent
radical consisting solely of carbon and hydrogen, containing no
unsaturation and having from one to ten carbon atoms, e.g., methyl,
ethyl, n-propyl, 2-methylpropyl (iso-butyl), 1-methylethyl
(iso-propyl), n-butyl, and 1,1-dimethylethyl (t-butyl), heptyl and
the like, which can be optionally substituted by cycloalkyl,
hydroxy, mercapto, alkylthio, aralkoxy, carboxy, amino, mono- and
di-alkylamino, guanidino, N,N-dialkylguanidino, carbamoyl, aryl,
and heteroaryl.
[0080] "Alkanyl" or "alkylene" refers to a straight chain divalent
radical consisting solely of carbon and hydrogen, containing no
unsaturation and having from one to five carbon atoms, e.g.,
methylene, ethylene, propylene (or propanyl) and the like.
[0081] "Lower alkenyl" refers to a straight chain univalent
hydrocarbon radical having from two to six carbon atoms and
containing at least one unsaturated bond, e.g., prop-2-enyl,
pent-4-enyl and the like.
[0082] "Alkylamino" refers to a radical of the formula --NHR.sub.a
where R.sub.a is alkyl as defined above, e.g., methylamino,
ethylamino, iso-propylamino, n-butylamino, and the like.
[0083] "Haloalkyl" refers to a radical of the formula
--R.sub.aR.sub.d where R.sub.a is alkyl as defined above
substituted by one or more halo groups (R.sub.d) as defined above,
e.g., 2-chloroethyl, 2-bromoethyl, trifluoromethyl, and the
like.
[0084] "Dialkylaminoalkyl" refers to a radical of the formula
--R.sub.aN(R.sub.a).sub.2 where each R.sub.a is independently an
alkyl radical as defined above, e.g., dimethyl-aminoethyl,
diethylamino-n-propyl, dimethylamino-n-propyl, and the like.
[0085] "Aminosulfonyl" refers to --S(O).sub.2NH.sub.2.
[0086] "Alkylsulfonyl" refers to a radical of the formula
--S(O).sub.2R.sub.a where R.sub.a is alkyl as defined above, e.g.,
methylsulfonyl, ethylsulfonyl, iso-propylsulfonyl, and the
like.
[0087] "Alkylsulfinyl" refers to a radical of the formula
--S(O)R.sub.a where R.sub.a is alkyl as defined above.
[0088] "Alkylthio" refers to a radical of the formula --SR.sub.a
where R.sub.a is optionally-substituted alkyl as defined above,
e.g., methylthio, ethylthio, iso-propylthio, n-butylthio, and the
like.
[0089] "Alkoxy" refers to a radical of the formula --OR.sub.a
wherein R.sub.a is alkyl as defined above, e.g., methoxy, ethoxy,
n-propoxy, 1-methylethoxy, n-butoxy, t-butoxy, and the like.
[0090] "Alkoxycarbonylalkyl" refers to a radical of the formula
--R.sub.aC(O)R.sub.b where R.sub.a is alkyl as defined above and
R.sub.b is alkoxy as defined above, e.g., methoxycarbonylethyl,
ethoxycarbonylethyl, methoxycarbonyl-iso-propyl, and the like.
[0091] "Aryl" refers to a monovalent unsaturated aromatic
carbocyclic radical having a single ring (e.g., phenyl), two
condensed rings (e.g., naphthyl) or three condensed rings (e.g.,
phenanthrenyl or fluorenyl) which can be optionally substituted by
one or more substituents independently selected from: alkyl,
hydroxy, carboxy, halo, cyano, amino, nitro, tetrazolyl,
heteroaryl, aminoalkoxy, alkylthio, haloalkyl, alkoxy,
alkoxycarbonyl, alkoxycarbonylalkyl, alkyl sulfonyl, alkyl
sulfinyl, aminosulfonyl optionally substituted by alkyl, carbamoyl
optionally substituted by alkyl or dialkylaminoalkyl, or the
substituent can be another aryl group as defined herein (e.g., to
form an optionally substituted biphenyl radical).
[0092] "Aryloxy" refers to a radical of the formula --OR.sub.b
wherein R.sub.b is aryl as defined above, e.g., phenoxy,
quinol-2-yloxy, naphth-1-yloxy, or naphth-2-yloxy.
[0093] "Aralkyl" refers to a radical of the formula
--R.sub.aR.sub.b wherein R.sub.a is alkyl as defined above and
R.sub.b is aryl as defined above, e.g., benzyl, phenylethylene,
3-phenylpropyl, and the like.
[0094] "Aralkoxy" refers to a radical of the formula
--OR.sub.aR.sub.b wherein R.sub.a is alkyl as defined above and
R.sub.b is aryl as defined above, e.g., benzyloxy,
3-naphth-2-ylpropoxy, and the like.
[0095] "Alkoxycarbonyl" refers to a radical of the formula
--C(O)R.sub.b wherein R.sub.b is alkoxy as defined above, e.g.,
methoxycarbonyl, ethoxycarbonyl, t-butoxycarbonyl, and the
like.
[0096] "Aralkoxycarbonyl" refers to a radical of the formula
--C(O)R.sub.o wherein R.sub.o is aralkoxy as defined above, e.g.,
benzyloxycarbonyl, and the like.
[0097] "Cycloalkyl" refers to a monovalent ring radical consisting
solely of carbon and hydrogen atoms, containing no unsaturation and
having from five to seven carbon atoms, e.g., cyclopentyl,
cyclohexyl and cycloheptyl.
[0098] "Cycloalkylalkyl" refers to a radical of the formula
--R.sub.oR.sub.a where R.sub.a is alkyl as defined above and
R.sub.o is cycloalkyl as defined above, e.g., cyclohexylmethyl,
cyclohexylethyl, cyclopentylmethyl, and the like.
[0099] "Heteroaryl" refers to a monovalent unsaturated aromatic
carbocyclic radical having a single ring or multiple condensed
rings with at least one heteroatom such as N,O,S, (e.g., pyridyl,
quinolyl, indolyl, carbazolyl, dibenzofuranyl, dibenzothiophenyl,
phenanthridinyl), which can be optionally substituted by one or
more substituents independently selected. from: alkyl, hydroxy,
carboxy, halo, cyano, amino, nitro, tetrazolyl, aryl, aminoalkoxy,
alkylthio, haloalkyl, alkoxy, alkoxycarbonyl, alkoxycarbonylalkyl,
alkyl sulfonyl, alkyl sulfinyl, aminosulfonyl optionally
substituted by alkyl, and carbamoyl optionally substituted by alkyl
or dialkylaminoalkyl.
[0100] "Heteroaralkyl" refers to a radical of the formula
--R.sub.aR.sub.b where R.sub.a is alkyl as defined above and
R.sub.b is heteroaryl as defined above.
[0101] "Heterocycloalkyl" refers to a monovalent saturated
carbocyclic radical having a single ring or multiple condensed
rings with at least one heteroatom such as N,O,S (e.g., morpholino,
piperazinyl, piperidinyl, pyrrolidinyl).
[0102] "Optional" or "optionally" means that the subsequently
described event of circumstances may or may not occur, and that the
description includes instances where said event or circumstance
occurs and instances in which it does not. For example, "optionally
substituted quinol-2-yl" means that the quinol-2-yl radical may or
may not be substituted and that the description includes both
substituted quinol-2-yl radicals and quinol-2-yl radicals having no
substitution.
[0103] "Amino-protecting group" as used herein refers to those
organic groups intended to protect nitrogen atoms against
undesirable reactions during synthetic procedures, and includes,
but is not limited to, benzyl, acyl, acetyl, benzyloxycarbonyl
(carbobenzyloxy), p-methoxybenzyloxy-carb- onyl,
p-nitrobenzyloxycarbonyl, c-butoxycarbonyl, trifluoroacetyl, and
the like.
[0104] "Base" as used here includes both strong bases such as
sodium hydroxide, lithium hydroxide, ammonium hydroxide, potassium
carbonate and the like, and organic bases such as pyridine,
diisopropylethylamine, N-methylmorpholine, triethylamine,
dimethylaminopyridine and the like.
[0105] "Pharmaceutically acceptable salt" refers to those salts
which retain the biological effectiveness and properties of the
free bases or free acids and which are not biologically or
otherwise undesirable. If the compound exists as a free base, the
desired salt may be prepared by methods known to those of ordinary
skill in the art, such as treatment of the compound with an
inorganic acids such as hydrochloric acid, hydrobromic acid,
sulfuric acid, nitric acid, phosphoric acid and the like; or with
an organic acids such as acetic acid, propionic acid, glycolic
acid, pyruvic acid, oxalic acid, maleic acid, malonic acid,
succinic acid, fumaric acid, tartaric acid, citric acid, benzoic
acid, cinnamic acid, mandelic acid, methanesulfonic acid,
ethanesulfonic acid, p-toluenesulfonic acid, salicylic acid, and
the like. If the compound exists as a free acid, the desired salt
may also be prepared by methods known to those of ordinary skill in
the art, such as the treatment of the compound with an inorganic
base or an organic base. Salts derived from inorganic bases
include, but are not limited to, the sodium, potassium, lithium,
ammonium, calcium, magnesium, iron, zinc, copper, manganese,
aluminum salts and the like. Salts derived from organic bases
include, but are not limited to, salts of primary, secondary, and
tertiary amines, substituted amines including naturally occurring
substituted amines, cyclic amines and basic ion exchange resins,
such as isopropylamine, trimethylamine, diethylamine,
triethylamine, tripropylamine, ethanolamine,
2-dimethylaminoethanol, 2-diethylaminoethanol, trimethamine,
dicyclohexylamine, lysine, arginine, histidine, caffeine, procaine,
hydrabamine, choline, betaine, ethylenediamine, glucosamine,
methylglucamine, theobromine, purines, piperazine, piperidine,
N-ethylpiperidine, polyamine resins and the like.
[0106] "Mammal" includes humans and all domestic and wild animals,
including, without limitation, cattle, horses, swine, sheep, goats,
dogs, cats, and the like.
[0107] "Therapeutically effective amount" refers to that amount of
a compound of formula (I) which, when administered to a mammal in
need thereof, is sufficient to effect treatment, as defined below,
for disease-states alleviated by the inhibition of matrix
metalloprotease activity, such as the activity of stromelysin,
gelatinase, matrilysin and/or collagenase. The amount of a compound
of formula (I) which constitutes a
[0108] "Therapeutically effective amount" will vary depending on
the compound, the disease-state and its severity, and the mammal to
be treated, but can be determined routinely by one of ordinary
skill in the art having regard to his own knowledge and to this
disclosure.
[0109] "Treating" or "treatment" as used herein cover the treatment
of a disease-state in a mammal, particularly in a human, which
disease-state is alleviated by the inhibition of matrix
metalloprotease activity, such as the activity of stromelysin,
gelatinase, matrilysin and/or collagenase, and include:
[0110] (i) preventing the disease-state from occurring in a mammal,
in particular, when such mammal is predisposed to the disease-state
but has not yet been diagnosed as having it;
[0111] (ii) inhibiting the disease-state, i.e., arresting its
development; or
[0112] (iii) relieving the disease-state, i.e., causing regression
of the disease-state.
[0113] "Stereoisomers" refers to compounds having identical
molecular formulae and nature or sequence of bonding but differing
in the arrangement of their atoms in space.
[0114] The compounds of formula (I), or their pharmaceutically
acceptable salts, have at least two asymmetric carbon atoms in
their structure, and may therefore exist as single stereoisomers,
racemates, and as mixtures of enantiomers and diastereomers. All
such single stereoisomers, racemates and mixtures thereof are
intended to be within the scope of this invention.
[0115] When naming the single stereoisomers of compounds of formula
(I) an absolute descriptor, R or S, may be assigned to the chiral
carbon atoms therein according to the "Sequence Rule" procedure of
Cahn, Ingold and Prelog.
[0116] Nomenclature
[0117] The nomenclature used herein is a modified form of
I.U.P.A.C. nomenclature wherein the compounds of the invention are
named as peptide derivatives. Where R.sup.3 of Formula (I)
comprises the side chain of an amino acid residue, that portion of
the chemical structure which includes R.sup.3 together with the
adjacent nitrogen atom (illustrated below and named as the N
nitrogen, as opposed to the N' nitrogen) and carbonyl group is
given the name of the corresponding amino acid. The naming and
numbering of the compounds of the present invention is illustrated
below for representative compounds of formula (I).
[0118] For example, the following compound of formula (I) 12
[0119] wherein R.sup.1 is carboxy; R.sup.2 is biphenyl; R.sup.3 is
4-(cyano)benzylthioisopropyl; R.sup.7 is phenyl; X is propanyl; and
p is 0, is named
N-(2R-carboxymethyl-5-(biphen-4-yl)pentanoyl)-L-S-((4-cyanoph-
enyl)methyl)-penicillamine-N'-(phenyl)carboxamide. Another name for
this compound is
N-(5-(biphen-4-yl)-2R-carboxymethylpentanoyl)-L-S-((4-cyanoph-
enyl)methyl)-penicillamine-N'-(phenyl)carboxamide. For ease of
reference, the portions of the structure are associated with their
corresponding nomenclature.
[0120] The structures and names of several other representative
compounds of formula (I) follow. 13
[0121] The above compound is named
N-(2R-carboxymethyl-5-(biphen-4-yl)pent-
anoyl)-L-t-butylglycine-N'-(pyrid-4-yl)carboxamide. The term
t-leucine can be interchanged with t-butylglycine, and the term
pyridinyl can be interchanged with pyridyl. Another name for the
above compound is:
N-(5-biphen-4-yl-2R-carboxymethylpentanoyl)-L-t-leucine-N'-(pyridin-4-yl)-
carboxamide. 14
[0122] The above compound is named
N-(2R-carboxymethyl-5-(7-(glycyl)aminof-
luoren-2-yl)pentanoyl)-L-leucine-N'(4-(methoxycarbonyl)phenyl)carboxamide.
Another name for this compound is
N-(5-(7-(glycyl)aminofluoren-2-yl)-2R-c-
arboxymethylpentanoyl)-L-leucine-N'-(4-methoxycarbonylphenyl)carboxamide.
15
[0123] The above compound is named
N-(2R-carboxymethyl-5-(phenyl)pentanoyl-
-L-6-(N,N'-diethylguanido)lysyl-N'-(4-(ethoxycarbonyl)phenyl)carboxamide.
The term guanidino can be used interchangeably with guanido.
Another name for this compound is
N-(5-phenyl-2R-carboxymethylpentanoyl)-L-6-(N,N'-die-
thylguanidino)-lysyl-N'-(4-ethoxycarbonylphenyl)carboxamide. 16
[0124] The above compound is named
N-(2R-(N"-formyl-N"-hydroxyamino)methyl- -4-((3-chloro,
5-morpholino)phen-1-yl) oxybutanoyl)-L-cyclohexylglycine-N'-
(4-(indol-5-yl)butyl)carboxamide. Another name for this compound is
N-(4-((3-chloro,
5-morpholino)phen-1-yl)-2R-(N"-formyl-N"-hydroxyamino)me-
thyloxy-butanoyl)-L-cyclohexylglycine-N'-(4-(indol-5-yl)butyl)carboxamide.
[0125] Utility, Testing and Administration
[0126] Utility
[0127] The compounds of formula (I) inhibit mammalian matrix
metalloproteases, such as the stromelysins, gelatinases, matrilysin
and collagenases, and are therefore useful for treating diseases
associated with the MMP-induced excessive degradation of matrix and
connective tissue within the mammal, for example, arthritic
diseases (rheumatoid arthritis and osteoarthritis), bone resorptive
diseases (such as osteoporosis), the enhanced collagen destruction
associated with diabetes, periodontal disease, corneal ulceration,
ulceration of the skin, tumor invasion and metastasis, and aberrant
angiogenesis.
[0128] Testing
[0129] The ability of the compounds of formula (I) to inhibit
matrix metalloprotease activity, such as the activity of
stromelysin, gelatinase, matrilysin and/or collagenase may be
demonstrated by a variety of in vitro and in vivo assays known to
those of ordinary skill in the art, such as the assay described in
Anal. Biochem. (1985), Vol. 147, p. 437, and the MMP Enzymatic
Assay described in FEBS (1992), Vol. 296(3), p. 263, or
modifications thereof.
[0130] Administration
[0131] Administration of the compounds of formula (I), or their
pharmaceutically acceptable salts, in pure form or in an
appropriate pharmaceutical composition, can be carried out via any
of the accepted modes of administration or agents for serving
similar utilities. Thus, administration can be, for example,
orally, nasally, parenterally, topically, transdermally, or
rectally, in the form of solid, semi-solid, lyophilized powder, or
liquid dosage forms, such as for example, tablets, suppositories,
pills, soft elastic and hard gelatin capsules, powders, solutions,
suspensions, or aerosols, or the like, preferably in unit dosage
forms suitable for simple administration of precise dosages. The
compositions will include a conventional pharmaceutical carrier or
excipient and a compound of formula (I) as the/an active agent,
and, in addition, may include other medicinal agents,
pharmaceutical agents, carriers, adjuvants, etc.
[0132] Generally, depending on the intended mode of administration,
the pharmaceutically acceptable compositions will contain about 1%
to about 99% by weight of a compound(s) of formula (I), or a
pharmaceutically acceptable salt thereof, and 99% to 1% by weight
of a suitable pharmaceutical excipient. Preferably, the composition
will be about 5% to 75% by weight of a compound(s) of formula (I),
or a pharmaceutically acceptable salt thereof, with the rest being
suitable pharmaceutical excipients.
[0133] The preferred route of administration is oral, using a
convenient daily dosage regimen which can be adjusted according to
the degree of severity of the disease-state to be treated. For such
oral administration, a pharmaceutically acceptable composition
containing a compound(s) of formula (I), or a pharmaceutically
acceptable salt thereof, is formed by the incorporation of any of
the normally employed excipients, such as, for example,
pharmaceutical grades of mannitol, lactose, starch, pregelatinized
starch, magnesium stearate, sodium saccharine, talcum, cellulose
ether derivatives, glucose, gelatin, sucrose, citrate, propyl
gallate, and the like. Such compositions take the form of
solutions, suspensions, tablets, pills, capsules, powders,
sustained release formulations and the like.
[0134] Preferably such compositions will take the form of capsule,
caplet or tablet and therefore will also contain a diluent such as
lactose, sucrose, dicalcium phosphate, and the like; a disintegrant
such as croscarmellose sodium or derivatives thereof; a lubricant
such as magnesium stearate and the like; and a binder such as a
starch, gum acacia, polyvinylpyrrolidone, gelatin, cellulose ether
derivatives, and the like.
[0135] The compounds of formula (I), or their pharmaceutically
acceptable salts, may also be formulated into a suppository using,
for example, about 0.5% to about 50% active ingredient disposed in
a carrier that slowly dissolves within the body, e.g.,
polyoxyethylene glycols and polyethylene glycols (PEG), e.g., PEG
1000 (96%) and PEG 4000 (4%).
[0136] Liquid pharmaceutically administrable compositions can, for
example, be prepared by dissolving, dispersing, etc., a compound(s)
of formula (I) (about 0.5% to about 20%), or a pharmaceutically
acceptable salt thereof, and optional pharmaceutical adjuvants in a
carrier, such as, for example, water, saline, aqueous dextrose,
glycerol, ethanol and the like, to thereby form a solution or
suspension.
[0137] If desired, a pharmaceutical composition of the invention
may also contain minor amounts of auxiliary substances such as
wetting or emulsifying agents, Ph buffering agents, antioxidants,
and the like, such as, for example, citric acid, sorbitan
monolaurate, triethanolamine oleate, butylated hydroxytoluene,
etc.
[0138] Actual methods of preparing such dosage forms are known, or
will be apparent, to those skilled in this art; for example, see
Remington's Pharmaceutical Sciences, 18th Ed., (Mack Publishing
Company, Easton, Pa., 1990). The composition to be administered
will, in any event, contain a therapeutically effective amount of a
compound of formula (I), or a pharmaceutically acceptable salt
thereof, for treatment of a disease-state alleviated by the
inhibition of matrix metalloprotease activity in accordance with
the teachings of this invention.
[0139] The compounds of formula (I), or their pharmaceutically
acceptable salts, are administered in a therapeutically effective
amount which will vary depending upon a variety of factors
including the activity of the specific compound employed, the
metabolic stability and length of action of the compound, the age,
body weight, general health, sex, diet, mode and time of
administration, rate of excretion, drug combination, the severity
of the particular disease-state, and the host undergoing therapy.
Generally, a therapeutically effective daily dose is from about
0.14 mg to about 14.3 mg/kg of body weight per day of a compound of
formula (I), or a pharmaceutically acceptable salt thereof;
preferably, from about 0.7 mg to about 10 mg/kg of body weight per
day; and most preferably, from about 1.4 mg to about 7.2 mg/kg of
body weight per day. For example, for administration to a 70 kg
person, the dosage range would be from about 10 mg to about 1.0
gram per day of a compound of formula (I), or a pharmaceutically
acceptable salt thereof, preferably from about 50 mg to about 700
mg per day, and most preferably from about 100 mg to about 500 mg
per day.
[0140] Preferred Embodiments
[0141] Preferred are the compounds of formula (I) where X is
alkanyl and where p is zero, 2 or 3.
[0142] Of the compounds where p is 2 or 3, particularly preferred
are those compounds where R.sup.1 is carboxy, R.sup.2 is biphenyl,
R.sup.3 is cyclohexyl, and R.sup.7 is optionally substituted phenyl
[especially 4-(aminosulfonyl)phenyl] or N-morpholino.
[0143] Of the compounds where p is zero, particularly preferred are
the group of compounds where R.sup.2 is alkyl, optionally
substituted phenyl, or a group of the formula: 17
[0144] and R.sup.7 is 4-pyridyl or optionally substituted
phenyl.
[0145] Within this group, a preferred subgroup of compounds are
those where R.sup.1 is carboxy, hydroxycarbamoyl, or
N-hydroxyformamide; R.sup.2 is phenyl, biphenyl, 4-(pyridyl)phenyl,
or 2-methylpropyl; R.sup.3 is t-butyl, 4-aminobutyl,
alkylaminobutyl, dialkylaminobutyl, 4-(N,N'diethyl-guanidino)butyl,
propyl, 2-methylpropyl, 1-hydroxyisopropyl, 1-hydroxyethyl, or
cyclohexyl; and X is a single bond, ethylene or propanyl.
[0146] Within this subgroup, a preferred class of compounds are
those where R.sup.2 is biphenyl, R.sup.3 is t-butyl and R.sup.7 is
4-pyridyl, particularly where R.sup.1 is carboxy,
N-hydroxyformamide, or hydroxycarbamoyl.
[0147] Also preferred is the subgroup of compounds where R.sup.2 is
a group of the formula: 18
[0148] where A is CH.sub.2; R.sup.10 is H or acylamide; R.sup.11 is
H; R.sup.7 is optionally substituted phenyl; and X is propanyl.
[0149] Within this subgroup, preferred are the compounds where
R.sup.1 is carboxy, hydroxycarbamoyl, or N-hydroxyformamide;
R.sup.3 is alkyl (especially 2-methylpropyl); and R.sup.7 is
alkoxycarbonylphenyl [especially 4-(methoxycarbonyl)phenyl].
[0150] Also within this group, preferred are the subgroup of
compounds where R.sup.1 is carboxy; R.sup.2 is phenyl; R.sup.3 is
alkyl [especially 4-(amino)butyl and 4-(diethylguanidino)N-butyl]
or cycloalkyl (especially cyclohexyl); and R.sup.7 is optionally
substituted phenyl [especially 4-(ethoxycarbonyl)phenyl or
4-(dialkylaminoethylaminosulfonyl)phenyl]; and X is ethylene or
propanyl.
[0151] Also within this group, preferred are the subgroup of
compounds where R.sup.1 is mercapto, carboxy, hydroxycarbamoyl, or
N-hydroxyformamide; R.sup.2 is 2-methylpropyl; R.sup.3 is alkyl
[especially propyl, 2-methylpropyl], cycloalkyl [especially
cyclohexyl] or heteroaralkyl [especially 3-methylindolyl]; R.sup.7
is optionally substituted phenyl [especially 4-(methoxy)phenyl,
4-(carboxy)phenyl, 4-(methoxycarbonyl)phenyl or
4-(dimethylaminoethylcarbamoyl)phenyl]; and X is a single bond.
[0152] Also within this group, preferred are the subgroup of
compounds where R.sup.1 is carboxy; R.sup.2 is
4-(2-hydroxyethyl)phenyl, 4-(2-hydroxypropyl)-phenyl,
4-(2-hydroxybutyl)phenyl, 4-(pyridyl)phenyl, biphenyl,
4'-(aminoethoxy)biphenyl, 4'-(cyano)biphenyl, or
4'-(hydroxy)biphenyl; R.sup.3 is 2-methylpropyl; R.sup.7 is
4-(methoxycarbonyl)phenyl; and X is propanyl.
[0153] Particularly preferred is the subgroup of compounds where
R.sup.2 is biphenyl, especially where R.sup.7 is optionally
substituted phenyl.
[0154] Within this particularly preferred subgroup, preferred are
the compounds where R.sup.1 is carboxy; R.sup.3 is alkyl or
cycloalkyl [especially cyclohexyl, 4-(amino)butyl,
4-(isopropylamino)butyl, 1-hydroxyisopropyl or t-butyl]; X is
propanyl, and R.sup.7 is phenyl,
4-(hydroxyethylaminosulfonyl)-phenyl,
4-(dimethylaminoethyl-aminosulfonyl- )phenyl,
4-(ethoxycarbonyl)phenyl, 4-(N-morpholinopropylaminosulfonyl)phen-
yl, 4-(methylaminosulfonyl)phenyl,
4-(hydroxyethylaminosulfonyl)phenyl, or
4-(methylsulfinyl)phenyl.
[0155] Another preferred group, particularly for matrilysin
inhibition, are the compounds of formula (II), particularly those
compounds wherein R.sup.1 is mercapto or acetylthio.
[0156] Within this second group, a preferred subgroup of compounds
are those compounds wherein R.sup.2 is alkyl, aralkyl,
cycloalkylalkyl; R.sup.3 is cycloalkyl or alkyl (optionally
substituted by cycloalkyl, hydroxy, aralkoxy, alkylthio, pyridinyl
or indolyl); R.sup.4 is cyano, carboxy, hydroxy, alkoxy,
alkoxycarbonyl, alkoxycarbonylalkyl, carbamoyl (optionally
substituted by aralkylaminoalkyl), or aminosulfonyl (optionally
substituted by alkyl); and R.sup.5 is hydrogen.
[0157] Within this subgroup, a preferred class of compounds are
those compounds wherein R.sup.2 is alkyl; R.sup.3 is cyclohexyl,
alkyl (optionally substituted by cyclohexyl, hydroxy, benzyloxy,
methylthio, pyridinyl or indolyl); and R.sup.4 is carboxy,
alkoxycarbonyl and aminosulfonyl.
[0158] Within this class of compounds, compounds wherein R.sup.2 is
2-methylpropyl are preferred. Particularly preferred are those
compounds wherein R.sup.3 is 2-methylpropyl.
[0159] A third group preferred for matrilysin inhibition, are the
compounds of formula (II) wherein R.sup.1 is carboxy.
[0160] Within this third group, a preferred subgroup of compounds
are those compounds wherein R.sup.2 is alkyl, aralkyl,
cycloalkylalkyl; R.sup.3 is cycloalkyl or alkyl (optionally
substituted by cycloalkyl, hydroxy, aralkoxy, alkylthio, pyridinyl
or indolyl); R.sup.4 is cyano, hydroxy, alkoxy, carboxy,
alkoxycarbonyl, alkoxycarbonylalkyl, carbamoyl (optionally
substituted by aralkylaminoalkyl), or aminosulfonyl (optionally
substituted by alkyl); and R.sup.5 is hydrogen.
[0161] Within this subgroup, a preferred class of compounds are
those compounds wherein R.sup.2 is alkyl; R.sup.3 is cyclohexyl,
alkyl (optionally substituted by cyclohexyl, hydroxy, benzyloxy,
methylthio, pyridinyl or indolyl); and R.sup.4 is carboxy,
alkoxycarbonyl and aminosulfonyl.
[0162] Within this class of compounds, preferred compounds are
those compounds wherein R.sup.2 is 2-methylpropyl. Particularly
preferred are those compounds wherein R.sup.3 is cyclohexyl,
2-methylpropyl, pyridin-3-ylmethyl, 1-benzyloxyethyl,
1-methylpropyl, 1,1-dimethylethyl, 1-hydroxyethyl, and
indol-2-ylmethyl; and R.sup.4 is methoxycarbonyl.
[0163] A fourth group preferred for matrilysin inhibition, are the
compounds of formula (II) wherein R.sup.1 is hydroxycarbamoyl.
[0164] Within this fourth group, a preferred subgroup of compounds
are those compounds wherein R.sup.2 is alkyl, aralkyl,
cycloalkylalkyl; R.sup.3 is cycloalkyl or alkyl (optionally
substituted by cycloalkyl, hydroxy, aralkoxy, alkylthio, pyridinyl
or indolyl); R.sup.4 is cyano, hydroxy, alkoxy, carboxy,
alkoxycarbonyl, alkoxycarbonylalkyl, carbamoyl (optionally
substituted by aralkylaminoalkyl), or aminosulfonyl (optionally
substituted by alkyl); and R.sup.5 is hydrogen.
[0165] Within this subgroup, a preferred class of compounds are
those compounds wherein R.sup.2 is alkyl; R.sup.3 is cyclohexyl,
alkyl (optionally substituted by cyclohexyl, hydroxy, benzyloxy,
methylthio, pyridinyl or indolyl); and R.sup.4 is carboxy,
alkoxycarbonyl and aminosulfonyl.
[0166] Within this class, preferred compounds are those compounds
wherein R.sup.2 is 2-methylpropyl. Particularly preferred are those
compounds wherein R.sup.3 is cyclohexyl, 2-methylpropyl,
pyridin-3-ylmethyl, 1-benzyloxyethyl, 1-methylpropyl,
1,1-dimethylethyl, 1-hydroxyethyl, and indol-2-ylmethyl.
[0167] Presently, the most preferred compounds of formula (I) are
the following:
[0168]
N-(2R-(N"-hydroxycarbamoyl)methyl-4-(methyl)pentanoyl)-L-tryptophan-
-N'-(4-(carboxy)phenyl)carboxamide;
[0169]
N-(2R-(N"-hydroxycarbamoyl)methyl-4-(methyl)pentanoyl)-L-leucine-N'-
-(4-(methoxycarbonyl)phenyl)carboxamide;
[0170]
N-(2R-(N"-hydroxycarbamoyl)methyl-4-(methyl)pentanoyl)-L-leucine-N'-
-(4-(carboxy)phenyl)carboxamide;
[0171]
N-(2R-mercaptomethyl-4-(methyl)pentanoyl)-L-leucine-N'-(4-(methoxyc-
arbonyl)phenyl)carboxamide;
[0172]
N-(2R-acetylthiomethyl-4-(methyl)pentanoyl)-L-leucine-N'-(4-methoxy-
carbonylphenyl)carboxamide;
[0173]
N-(2R-carboxymethyl-4-(methyl)pentanoyl)-L-leucine-N'-(4-(methoxyca-
rbonyl)phenyl)carboxamide;
[0174]
N-(2R-(N"-hydroxycarbamoyl)methyl-4-(methyl)pentanoyl)-L-cyclohexyl-
glycine-N'-(4-(methoxycarbonyl)phenyl)carboxamide;
[0175]
N-(2R-(N"-hydroxycarbamoyl)methyl-4-(methyl)pentanoyl)-L-t-leucine--
N'-(4-(methoxycarbonyl)phenyl)carboxamide;
[0176]
N-(2R-(N-hydroxycarbamoyl)methyl-5-(biphen-4-yl)pentanoyl-L-t-leuci-
ne-N'-(pyrid-4-yl)carboxamide;
[0177]
N-(2R-carboxymethyl-5-(biphen-4-yl)pentanoyl)-L-t-leucine-N'-(pyrid-
in-4-yl)carboxamide;
[0178]
N-(2R-carboxymethyl-5-(biphen-4-yl)pentanoyl)-L-t-leucine-N'-(4-((2-
-hydroxyethyl)aminosulfonyl)phenyl)carboxamide;
[0179]
N-(2R-carboxymethyl-5-(4-(pyrid-4-yl)phenyl)pentanoyl)-L-leucine-N'-
-(4-(methoxycarbonyl)phenyl)carboxamide;
[0180]
N-(2R-carboxymethyl-5-(biphen-4-yl)pentanoyl)-L-.beta.-hydroxyvalin-
e-N'-(phenyl)carboxamide;
[0181]
N-(N"-formyl-N"-hydroxyamino)methyl-5-(biphen-4-yl)pentanoyl)-L-t-l-
eucine-N'-(pyridin-4-yl)carboxamide;
[0182]
N-(2R,S)-(N"-formyl-N"-hydroxyamino)methyl-4-(methyl)pentanoyl)-L-t-
-leucine-N'-(4-(methoxycarbonyl)phenyl)carboxyamide,
[0183]
N-(2R-carboxymethyl-5-(biphen-4-yl)pentanoyl)-L-t-leucine-N'-(4R,S--
(methylsulfinyl)phenyl)carboxamide;
[0184]
N-(2R-carboxymethyl-5-(biphen-4-yl)pentanoyl)-L-t-leucine-N'-(4-(me-
thylaminosulfonyl)phenyl)carboxamide;
[0185]
N-(2R-carboxymethyl-5-(biphen-4-yl)pentanoyl)-L-t-leucine-N'-(4-(3--
(morpholin-4-yl)propylaminosulfonyl)phenyl)carboxamide;
[0186]
N-(2R-carboxymethyl-5-(biphen-4-yl)pentanoyl)-L-cyclohexylglycine-N-
'-(4-((2-hydroxyethyl)aminosulfonyl)phenyl)carboxamide;
[0187]
N-(2R-carboxymethyl-5-(biphen-4-yl)pentanoyl)-L-cyclohexylglycine-N-
'-(4((2-(dimethylamino)ethyl)aminosulfonyl)phenyl)carboxamide;
[0188]
N-(2R-(N"-hydroxycarbamoyl)methyl-4-(methyl)pentanoyl)D,L-norvaline-
-N'-(4-(dimethylaminoethylcarbamoyl)phenyl)carboxamide;
[0189]
N-(2R-carboxymethyl-5-(biphen-4-yl)pentanoyl)-L-lysine-N'-(4-(ethox-
ycarbonyl)phenyl)carboxamide;
[0190]
N-(2R-carboxymethyl-5-(phenyl)pentanoyl)-L-lysine-N'-(4-(ethoxycarb-
onyl)phenyl)carboxamide;
[0191]
N-(2R-carboxymethyl-5-(biphen-4-yl)pentanoyl)-L-(N.epsilon.-isoprop-
yl)lysine-N'-(4-(ethoxycarbonyl)phenyl)carboxamide;
[0192]
N-(2R-carboxymethyl)-4-(phenyl)butanoyl)-L-cyclohexylglycine-N'-(4--
(N",N"-dimethylaminoethylaminosulfonyl)-phenyl)carboxamide; and
[0193]
N-(2R-carboxymethyl-5-(phenyl)pentanoyl)-L-(N,N'-diethylguanido)lys-
ine-N'-(4-(ethoxycarbonyl)phenyl)carboxamide.
SYNTHESIS OF COMPOUNDS OF FORMULA (I)
[0194] The compounds of formula (I) are prepared as described
below, for example with reference to Reaction Schemes 1-7, in which
the substituent groups illustrated (e.g., R.sup.1, R.sup.2, etc.)
have the same meanings as described in the Summary of the
Invention, unless otherwise specified. Certain of the reaction
schemes illustrate structures of formula (I) where p is zero and
R.sup.7 is an optionally substituted phenyl group [the substituents
R.sup.4 and R.sup.5 having been described in connection with
formula (II) in the Summary of the Invention]. As those skilled in
the art will appreciate, while the corresponding compounds where p
is 1-4 and where R.sup.7 is as otherwise defined can be analogously
prepared, combinations of substituents and/or variables in
compounds of formula (I) and intermediates thereof are permissible
only when such combinations result in stable compounds.
[0195] Compounds of formula (I) and their pharmaceutically
acceptable salts, as single stereoisomers or as mixtures thereof,
are peptide derivatives all or portions of which can be prepared
from the constituent .alpha.-amino acid derivative(s). Standard
methods for the formation of peptide bonds are illustrated by M.
Bodanszky et al., The Practice of Peptide Synthesis (1984),
Springer-Verlag; M. Bodanszky, Principles of Peptide Synthesis
(1984), Springer-Verlag; J. P. Greenstein et al., Chemistry of the
Amino Acids (1961), Vol. 1-3, John Wiley and Sons Inc.; G. R.
Pettit, Synthetic Peptides (1970), Vol. 1-2, Van Nostrand Reinhold
Company.
[0196] Synthetic Reaction Parameters
[0197] The terms "solvent", "inert organic solvent" or "inert
solvent" mean a solvent inert under the conditions of the reaction
being described in conjunction therewith [including, for example,
benzene, toluene, acetonitrile, tetrahydrofuran ("THF"),
dimethylformamide ("DMF"), chloroform, methylene chloride (or
dichloromethane), diethyl ether, methanol, pyridine and the like].
Unless specified to the contrary, the solvents used in the
reactions of the present invention are inert organic solvents.
[0198] The term "q.s." means adding a quantity sufficient to
achieve a stated function, such as to bring a solution to a desired
volume.
[0199] Unless specified to the contrary, the reactions described
herein take place at atmospheric pressure within a temperature
range from 5.degree. C. to 100.degree. C. (preferably from
10.degree. C. to 50.degree. C.; most preferably at "room" or
"ambient" temperature, e.g., 20.degree. C.). Further, unless
otherwise specified, the reaction times and conditions are intended
to be approximate, e.g., taking place at about atmospheric pressure
within a temperature range of about 5.degree. C. to about
100.degree. C. (preferably from about 10.degree. C. to about
50.degree. C.; most preferably about 20.degree. C.) over a period
of about 1 to about 10 hours (preferably about 5 hours). Parameters
given in the Examples are intended to be specific, not
approximate.
[0200] Amide couplings used to form the compounds of formula (I)
are generally performed by the carbodiimide method with reagents
such as dicyclohexylcarbodiimide or
N'-ethyl-N'-(3-dimethylaminopropyl)-carbodiim- ide (EDCI) in the
presence of 1-hydroxybenzotriazole (HOBT) in an inert solvent such
as dimethylformamide (DMF). Other methods of forming the amide or
peptide bond include, but are not limited to synthetic routes via
an acid chloride, acyl azide, mixed anhydride or activated ester
such as nitrophenyl ester. Typically, solution phase amide
couplings with or. without peptide fragments are performed.
[0201] The selection of protecting groups for the terminal amino or
carboxy groups of compounds used in the preparation of the
compounds of formula (I) is dictated in part by the particular
amide or peptide coupling conditions, and in part by the amino acid
and/or peptide components involved in the coupling.
Amino-protecting groups commonly used include those which are well
known in the art, e.g., p-methoxybenzyloxycarbonyl,
benzyloxycarbonyl (also referred to as carbobenzyloxy or CBZ),
p-nitrobenzyloxycarbonyl, t-butoxycarbonyl (BOC), and the like. It
is preferred to use either BOC or CBZ as the protecting group for
the .alpha.-amino group because of the relative ease of its removal
by mild acids [e.g., by trifluoroacetic acid (TFA) or hydrochloric
acid in ethyl acetate] or by catalytic hydrogenation.
[0202] Isolation and purification of the compounds and
intermediates described herein can be effected, if desired, by any
suitable separation or purification procedure such as, for example,
filtration, extraction, crystallization, column chromatography,
thin-layer chromatography or thick-layer chromatography, or a
combination of these procedures. Specific illustrations of suitable
separation and isolation procedures can be had by reference to the
examples hereinbelow. However, other equivalent separation or
isolation procedures can, of course, also be used.
[0203] The individual stereoisomers of compounds of formula (I) may
be separated from each other by methods known to those of ordinary
skill in the art, e.g., by selective crystallization or by
chromatography, and/or by the methods disclosed herein.
[0204] Preparation of Formula (E)
[0205] Compounds of formula (E) are intermediates used in the
preparation of compounds of formula (I), and are prepared as shown
in Reaction Scheme 1 wherein R.sup.12 is mesyl or tosyl: 19
[0206] Starting Materials--Compounds of formula (Ea) may be
prepared according to methods known to those of ordinary skill in
the art (e.g., see European Published Patent Application 0 276 436)
or may be prepared according to the method described in Example 1
below. Compounds of formula (Ed) are commercially available or may
be prepared according to methods known to those of ordinary skill
in the art.
[0207] Formula (Eb)--In general, compounds of formula (R) are
prepared by first treating a compound of formula (Ea) in an aprotic
solvent, preferably tetrahydrofuran and methylene chloride, at
0-15.degree. C., preferably at 0.degree. C., in the presence of a
base, preferably diisopropylethylamine and
bis-(trimethylsilyl)-acetamide, with paraformaldehyde. The
resulting solution is brought to 25-37.degree. C., preferably to
37.degree. C., for 18 hours. The alcohol of formula (Eb) is then
isolated by standard methods, preferably by evaporation of solvent,
extraction and filtration.
[0208] Formula (Ec)--An alcohol of formula (Eb) in an aprotic
solvent, preferably methylene chloride, is then cooled to
-20.degree. C. to about 0.degree. C., preferably to about
-20.degree. C., and is then esterified by the standard procedure of
treating the alcohol with at least a stoichiometric amount to about
a 100% excess of either mesyl chloride or tosyl chloride. The
esterification takes place over an initial period of time
(preferably 15 minutes) at -20.degree. C., followed by second
period of time (preferably 3.5 hours) at room temperature. The
ester of formula (Ec) is then isolated from the reaction mixture by
standard isolation procedures, preferably by extraction, filtration
and evaporation.
[0209] Formula (Ee)--An ester of formula (Ec) in an aprotic
solvent, preferably DMF, is then reacted with a salt of a compound
of formula (Ed) (preferably the sodium salt formed from the
reaction of the compound of formula (Ed) with sodium hydride in an
aprotic solvent, preferably DMF), for about 16-20 hours, preferably
for about 18 hours, at temperatures beginning at about 0.degree. C.
and slowly warming to room temperature. The resulting mercapto
compound of formula (Ee) is isolated from the reaction mixture by
standard isolation techniques, such as by extraction, evaporation,
and flash chromatography.
[0210] Formula (E)--A compound of formula (Ee) is then hydrolyzed
under basic conditions, preferably in the presence of sodium
hydroxide, to form a compound of formula (E), which is isolated
from the reaction mixture by standard isolation techniques.
[0211] Preparation of Formula (Ia)
[0212] Compounds of formula (Ia) are compounds of formula (I)
wherein R.sup.1 is a group of the formula 20
[0213] (where, when R.sup.6 is aryl it is preferably naphth-1-yl,
naphth-2-yl or phenyl, and when R.sup.6 is heteroaryl it is
preferably pyridyl or quinol-2-yl; R.sup.2 is preferably alkyl; and
R.sup.5 is preferably hydrogen) are prepared as described in
Reaction Scheme 2. 21
[0214] Starting Materials--N-protected amino acids of formula (A)
and compounds of formula (B) are commercially available or may be
prepared according to methods known to those of ordinary skill in
the art. Compounds of formula (E) are prepared as described with
reference to Reaction Scheme 1.
[0215] Formula (C)--In general, compounds of formula (Ia) are
prepared by first coupling a compound of formula (A) with a
compound of formula (B) (or with another compound of the formula
H.sub.2N--(CH.sub.2).sub.p--R.su- p.7, under standard amide
coupling conditions to form a compound of formula (C). For example,
to a cold (0-5.degree. C.) solution of the compound of formula (A)
and an excess molar amount of HOBT in DMF is added an excess molar
amount of EDCI. The resulting solution is stirred from about 1 to
about 2 hours, preferably for about 1 hour, at 0-5.degree. C.,
preferably at 0.degree. C. To the cold solution is then added a
solution of an equimolar amount of a compound of formula (B) in the
presence of a base, preferably DMAP. The resulting mixture is
stirred from 12 to 24 hours, preferably for 24 hours, at room
temperature, preferably at 25.degree. C. The compound of formula
(C) is then isolated from the reaction mixture by standard peptide
isolation techniques.
[0216] Formula (D)--The amino-protecting group of the compound of
formula (C) is then removed under mild acidic conditions,
preferably in the presence of trifluoroacetic acid, to yield a
compound of formula (D).
[0217] Alternative Preparations of Formula (D)--Another method of
preparing a compound of formula (D) particularly when R.sup.3 is
t-butyl, other .beta.-branched amino acid side chains, or
cyclohexyl, p is zero, and R.sup.7 is aryl or heteroaryl, employs
the intermediate (A-1), the preparation of which is illustrated in
Reaction Scheme 2A. Another alternative method of preparing a
compound of formula (D) particularly when R.sub.3 is
1-hydroxyisopropyl or another .beta.-hydroxy amino acid side chain,
and R.sup.7 is aryl or heteroaryl, is illustrated in Reaction
Scheme 2B. 22
[0218] As illustrated in Reaction Scheme 2A, a compound of formula
(A) is coupled with about one molar equivalent of
N-hydroxysuccinimide in acetonitrile at 0.degree. C. in the
presence of DCC. The reaction takes place with stirring at
0.degree. C. to 25.degree. C., for 8 to 16 hours to give the
corresponding N-hydroxysuccinimide ester of formula (A-1). This
ester is then reacted with a compound of formula B or another
compound of the formula H.sub.2N--(CH.sub.2).sub.p--R.sup.7 in an
inert solvent at 100.degree. C. preferably for 3 hours; the
resultant compound of formula (C) is isolated and deprotected to
yield a compound of formula (D) as described above in Reaction
Scheme 2. 23
[0219] As illustrated in Reaction Scheme 2B, a compound of formula
(C-1) in an inert anhydrous solvent such as THF is stirred with
n-butyllithium at a temperature below 10.degree. C., preferably
0.degree. C., for about 1 hour, then cooled to about -70.degree. C.
and reacted with 3 molar equivalents of acetone. A compound of
formula (C-2), as a racemate, is isolated and purified by standard
procedures. Following hydrogenolytic removal of the CBZ protecting
group, a compound of formula (D-1) is obtained.
[0220] Formula (Ia)--As illustrated in Reaction Scheme 2, a
compound of formula (D) is coupled with a compound of formula (E)
under standard peptide coupling conditions. For example, to a cold
(0-5.degree. C., preferably 0.degree. C.) solution of the compound
of formula (D) in an inert solvent, preferably THF, is added
1,1'-carbonyldiimidazole. The resulting mixture is stirred from 60
to 90 minutes, preferably for 75 minutes, at 0-5.degree. C.,
preferably at 0.degree. C., and then reacted with the compound of
formula (E) for about 12 to 17 hours, preferably for about 15
hours. The resulting compound of formula (Ia) is then isolated from
the reaction mixture by standard peptide isolation techniques, for
example, extraction and reverse phase HPLC.
[0221] Preparation of Formula (F)
[0222] Compounds of formula (F): 24
[0223] where R.sup.8 is t-butyl, are intermediates used in the
preparation of compounds of formula (I) as illustrated below in
Reaction Scheme 4. The compounds of formula (F) are prepared as
shown in Reaction Scheme 3. 25
[0224] Starting Materials--Compounds of formula (Fa) are
commercially available or may be prepared according to methods
known to those of ordinary skill in the art, for example, by the
method described in Example. 11 below. L-(+)-2,10-Camphor sultam
and D-(-)-2,10-camphor sultam are commercially available, for
example, from Aldrich.
[0225] Formula (Fb)--In general, compounds of formula (F)
(illustrated as one of the two isomers obtainable by this
synthesis) are prepared by first condensing a compound of formula
(Fa) [where the group R.sup.2 encompasses the group "X" of Formula
(I) and can be, e.g., a biphenylpropylene or fluorenyl propylene
group] with L-(+)-2,10-camphor sultam to form a compound of formula
(Fb).
[0226] Formula (Fc)--Using sodium hexamethyldisilazide to generate
the anion for 1 hour, the reaction is quenched with
t-butylbromoacetate to form the corresponding ester of formula
(Fc).
[0227] Formula (F)--The camphor group is then removed under basic
conditions, such as lithium hydroperoxide (formed in situ from
lithium hydroxide and hydrogen peroxide) initially at reduced
temperature (preferably 0.degree. C.) for 15 minutes and warmed to
room temperature for 2 hours. The mixture is cooled back to
0.degree. C. and an aqueous mixture of sodium sulfite and sodium
bicarbonate is added with stirring, after which the mixture is
allowed to return to room temperature, and the pH is neutralized to
yield an individual stereoisomer of a compound of formula (F)
wherein the carbon to which the --X--R.sup.2 group is attached is
in the (R) configuration. In a similar manner, but substituting
D-(-)-2,10-camphor sultam for L-(+)-2,10-camphor sultam, the
corresponding individual stereoisomers in the (S) configuration can
be prepared.
[0228] Alternative Preparation of Formula (F)--Another method of
preparing stereoisomers of formula (F) utilizes the commercially
available chiral compound, 4S-phenylmethyloxazolidinone, as shown
below in Reaction Scheme 3A (following the Starting Materials
section, where preparation of the compounds of starting material of
formula (Fa') are illustrated).
[0229] Starting Materials--
[0230] Compounds of formula (Fa') where X is
--O--CH.sub.2--CH.sub.2-- are prepared as illustrated in Reaction
Scheme 3A-1. 26
[0231] A commercially-available alcohol (a) is reacted with
ethyl-4-bromocrotonate (b) in the presence of stoichiometric sodium
hydride in a solvent such as DMF at 0.degree. C. to room
temperature, or in the case of a phenol (a), by refluxing with (b)
in acetone in the presence of excess potassium carbonate for
several hours. The resulting unsaturated ester (c) is converted by
hydrogenation in the presence of platinum on carbon to the
saturated ester (d), which is then saponified with aqueous sodium
hydroxide in ethanol to the acid (e). The acid (e) is converted to
the acid chloride (Fa'-1) through the action of oxalylchloride at
between room temperature and 50.degree. C.
[0232] Compounds of formula (Fa') where X is
--S--CH.sub.2CH.sub.2-- are prepared as illustrated in Reaction
Scheme 3A-2. 27
[0233] A commercially-available thiol (f) is reacted with lithium
hydride in DMF at room temperature for several hours to form the
lithium thiolate. Excess butyrolactone (g) is added and heated to
reflux under argon to give the acid (h). Acid (h) is then converted
to the acid chloride (Fa'-2) with oxalyl chloride, as before.
[0234] Compounds of formula (Fa') where X is
--CH.sub.2CH.sub.2--O-- are prepared as illustrated in Reaction
Scheme 3A-3. 28
[0235] Compounds of formula (l) and (k) are in many cases
commercially available. When not, they are prepared as follows.
Compounds of formula (j), where R.sup.2 is aryl or heteroaryl, are
converted to alkenes (k) by treatment for several hours with
vinyl-tributylstannane (commercially available from Aldrich
Chemical Co.) in the presence of catalytic tetrakis
(triphenylphosphine)palladium at reflux in toluene. The alkenes (k)
may be further converted to the alcohols (l) by hydroboration with
borane in THF at 0.degree. C. to room temperature, over a period of
several hours, followed by oxidation with alkaline hydrogen
peroxide. The alcohols (l) are converted to the acids (m) by
treatment with chloroacetic acid and excess sodium hydride in DMF
at elevated temperature, preferably 60.degree. C. The acids (m) are
converted to the acid chlorides (Fa'-3) with oxalyl chloride, as
before.
[0236] Compounds of formula (Fa') where X is
--CH.sub.2CH.sub.2--S-- are prepared according to Reaction Scheme
3A-4. 29
[0237] The alcohols (l) are converted to thioacetates (n) by
addition of thioacetic acid to the reagent generated from
triphosphine and diethyl azodicarboxylate in THF at 0.degree. C.
The thioacetates (n) are converted to the acids (p) by treatment
with potassium carbonate in methanol in the presence of
chloroacetic acid. The acids (p) are converted to the acid chloride
(Fa'-4) with oxalyl chloride, as before. 30
[0238] Formula (Fb')--A compound of formula (Fa') is first
condensed with 4S-phenylmethyloxazolidinone under standard
conditions to give the corresponding compound of formula (Fb').
[0239] Formula (Fc')--An approximately equimolar amount of sodium
hexamethyldisilazide is added to a compound of formula (Fb') in an
inert solvent such as THF. The reaction takes place at -80.degree.
C. to -95.degree. C., for about 15 minutes. t-Butylbromoacetate is
added in excess to this mixture and the solution is stirred for
about 2 hours at -90.degree. C. to -60.degree. C. to yield a single
stereoisomer of formula (Fc'), which is purified by standard
organic chemistry procedures.
[0240] Formula (F')--The oxazolidinone group of a compound of
formula (Fc') is removed under basic conditions to yield an
individual stereoisomer of formula (F'), for example as described
with reference to the preparation of formula (F) in Reaction Scheme
3. The compounds of formula (F') can be used interchangeably with
those of formula (F) in the syntheses that follow.
[0241] Alternative Preparation of Formula (F)--Formula F can also
be prepared as described with reference to Reaction Scheme 3B.
31
[0242] Starting Materials--The compound illustrated as formula
(Fc") can be prepared analogously to the preparation of formula
(Fc') as described with reference to Reaction Scheme 3A, by
substituting for the compound of formula (Fa') the corresponding
allyl compound where the group shown as X is prop-2-enyl and
R.sup.2 is H.
[0243] Formula (Fc"-1)--Arylation or heteroarylation of (Fc") is
carried out in the presence of a base and a palladium catalyst by
adding aryl- or heteroaryl-halide, preferably bromide or iodide,
and heating the reaction mixture for about 2 to 4 hours, preferably
4 hours, at about 100.degree. C. to form a compound of the formula
(Fc"-1).
[0244] Formula (Fc"-2)--Catalytic hydrogenation (Pd/C) of an allyl
compound of formula (Fc"-1) yields the corresponding alkyl compound
of the formula (Fc"-2).
[0245] Formula (F)--A compound of formula (Fc"-2) is subjected to
basic conditions, such as lithium hydroperoxide (formed in situ
from lithium hydroxide and hydrogen peroxide) initially at reduced
temperature (preferably 0.degree. C.) for 15 minutes and warmed to
room temperature for 2 hours. The mixture is cooled back to
0.degree. C. and an aqueous mixture of sodium sulfite and sodium
bicarbonate is added with stirring, after which the mixture is
allowed to return to room temperature, the pH neutralized, and the
compound of formula (F) is obtained by standard isolation.
[0246] Preparation of Formulae (Ib), (Ic), (Id) and (Ie)
[0247] The compounds of formulae (Ib), (Ic), (Id) and (Ie) each
represent sub-genuses of formula I in which the R.sup.1 substituent
varies, prepared sequentially as described in Reaction Scheme 4,
where R.sup.8 is t-butyl. In compounds of formula (Ib) R.sup.1 is
alkoxycarbonyl or aralkoxycarbonyl. In compounds of formula (Ic)
R.sup.1 is carboxy. In compounds of formula (Id) R.sup.1 is
benzyloxycarbamoyl. In compounds of formula (Ie) R.sup.1 is
hydroxycarbamoyl. 32
[0248] Starting Materials--Formula (D) is prepared as described
with reference to Reaction Schemes 2, 2A and 2B. Formula (F) is
prepared as described with reference to Reaction Schemes 3 and 3A.
O-Benzylhydroxylamine is commercially available, for example, as
the hydrochloride salt from Aldrich Co.
[0249] Formula (Ib)--A compound of formula (F) is coupled with a
compound of formula (D) under standard amide coupling conditions to
form a compound of formula (Ib). For example, to a solution of a
compound of formula (F) in an aprotic solvent, preferably DMF,
containing a slightly excess molar amount of HOBT, is added an
excess molar amount of EDCI. The resulting mixture is stirred from
1 to 2 hours (preferably 1 hour) at 0-5.degree. C. (preferably at
0.degree. C.). To the cold solution is then added an equimolar
amount of a compound of formula (D) in the presence of a base,
preferably DMAP. The resulting mixture is then stirred from 12 to
24 hours (preferably 24 hours) at room temperature (preferably at
25.degree. C.). The compound of formula (Ib) is then isolated from
the reaction mixture by standard peptide isolation techniques, for
example, evaporation of solvents, extraction, flash chromatography
and/or HPLC.
[0250] Formula (Ic)--A compound of formula (Ib) is hydrolyzed under
mild acidic conditions, preferably with trifluoroacetic acid, to
yield a compound of formula (Ic).
[0251] Formula (Id)--A compound of formula (Ic) is then treated
with O-benzylhydroxylamine under standard amide coupling conditions
to form a compound of formula (Id). For example, a cold
(0-5.degree. C.) solution of the compound of formula (Ic) and HOBT
in an inert solvent, preferably DMF, is treated with an excess
molar amount of EDCI. After stirring the resulting mixture for 30
minutes to an hour at 0-5.degree. C. (preferably at 0.degree. C.),
an equimolar amount of O-benzylhydroxyamine is added. The reaction
mixture is allowed to warm and remain at room temperature overnight
for 8 to 16 hours. The compound of formula (Id) is then isolated
from the reaction mixture by standard isolation techniques, for
example, by extraction and flash chromatography.
[0252] Formula (Ie)--The hydroxyl-protecting group (benzyl) of a
compound of formula (Id) is removed under catalytic hydrogenation
conditions (Pd/C) to yield a compound of formula (Ie).
[0253] Alternative Preparation of Formula (Ie)--An alternative
method for preparing formula (Ie) (particularly where R.sup.4 is a
sulfur-containing moiety, such as alkylsulfinyl) is to treat the
corresponding compound of formula (1c) with hydroxylamine
hydrochloride and a peptide coupling reagent, preferably
benzotriazol-1-yloxy-tris(dimethylamino)phosphonium
hexafluorophosphate, in the presence of a tertiary amine base such
as N-methylmorpholine in DMF solvent. The resulting compound of
formula (1e) is isolated from the reaction mixture by standard
isolation techniques, for example, by extraction and
concentration.
[0254] Alternative Preparation of Formula (Ib)--A particularly
preferred method of preparing compounds of formula (Ib) when
R.sup.2 is an aryl or heteroaryl, and where X (not shown) is
propanyl and p (not shown) is zero is shown in Reaction Scheme 4A.
33
[0255] Starting Materials--The compound illustrated as formula
(Fc") can be prepared analogously to the preparation of formula
(Fc') as described with reference to Reaction Scheme 3A, by
substituting for the compound of formula (Fa') the corresponding
allyl compound where R.sup.2 is prop-2-enyl. The compound of
formula (D') is a compound of formula (D) and can be as described
in Reaction Scheme 2. The halo-aryl or halo-heteroaryl reactants
used in the preparation of compounds of formula (D'-2) are
commercially available, or can be prepared according to methods
known to those of ordinary skill in the art, e.g., as described in
Example 41C.
[0256] A compound of formula (F") is prepared by alkaline
hydrolysis of the oxazolidinone group from a compound of formula
(Fc"). After isolation by standard procedures, (F") is coupled with
a compound of formula (D') under standard peptide coupling
conditions as described above with reference to Reaction Scheme 2,
to form a compound of formula (D'-1). Arylation or heteroarylation
of (D'-1) is accomplished by adding aryl- or heteroaryl-halide
(preferably aryl- or heteroaryl bromide, iodide or triflate) and
heating the reaction mixture for about 2 hours at about 100.degree.
C. to form a compound of the formula (D'-2). Catalytic
hydrogenation (Pd/C) of (D'-2) yields a compound of formula
(Ib').
[0257] Preparation of Formula (G)
[0258] Compounds of formula (G): 34
[0259] are intermediates in the preparation of compounds of formula
(I) and are prepared as illustrated below in Reaction Scheme 6. The
compounds of formula (G) are prepared as shown in Reaction Scheme
5. 35
[0260] Starting Materials--Compounds of formula (Ga) and thioacetic
acid are commercially available, for example, from TCI America
Organic Chemicals and the Aldrich Company, respectively.
[0261] Formula (Gb)--A compound of formula (Ga) is hydrolyzed with
an equimolar amount of a base, for example, potassium hydroxide, to
yield a compound of formula (Gb).
[0262] Formula (Gc)--A compound of formula (Gb) is deprotonated
under basic conditions, for example, in the presence of
triethylamine, at 0-5.degree. C. (preferably at 0.degree. C.) and
then reacted with formaldehyde, followed by treatment with aqueous
base, preferably potassium carbonate, to yield a compound of
formula (Gc), which is isolated from the reaction mixture by
standard isolation procedures.
[0263] Formula (Gd)--A compound of formula (Gc) is hydrolyzed under
basic conditions, preferably in the presence of lithium hydroxide,
to yield a compound of formula (Gd).
[0264] Formula (G)--A compound of formula (Gd) is reacted with an
excess molar amount of thioacetic acid at 90-100.degree. C.
(preferably at 95.degree. C.) under an inert atmosphere. The
compound of formula (G) is then isolated from the reaction mixture
by standard isolation techniques, for example, by extraction and
evaporation.
[0265] Preparation of Formulae (If) and (Ig)
[0266] The compounds of formulae (If) and (Ig) each represent
sub-genuses of formula I in which the R.sup.1 substituent is
sulfur-containing, prepared sequentially as described in Reaction
Scheme 6. In compounds of formula (If), R.sup.1 is acetylthio. In
compounds of formula (Ig), R.sup.1 is mercapto. 36
[0267] Formula (If)--A compound of formula (G) is coupled with a
compound of formula (D) under standard amide coupling conditions to
yield a compound of formula (If). For example, to a solution of the
compound of formula (G) and HOBT in an aprotic solvent, preferably
DMF, is added an excess molar amount of EDCI. Subsequently, the
compound of formula (D) is added and the resulting mixture is
stirred overnight at room temperature. The resulting compound of
formula (If) is then isolated from the reaction mixture by standard
isolation techniques, for example, by evaporation of solvent,
extraction, and flash chromatography.
[0268] Formula (Ig)--A compound of formula (If) is hydrolyzed under
basic conditions, preferably in a protic solvent such as methanol
in the presence of ammonium hydroxide, to form a compound of
formula (Ig).
[0269] Preparation of Formula (Ih)
[0270] Compounds of formula (Ih) are a sub-genus of formula (I)
where R.sup.1 is N-hydroxyformamide, and are prepared as shown in
Reaction Scheme 7. 37
[0271] Starting Materials--The compound illustrated as formula
(Pb") can be prepared analogously to the preparation of formula
(Fb') as described with reference to Reaction Scheme 3A, by
substituting for the compound of formula (Pa') the corresponding
allyl compound where R.sup.1 is prop-2-enyl.
[0272] Formula (P-1)--The compound of formula (Fb") is
hydroxymethylated by incubation with titanium tetrachloride at
reduced temperature, preferably 0.degree. C., under basic
conditions for one to three hours, preferably 1 hour followed by
addition of S-trioxane and titanium tetrachloride with continued
incubation at 0.degree. C. for 3 to 5 hours, preferably 4 hours.
The compound of formula (P-1) is then isolated by standard methods,
e.g., extraction and column chromatography.
[0273] Formula (P-2)--The compound of formula (P-1) is reacted with
an excess molar amount of 0-benzylhydroxylamine and of
trimethylaluminum at reduced temperature, preferably 0.degree. C.
The reaction is allowed to proceed with stirring for 5 to 7 hours,
preferably 6 hours, at 0.degree. C. under argon. The resulting
compound of formula (P-2) is isolated by standard procedures.
[0274] Formula (P-3)--Excess mesyl chloride is reacted with the
compound of formula (P-2) in pyridine at 0.degree. C. for several
hours, preferably 3 hours. The reaction mixture is cooled on ice,
organic solvent-extracted, and concentrated. The concentrated
extract is refluxed under basic conditions for several hours,
preferably 3 hours, thus yielding the azetidinone compound of
formula (P-3), which is purified by standard procedures.
[0275] Formula (P-4)--The compound of formula (P-3) is reacted with
a desired halogenated R.sup.2 group ( e.g., an aryl- or heteroaryl
halide, preferably bromide or iodide) in an inert solvent in the
presence of a base, such as triethylamine, and a palladium
catalyst, preferably formed from palladium (II) acetate and about 2
molar equivalents of tri-o-tolylphosphine. After heating the
reaction mixture for 15 to 20 hours, preferably 18 hours at
100.degree. C., the corresponding compound of formula (P-4) is
isolated and purified by standard procedures.
[0276] Formula (P-5)--Cleavage of the azetidinone ring of a
compound of formula (P-4) is carried out under basic conditions at
room temperature for 1 to 3 hours, preferably 1 hour. The resultant
compound is extracted into organic solvent, concentrated,
redissolved in a base-containing solvent (e.g., pyridine), and
carboxylated with formic anhydride at reduced temperature,
preferably 0.degree. C., for 30 minutes, to yield the corresponding
compound of formula (P-5), which is isolated by standard
procedures.
[0277] Formula (P-6)--A compound of formula (P-5) is coupled with a
compound of formula (D') under standard amide coupling conditions
to form the corresponding compound of formula (P-6), which is
isolated by standard procedures.
[0278] Formula (Ih)--Catalytic hydrogenation of a compound of
formula (P-6) with Pd/C, followed by removal of the catalyst by
filtration yields the corresponding compound of formula (Ih).
[0279] Preparation of Salts
[0280] In addition, all compounds of formula (I) that exist in
either the free acid or the free base form may be converted to
their pharmaceutically acceptable salts by treatment with the
appropriate inorganic or organic base or with the appropriate
inorganic or organic acid, respectively. Salts of the compounds of
formula (I) can also be converted to the free acid or free base
form or to another salt. For example, a compound of formula (I)
having a carboxylic acid moiety can be converted to the carboxylate
form by addition of 1 equivalent of NaOH or KOH in an alcoholic
solvent followed by evaporation of solvent. A compound of formula
(I) in the form of a free base can be converted to the chloride
salt, for example, by addition of 1 equivalent of HCl in an organic
solvent, followed by concentration.
[0281] Preferred Synthesis and Last Steps
[0282] In summary, compounds of formula (I) are prepared by:
[0283] (A) contacting a compound of formula (D) 38
[0284] with a compound of formula (F) 39
[0285] where R.sup.1 is alkoxycarbonyl, aralkoxycarbonyl, aryl- or
heteroaryl-thiomethylphosphinoyl, or acetylthio;
[0286] in the presence of a base and an amide coupling reagent to
give the corresponding compound of formula (I); or
[0287] (B) catalytically hydrogenating the corresponding compound
where X and R.sup.2 together are optionally aryl- or
heteroaryl-substituted alkenyl; or
[0288] (C) treating a compound of formula (I), where R.sup.1 is
alkoxycarbonyl or aralkoxycarbonyl, under mild acidic conditions to
give the corresponding compound of formula (I) where R.sup.1 is
carboxy; or
[0289] (D) contacting a compound of formula (I), where R.sup.1 is
carboxy, with O-benzylhydroxylamine to give the corresponding
compound of formula (I) where R.sup.1 is benzyloxycarbamoyl, or
[0290] (E) catalytically hydrogenating a compound of formula (I),
where R.sup.1 is benzyloxycarbamoyl, to give the corresponding
compound of formula (I) where R.sup.1 is hydroxycarbamoyl; or
[0291] (F) contacting a compound of formula (I), where R.sup.1 is
carboxy, with hydroxylamine to give the corresponding compound of
formula (I) where R.sup.1 is hydroxycarbamoyl; or
[0292] (G) catalytically hydrogenating a compound of the formula
40
[0293] where BnO is benzyloxy, to give the corresponding compound
of formula (I) where R.sup.1 is N-hydroxyformamide; or
[0294] (H) treating a compound of formula (I), wherein R.sup.1 is
acetylthio, with ammonium hydroxide in a protic solvent to give the
corresponding compound of formula (I) where R.sup.1 is
mercapto.
[0295] A preferred method of making compounds of formula (I) where
R.sup.1 is N-hydroxyformamide entails converting a compound of
formula (P-4) 41
[0296] wherein R.sup.2 is aryl or heteroaryl, by basic hydrolysis
followed by formylation to give a compound of formula (P-5) 42
[0297] reacting the compound of formula (P-5) with a compound of
formula (D) to give a compound of formula (P-6) 43
[0298] and catalytically hydrogenating the compound of formula
(P-6).
[0299] Compounds prepared by the above-described process of the
invention may be identified by the presence of a detectable amount
of one or more compounds of formulae (P-3), (P-4) or (P-6). While
it is well known that pharmaceuticals must meet pharmacopoeia
standards before approval and/or marketing, and that synthetic
reagents (such as O-benzylhydroxylamine) or precursors [such as
(P-3), (P-4), or (P-6)] should not exceed the limits prescribed by
pharmacopoeia standards, final compounds prepared by a process of
the present invention may have minor, but detectable, amounts of
such materials present, for example at levels in the range of 50
ppm or lower. These levels of (P-3) can be detected, e.g., by
GC-MS, or of (P-4) can be detected, e.g., by HPLC-MS or by HPLC
with fluorescence detection, or of (P-6) can be detected, e.g., by
HPLC with fluorescence detection. It is important to monitor the
purity of pharmaceutical compounds for the presence of such
materials, which presence is additionally disclosed as a method of
detecting use of a process of the invention.
EXAMPLES
[0300] The following preparations and examples are given to enable
those skilled in the art to more clearly understand and to practice
the present invention. They should not be considered as limiting
the scope of the invention, but merely as being illustrative and
representative thereof.
Example 1
[0301] Compounds of Formula (Ea)
[0302] 1A. Crystalline phosphinic acid (8.4 g, 0.13 mol) was
stirred in neat triethylorthoformate (22 mL, 0.20 mL) for 90
minutes at room temperature. This was then transferred via cannula
to a stirred solution of ethylisobutylacrylate (8 g, 0.036 mol) and
tetramethylguanidine (4.5 mL, 0.036 mol) that had been cooled to
0.degree. C. for 10 minutes. The ice bath was removed and the
reaction stirred for 4 hours. The mixture was diluted with 200 mL
of diethyl ether and the solution washed with 1 N HCl (100 mL),
water (4.times.100 mL) brine (100 mL), and dried over magnesium
sulfate. This was rotary-evaporated to yield 8.15 g of
2-(ethoxy)phosphinoylmethyl-4-methylpentanoic acid ethyl ester as a
slightly yellow colored oil, MS: 349 (M-H.sub.2O).sup.+.
[0303] 1B. In a similar manner, the following compounds of formula
(Ea) are prepared:
[0304] 2-(ethoxy)phosphinoylmethyl-5-phenylpentanoic acid ethyl
ester;
[0305] 2-(ethoxy)phosphinoylmethyl-4-phenylbutanoic acid ethyl
ester;
[0306] 2-(ethoxy)phosphinoylmethyl-3-phenylpropanoic acid ethyl
ester;
[0307] 2-(ethoxy)phosphinoylmethyl-3-cyclohexylpropanoic acid ethyl
ester; and
[0308] 2-((ethoxy)phosphinoylmethyl)pentanoic acid ethyl ester.
Example 2
[0309] Compound of Formula (Eb)
[0310] 2A. Crude 2-(ethoxy)phosphinoylmethyl-4-methylpentanoic acid
ethyl ester (26 g) was dissolved in 600 mL THF/CH.sub.2Cl.sub.2
(50/50) and cooled to 0.degree. C. Diisopropyl-ethylamine (32 mL)
and 90.8 mL of bis-(trimethylsilyl)acetamide were then added to the
solution and the resulting mixture was stirred for 20 minutes
before paraformaldehyde (5.5 g) was added. The solution was brought
to room temperature and heated at 37.degree. C. for 18 hours. The
solvent was removed by evaporation, and the resulting oil dissolved
in 200 mL ethyl acetate. The solution was washed with 50 mL of 1N
HCl (2.times.), 50 mL of brine (2.times.), dried over MgSO.sub.4,
filtered and evaporated to yield 19.3 g of
2-(ethoxy)(hydroxymethyl)phosphinoylmethyl-4-methylpentanoic acid
ethyl ester as a faintly yellow oil, MS: 281.2 (MH.sup.+).
[0311] 2B. In a similar manner, the following compounds of formula
(Eb) are prepared:
[0312] 2-(ethoxy)(hydroxymethyl)phosphinoylmethyl-5-phenylpentanoic
acid ethyl ester;
[0313] 2-(ethoxy)(hydroxymethyl)phosphinoylmethyl-4-phenylbutanoic
acid ethyl ester;
[0314] 2-(ethoxy)(hydroxymethyl)phosphinoylmethyl-3-phenylpropanoic
acid ethyl ester;
[0315]
2-(ethoxy)(hydroxymethyl)phosphinoylmethyl-3-cyclohexylpropanoic
acid ethyl ester; and
[0316] 2-((ethoxy)(hydroxymethyl)phosphinoylmethyl)pentanoic acid
ethyl ester.
Example 3
[0317] Compounds of Formula (Ec)
[0318] 3A.
2-(Ethoxy)(hydroxymethyl)phosphinoylmethyl-4-methylpentanoic acid
ethyl ester (5 g) was dissolved in 20 mL of CH.sub.2Cl.sub.2 and
cooled to -20.degree. C. (in duplicate). Methanesulfonyl chloride
(1.5 mL) and triethylamine (3.0 mL) were added to solution
dropwise. After 15 minutes the bath was removed and the reaction
left at room temperature for 31/2 hours. Each solution was then
washed with 10 mL cold 2% HCl, 10 mL NaHCO.sub.3 (sat), 10 mL
brine, dried with MgSO.sub.4, filtered and evaporated to yield 12.8
g (combined yield) of 2-(ethoxy)(methane-sulfony-
loxymethyl)phosphinoylmethyl-4-methylpentanoic acid ethyl
ester.
[0319] 3B. In a similar manner, but replacing methanesulfonyl
chloride with p-toluenesulfonyl chloride,
2-(ethoxy)-(p-toluenesulfonyloxymethyl)--
phosphinoylmethyl-4-methyl-pentanoic acid ethyl ester is
prepared.
[0320] 3C. In a similar manner, the following compounds of formula
(Ec) are prepared:
[0321]
2-(ethoxy)(methanesulfonyloxymethyl)phosphinoylmethyl-5-phenylpenta-
noic acid ethyl ester;
[0322]
2-(ethoxy)(methanesulfonyloxymethyl)phosphinoylmethyl-4-phenylbutan-
oic acid ethyl ester;
[0323]
2-(ethoxy)(methanesulfonyloxymethyl)phosphinoylmethyl-3-phenylpropa-
noic acid ethyl ester;
[0324]
2-(ethoxy)(methanesulfonyloxymethyl)phosphinoylmethyl-3-cyclohexylp-
ropanoic acid ethyl ester;
[0325]
2-((ethoxy)(methanesulfonyloxymethyl)phosphinoylmethyl)-pentanoic
acid ethyl ester;
[0326]
2-(ethoxy)(p-toluenesulfonyloxymethyl)phosphinoylmethyl-5-phenylpen-
tanoic acid ethyl ester;
[0327]
2-(ethoxy)(p-toluenesulfonyloxymethyl)phosphinoylmethyl-4-phenylbut-
anoic acid ethyl ester;
[0328]
2-(ethoxy)(p-toluenesulfonyloxymethyl)phosphinoylmethyl-3-phenylpro-
panoic acid ethyl ester;
[0329]
2-(ethoxy)(p-toluenesulfonyloxymethyl)phosphinoylmethyl-3-cyclohexy-
lpropanoic acid ethyl ester; and
[0330] 2-(
(ethoxy)(p-toluenesulfonyloxymethyl)phosphinoylmethyl)-pentanoi- c
acid ethyl ester.
Example 4
[0331] Compounds of Formula (Ee)
[0332] 4A. Sodium hydride (1.52 g, (60%)) and 2-quinolinethiol (6
g) were stirred together at 0.degree. C. in 50 mL DMF. After the
initial H.sub.2 evolution had subsided, the mixture was stirred at
room temperature for 2.5 hours. The mixture was then cooled to
0.degree. C. and
2-(ethoxy)(methanesulfonyloxymethyl)-phosphinoylmethyl-4-methylpentanoic
acid ethyl ester (12.8 g) in 10 mL DMF was added via cannula and
the resulting mixture was then stirred for 18 hours, slowly warming
to room temperature. The DMF was removed by evaporation, the
residue dissolved in 50 mL ethyl acetate and washed with 50 mL
H.sub.2O (2.times.), brine (50 mL), dried with MgSO.sub.4 and
evaporated to a yellow semi-solid. Purification by flash
chromatography using 10% ethyl acetate/hexane to 80% ethyl
acetate/hexane for the elution yielded 10 g of
2-(ethoxy)(quinolin-2-ylthiomethyl)-phosphinoyl-methyl-4-methylpentanoic
acid ethyl ester (Rf 0.35 80% ethyl acetate/hexane), MS: 424.1
(MH.sup.+).
[0333] 4B. In a similar manner, but replacing 2-quinolinethiol with
1-naphthalenethiol, 2-naphthalenethiol or thiophenol, the following
compounds of formula (Ee) are prepared:
[0334]
2-(ethoxy)(naphth-1-ylthiomethyl)phosphinoylmethyl-4-methylpentanoi-
c acid ethyl ester;
[0335]
2-(ethoxy)(naphth-2-ylthiomethyl)phosphinoylmethyl-4-methylpentanoi-
c acid ethyl ester; and
[0336]
2-(ethoxy)(phenylthiomethyl)phosphinoylmethyl-4-methylpentanoic
acid ethyl ester.
[0337] 4C. In a similar manner, the following compounds of formula
(Ee) are prepared:
[0338]
2-(ethoxy)(quinolin-2-ylthiomethyl)phosphinoylmethyl-5-phenylpentan-
oic acid ethyl ester;
[0339]
2-(ethoxy)(quinolin-2-ylthiomethyl)phosphinoylmethyl-4-phenylbutano-
ic acid ethyl ester;
[0340]
2-(ethoxy)(quinolin-2-ylthiomethyl)phosphinoylmethyl-3-phenylpropan-
oic acid ethyl ester;
[0341]
2-(ethoxy)(quinolin-2-ylthiomethyl)phosphinoylmethyl-3-cyclohexylpr-
opanoic acid ethyl ester;
[0342]
2-((ethoxy)(quinolin2-ylthiomethyl)phosphinoylmethyl)-pentanoic
acid ethyl ester;
[0343]
2-(ethoxy)(naphth-1-ylthiomethyl)phosphinoylmethyl-5-phenylpentanoi-
c acid ethyl ester;
[0344]
2-(ethoxy)(naphth-1-ylthiomethyl)phosphinoylmethyl-4-phenylbutanoic
acid ethyl ester;
[0345]
2-(ethoxy)(naphth-1-ylthiomethyl)phosphinoylmethyl-3-phenylpropanoi-
c acid ethyl ester;
[0346]
2-(ethoxy)(naphth-1-ylthiomethyl)phosphinoylmethyl-3-cyclohexylprop-
anoic acid ethyl ester;
[0347]
2-((ethoxy)(naphth-l-ylthiomethyl)phosphinoylmethyl)-pentanoic acid
ethyl ester;
[0348]
2-(ethoxy)(naphth-2-ylthiomethyl)phosphinoylmethyl-5-phenylpentanoi-
c acid ethyl ester;
[0349]
2-(ethoxy)(naphth-2-ylthiomethyl)phosphinoylmethyl-4-phenylbutanoic
acid ethyl ester;
[0350]
2-(ethoxy)(naphth-2-ylthiomethyl)phosphinoylmethyl-3-phenylpropanoi-
c acid ethyl ester;
[0351]
2-(ethoxy)(naphth-2-ylthiomethyl)phosphinoylmethyl-3-cyclohexylprop-
anoic acid ethyl ester;
[0352]
2-((ethoxy)(naphth-2-ylthiomethyl)phosphinoylmethyl)-pentanoic acid
ethyl ester;
[0353]
2-(ethoxy)(phenylthiomethyl)phosphinoylmethyl-5-phenylpentanoic
acid ethyl ester;
[0354]
2-(ethoxy)(phenylthiomethyl)phosphinoylmethyl-4-phenylbutanoic acid
ethyl ester;
[0355]
2-(ethoxy)(phenylthiomethyl)phosphinoylmethyl-3-phenylpropanoic
acid ethyl ester;
[0356]
2-(ethoxy)(phenylthiomethyl)phosphinoylmethyl-3-cyclohexylpropanoic
acid ethyl ester; and
[0357] 2-((ethoxy)(phenylthiomethyl)phosphinoylmethyl)-pentanoic
acid ethyl ester.
Example 5
[0358] Compounds of formula (E)
[0359] 5A.
2-(Ethoxy)(quinolin-2-ylthiomethyl)phosphinoyl-methyl-4-methylp-
entanoic acid ethyl ester (4.5 g) was dissolved in 100 mL THP and
12.5 mL of 2N NaOH was added together with enough methanol to make
the solution homogeneous. After 18 hours the THF was removed by
evaporation, the residue diluted with 50 mL H.sub.2O and washed
with 50 mL ethyl acetate. The aqueous phase was then acidified to
pH 4, and the product extracted with 50 mL ethyl acetate
(2.times.). The ethyl acetate was washed with 20 mL brine, dried
with MgSO.sub.4 and evaporated to yield 3.8 g of
2-(hydroxy)(quinolin-2-ylthiomethyl)phosphinoylmethyl-4-methylpentanoic
acid as a yellow oil, MS: 368 (MH.sup.+).
[0360] 5B. In a similar manner, the following compounds of formula
(E) are prepared:
[0361]
2-(hydroxy)(naphth-1-ylthiomethyl)phosphinoylmethyl-4-methylpentano-
ic acid;
[0362]
2-(hydroxy)(naphth-2-ylthiomethyl)phosphinoylmethyl-4-methylpentano-
ic acid; and
[0363]
2-(hydroxy)(phenylthiomethyl)phosphinoylmethyl-4-methylpentanoic
acid.
[0364] 5C. In a similar manner, the following compounds of formula
(E) are prepared:
[0365]
2-(hydroxy)(quinolin-2-ylthiomethyl)phosphinoylmethyl-5-phenylpenta-
noic acid;
[0366]
2-(hydroxy)(quinolin-2-ylthiomethyl)phosphinoylmethyl-4-phenylbutan-
oic acid;
[0367]
2-(hydroxy)(quinolin-2-ylthiomethyl)phosphinoylmethyl-3-phenylpropa-
noic acid;
[0368]
2-(hydroxy)(quinolin-2-ylthiomethyl)phosphinoylmethyl-3-cyclohexylp-
ropanoic acid;
[0369]
2-((hydroxy)(quinolin-2-ylthiomethyl)phosphinoylmethyl)-pentanoic
acid;
[0370]
2-(hydroxy)(naphth-1-ylthiomethyl)phosphinoylmethyl-5-phenylpentano-
ic acid;
[0371]
2-(hydroxy)(naphth-1-ylthiomethyl)phosphinoylmethyl-4-phenylbutanoi-
c acid;
[0372]
2-(hydroxy)(naphth-1-ylthiomethyl)phosphinoylmethyl-3-phenylpropano-
ic acid;
[0373]
2-(hydroxy)(naphth-1-ylthiomethyl)phosphinoylmethyl-3-cyclohexylpro-
panoic acid;
[0374]
2-((hydroxy)(naphth-1-ylthiomethyl)phosphinoylmethyl)-pentanoic
acid;
[0375]
2-(hydroxy)(naphth-2-ylthiomethyl)phosphinoylmethyl-5-phenylpentano-
ic acid;
[0376]
2-(hydroxy)(naphth-2-ylthiomethyl)phosphinoylmethyl-4-phenylbutanoi-
c acid;
[0377]
2-(hydroxy)(naphth-2-ylthiomethyl)phosphinoylmethyl-3-phenylpropano-
ic acid;
[0378]
2-(hydroxy)(naphth-2-ylthiomethyl)phosphinoylmethyl-3-cyclohexylpro-
panoic acid;
[0379]
2-((hydroxy)(naphth-2-ylthiomethyl)phosphinoylmethyl)-pentanoic
acid;
[0380]
2-(hydroxy)(phenylthiomethyl)phosphinoylmethyl-5-phenylpentanoic
acid;
[0381]
2-(hydroxy)(phenylthiomethyl)phosphinoylmethyl-4-phenylbutanoic
acid;
[0382]
2-(hydroxy)(phenylthiomethyl)phosphinoylmethyl-3-phenylpropanoic
acid;
[0383]
2-(hydroxy)(phenylthiomethyl)phosphinoylmethyl-3-cyclohexylpropanoi-
c acid; and
[0384] 2-((hydroxy)(phenylthiomethyl)phosphinoylmethyl)pentanoic
acid.
Example 6
[0385] Resolution of a Compound of Formula (E)
[0386]
2-(Hydroxy)(quinolin-2-ylthiomethyl)phosphinoyl-methyl-4-methylpent-
anoic acid (5.3 g) was dissolved in 50 mL of warm ethanol (abs) and
4.2 g of (-)-cinchonidine was added. After 30 minutes at room
temperature the salt began to precipitate out. The flask was
covered in foil and allowed to stand for 2 days. The salt was then
removed by suction filtration, and the filtrate evaporated to a
yellow foam. The salt and the filtrate were each dissolved in 100
mL ethyl acetate and washed successively with 1% HCl to remove the
cinchonidine while keeping the pH above 4. Both solutions were each
dried over MgSO.sub.4 and evaporated to yield 2.4 g of a single
stereoisomer, [.alpha.].sub.D.sup.24=+10.68.degree. (9.73 mg in
methanol (2 mL)) and 2.5 g of the other single stereoisomer,
[.alpha.].sub.D.sup.24=-8.70.degree. (9.88 mg in methanol (2
mL)).
Example 7
[0387] Compounds of Formula (B)
[0388] 7A. To a cold (0.degree. C.) suspension of
4-acetamido-benzenesulph- onyl chloride (4.0 g, 17 mmol) in
CH.sub.2Cl.sub.2 (40 mL) was added pyridine (1.7 mL, 20 mmol) and
DMAP (209 mg, 1.7 mmol). (A clear solution resulted). Anhydrous
methylamine was bubbled into the solution for 1 hour at 0.degree.
C., and then the solution was allowed to stir at 25.degree. C. for
2 hours. The solution was extracted with 1M NaOH (3.times.15 mL)
and the combined extracts were adjusted to pH 6 at 0.degree. C.
with 3M HCl. The product, which precipitated as fluffy white
crystals, was filtered and washed with cold water to afford 3.2 g
(82%) of 4-acetamido-N-methylbenzenesulphonamide: .sup.1H NMR (300
MHz, MeOH) .delta. 2.35 (s,3H), 2.70 (s,3H), 7.96 (s,4H).
[0389] 7B. A mixture of 4-acetamido-N-methylbenzenesulphonamide
(3.2-g, 14 mmol) and 100 mL of 1M HCl was refluxed under argon for
3 hours. After cooling to 25.degree. C., CH.sub.2Cl.sub.2 (10 mL)
was added and the aqueous phase was neutralized with 1M NaOH at
0.degree. C. The aqueous phase was separated and extracted with
CH.sub.2Cl.sub.2 (2.times.25 mL). The combined organic phases were
washed with brine (10 mL), dried (NaSO.sub.4) and concentrated to
afford 1.5 g (58%) of a compound of formula (B) where R.sup.4 is
N-methylsulfonamide as a colorless solid: .sup.1H NMR (300 MHz,
MeOH) .delta. 2.46 (s,3H), 6.67-6.72 (AA.sup.1 part of
AA.sup.1XX.sup.1, 2H), 7.48-7.52 (XX.sup.1 part of
AA.sup.1XX.sup.1, 2H).
Example 8
[0390] Compounds of Formula (C)
[0391] 8A. To a cold (0.degree. C.) solution of
N-tert-butoxycarbonyl-L-le- ucine (1.4 g, 6.3 mmol) and HOBT (1.5
g, 9.8 mmol) in DMF (30 mL) was added EDCI (2.5 g, 14 mmol) in
portions. After stirring for 1 hour at 0.degree. C., the resulting
solution was treated with methyl 4-aminobenzoate (1.09 mL, 6.8
mmol) and DMAP (0.32 g, 2.6 mmol). After stirring for 24 hours at
25.degree. C., the DMF was removed in vacuo. The residue was
dissolved in CH.sub.2Cl.sub.2 and washed with saturated NaHCO.sub.3
solution, 1M HCl (twice), and brine. Drying over Na.sub.2SO.sub.4
and concentration in vacuo afforded the crude product which was
purified by flash chromatography on SiO.sub.2 (20% ethyl
acetate/hexenes eluent). There was obtained 1.0 g (85%) of
N-t-butoxycarbonyl-L-leucine-N'-(4-methoxycarbonylphenyl)carboxamide
as a foamy solid, MS (FAB) 363 (M-H).sup.-.
[0392] 8B. In a similar manner, the following compounds of formula
(C) were prepared:
[0393]
N-t-butoxycarbonyl-L-tryptophan-N'-phenylmethylcarboxamide;
[0394] N-t-butoxycarbonyl-L-tryptophan-N'-phenylcarboxamide;
[0395]
N-t-butoxycarbonyl-L-tryptophan-N'-(4-methoxycarbonylphenyl)carboxa-
mide;
[0396]
N-t-butoxycarbonyl-L-tryptophan-N'-(4-ethoxycarbonylphenyl)carboxam-
ide;
[0397]
N-t-butoxycarbonyl-L-leucine-N'-(4-(N"-methylaminosulfonyl)phenyl)--
carboxamide;
[0398]
N-t-butoxycarbonyl-L-alanine-N'-(4-methoxycarbonylphenyl)carboxamid-
e;
[0399]
N-t-butoxycarbonyl-L-methionine-N'-(4-methoxycarbonylphenyl)carboxa-
mide;
[0400]
N-t-butoxycarbonyl-L-leucine-N'-(3-ethoxycarbonylphenyl)carboxamide-
;
[0401]
N-t-butoxycarbonyl-L-leucine-N'-(2-methoxycarbonylphenyl)carboxamid-
e;
[0402]
N-t-butoxycarbonyl-L-leucine-N'-(4-(1-methylethyloxy)carbonyl)pheny-
l)-carboxamide;
[0403]
N-t-butoxycarbonyl-L-leucine-N'-(aminosulfonyl)phenyl)carboxamide;
[0404]
N-t-butoxycarbonyl-L-leucine-N'-(4-methoxycarbonylmethyl-phenyl)car-
boxamide;
[0405]
N-t-butoxycarbonyl-L-pyridin-3-ylalanine-N'-(4-methoxycarbonylpheny-
l)carboxamide;
[0406]
N-t-butoxycarbonyl-L-cyclohexylglycine-N'-(4-methoxy-carbonylphenyl-
)carboxamide;
[0407]
N-t-butoxycarbonyl-L-isoleucine-N'-(4-methoxycarbonylphenyl)carboxa-
mide;
[0408]
N-t-butoxycarbonyl-L-O-benzylthreonine-N'-(4-methoxy-carbonylphenyl-
)carboxamide;
[0409]
N-t-butoxycarbonyl-L-t-leucine-N'-(4-methoxycarbonylphenyl)carboxam-
ide;
[0410]
N-t-butoxycarbonyl-L-leucine-N'-(4-cyanophenyl)carboxamide;
[0411]
N-t-butoxycarbonyl-L-leucine-N'-(4-(N"-(2-dimethylaminoethyl-carbam-
oyl)carboxamide; and
[0412]
N-t-butoxycarbonyl-L-leucine-N'-(4-(N"-(3-dimethylamino-propyl)carb-
amoyl)phenyl)carboxamide.
[0413] 8C. In a similar manner, the following compounds of formula
(C) are prepared:
[0414]
N-t-butoxycarbonyl-L-tryptophan-N'-(4-nitrophenyl)carboxamide;
[0415]
N-t-butoxycarbonyl-L-tryptophan-N'-(4-aminophenyl)carboxamide;
[0416]
N-t-butoxycarbonyl-L-leucine-N'-(4-methylsulfonylphenyl)carboxamide-
;
[0417]
N-t-butoxycarbonyl-L-leucine-N'-(4-ethylsulfonylphenyl)carboxamide;
and
[0418]
N-t-butoxycarbonyl-L-leucine-N'-(4-tetrazolylphenyl)carboxamide.
Example 9
[0419] Compounds of Formula (D)
[0420] 9A. To a cold (0.degree. C.) solution of
N-t-butoxycarbonyl-L-leuci- ne-N'-phenylcarboxamide (3.4 g, 11
mmol) in dry CH.sub.2Cl.sub.2 (10 mL) was added TFA (2 mL). The
solution was allowed to stir at 25.degree. C. for 6 hours and was
then concentrated in vacua. The residue was partitioned between
CH.sub.2Cl.sub.2. and H.sub.2O and the aqueous layer was basified
at 0.degree. C. with saturated K.sub.2CO.sub.3 solution. The
organic phase was separated and the aqueous layer was extracted
three times with CH.sub.2Cl.sub.2. The combined organic layers were
washed with brine and dried over Na.sub.2SO.sub.4. Concentration
afforded L-leucine-N'-phenylcarboxamide.
[0421] 9B. In a similar manner, the following compounds are
prepared:
[0422] L-leucine-N'-(4-methoxycarbonylphenyl)carboxamide;
[0423] L-tryptophan-N'-phenylmethylcarboxamide;
[0424] L-tryptophan-N'-phenylcarboxamide;
[0425] L-tryptophan-N'-(4-methoxycarbonylphenyl)carboxamide;
[0426] L-tryptophan-N'-(4-ethoxycarbonylphenyl)carboxamide;
[0427]
L-leucine-N'-(4-(N"-methylaminosulfonyl)phenyl)carboxamide;
[0428] L-alanine-N'-(4-methoxycarbonylphenyl)carboxamide;
[0429] L-methionine-N'-(4-methoxycarbonylphenyl)carboxamide;
[0430] L-leucine-N'-(3-ethoxycarbonylphenyl)carboxamide;
[0431] L-leucine-N'-(2-methoxycarbonylphenyl)carboxamide;
[0432]
L-leucine-N'-(4-(1-methylethyloxy)carbonyl)phenyl)carboxamide;
[0433] L-leucine-N'-(aminosulfonyl)phenyl)carboxamide;
[0434] L-leucine-N'-(4-methoxycarbonylmethylphenyl)carboxamide;
[0435]
L-pyridin-3-ylalanine-N'-(4-methoxycarbonylphenyl)carboxamide;
[0436]
L-spirocyclopentylglycine-N'-(4-methoxycarbonylphenyl)carboxamide;
[0437]
L-cyclohexylglycine-N'-(4-methoxycarbonylphenyl)carboxamide;
[0438] L-isoleucine-N'-(4-methoxycarbonylphenyl)carboxamide;
[0439]
L-O-benzylthreonine-N'-(4-methoxycarbonylphenyl)carboxamide;
[0440] L-t-leucine-N'-(4-methoxycarbonylphenyl)carboxamide;
[0441] L-leucine-N'-(4-cyanophenyl)carboxamide;
[0442]
L-leucine-N'-(4-(N"-(2-dimethylaminoethyl)carbamoyl)phenyl)carboxam-
ide; and
[0443]
L-leucine-N'-(4-(N"-(3-dimethylaminopropyl)carbamoyl)phenyl)carboxa-
mide.
[0444] 9c. In a similar manner, the following compounds of formula
(D) are prepared:
[0445] L-tryptophan-N'-(4-nitrophenyl)carboxamide;
[0446] L-tryptophan-N'-(4-aminophenyl)carboxamide;
[0447] L-leucine-N'-(4-methylsulfonylphenyl)carboxamide;
[0448] L-leucine-N'-(4-ethylsulfonylphenyl)carboxamide; and
[0449] L-leucine-N'-(4-tetrazolylphenyl)carboxamide.
Example 10
[0450] Compounds of Formula (Ia)
[0451] 10A. To a cold (0.degree. C.) solution of
2-(hydroxy)(quinolin-2-yl-
thiomethyl)-phosphinoylmethyl-4-methylpentanoic acid (0.20 g, 0.54
mmol) in THF (6 mL) was added 1,1'-carbonyldiimidazole (0.12 g, 0.7
mmol). The mixture was stirred for 75 minutes at 0.degree. C. and
was then treated with
L-tryptophan-N'-(4-ethoxycarbonylphenyl)carboxamide (0.22 g, 0.62
mmol) and stirred at 25.degree. C. for 15 hours. The THF was
evaporated and the residue was dissolved in ethyl acetate (60 mL).
The solution was washed with H.sub.2O (10 mL), brine (10 mL), and
dried over MgSO.sub.4. Concentration was followed by reverse phase
HPLC using a gradient of acetonitrile and 50 mM NH.sub.4OAc buffer
afforded 30 mg of
N-(2-(hydroxy)(quinolin-2-ylthiomethyl)phosphinoyl-methyl-4-methylpentano-
yl)-L-tryptophan-N'-(4-ethoxycarbonyl-phenyl)carboxamide as an off
white solid, MS(FAB) 701 (M-H).sup.+ (mixture of
diastereomers).
[0452] 10B. In a similar manner, the following compounds of formula
(Ia) were prepared:
[0453]
N-(2-(hydroxy)(quinolin-2-ylthiomethyl)phosphinoylmethyl-4-methylpe-
ntanoyl)-L-tryptophan-N'-(4-methoxy-carbonylphenyl)carboxamide,
[0454] MS(FAB) 687 (M+H).sup.+;
[0455]
N-(2-(hydroxy)(quinolin-2-ylthiomethyl)phosphinoylmethyl-4-methylpe-
ntanoyl)-L-alanine-N'-(4-methoxy-carbonylphenyl)carboxamide,
[0456] MS(FAB) 572 (M+H).sup.+;
[0457]
N-(2-(hydroxy)(quinolin-2-ylthiomethyl)phosphinoylmethyl-4-methylpe-
ntanoyl)-L-methionine-N'-(4-methoxy-carbonylphenyl)carboxamide,
[0458] MS(FAB) 632 (M+H).sup.+;
[0459]
N-(2-(hydroxy)(quinolin-2-ylthiomethyl)phosphinoylmethyl-4-methylpe-
ntanoyl)-L-leucine-N'-(4-methoxy-carbonylphenyl)carboxamide,
[0460] MS(FAB) 614 (M+H).sup.+;
[0461]
N-(2-(hydroxy)(quinolin-2-ylthiomethyl)phosphinoylmethyl-4-methylpe-
ntanoyl)-L-leucine-N'-(3-ethoxy-carbonylphenyl)carboxamide,
[0462] .sup.1H NMR (300 MHz, MeOH) .delta. 0.73-1.01 (m, 12H),
1.28-2.00 (m, 14H), 2.4-3.61 (m, 2H), 4.27-4.45 (m, 3H), 7.23-7.44
(m, 3H), 7.65-7.98 (m, 6H), 8.29 (s, 0.5H), 8.50 (8, 0.5H);
[0463]
N-(2-(hydroxy)(quinolin-2-ylthiomethyl)phosphinoylmethyl-4-methylpe-
ntanoyl)-L-leucine-N'-(2-methoxy-carbonylphenyl)carboxamide,
[0464] .sup.1H NMR (300 MHz, MeOH) .delta.0.78-0.99 (m, 13H),
1.3-2.4 (m, 7H), 2.90-3.05 (m, 1H), 3.5-3.75 (m, 2H), 3.89, 3.90,
3.94 (3s, 3H total), 4.35-3.50 (m, 1H), 7.05-8.10 (m, 11H), 8.32,
8.55, 8.60 (3d, J=8.7, 1H);
[0465]
N-(2-(hydroxy)(quinolin-2-ylthiomethyl)phosphinoylmethyl-4-methylpe-
ntanoyl)-L-leucine-N'-(4-(1,1-dimethyl-ethoxycarbonylphenyl)carboxamide,
[0466] MS(FAB) 642 (MH).sup.+;
[0467]
N-(2-(hydroxy)(quinolin-2-ylthiomethyl)phosphinoylmethyl-4-methylpe-
ntanoyl)-L-leucine-N'-(4-aminosulfonyl-phenyl)carboxamide,
[0468] .sup.1H NMR (300 MHz, MeOH) .delta. 0.76 (d, J=6.5, 3H),
0.81 (d, J=6.5, 3H), 0.85-1.1 (m, 7H), 1.2-2.1 (m, 7H), 2.92-2.95
(m, 1H), 3.45-3.70 (m, 2H), 4.35-4.45 (m, 1H), 7.28 (d, J=8.7, 1H),
7.45 (t, J=8.7, 1H), 7.68 (t, J=8.7, 1H), 7.7-7.8 (m, 3H), 7.87 (d,
J=8.7, 1H), 7.95-8.1 (m, 3H);
[0469]
N-(2-(hydroxy)(quinolin-2-ylthiomethyl)phosphinoylmethyl-4-methylpe-
ntanoyl)-L-leucine-N'-(4-methoxycarbonylmethyl-phenyl)carboxamide,
[0470] MS(FAB) 628 (MH).sup.+.
[0471] 10C. In a similar manner, the following compounds of formula
(Ia) are prepared:
[0472]
N-(2-(hydroxy)(quinolin-2-ylthiomethyl)phosphinoylmethyl-5-phenylpe-
ntanoyl)-L-leucine-N'-(4-methoxycarbonyl-phenyl)carboxamide;
[0473]
N-(2-(hydroxy)(quinolin-2-ylthiomethyl)phosphinoylmethyl-4-phenylbu-
tanoyl)-L-leucine-N'-(4-methoxycarbonyl-phenyl)carboxamide;
[0474]
N-(2-(hydroxy)(quinolin-2-ylthiomethyl)phosphinoylmethyl-3-phenylpr-
opanoyl)-L-leucine-N'-(4-methoxycarbonyl-phenyl)carboxamide;
[0475]
N-(2-(hydroxy)(quinolin-2-ylthiomethyl)phosphinoylmethyl-3-cyclohex-
ylpropanoyl)-L-leucine-N'-(4-methoxycarbonyl-phenyl)carboxamide;
[0476]
N-(2-((hydroxy)(quinolin-2-ylthiomethyl)phosphinoylmethyl)-pentanoy-
l)-L-leucine-N'-(4-methoxycarbonyl-phenyl)carboxamide;
[0477]
N-(2-(hydroxy)(naphth-1-ylthiomethyl)phosphinoylmethyl-5-phenylpent-
anoyl)-L-leucine-N'-(4-methoxycarbonyl-phenyl)carboxamide;
[0478]
N-(2-(hydroxy)(naphth-1-ylthiomethyl)phosphinoylmethyl-4-phenylbuta-
noyl)-L-leucine-N'-(4-methoxycarbonyl-phenyl)carboxamide;
[0479]
N-(2-(hydroxy)(naphth-1-ylthiomethyl)phosphinoylmethyl-3-phenylprop-
anoyl)-L-leucine-N'-(4-methoxycarbonyl-phenyl)carboxamide;
[0480]
N-(2-(hydroxy)(naphth-1-ylthiomethyl)phosphinoylmethyl-3-cyclohexyl-
propanoyl)-L-leucine-N'-(4-methoxycarbonyl-phenyl)carboxamide;
[0481]
N-(2-((hydroxy)(naphth-1-ylthiomethyl)phosphinoylmethyl)-pentanoyl)-
-L-leucine-N'-(4-methoxycarbonyl-phenyl)carboxamide;
[0482]
N-(2-(hydroxy)(naphth-2-ylthiomethyl)phosphinoylmethyl-5-phenylpent-
anoyl)-L-leucine-N'-(4-methoxycarbonyl-phenyl)carboxamide;
[0483]
N-(2-(hydroxy)(naphth-2-ylthiomethyl)phosphinoylmethyl-4-phenylbuta-
noyl)-L-leucine-N'-(4-methoxycarbonyl-phenyl)carboxamide;
[0484]
N-(2-(hydroxy)(naphth-2-ylthiomethyl)phosphinoylmethyl-3-phenylprop-
anoyl)-L-leucine-N'-(4-methoxycarbonyl-phenyl)carboxamide;
[0485]
N-(2-(hydroxy)(naphth-2-ylthiomethyl)phosphinoylmethyl-3-cyclohexyl-
propanoyl)-L-leucine-N'-(4-methoxycarbonyl-phenyl)carboxamide;
[0486]
N-(2-((hydroxy)(naphth-2-ylthiomethyl)phosphinoylmethyl)-pentanoyl)-
-L-leucine-N'-(4-methoxycarbonyl-phenyl)carboxamide;
[0487]
N-(2-(hydroxy)(phenylthiomethyl)phosphinoylmethyl-5-phenylpentanoyl-
)-L-leucine-N'-(4-methoxycarbonyl-phenyl)carboxamide;
[0488]
N-(2-(hydroxy)(phenylthiomethyl)phosphinoylmethyl-4-phenylbutanoyl)-
-L-leucine-N'-(4-methoxycarbonyl-phenyl)carboxamide;
[0489]
N-(2-(hydroxy)(phenylthiomethyl)phosphinoylmethyl-3-phenylpropanoyl-
)-L-leucine-N'-(4-methoxycarbonyl-phenyl)carboxamide;
[0490]
N-(2-(hydroxy)(phenylthiomethyl)phosphinoylmethyl-3-cyclohexylpropa-
noyl)-L-leucine-N'-(4-methoxycarbonyl-phenyl)carboxamide; and
[0491]
N-(2-((hydroxy)(phenylthiomethyl)phosphinoylmethyl)-pentanoyl)-L-le-
ucine-N'-(4-methoxycarbonyl-phenyl)carboxamide.
[0492] 10D. A solution of
N-(2-(hydroxy)(quinolin-2-ylthiomethyl)-phosphin-
oylmethyl-4-methylpentanoyl)-L-tryptophan-N'-(4-methoxycarbonyl-phenyl)car-
boxamide in THF (2 mL) and 1M NaOH (1 mL) was stirred for 24 hours
at 25.degree. C. The organic solvents were evaporated, and the
residue dissolved in ethyl acetate/H.sub.2O. The aqueous phase was
acidified with 1M HCl and the separated aqueous phase was extracted
twice with ethyl acetate. The combined organic layers were washed
with brine, dried (MgSO.sub.4) and concentrated to 27 mg of
N-(2-(hydroxy)(quinolin-2-ylthi-
omethyl)-phosphinoylmethyl-4-methylpentanoyl)-L-tryptophan-N'-(4-carboxyph-
enyl)-carboxamide as a yellow powder.
[0493] 10E. In a similar manner, but starting with
N-(2-(hydroxy)(quinolin-
-2-ylthiomethyl)phosphinoylmethyl-4-methylpentanoyl)-L-leucine-N'-(4-metho-
xy-carbonylphenyl)carboxamide (30 mg, 0.048 mmol) there was
obtained 10 mg of
N-(2-(hydroxy)(quinolin-2-ylthiomethyl)phosphinoylmethyl-4-methylpenta-
noyl)-L-leucine-N'-(4-carboxyphenyl)carboxamide as a semisolid
after trituration with ethyl acetate;
[0494] .sup.1H NMR.sup.1 (300 MHZ, MeOH) 0.81-1.02 (m, 12H),
1.1-2.3 (m, 10H), 2.82-3.00 (m, 1H), 3.49, 3.56 (2s,2H), 3.5-3.8
(m,2H), 4.45-4.55 (m,1H), 7.09 (d, J=8.2, 1H), 7.19 (d, J=8.2, 1H),
7.45 (t, J=8.2, 1H) 7.45-7.6 (m, 3H), 7.65-7.80 (m, 1H), 7.82-7.98
(m, 2H), 8.10-8.20 (m, 1H).
Example 11
[0495] Compounds of Formula (Fa)
[0496] 11A. To 4-methylpentanoic acid (25 g, 0.215 mmol) in a
25.degree. C. water bath, thionyl chloride (20.4 mL, 1.3 g) was
slowly added. Then the mixture was heated at 50.degree. C. under
argon for 3 hours (until the evolution of gas had stopped). The
crude reaction mixture was distilled at atmospheric pressure to
give 4-methylpentanoyl chloride (25.3 g, 87.3%), b.p. 143.degree.
C.
[0497] 11B. In a similar manner, but replacing 4-methylpentanoic
acid with 5-phenylpentanoic acid (5 g), 5-phenylpentanoyl chloride
was prepared (4.4 g), as a colorless liquid, b.p.
91.degree.-93.degree. C.
Example 12
[0498] Compounds of Formula (Fb)
[0499] 12A. To a suspension of 60% NaH (836 mg, 1.5 eq.) in toluene
(200 mL) at room temperature under argon was added
L-(+)-2,10-camphor sultam (3.0 g, 13.9 mmol) portion-wise. The
mixture was stirred vigorously at room temperature for one hour.
Then 4-methylpentanoyl chloride was carefully added dropwise to the
solution at 0.degree. C. After stirring the reaction at room
temperature for 3 hours, the reaction was quenched with 10 mL of
water, and 70 mL of ether was added. The reaction mixture was first
washed with 0.5N HCl (2.times.50 mL), then 5% K.sub.2CO.sub.3
(3.times.50 mL) and finally with brine (1.times.50 mL). The organic
layer was dried over MgSO.sub.4, filtered and evaporated to
dryness. Purification by column chromatography (1:6 ethyl
acetate/petroleum ether as eluant) gave
N-4-methylpentanoyl-L-(+)-2,10-camphor sultam (3.39 g, 78%).
[0500] 12B. In a similar manner, but replacing 4-methylpentanoyl
chloride with the appropriate chloride, the following compounds of
formula (Fb) were prepared:
[0501] N-3-phenylpropanoyl-L-(+)-2,10-camphor sultam, MS: 347
(M.sup.+);
[0502] N-5-phenylpentanoyl-L-(+)-2,10-camphor sultam, MS: 375
M.sup.+;
[0503] N-pentanoyl-L-(+)-2,10-camphor sultam, MS: 300
(M+H).sup.+.
Example 13
[0504] Compounds of Formula (Fc)
[0505] 13A. To a solution of N-4-methylpentanoyl-L-(+)-2,10-camphor
sultam (3.39 g, 10.8 mmol) in 75 mL of dry THF at -78.degree. C.
under argon was added NaN(TMS).sub.2 (1.0 M in THF, 11.34 mL, 1.05
eq.) dropwise over five minutes. After stirring at -78.degree. C.
for 1 hour, hexamethylphosphoramide (5 mL) was added to the
mixture, followed by t-butylbromoacetate (5.2 ml, 3 eq), then 400
mg of tetra n-butyl ammonium iodide was added in one portion. The
resulting solution was kept at -78.degree. C. under argon
overnight. The next morning, the reaction was quenched with water
(100 mL), and then it was extracted with ether (3.times.100 mL).
The combined ether layers were washed with brine, then dried over
Na.sub.2SO.sub.4, filtered and concentrated. Purification by column
chromatography (5:95 ethyl acetate/petroleum ether to 10:90 ethyl
acetate/petroleum ether as eluant) gave
N-(4-methyl-2-t-butoxycarbonylmet- hyl)pentanoyl-L-(+)-2,10-camphor
sultam (4 g, 86.5%).
[0506] 13B. In a similar manner, but replacing
N-4-methylpentanoyl-L-(+)-2- ,10-camphor sultam with the
appropriate compound of formula (Fb), the following compounds of
formula (Fc) were prepared:
[0507]
N-(3-phenyl-2-t-butoxycarbonylmethyl)propanoyl-L-(+)-2,10-camphor
sultam, MS: 461 (M.sup.+);
[0508]
N-(5-phenyl-2-t-butoxycarbonylmethyl)pentanoyl-L-(+)-2,10-camphor
sultam, MS: 490.1 (M+H).sup.+;
[0509] N-(2-t-butoxycarbonylmethyl)pentanoyl-L-(+)-2,10-camphor
sultam, MS: 414 (M+H).sup.+.
Example 14
[0510] Compounds of Formula (F)
[0511] 14A. To a stirred solution of
N-(4-methyl-2-t-butoxycarbonylmethyl)-
-pentanoyl-L-(+)-2,10-camphor sultam (5.45 g, 12.7 mmol) in 50%
aqueous THF (150 mL) at 0.degree. C. under argon was added
LiOH.H.sub.2O crystals (2.14 g, 4 eq.) followed by 30%
H.sub.2O.sub.2 (11.5 mL). Then the ice-bath was removed and the
resulting emulsion was stirred for 3 hours before it had turned
clear. Most of the THF was removed under reduced pressure at
35.degree. C. Then CH.sub.2Cl.sub.2 (150 mL) was added and with
stirring 4N HCl was added to pH=2. After adding NaCl, the aqueous
layer was further extracted with CH.sub.2Cl.sub.2 (3.times.150 mL).
The CH.sub.2Cl.sub.2 was removed under reduced pressure at
35.degree. C. and then the residue was taken up in ethyl acetate
(150 mL). This solution was then extracted with 5% K.sub.2CO.sub.3
(3.times.50 mL) and the combined extracts were washed with ether
(50 mL). Then CH.sub.2Cl.sub.2 was added to the aqueous layer and
with stirring with NaCl, the aqueous layer was extracted with
CH.sub.2Cl.sub.2 (3.times.70 mL) and the combined extracts were
dried over Na.sub.2SO.sub.4, filtered and concentrated to give
(2R)-4-methyl-2-t-butoxycarbonylmethyl-pentanoic acid as a
colorless oil (2.95 g, quantitative yield).
[0512] 14B. In a similar manner, but replacing
N-(4-methyl-2-t-butoxycarbo- nylmethyl)pentanoyl-L-(+)-2,10-camphor
sultam with the appropriate compound of formula (Fc), the following
compounds of formula (F) were prepared:
[0513] (2R)-3-phenyl-2-t-butoxycarbonylmethyl-propanoic acid,
[0514] MS: 265 (M+H).sup.+;
[0515] (2R)-5-phenyl-2-t-butoxycarbonylmethyl-pentanoic acid,
[0516] MS: 293.1 (M+H).sup.+;
[0517] (2R)-2-t-butoxycarbonylmethyl-pentanoic acid, (colorless
oil, 1.09 g).
[0518] 14C. (2R)-3-Phenyl-2-t-butoxycarbonylmethyl-propanoic acid
(55 mg) was taken up in glacial acetic acid (20 mL) and PtO.sub.2
(25 mg) was added in acetic acid. Then the beaker was placed in a
Parr bomb, it was evacuated and charged with 100 psi of H.sub.2.
After stirring for 3 days, the mixture was suction filtered through
a 1 cm bed of celite. The filtrate was then concentrated to a
yellow oil, (2R)-3-cyclohexyl-2-t-but- oxycarbonylmethyl-propanoic
acid (56 mg), MS: 269.5 (M-H).sup.-.
Example 15
[0519] Compounds of Formula (Ib)
[0520] 15A. To a solution of
4-methyl-2-t-butoxycarbonyl-methylpentanoic acid (0.28 g, 1.2 mmol)
in DMF (5 mL) containing HOBT (0.22 g, 1.8 mmol) was added EDCI
(0.31 g, 1.8 mmol). The mixture was stirred at 0.degree. C. for 1
hour and was then treated with L-cyclohexylglycine-N'-(4-methoxy-
carbonyl-phenyl)carboxamide (1.2 mmol) and DMAP (27 mg, 0.24 mmol).
Stirring was continued for 24 hours at 25.degree. C. and then the
DMF was evaporated. The residue was dissolved in CH.sub.2Cl.sub.2
(20 mL) and the solution was washed with 1M HCl (10 mL), saturated
NaHCO.sub.3 (10 mL), brine (10 mL) and was dried over
Na.sub.2SO.sub.4. Concentration in vacuo afforded an oil which was
purified by flash chromatography on SiO.sub.2 using 20% ethyl
acetate/hexenes as element. There was obtained 0.22 g (22%) of
N-(4-methyl-2-t-butoxycarbonylmethyl-pentanoyl)-L-cyclohexylglyc-
ine-N'-(4-methoxycarbonylphenyl)carboxamide as a solid, MS(FAB) 503
(MH).sup.+.
[0521] 15B. In a similar manner, the-following compounds were
prepared:
[0522]
N-(4-methyl-2-t-butoxycarbonylmethylpentanoyl)-L-pyridin-3-ylalanin-
e-N'-(4-methoxycarbonylphenyl)carboxamide;
[0523]
N-(4-methyl-2-t-butoxycarbonylmethylpentanoyl)-L-O-benzylthreonine--
N'-(4-methoxycarbonylphenyl)carboxamide;
[0524]
N-(4-methyl-2-t-butoxycarbonylmethylpentanoyl)-L-isoleucine-N'-(4-m-
ethoxycarbonylphenyl)carboxamide;
[0525]
N-(4-methyl-2-t-butoxycarbonylmethylpentanoyl)-L-leucine-N'-(4-meth-
oxycarbonylphenyl)carboxamide;
[0526]
N-(4-methyl-2-t-butoxycarbonylmethylpentanoyl)-L-t-leucine-N'-(4-me-
thoxycarbonylphenyl)carboxamide;
[0527]
N-(4-methyl-2-t-butoxycarbonylmethylpentanoyl)-L-leucine-N'-(4-cyan-
ophenyl)carboxamide;
[0528]
N-(4-methyl-2-t-butoxycarbonylmethylpentanoyl)-L-leucine-N'-(4-(N"--
(3-dimethylaminopropyl)carbamoyl)phenyl)-carboxamide;
[0529]
N-(4-methyl-2-t-butoxycarbonylmethylpentanoyl)-L-leucine-N'-(4-(N"--
(2-dimethylaminoethyl)carbamoyl)phenyl)-carboxamide;
[0530]
N-(4-methyl-2-t-butoxycarbonylmethylpentanoyl)-L-leucine-N'-(4-amin-
osulfonylphenyl)carboxamide;
[0531]
N-(4-methyl-2-t-butoxycarbonylmethylpentanoyl)-L-leucine-N'-(4-meth-
ylaminosulfonylphenyl)carboxamide;
[0532]
N-(2-t-butoxycarbonylmethylpentanoyl)-L-leucine-N'-(4-methoxycarbon-
ylphenyl)carboxamide;
[0533]
N-(3-phenyl-2-t-butoxycarbonylmethylpropanoyl)-L-leucine-N'-(4-meth-
oxycarbonylphenyl)carboxamide;
[0534]
N-(3-cyclohexyl-2-t-butoxycarbonylmethylpropanoyl)-L-leucine-N'-(4--
methoxycarbonylphenyl)carboxamide;
[0535]
N-(4-phenyl-2-t-butoxycarbonylmethylbutanoyl)-L-leucine-N'-(4-metho-
xycarbonylphenyl)carboxamide; and
[0536]
N-(5-phenyl-2-t-butoxycarbonylmethylpentanoyl)-L-leucine-N'-(4-meth-
oxycarbonylphenyl)carboxamide.
Example 16
[0537] Compounds of Formula (Ic)
[0538] 16A. To a cold (0.degree. C.) solution of
N-(4-methyl-2-t-butoxycar-
bonylmethyl-pentanoyl)-L-cyclohexylglycine-N'-(4-methoxycarbonylphenyl)-ca-
rboxamide (70 mg, 0.14 mmol) in CH.sub.2Cl.sub.2 (2 mL) was added
TFA (0.5 mL). After stirring for 5 hours at 25.degree. C., the
solution was concentrated in vacuo and the product was purified by
reverse phase HPLC using a gradient of acetonitrile and 50 mM
NH.sub.4OAc buffer to provide 44 mg (71%) of
N-(4-methyl-2-carboxymethylpentanoyl)-L-cyclohexylglycine--
N'-(4-methoxycarbonylphenyl)-carboxamide as a white solid, MS(FAB)
445 (M-H).sup.-.
[0539] 16B. In a similar manner, the following compounds were
prepared:
[0540]
N-(4-methyl-2-carboxymethylpentanoyl)-L-isoleucine-N'-(4-methoxycar-
bonylphenyl)carboxamide,
[0541] MS(FAB) 419 (M-H).sup.-;
[0542]
N-(4-methyl-2-carboxymethylpentanoyl)-L-leucine-N'-(4-methoxy-carbo-
nylphenyl)carboxamide, MS(FAB) 419 (M-H).sup.-;
[0543]
N-(4-methyl-2-carboxymethylpentanoyl)-L-t-leucine-N'-(4-methoxycarb-
onylphenyl)carboxamide;
[0544]
N-(4-methyl-2-carboxymethylpentanoyl)-L-leucine-N'-(4-cyano-phenyl)-
carboxamide,
[0545] .sup.1H NMR (300 MHz, MeOH) .delta.0.84-0.99 (m, 12H),
1.15-1.82 (m, 6H), 2.36-2.41 (m, 1H), 2.52-2.65 (m, 1H), 2.8-2.95
(m, 1H), 4.49-4.54 (m, 1H), 7.4-7.9 (m, 4H);
[0546]
N-(4-methyl-2-carboxymethylpentanoyl)-L-leucine-N'-(4-aminosulfonyl-
phenyl)carboxamide,
[0547] .sup.1H NMR (300 MHZ, MeOH) .delta.0.85-1.00 (m, 12H),
1.1-1.3 (m, 2H), 1.52-1.85 (m, 4H), 2.31-2.95 (m, 3H), 4.49-4.55
(m, 1H), 7.75-7.91 (m, 4H);
[0548]
N-(4-methyl-2-carboxymethylpentanoyl)-L-leucine-N'-(4-methyl-aminos-
ulfonylphenyl)carboxamide, MS(FAB) 459 (M-H).sup.-;
[0549]
N-(2-carboxymethylpentanoyl)-L-leucine-N'-(4-methoxy-carbonylphenyl-
)carboxamide, MS(FAB) 405 (M-H).sup.-;
[0550]
N-(3-phenyl-2-carboxymethylpropanoyl)-L-leucine-N'-(4-methoxy-carbo-
nylphenyl)carboxamide, MS(FAB) 455 (M+H).sup.+;
[0551]
N-(3-cyclohexyl-2-carboxymethylpropanoyl)-L-leucine-N'-(4-methoxyca-
rbonylphenyl)carboxamide,
[0552] MS(FAB) 459 (M-H).sup.-;
[0553]
N-(4-phenyl-2-carboxymethylbutanoyl)-L-leucine-N'-(4-methoxy-carbon-
ylphenyl)carboxamide, MS(FAB) 467 (M-H).sup.-;
[0554]
N-(4-phenyl-2-carboxymethylbutanoyl)-L-cyclohexylglycine-N'-(4-meth-
oxycarbonylphenyl)carboxamide;
[0555]
N-(4-phenyl-2-carboxymethylbutanoyl)-L-t-leucine-N'-(4-methoxycarbo-
nylphenyl)carboxamide;
[0556]
N-(5-phenyl-2-carboxymethylpentanoyl)-L-leucine-N'-(4-methoxy-carbo-
nylphenyl)carboxamide, MS(FAB) 481 (M-H).sup.-; and
[0557]
N-(4-methyl-2-carboxymethylpentanoyl)-L-O-benzylthreonine-N'-(4-met-
hoxycarbonylphenyl)carboxamide,
[0558] MS(FAB) 497 (M-H).sup.-.
[0559] 16C. In a similar manner, but triturating the crude product
with ether and then decanting the ether to yield the following
compounds as TFA salts:
[0560]
N-(4-methyl-2-carboxymethylpentanoyl)-L-pyridin-3-ylalanine-N'-(4-m-
ethoxycarbonylphenyl)carboxamide,
[0561] MS(FAB) 456 (M.sup.+H).sup.+;
[0562]
N-(4-methyl-2-carboxymethylpentanoyl)-L-leucine-N-(4-(N"-(3-dimethy-
laminopropyl)carbamoyl)phenyl)-carboxamide, MS(FAB) 491
(M+H).sup.+; and
[0563]
N-(4-methyl-2-carboxymethylpentanoyl)-L-leucine-N'-(4-(N"-(2-dimeth-
ylaminoethyl)carbamoyl)phenyl)-carboxamide, MS(FAB) 491
(M+H).sup.+.
[0564] 16D. A mixture of
N-(4-methyl-2-t-butoxycarbonylmethyl-pentanoyl)-L-
-O-benzylthreonine-N'-(4-methoxycarbonylphenyl)-carboxamide (60 mg)
and Pd/C in ethyl acetate/THF (1:1, 25 mL) was hydrogenated
overnight at 1 atm pressure. Filtration through Celite,
concentration of the filtrate, and trituration of the residue with
ether/hexenes produced
N-(4-methyl-2-t-butoxycarbonylmethyl-pentanoyl)-L-threonine-N'-(4-methoxy-
carbonyl-phenyl)carboxamide, MS(FAB) 407 (M-H).sup.-.
[0565] 16E. By following the procedure of part A and substituting
N-(4-methyl-2-t-butoxycarbonylmethyl-pentanoyl)-L-cyclohexylglycine-N'-(4-
-methoxycarbonylphenyl)-carboxamide with the following:
[0566]
N-(2R-(t-butoxycarbonyl)methyl-5-(biphen-4-yl)pentanoyl)-L-lysine-N-
'-(4-(ethoxycarbonyl)phenyl)carboxamide;
[0567]
N-(2R-(t-butoxycarbonyl)methyl-5-(phenyl)pentanoyl)-L-lysine-N'-(4--
(ethoxycarbonyl)phenyl)carboxamide;
[0568]
N-(2R-(t-butoxycarbonyl)methyl-5-(biphen-4-yl)pentanoyl)-L-(N.epsil-
on.-isopropyl)lysine-N'-(4-(ethoxycarbonyl)phenyl)carboxamide;
[0569]
N-(2R-(t-butoxycarbonyl)methyl-4-(phenyl)butanoyl)-L-cyclohexylglyc-
ine-N'-(4-(N",N"-dimethylaminoethylaminosulfonyl)-phenyl)carboxamide;
[0570]
N-(2R-(t-butoxycarbonyl)methyl-5-(phenyl)pentanoyl)-L-(N,N'-diethyl-
guanido)lysine-N'-(4-(ethoxycarbonyl)phenyl)carboxamide;
[0571]
N-(2R-(t-butoxycarbonyl)methyl-5-(biphen-4-yl)pentanoyl)-L-t-leucin-
e-N'-(4-(methylthio)phenyl)carboxamide;
[0572]
N-(2R-(t-butoxycarbonyl)methyl-5-biphen-4-yl)pentanoyl)-L-t-leucine-
-N'-(3-(2-hydroxyethyl)phenyl)carboxamide;
[0573]
N-(2R-(t-butoxycarbonyl)methyl-5-biphen-4-yl)pentanoyl)L-S-((4-cyan-
ophenyl)methyl)penicillamine-N'-(phenyl)carboxamide;
[0574]
N-(2R-(t-butoxycarbonyl)methyl-5-(biphen-4-yl)pentanoyl)-L-cyclohex-
ylglycine-N'-(2-(4-aminosulfonyl)phenylethyl)carboxamide;
[0575]
N-(2R-(t-butoxycarbonyl)methyl-5-(biphen-4-yl)pentanoyl)-L-cyclohex-
ylglycine-N'-(3-(morpholin-4-yl)propyl)carboxamide;
[0576]
N-(2R-(t-butoxycarbonyl)methyl-5-(biphen-4-yl)pentanoyl)-L-t-leucin-
e-N'-(4-(methylaminosulfonyl)phenyl)carboxamide;
[0577]
N-(2R-(t-butoxycarbonyl)methyl-5-(biphen-4-yl)pentanoyl)-L-cyclohex-
ylglycine-N'-(4-((2-hydroxyethyl)aminosulfonyl)phenyl)carboxamide;
[0578]
N-(2R-(t-butoxycarbonyl)methyl-5-(biphen-4-yl)pentanoyl)-L-cyclohex-
ylglycine-N'-(4-(N",N"-dimethylaminoethylaminosulfonyl)phenyl)carboxamide;
and
[0579]
N-(2R-(t-butoxycarbonyl)methyl-5-(biphen-4-yl)pentanoyl)-L-t-leucin-
e-N'-(4-((3-(morpholin-4-yl)propyl)aminosulfonyl)phenyl)carboxamide,
there are obtained:
[0580]
N-(2R,S)-(N"-formyl-N"-hydroxyamino)methyl-4-(methyl)pentanoyl)-L-l-
eucine-N'-(4-(methoxycarbonyl)phenyl)carboxyamide: MS: 434.2
(M-H).sup.-388 (M-HCO--OH);
[0581]
N-(2R-(N"-hydroxycarbamoyl)methyl-4-(methyl)pentanoyl)D,L-norvaline-
-N'-(4-((2-(dimethylamino)ethyl)carbamoyl)phenyl)carboxamide: MS:
478 (M.sup.+H).sup.+;
[0582]
N-(2R-carboxymethyl-5-(biphen-4-yl)pentanoyl)-L-lysine-N'-(4-(ethox-
ycarbonyl)phenyl)carboxamide: MS: 588.3 (M+H).sup.+;
[0583]
N-(2R-carboxymethyl-5-(phenyl)pentanoyl)-L-lysine-N'-(4-(ethoxycarb-
onylphenyl)carboxamide: Ms: 512.3 (M+H).sup.+;
[0584]
N-(2R-carboxymethyl-5-(biphen-4-yl)pentanoyl)-L-(N.epsilon.-isoprop-
yl)lysine-N'-(4-(ethoxycarbonyl)phenyl)carboxamide: MS: 630
(M+H).sup.+;
[0585]
N-(2R-carboxymethyl)-4-(phenyl)butanoyl)-L-cyclohexylglycine-N'-(4--
(N",N"-dimethylaminoethylaminosulfonyl)-phenyl)carboxamide: MS: 586
(M+H).sup.+;
[0586]
N-(2R-carboxymethyl-5-(phenyl)pentanoyl)-L-(N,N'-diethylguanido)lys-
ine-N'-(4-(ethoxycarbonyl)phenyl)carboxamide; MS: 610.4
(M+H).sup.+;
[0587]
N-(2R-carboxymethyl)-5-(biphen-4-yl)pentanoyl)-L-t-leucine-N'-(4-(m-
ethylthio)phenyl)carboxamide: FAB-MS (M+Na)+ calc. 569.2450; found:
569.2461;
[0588]
N-(2R-carboxymethyl-5-biphen-4-yl)pentanoyl)-L-t-leucine-N'-(3-(2-h-
ydroxyethyl)phenyl)carboxamide: FAB-MS (M+H).sup.+calc. 545.3015;
found: 545.3021;
[0589]
N-(2R-carboxymethyl-5-biphen-4-yl)pentanoyl)L-S-((4-cyanophenyl)met-
hyl)penicillamine-N'-(phenyl)carboxamide: FAB-MS (M+H).sup.+calc.
634.2740; found: 634.2749;
[0590]
N-(2R-carboxymethyl-5-(biphen-4-yl)pentanoyl)-L-cyclohexylglycine-N-
'-(2-(4-aminosulfonyl)phenylethyl)carboxamide: FAB-MS
(M+H).sup.+calc. 634.2951; found: 634.2963;
[0591]
N-(2R-carboxymethyl-5-(biphen-4-yl)pentanoyl)-L-cyclohexylglycine-N-
'-(3-(morpholin-4-yl)propyl)carboxamide: FAB-MS (M+H).sup.+calc.
578.3594; found: 578.3583;
[0592]
N-(2R-carboxymethyl-5-(biphen-4-yl)pentanoyl)-L-t-leucine-N'-(4-(me-
thylaminosulfonyl)phenyl)carboxamide: .sup.1H NMR (300 MHz,
acetone-d6) .delta. 9.73 (br,s, 1H), 7.91 (d, 2H, J=9 Hz), 7.79 (d,
2H, J=9 Hz), 7.56 (d, 2H, J=8 Hz), 7.25-7.45 (m, 6H), 7.19 (d, 2H,
J=8 Hz), 4.48 (d, 1H, J=9 Hz), 2.38-3.00 (m, 9H), 1.43-1.72 (m,
4H), 1.04 (s, 9H);
[0593]
N-(2R-carboxymethyl-5-(biphen-4-yl)pentanoyl)-L-cyclohexylglycine-N-
'-(4-((2-hydroxyethyl)aminosulfonyl)phenyl)carboxamide: FAB-MS
(M+Cs).sup.+calc. 782.1876; found: 782.1896;
[0594]
N-(2R-carboxymethyl-5-(biphen-4-yl)pentanoyl)-L-cyclohexylglycine-N-
'-(4-((2-dimethylamino)ethyl)aminosulfonyl)phenyl)carboxamide:
FAB-MS (M+Cs).sup.+calc. 809.2349; found: 809.2369; and
[0595]
N-(2R-carboxymethyl-5-(biphen-4-yl)pentanoyl)-L-t-leucine-N'-(4-((3-
-(morpholin-4-yl)propyl)aminosulfonyl)phenyl)carboxamide: FAB-MS
(M+H).sup.+calc. 707.3478; found: 707.3489.
Example 17
[0596] Compounds of Formula (Id)
[0597] 17A. A solution of
N-(4-methyl-2-carboxymethylpentanoyl)-L-leucine--
N'-(4-methoxycarbonylphenyl)carboxamide (0.28 g, 0.66 mmol) and
HOBT (0.12 g) in dry DMF (20 mL) was cooled to 0.degree. C. and
treated with EDCI (0.32 g). After stirring 0.5 hours at 0.degree.
C., O-benzylhydroxylamine (0.30 mL) was added and the reaction was
allowed to warm to 25.degree. C. overnight. The DMF was removed in
vacuo and the residue was taken up in CH.sub.2Cl.sub.2 and washed
with 5% HCl/5% NaHCO.sub.3 and brine and the solution was dried
over Na.sub.2SO.sub.4. After concentration, the product was
purified by flash chromatography (SiO.sub.2,
[0598] R.sub.f=0.6, 10% MeOH/CH.sub.2Cl.sub.2). The product
containing fractions were further purified by trituration with
CH.sub.2Cl.sub.2 to give
N-(4-methyl-2-(N"-benzyloxycarbamoyl)methylpentanoyl)-L-leucine-N'-(-
4-methoxycarbonylphenyl)carboxamide as a solid, mp 198-199.degree.
C.
[0599] 17B. In a similar manner, the following compounds were
prepared:
[0600]
N-(2-(N"-benzyloxycarbamoyl)methylpentanoyl)-L-leucine-N'-(4-methox-
ycarbonylphenyl)carboxamide;
[0601]
N-(4-phenyl-2-(N"-benzyloxycarbamoyl)methylbutanoyl)-L-leucine-N'-(-
4-methoxycarbonylphenyl)carboxamide; and
[0602]
N-(4-methyl-2-(N"-benzyloxycarbamoyl)methylpentanoyl)-L-tryptophan--
N'-(4-methoxycarbonylphenyl)carboxamide.
[0603] 17C.
N-(4-Methyl-2-(N"-benzyloxycarbamoyl)-methylpentanoyl)-L-leuci-
ne-N'-(4-methoxycarbonylphenyl)-carboxamide (210 mg) was hydrolyzed
with 1M NaOH (1.4 mL) at 50-60.degree. C. for 2 hour in THF (20 mL)
and MeOH (5 mL). The organic solvents were evaporated and the
residue was taken up in 10 mL H.sub.2O and washed with ether
(2.times.10 mL). The aqueous phase was acidified to pH 2 with 10%
HCl and extracted with ethyl acetate (3.times.10 mL). The combined
extracts were washed with brine dried (Na.sub.2SO.sub.4), and
concentrated to afford N-(4-methyl-2-(N"-benzylox-
ycarbamoyl)-methylpentanoyl)-L-leucine-N'-(4-carboxyphenyl)-carboxamide
(110 mg).
Example 18
[0604] Compounds of Formula (Ie)
[0605] 18A. To a solution of
N-(4-phenyl-2-(N"-benzyloxycarbamoyl)methyl-b-
utanoyl)-L-leucine-N'-(4-methoxycarbonylphenyl)carboxamide (25 mg)
in 20 mL MeOH and 10 mL THF was added 10% Pd/C (20 mg). The
suspension was hydrogenated for 1 hour and then suction filtered
through Celite. Concentration afforded the product which was
purified on silica (2.5% MeOH/CH.sub.2Cl.sub.2) to give 8 mg of
N-(4-phenyl-2-(N"-hydroxycarbamoyl-
)methyl-butanoyl)-L-leucine-N'-(4-methoxycarbonylphenyl)carboxamide,
MS(FAB) 482 (M-H).sup.-.
[0606] 18B. In a similar manner, the following compounds were
prepared:
[0607]
N-(4-methyl-2-(N"-hydroxycarbamoyl)methylpentanoyl)-L-leucine-N'-(4-
-carboxyphenyl)carboxamide;
[0608]
N-(4-methyl-2-(N"-hydroxycarbamoyl)methylpentanoyl)-L-leucine-N'-(4-
-methoxycarbonylphenyl)carboxamide,
[0609] MS(FAB) 436 (M+H).sup.+;
[0610]
N-(2-(N"-hydroxycarbamoyl)methylpentanoyl)-L-leucine-N'-(4-methoxyc-
arbonylphenyl)carboxamide,
[0611] MS(FAB) 420 (M-H).sup.-;
[0612]
N-(4-phenyl-2-(N"-hydroxycarbamoyl)methylbutanoyl)-L-t-leucine-N'-(-
4-methoxycarbonylphenyl)carboxamide;
[0613]
N-(4-phenyl-2-(N"-hydroxycarbamoyl)methylbutanoyl)-L-cyclohexylglyc-
ine-N'-(4-methoxycarbonyl-phenyl)carboxamide;
[0614]
N-(4-phenyl-2-(N"-hydroxycarbamoyl)methylbutanoyl)-L-leucine-N'-(4--
methoxycarbonylphenyl)carboxamide;
[0615]
N-(4-methyl-2-(N"-hydroxycarbamoyl)methylpentanoyl)-L-t-leucine-N'--
(4-methoxycarbonylphenyl)carboxamide;
[0616]
N-(4-methyl-2-(N"-hydroxycarbamoyl)methylpentanoyl)-L-tryptophan-N'-
-(4-methoxycarbonylphenyl)carboxamide,
[0617] MS(FAB) 507 (M-H).sup.-; and
[0618]
N-(4-phenyl-2-(N"-hydroxycarbamoyl)methylbutanoyl)-L-leucine-N'-(4--
methoxycarbonylphenyl)carboxamide.
[0619] 18C. In a similar manner, the following compounds are
prepared:
[0620]
N-(3-phenyl-2-(N"-hydroxycarbamoyl)methylpropanoyl)-L-leucine-N'-(4-
-methoxycarbonylphenyl)carboxamide;
[0621]
N-(5-phenyl-2-(N"-hydroxycarbamoyl)methylpentanoyl)-L-leucine-N'-(4-
-methoxycarbonylphenyl)carboxamide; and
[0622]
N-(3-cyclohexyl-2-(N"-hydroxycarbamoyl)methylpropanoyl)-L-leucine-N-
'-(4-methoxycarbonylphenyl)carboxamide.
Example 19
[0623] Compounds of Formula (Gb)
[0624] 19A. To a cold (0.degree. C.) solution of diethyl
isobutylmalonate (21.6 g, 0.1 mol) in 150 mL of ethanol was added a
solution of KOH (5.89 g, 0.1 mol) slowly over 30 minutes. The clear
solution was stirred at 25.degree. C. for 60 hours. The ethanol was
removed under reduced pressure and the solid residue was dissolved
in 50 mL of H.sub.2O. The aqueous solution was acidified to pH 2
with 4M HCl and extracted with ether (2.times.50 mL). The combined
extracts were dried over MgSO.sub.4 and evaporated to provide 19.0
g (100%) of ethyl isobutylmalonate as a colorless oil.
[0625] 19B. In a similar manner, the following compounds of formula
(Gb) are prepared: ethyl tert-butylmalonate; ethyl propylmalonate;
ethyl benzylmalonate; and ethyl cyclohexymethylmalonate.
Example 20
[0626] Compounds of Formulae (Gc) and (Gd)
[0627] 20A. To neat ethyl isobutylmalonate (25 g, 0.13 mol) at
0.degree. C. was slowly added ice cold diethylamine (15.1 mL, 0.15
mol). After stirring for 15 minutes, formalin (11.1 mL of 37%
aqueous formaldehyde) was added dropwise and the mixture was
allowed to stir at 25.degree. C. for 3 days. The reaction was
treated with a solution of 20 g of K.sub.2CO.sub.3 in 40 mL of
H.sub.2O and extracted with ether (2.times.100 mL). The combined
ether layers were washed with brine, dried over MgSO.sub.4, and
evaporated at 20.degree. C. on a rotary evaporator. The crude
product ethyl 4-methyl-2-methylenepentanoate (containing some
ether) was dissolved in 250 mL of absolute ethanol and treated with
acetonitrile (250 mL), 1M LiOH (9.7 g in 250 mL of H.sub.2O, 0.23
mol) . After stirring overnight, the organic solvents were
evaporated and the aqueous residue was extracted with ethyl acetate
(2.times.150 mL). The combined extracts were washed with brine,
dried (MgSO.sub.4), and evaporated to afford 10.5 g of
4-methyl-2-methylenepentanoic acid as a colorless oil.
[0628] 20B. In a similar manner, the following compounds of formula
(Gd) are prepared: 4-phenyl-2-methylenebutanoic acid;
3-cyclohexyl-2-methylene- propanoic acid;
5-phenyl-2-methylenepentanoic acid; 2-methylenepentanoic acid; and
3,3-dimethyl-2-methylenebutanoic acid.
Example 21
[0629] Compounds of Formula (G)
[0630] A mixture of 4-methyl-2-methylenepentanoic acid (5.0 g) and
thioacetic acid (25 mL) was heated at 95.degree. C. under argon for
3 days. The excess thioacetic acid was evaporated and the residual
oil was dissolved in ethyl acetate (40 mL) and extracted with
saturated NaHCO.sub.3 (3.times.40 mL). The combined NaHCO.sub.3
extracts were combined and acidified at 0.degree. C. to pH 2 with
1M HCl. The aqueous layer was extracted with CH.sub.2Cl.sub.2
(3.times.40 mL), the combined organic phases were dried
(MgSO.sub.4) and evaporated to give 3.0 g of
4-methyl-2-acetylthiomethyl-pentanoic acid; .sup.1H NMR (80 MHz,
CDCl.sub.3) .delta. 0.95 (d, J=8.0, 6H), 1.20-1.90 (m, 4H), 2.35
(s, 3H), 2.50-3.20 (m, 3H), 6.7 (br s, 1H).
Example 22
[0631] Compounds of Formula (If)
[0632] 22A. To a solution of 4-methyl-2-acetylthiomethyl-pentanoic
acid (204 mg, 1.0 mmol) in dry DMF (15 mL) containing HOBT (92 mg,
0.6 mmol) and L-leucine-N'-(4-methoxycarbonylphenyl)carboxamide
(0.6 mmol) was added EDCI (345 mg, 1.8 mmol). The solution was
stirred overnight at 25.degree. C. and then the DMF was removed in
vacuo. The residue was dissolved in ethyl acetate (35 mL) and
washed with 1M HCl, 1M NaOH, and brine. Drying over MgSO.sub.4 and
evaporation afforded a semisolid which was flash chromatographed on
silica gel (ethyl acetate 1:petroleum ether 2) to give
N-(4-methyl-2-acetylthio-methylpentanoyl)-L-leucine-N'-(4-meth-
oxycarbonylphenyl)carboxamide (190 mg) as a white solid.
[0633] 22D. In a similar manner, the following compounds of formula
(If) are prepared:
[0634]
N-(5-phenyl-2-acetylthiomethylpentanoyl)-L-leucine-N'-(4-methoxycar-
bonylphenyl)carboxamide;
[0635]
N-(4-phenyl-2-acetylthiomethylbutanoyl)-L-leucine-N'-(4-methoxycarb-
onylphenyl)carboxamide;
[0636]
N-(3-phenyl-2-acetylthiomethylpropanoyl)-L-leucine-N'-(4-methoxycar-
bonylphenyl)carboxamide;
[0637]
N-(3-cyclohexyl-2-acetylthiomethylpropanoyl)-L-leucine-N'-(4-methox-
ycarbonylphenyl)carboxamide;
[0638]
N-(2-acetylthiomethylpentanoyl)-L-leucine-N'-(4-methoxycarbonylphen-
yl)carboxamide;
[0639]
N-(5-phenyl-2-acetylthiomethylpentanoyl)-L-leucine-N'-(4-aminocarbo-
nylphenyl)carboxamide;
[0640]
N-(4-phenyl-2-acetylthiomethylbutanoyl)-L-leucine-N'-(4-carboxyphen-
yl)carboxamide;
[0641]
N-(3-phenyl-2-acetylthiomethylpropanoyl)-L-leucine-N'-(4-methylsulf-
onylphenyl)carboxamide;
[0642]
N-(3-cyclohexyl-2-acetylthiomethylpropanoyl)-L-leucine-N'-(4-carbam-
oylphenyl)carboxamide;
[0643]
N-(2-acetylthiomethylpentanoyl)-L-leucine-N'-(4-cyanophenyl)carboxa-
mide;
[0644]
N-(5-phenyl-2-acetylthiomethylpentanoyl)-L-tryptophan-N'-(4-methoxy-
carbonylphenyl)carboxamide;
[0645] N-(4-phenyl-2-acetylthiomethylbutanoyl)-L-tryptophan-
N'-(4-methoxycarbonylphenyl)carboxamide;
[0646]
N-(3-phenyl-2-acetylthiomethylpropanoyl)-L-tryptophan-N'-(4-methoxy-
carbonylphenyl)carboxamide;
[0647]
N-(3-cyclohexyl-2-acetylthiomethylpropanoyl)-L-tryptophan-N'-(4-met-
hoxycarbonylphenyl)carboxamide; and
[0648]
N-(2-acetylthiomethylpentanoyl)-L-tryptophan-N'-(4-methoxycarbonylp-
henyl)carboxamide.
Example 23
[0649] Compounds of formula (Ig)
[0650] 23A. To a solution of
N-(4-methyl-2-acetylthiomethylpentanoyl)-L-le-
ucine-N'-(4-methoxycarbonyl-phenyl)carboxamide (85 mg, 0.19 mmol)
in MeOH (8 mL) at 0.degree. C. was added concentrated HN.sub.4OH
(0.4 mL) . After stirring at 0.degree. C. for 5 hours, the methanol
was evaporated and ether (30 mL) was added. The ether solution was
washed with 0.5 M HCl, brine, and was dried over MgSO.sub.4.
Concentration afforded
N-(4-methyl-2-mercaptomethylpentanoyl)-L-leucine-N'-(4-methoxycarbonyl-ph-
enyl)carboxamide in quantitative yield as a white foam, MS(FAB) 407
(M-H).sup.-.
[0651] 23B. In a similar manner, the following compounds of formula
(Ig) are prepared:
[0652]
N-(5-phenyl-2-mercaptomethylpentanoyl)-L-leucine-N'-(4-methoxycarbo-
nylphenyl)carboxamide;
[0653]
N-(4-phenyl-2-mercaptomethylbutanoyl)-L-leucine-N'-(4-methoxycarbon-
ylphenyl)carboxamide;
[0654]
N-(3-phenyl-2-mercaptomethylpropanoyl)-L-leucine-N'-(4-methoxycarbo-
nylphenyl)carboxamide;
[0655]
N-(3-cyclohexyl-2-mercaptomethylpropanoyl)-L-leucine-N'-(4-methoxyc-
arbonylphenyl)carboxamide;
[0656]
N-(2-mercaptomethylpentanoyl)-L-leucine-N'-(4-methoxycarbonylphenyl-
)carboxamide;
[0657]
N-(5-phenyl-2-mercaptomethylpentanoyl)-L-leucine-N'-(4-aminocarbony-
lphenyl)carboxamide;
[0658]
N-(4-phenyl-2-mercaptomethylbutanoyl)-L-leucine-N'-(4-carboxyphenyl-
)carboxamide;
[0659]
N-(3-phenyl-2-mercaptomethylpropanoyl)-L-leucine-N'-(4-methylsulfon-
ylphenyl)carboxamide;
[0660]
N-(3-cyclohexyl-2-mercaptomethylpropanoyl)-L-leucine-N'-(4-carbamoy-
lphenyl)carboxamide;
[0661]
N-(2-mercaptomethylpentanoyl)-L-leucine-N'-(4-cyanophenyl)carboxami-
de;
[0662]
N-(5-phenyl-2-mercaptomethylpentanoyl)-L-tryptophan-N'-(4-methoxyca-
rbonylphenyl)carboxamide;
[0663]
N-(4-phenyl-2-mercaptomethylbutanoyl)-L-tryptophan-N'-(4-methoxycar-
bonylphenyl)carboxamide;
[0664]
N-(3-phenyl-2-mercaptomethylpropanoyl)-L-tryptophan-N'-(4-methoxyca-
rbonylphenyl)carboxamide;
[0665]
N-(3-cyclohexyl-2-mercaptomethylpropanoyl)-L-tryptophan-N'-(4-metho-
xycarbonylphenyl)carboxamide; and
[0666]
N-(2-mercaptomethylpentanoyl)-L-tryptophan-N'-(4-methoxycarbonylphe-
nyl)carboxamide.
Example 24
[0667] Formula (Fc")
[0668] To a stirred solution of 6.48 g (25.0 mmo) of
N-(4-pentenoyl)-4S-phenylmethyl-2-oxazolidinone in 50 mL of dry THF
under argon at -95.degree. C. was added 27.5 mL (27.5 mmol) of 1.0
M sodium hexamethyldisilazide in THF via syringe at a rate to
maintain the reaction temperature at less than -75.degree. C. After
15 min at -80.degree. C. to -95.degree. C., 5.65 mL (6.83 g, 35
mmol) of t-butyl bromoacetate, which had been filtered through
basic alumina immediately prior to use, was added via syringe over
a 1 min period. The solution was stirred at -90.degree. C. to
-60.degree. C. for 2 h, and then partitioned between hexane (100
mL) and dilute aq. NaHSO.sub.4. The organic layer was washed with
sat. aq. NaCl containing a little 1 M phosphate buffer (pH 7),
dried over Na.sub.2SO.sub.4, and concentrated. The residue was
recrystallized from 75 mL of hexane to give 5.56 g (60%) of
N-(2R-(t-Butoxycarbonyl)methyl-4-pentenoyl)-4S-phenylmethyl-2-oxazolidino-
ne as pale yellow needles: mp 75-76.degree. C.
[0669] El. Anal. Calc. for C.sub.21H.sub.27NO.sub.5: C, 67.54; H,
7.29, N,. 3.75. Found: C, 67.76; H, 7.34; N, 3.87.
Example 25
[0670] Formula (Fc"-1) Where R.sup.2 is Biphenyl
[0671] To a solution of 4.75 g (12.7 mmol) of
N-(2R-(t-Butoxycarbonyl)-met-
hyl-4-pentenoyl)-4S-phenylmethyl-2-oxazolidinone, 3.73 g (16.0
mmol) of 4-bromobiphenyl, 0.234 g (0.77 mmol) of
tri-o-tolylphosphine, and 2.22 mL (1.62 g, 16.0 mmol) of
triethylamine in 10 mL of anhydrous DMF under argon was added 0.086
g (0.385 mmol) of palladium(II) acetate. The solution was heated at
100.degree. C. for 4 h, cooled to room temperature, diluted with
ethyl acetate. The precipitate was removed by filtration, and the
filtrate was partitioned between 150 mL of 2:1 ethyl acetate:hexane
and 50 mL of pH 7 phosphate buffer (0.5 M) containing a little
sodium sulfite. The organic layer was washed with 0.2 N aq. sodium
bisulfate and brine/pH 7 phosphate buffer, dried over sodium
sulfate, and concentrated. The residue was dissolved in 50 mL of
ethyl acetate, diluted with 250 mL of isooctane, and seeded with a
few crystals of the product. The solid was removed by filtration,
and recrystallized from 250 mL of 4:1 isooctane: ethyl acetate to
give 4.20 g (63%) of the product,
N-(2R-(t-Butoxycarbonyl)methyl-(5-(biphen-4-yl)-4-pentenoyl)-4S-phenylmet-
hyl-2-oxazolidinone, as fine white needles: mp 118-119.degree. C.;
.sup.1H NMR (300 MHz, CDCl.sub.3) 7.25-7.60 (m, 14H), 6.47 (d, 1H,
J=16 Hz), 6.25 (dt, 1H, J=16 and 8 Hz), 4.65-4.70 (m, 1H),
4.34-4.44 (m, 1H), 4.11 (dd, 1H, J=9 and 2 Hz), 4.01 (t, 1H, J=8
Hz), 3.33 (dd, 1H, J=14 and 3 Hz), 2.89 (dd, 1H, J=17 and 11 Hz),
2.76 (dd, 1H, J=14 and 10 Hz), 2.40-2.57 (m, 3H), 1.43 (s, 9H).
[0672] El. Anal. Calc. for C.sub.33H.sub.35NO.sub.5: C, 75.40; H,
6.77, N, 2.66. Found: C, 75.17; H, 6.84; N, 2.58.
Example 26
[0673] Formula (Fc"-2) Where R.sup.2 in Biphenyl
[0674] A solution of 5.23 g (10.00 mmol) of
N-(2R-(t-butoxycarbonyl)methyl-
-(5-(biphen-4-yl)-4-pentenoyl)-4S-phen ylmethyl-2-oxazolidinone in
50 mL of ethyl acetate was hydrogenated at 1 atm of hydrogen over
500 mg of 10% Pd/C for 2 h at room temperature. The catalyst was
removed by filtration through Celite, and the filtrate was
concentrated to about 20 mL, then diluted with about 75 mL of
isooctane. The solution was seeded with a few crystals of the
product, and the mixture was concentrated to about 50 mL, then
cooled to -20.degree. C. Filtration of the precipitate gave 4.91 g
(94%) of
N-(2R-(t-butoxycarbonyl)methyl-(5-(biphen-4-yl)pentanoyl)-4S-phe-
nylmethyl-2-oxazolidinone as a white powder: mp: 75-76.degree. C.;
.sup.1H NMR (300 MHz, CDCl.sub.3) 7.22-7.59 (m, 14H), 4.61-4.70 (m,
1H), 4.18-4.24 (m, 1H), 4.14 (d, 2H, J=5 Hz), 3.34 (dd, 1H, J=13
and 3 Hz), 2.57-2.89 (m, 4H), 2.48 (dd, 1H, J=13 and 5 Hz),
1.65-1.83 (m, 3H), 1.50-1.60 (m, 1H), 1.42 (s, 9H).
[0675] El. Anal. Calc. for C.sub.33H.sub.37NO.sub.5: C, 75.11; H,
7.07, N, 2.65. Found: C, 75.34; H, 7.11; N, 2.69.
Example 27
[0676] Formula F Where R.sup.2 is Biphenyl
[0677] To a solution of 4.02 g (7.62 mmol) of
N-(2R-(t-butoxycarbonyl)-met-
hyl-(5-(biphen-4-yl)pentanoyl)-4S-phenylmethyl-2-oxazolidinone in
60 mL of THF at 0.degree. C. was added 2.8 mL of 30% aq. hydrogen
peroxide followed by 8.0 mL of 2 N aq. lithium hydroxide. The
mixture was stirred vigorously at 0.degree. C. for 15 min, and then
allowed to warm to room temperature. After 2 h, the mixture was
cooled to 0.degree. C., and 20 mL of 2 N aq. sodium sulfite and 30
mL of saturated aq. sodium bicarbonate were added. After 10 min at
0.degree. C., the mixture was stirred an additional 1 h at room
temperature and then poured into 1 M pH 7 phosphate buffer. The
aqueous phase was acidified to pH 6 by addition of solid sodium
bisulfate, and then the mixture was extracted with 1:1 ethyl
acetate:hexane (200 mL). The organic layer was washed with brine,
dried over sodium sulfate, and concentrated. The residue was
chromatographed on 125 g of silica gel, eluting with 20% to 30%
ethyl acetate:hexane containing 0.5% acetic acid. The
product-containing fractions were concentrated and then azeotroped
several times with toluene to give 2.93 (>100%) of the product,
2R-(t-butoxycarbonyl)methyl-(5-(biphen-4-yl)pe- ntanoic acid, as a
thick syrup which slowly solidified upon storage at -20.degree. C.:
mp 44-45.degree. C. (after drying solid in vacua); .sup.1H NMR (300
MHz, CDCl.sub.3) .delta. 7.22-7.59 (m, 9H), 2.82-2.90 (m, 1H),
2.59-2.75 (m, 3H), 2.40 (dd, 1H, J=14 and 5 Hz), 1.55-1.80 (m, 4H),
1.42 (s, 9H).
[0678] El. Anal. Calc. for C.sub.23H.sub.28O.sub.4: C, 74.97; H,
7.66. Found: C, 75.08; H, 7.76.
Example 28
[0679] Formula (A-1)
[0680] To a solution of 5.00 g (21.6 mmol) of
N-(t-butoxycarbonyl)-L-t-leu- cine and 2.50 g (21.7 mmol) of
N-hydroxysuccinimide in 40 mL of acetonitrile at 0.degree. C. was
added dropwise a solution of 4.12 g (20 mmol) of
dicyclohexylcarbodiimide in 40 mL of acetonitrile. The mixture was
stirred at 0.degree. C. to room temperature overnight, and then the
mixture was filtered to remove the precipitated dicyclohexylurea.
The filtrate was concentrated and the residue was triturated with
ethyl acetate/dichloromethane to give 5.06 g (80%) of
N-(t-Butoxycarbonyl)-L-t-- leucine, N-hydroxysuccinimide ester as a
white solid: mp 136-137.degree. C.; .sup.1H NMR (300 MHz,
CDCl.sub.3) .delta. 5.07 (br d, 1H), 4.43 (d, 1H, J=10 Hz), 2.84
(s, 4H), 1.46 (s, 9H), 1.10 (s, 9H).
Example 29
[0681] Formula (C) Where R.sup.3 is t-Butyl, and R.sup.7 is
4-Pyridine (in Place of the Illustrated Phenyl Group)
[0682] A solution of 2.00 g (6.32 mmol) of N-(
-butoxycarbonyl)-L-t-leucin- e, N-hydroxysuccinimide ester and 2.98
g (31.6 mmol) of 4-aminopyridine in 20 mL of dioxane was heated at
100.degree. C. for 3 h. The reaction was cooled to room temperature
and concentrated. The residue was purified by chromatography on
silica gel, eluting with 5% to 10% methanol in dichloromethane, to
give 1.06 g (54%) of N-(t-butoxycarbonyl)-L-t-leucine-
-N'-(pyrid-4-yl)carboxamide as a white solid: .sup.1H NMR (300 MHz,
CDCl.sub.3) .delta. 8.47 (d, 2H, J=6 Hz), 8.35 (br s, 1H), 7.50 (d,
2H, J=6 Hz), 5.23 (broad d, 1H), 4.00 (br d, 1H), 1.44 (s, 9H),
1.05 (s, 9H).
[0683] FAB-MS ((M+H).sup.+calculated: 308.1974 observed:
308.1970.
Example 30
[0684] 30A. Formula (D) Where R.sup.3 is t-Butyl, and R.sup.7 is
4-Pyridine (in Place of the Illustrated Phenyl Group)
[0685] To a solution of 132 mg (0.43 mmol) of
N-(t-butoxycarbonyl)-L-t-leu- cine-N'-(pyrid-4-yl)carboxamide in 2
mL of dichloromethane was added 1 mL of trifluoroacetic acid. After
1 h at room temperature, the solution was diluted with ca. 5 mL of
toluene and concentrated. Repeated dissolution in
toluene/dichloromethane/methanol and concentration eventually
provided 190 mg (100%) of L-t-leucine-N'-(pyrid-4-yl)carboxamide
bis(trifluoroacetate) as a white solid: .sup.1H NMR (300 MHz,
DMSO-d6) .delta. 11.58 (br s, 1H), 8.68 (d, 2H, J=6 Hz), 8.35 (br
s, 2H), 7.91 (d, 2H, J=6 Hz), 3.81 (s, 1H), 1.04 (s, 9H).
[0686] FAB-MS ((M+H).sup.+calculated: 208.1500 observed:
208.1496.
[0687] 30B. Formula (D) Where R.sup.3 is t-Butyl, and R.sup.7 is
4-(Methylthio)phenyl
[0688] In a manner analogous to that of part A was prepared
L-t-leucine-N'-(4-(methylthio)phenyl)carboxamide: .sup.1H NMR (300
MHz, CDCl.sub.3) .delta. 9.02 (s, 1H), 7.49 (d, 2H, J=6.5 Hz), 7.23
(d, 2H, J=6.5 Hz), 3.23 (a, 2H), 2.44 (s, 3H), 1.03 (s, 9H).
Example 31
[0689] 31A. Formula (Ib) Where R.sup.2 in Biphenyl (and X in
Propanyl), R.sup.3 and R.sup.8 are t-Butyl, and R.sup.7 is
4-Pyridine (in Place of the Illustrated Phenyl Group)
[0690] To a solution of 357 mg (0.97 mmol) of
2R-(t-butoxycarbonyl)methyl-- (5-(biphen-4-yl)pentanoic acid, 422
mg (0.97 mmol) of L-t-leucine-N'-(4-(pyrid-4-yl)carboxamide
bis(trifluoroacetate), and 0.50 mL (3.6 mmol) of triethylamine in 5
mL of DMF was added 442 mg (1.00 mmol) of
benzotriazol-1-yl-tris-(dimethylamino)phosphonium
hexafluorophosphate. After 4 h, the reaction was partitioned
between ethyl acetate and water. The aqueous layer was extracted
twice with ethyl acetate, and the combined organic layers were
washed with water and with brine, dried over sodium sulfate, and
concentrated. Purification of the residue by silica gel
chromatography, eluting with 25% to 75% ethyl acetate in hexane,
gave 360 mg (66%) of N-(2R-(t-butoxycarbonyl)methyl-5--
(biphen-4-yl)pentanoyl)-L-t-leucine-N'-(pyrid-4-yl)carboxamide:
.sup.1H NMR (300 MHz, CDCl.sub.3) .delta. 8.52 (s, 1H), 8.42 (d,
2H, J=6 Hz), 7.54 (d, 2H, J=7 Hz), 7.40-7.48 (m, 6 H), 7.32 (t, 1H,
J=7 Hz), 7.13 (d, 2H, J=8 Hz), 6.62 (d, 1H, J=9 Hz), 4.35 (d, 1H,
J=9 Hz), 2.58-2.68 (m, 4H), 2.40 (dd, 1H, J=16 and 3 Hz), 1.40-1.75
(s over m , obscured by H.sub.2O, 13 H), 1.08 (s, 9H).
[0691] 31B. Formula (Tb) Varying R.sup.7
[0692] By following the procedure of part A and substituting
L-t-leucine-N'-(4-(pyrid-4-yl)carboxamide bis(trifluoroacetate)
with the following:
L-t-leucine-N'-(4-((2-hydroxyethyl)aminosulfonyl)phenyl)carbox-
amide; L-t-leucine-N'-(4-(methylthio)phenyl)carboxamide; there are
obtained:
[0693]
N-(2R-(t-butoxycarbonyl)methyl-5-(biphen-4-yl)pentanoyl)-L-t-leucin-
e -N'-(4-((2-hydroxyethyl)aminosulfonyl)phenyl)carboxamide: mp
89-92.degree. C.;
[0694] .sup.1H NMR (300 MHz, methanol-d4) .delta. 7.80 (s, 4H),
7.51 (d, 2H, J=7 Hz), 7.41 (d, 2H, J=7 Hz), 7.36 (d, 2H, J=8 Hz),
7.28 (t, 1H, J=7 Hz), 7.14 (d, 2H, J=8 Hz), 4.47 (s, 1H), 3.47 (t,
2H, J=6 Hz), 2.85-2.95 (t overlapping m, 3H), 2.52-2.62 (m, 3H),
2.32 (dd, 1H, J=16.5 and 5 Hz), 1.48-1.62 (m, 4H), 1.41 (9, 9H),
1.09 (s, 9H)
[0695] El. Anal. Calc. for C.sub.37H.sub.49N.sub.3O.sub.7S: C,
65.37; H, 7.26; N, 6.18; S, 4.72.
[0696] Found: C, 65.13; H, 7.33; N, 6.22; S, 4.63;
[0697]
N-(2R-(t-butoxycarbonyl)methyl-5-(biphen-4-yl)pentanoyl)-L-t-leucin-
e -N'-(4-(methylthio)phenyl)carboxamide: .sup.1H NMR (300 MHz,
CDCl.sub.3) .delta. 7.79 (s, 1H), 7.53 (d, 2H, J=7 Hz), 7.31-7.44
(m, 7H), 7.19 (d, 2H, J=9 Hz), 7.13 (d, 2H, J=8 Hz), 6.58 (d, 1H,
J=9 Hz), 4.36 (d, 1H, J=9 Hz), 2.55-2.67 (m, 4H), 2.34-2.40 (a
overlapping m, 4H), 1.38-1.75 (s overlapping m, 13 H), 1.07 (s,
9H)
[0698] El. Anal. Calc. for C.sub.36H.sub.46NO.sub.4S. 0.25
H.sub.2O: C, 71.19; H, 7.72, N, 4.61, S, 5.28. Found: C, 71.20; H,
7.78, N, 4.58, S, 5.28;
Example 32
[0699] 32A. Formula (Ic) Where R.sup.2 is Biphenyl (and X is
Propanyl), R.sup.3 is t-Butyl, and R.sup.7 is 4-Pyridine (in Place
of the Illustrated Phenyl Group)
[0700] To a solution of 360 mg (0.64 mmol) of
N-(2R-(t-butoxycarbonyl)-met-
hyl-5-(biphen-4-yl)pentanoyl)-L-t-leucine-N'-(pyrid-4-yl)carboxamide
in 4 mL of dichloromethane was added 2 mL of trifluoroacetic acid.
After 1 h at room temperature, the solution was diluted with
toluene and concentrated. The residue was dissolved in ethyl
acetate (15 mL) and washed with 0.5 M pH 4 citrate buffer
(2.times.15 mL). The combined aq. layers were extracted with ethyl
acetate (2.times.15 mL), and the combined organic layers were
washed with brine, dried over sodium sulfate, and concentrated. The
residue was triturated with ethyl acetate/hexane to give 230 mg
(71%) of N-(2R-carboxymethyl-5-(biphen-4-yl-
)pentanoyl)-L-t-leucine-N'-(pyridin-4-yl)carboxamide as a white
solid: mp 198-201.degree. C.; .sup.1H NMR (300 MHz, MeOH-d4)
.delta. 8.32 (d, 2H, J=7 Hz), 7.64 (d, 2H, J=5 Hz), 7.45 (d, 2H,
J=7 Hz), 7.24-7.38 (m, 7H), 7.10 (d, 2H, J=8 Hz), 4.42 (s, 1H),
2.85-3.00 (m, 1H), 2.33-2.65 (m, 4H), 1.40-1.62 (m, 4H), 1.02 (S,
9H).
[0701] El. Anal. Calc. for C.sub.30H.sub.36N.sub.3O.sub.4.0.5
H2O.0.5 ethyl acetate (solvate): C, 69.29; H, 7.27, N, 7.58. Found:
C, 69.46; H, 7.09; N, 7.55.
[0702] 32B. Formula (Ic) Varying R.sup.7
[0703] By following the procedure of part A and substituting
N-(2R-(t-butoxycarbonyl)-methyl-5-(biphen-4-yl)pentanoyl)-L-t-leucine-N'--
(pyrid-4-yl)carboxamide with the following:
[0704]
N-(2R-(t-butoxycarbonyl)methyl-5-(biphen-4-yl)pentanoyl)-L-t-leucin-
e -N'-(4-((2-hydroxyethyl)aminosulfonyl)phenyl)carboxamide;
[0705]
N-(2R-(t-butoxycarbonyl)methyl-5-(biphen-4-yl)pentanoyl)-L-t-leucin-
e -N'-(4R/S-(methylsulfinyl)phenyl)carboxamide;
[0706]
N-(2R-(t-butoxycarbonyl)methyl-5-(fluoren-2-yl)pentanoyl)-L-leucine-
- N'-(4-(methoxycarbonyl)phenyl)carboxamide;
[0707]
N-(2R-(t-butoxycarbonyl)methyl-5-(7-(glycyl)aminofluoren-2-yl)penta-
noyl)-L-leucine-N'-(4-(methoxycarbonyl)phenyl)carboxamide;
[0708]
N-(2R-(t-butoxycarbonyl)methyl-5-(4-(pyrid-4-yl)phenyl)pentanoyl)-L-
-leucine-N'-(4-(methoxycarbonyl)phenyl)carboxamide;
[0709]
N-(2R-(t-butoxycarbonyl)methyl-5-(biphen-4-yl)pentanoyl)-L-.beta.-h-
ydroxyvaline-N'-(phenyl)carboxamide;
[0710]
N-(2R-(t-butoxycarbonyl)methyl-5-(biphen-4-yl)pentanoyl)-L-t-leucin-
e-N'-(4-(methyl-sulfonyl)phenyl)carboxamide;
[0711]
N-(2R-(t-butoxycarbonyl)methyl-5-(4-(2-hydroxyethyl)phenyl)pentanoy-
l)-L-leucine-N'-(4-(methoxycarbonyl)phenyl)carboxamide;
[0712]
N-(2R-(t-butoxycarbonyl)methyl-5-(4'-hydroxybiphen-4-yl)pentanoyl)--
L-leucine-N'-(4-(methoxycarbonyl)phenyl)carboxamide;
[0713]
N-(2R-(t-butoxycarbonyl)methyl-5-(4'-cyanobiphen-4-yl)pentanoyl)-L--
leucine-N'-(4-(methoxycarbonyl)phenyl)carboxamide;
[0714]
N-(2R-(t-butoxycarbonyl)methyl-5-(biphen-4-yl)pentanoyl)-L-leucine--
N'-(4-(methoxycarbonyl)phenyl)carboxamide;
[0715]
N-(2R-(t-butoxycarbonyl)methyl-5-(4'-(2-aminoethoxy)biphen-4-yl)pen-
tanoyl)-L-leucine-N'-(4-(methoxycarbonyl)phenyl)carboxamide;
[0716]
N-(2R-(t-butoxycarbonyl)methyl-5-(4-(pyridin-4-yl)phenyl)pentanoyl)-
-L-cyclohexylglycine-N'-(4-((2-hydroxyethyl)aminosulfonyl)-phenyl)carboxam-
ide;
[0717]
N-(2R-(t-butoxycarbonyl)methyl-5-(biphen-4-yl)pentanoyl)-L-threonin-
e-N'-(4S -(methylsulfinyl)phenyl)carboxamide;
[0718]
N-(2R-(t-butoxycarbonyl)methyl-5-(2-fluorobiphen-4-yl)pentanoyl)-L--
leucine-N'-(4-(methoxycarbonyl)phenyl)carboxamide;
[0719]
N-(2R-(t-butoxycarbonyl)methyl-4-((biphen-4-yl)thio)butanoyl)-L-t-l-
eucine-N'-(pyridin-4-yl)carboxamide;
[0720]
N-(2R-(t-butoxycarbonyl)methyl-5-(4-(2-aminopyridin-5-yl)phenyl)pen-
tanoyl)-L-threonine-N'-(4S-(methylsulfinyl)phenyl)carboxamide;
[0721]
N-(2R-(t-butoxycarbonyl)methyl-5-(2-hydroxybiphen-4-yl)pentanoyl)-L-
-threonine-N'-(4S-(methylsulfinyl)phenyl)carboxamide;
[0722]
N-(2R-(t-butoxycarbonyl)methyl-5-(4'-cyanobiphen-4-yl)pentanoyl)-L--
(trans-4-hydroxycyclohexyl)glycine-N'-(4S-(methylsulfinyl)phenyl)carboxami-
de;
[0723]
N-(2R-(t-butoxycarbonyl)methyl-5-(biphen-4-yl)pentanoyl)-L-(4-hydro-
xytetrahydropyran-4-yl)glycine-N'-(4S-(methylsulfinyl)phenyl)carboxamide;
and
[0724]
N-(2R-(t-butoxycarbonyl)methyl-5-(2R/S-hydroxy-3,3,3-trifluoropropy-
l)phenyl)pentanoyl)-L-(cyclohexyl)glycine-N'-(4S-(4-((2-hydroxyethyl)amino-
sulfonyl)phenyl)carboxamide, there are obtained:
[0725]
N-(2R-carboxymethyl-5-(biphen-4-yl)pentanoyl)-L-t-leucine,-N'-(4-((-
2-hydroxyethyl)aminosulfonyl)phenyl)carboxamide: .sup.1H NMR (300
MHz, MeOH-d4) .delta. 7.80 (s, 4H), 7.50 (d, 2H, J=7 Hz), 7.25-7.42
(m, 5H), 7.15 (d, 1 H, J=8 Hz), 4.47 (s, 1H), 3.47 (t, 2H, J=6 Hz),
2.89-3.00 (m, 1H), 2.87 (t, 2H, J=6 Hz), 2.49-2.70 (m, 3H), 2.39
(dd, 1H, J=16 and 5 Hz), 1.46-1.67 (m, 4H), 1.07 (s, 9H)
[0726] El. Anal. Calc. for C.sub.33H.sub.41N.sub.3O.sub.7S.0.5
H.sub.2O: C, 62.64; H, 6.69; N, 6.64; S, 5.07. Found: C, 62.61; H,
6.80; N, 6.31; S, 4.97;
[0727]
N-(2R-carboxymethyl-5-(biphen-4-yl)pentanoyl)-L-t-leucine-N'-(4R/S--
(methylsulfinyl)phenyl)carboxamide: .sup.1H NMR (300 MHz, MeOH-d4)
.delta. 8.03 (d, 1H, J=9 Hz), 7.76 (d, 2H, J=9 Hz), 7.57 (dd, 2H,
J=9 and 2 Hz), 7.44 (d, 2H, J=7 Hz), 7.36 (t, 2H, J=7 Hz),
7.23-7.28 (m, 3H), 7.10 (d, 2H, J=8 Hz), 4.45 (d, 1H, J=9 Hz),
2.85-2.98 (m, 1H, J=8 Hz), 2.44-2.64 (m, 7H), 2.35 (dd, 1H, J=16
and 5 Hz), 1.43-1.62 (m, 4H), 1.03 (s, 9H);
[0728]
N-(2R-carboxymethyl-5-(fluoren-2-yl)pentanoyl)-L-leucine-N'-(4-(met-
hoxycarbonyl)phenyl)carboxamide: mp 188-190.degree. C.; .sup.1H NMR
(300 MHz, CDCl.sub.3) .delta. 12.06 (s, 1H), 10.30 (s, 1H), 8.19
(d, 1H, J=8 Hz), 7.86 (d, 2H, J=9 Hz), 7.68-7.77 (m, 3H), 7.62 (d,
1H, 8 Hz), 7.48 (d, 1H, J=7 Hz), 7.20-7.35 (m, 3H), 7.10 (d, 1H,
J=7 Hz), 4.48 (m, 1H), 3.76 (s, 3H), 3.70 (s, 2H), 2.15-2.75 (m,
5H), 1.35-1.75 (m, 5H), 0.90-0.99 (m, 4H)
[0729] El. Anal. Calc. for C.sub.34H.sub.38N.sub.2O.sub.6: C,
71.56; H, 6.71, N, 4.91.
[0730] Found: C, 71.51; H, 6.97; N, 4.84;
[0731]
N-(2R-carboxymethyl-5-(7-(glycyl)aminofluoren-2-yl)pentanoyl)-L-leu-
cine-N'-(4-(methoxycarbonyl)phenyl)carboxamide: mp 222-224.degree.
C. FAB-MS (M+H).sup.+calculated for C.sub.40H.sub.51N.sub.4O.sub.7:
699.3758; observed: 699.3770;
[0732]
N-(2R-carboxymethyl-5-(4-(pyrid-4-yl)phenyl)pentanoyl)-L-leucine-N'-
-(4-(methoxycarbonyl)phenyl)carboxamide: .sup.1H NMR (300 MHz,
d4-MeOH) .delta. 8.52 (d, 2H, J=5.5 Hz), 7.92 (d, 2H, J=9.19 Hz),
7.67 (d, 2H, J=8.82 Hz), 7.58 (d, 2H, J=6.25 Hz), 7.46 (d, 2H,
J=8.45 Hz), 7.20 (d, 2H, J=8.46 Hz), 4.6-4.4 (m, 1H), 3.84 (s, 3H),
2.83-2.59 (m,4H), 2.38 (dd, 1H, J=16.7 and 5 Hz), 1.71-1.57 (m,7H),
0.96 (dd, 6H, J=9.92 and 6.3 Hz);
[0733]
N-(2R-carboxymethyl-5-(biphen-4-yl)pentanoyl)-L-.beta.-hydroxyvalin-
e-N'-(phenyl)carboxamide: .sup.1H NMR (300 MHz, CDCl.sub.3) .delta.
8.73 (s, 1H), 8.07 (d, 1H, J=8.09 Hz), 7.52-7.25 (m, 11H), 7.10 (t,
1H, J=7.54 Hz), 6.97 (d, 2H, J=8.08 Hz), 4.41(d, 1H, J=8.45 Hz),
3.02-3.00 (m, 1H), 2.75 (dd, 1H, J=16.55 and 8.45 Hz), 2.53-2.51
(m, 2H), 2.44 (dd, 1H, J=17.1 and 4.6 Hz), 1.85-1.47 (m,4H), 1.45
(s, 3H), 1.21 (s, 3H)
[0734] El. Anal. Calc. for C.sub.3H.sub.3N.sub.4O.sub.4: C, 71.69;
H, 6.82; N, 5.57. Found: C, 71.65; H, 6.86; N, 5.53;
[0735]
N-(2R-carboxymethyl-5-(biphen-4-yl)pentanoyl)-L-t-leucine-N'-(4-(me-
thyl-sulfonyl)phenyl)carboxamide: .sup.1H NMR (300 MHz, MeOH-d4)
.delta. 8.10 (d, 1H, J=9 Hz), 7.84 (s, 4H), 7.47 (d, 2H, J=8 Hz),
7.25-7.40 (m, 5H), 7.13 (d, 2H, J=8 Hz), 4.48 (d, 1H, J=9 Hz), 2.95
(s, 3H), 2.44-2.70 (m, 4H), 2.36 (dd, 1H, J=16 and 5 Hz), 1.47-1.63
(m, 4H), 1.06 (s, 9H);
[0736]
N-(2R-carboxymethyl-5-(4-(2-hydroxyethyl)phenyl)pentanoyl)-L-leucin-
e-N'-(4-(methoxycarbonyl)phenyl)carboxamide: .sup.1H NMR (300 MHz,
MeOH-d4) .delta. 7.91 (d, 2H, J=9 Hz), 7.64 (d, 2H, J=9 Hz), 6.96
(s, 4H), 4.47-4.51 (m, 1H), 3.84 (s, 3H), 3.63 (t, 2H, J=7 Hz),
2.68 (t, 2H, J=7 Hz), 2.46-2.75 (m, 4H), 2.37 (dd, 1H, J=16 and 5
Hz), 1.51-1.73 (m, 7H), 0.93 and 0.89 (2d, 6H, J=7 Hz);
[0737]
N-(2R-carboxymethyl-5-(4'-hydroxybiphen-4-yl)pentanoyl)-L-leucine-N-
'-(4-(methoxycarbonyl)phenyl)carboxamide: mp 195-197.degree. C.;
FAB-MS (M+H) expected: 575.2757; observed: 595.2750;
[0738]
N-(2R-carboxymethyl-5-(4'-cyanobiphen-4-yl)pentanoyl)-L-leucine-N'--
(4-(methoxycarbonyl)phenyl)carboxamide: .sup.1H NMR (300 MHz,
MeOH-d4) .delta. 10.02 s, 1H), 8.37 (d, 1H, J=7 Hz), 7.87 (d, 2H,
J=8 Hz), 7.71 (d, 2H, J=8.5 Hz), 7.64 (d, 4H, J=9 Hz), 7.35 (d, 2H,
J=8 Hz), 7.16 (d, 2H, J=8 Hz), 4.50-4.53 (m, 1H, 3.81 (s, 3H),
2.49-2.78 (m, 4H), 2.35 (dd, 1H, J=16 and 5 Hz), 1.46-1.72 (m, 7H),
0.88 and 0.90 (2d, 2H, J=7 Hz);
[0739]
N-(2R-carboxymethyl-5-(biphen-4-yl)pentanoyl)-L-leucine-N'-(4-(meth-
oxycarbonyl)phenyl)carboxamide: mp 191-193.degree. C.;
[0740]
N-(2R-carboxymethyl-5-(4'-(2-aminoethoxy)biphen-4-yl)pentanoyl)-L-l-
eucine-N'-(4-(methoxycarbonyl)phenyl)carboxamide: FAB-MS
(M+H).sup.+ calc. 618.3179; found: 618.3189;
[0741]
N-(2R-carboxymethyl-5-(4-(pyridin-4-yl)phenyl)pentanoyl)-L-cyclohex-
ylglycine-N'-(4-((2-hydroxyethyl)aminosulfonyl)-phenyl)carboxamide;
[0742]
N-(2R-carboxymethyl-5-(biphen-4-yl)pentanoyl)-L-threonine-N'-(4S-(m-
ethylsulfinyl)phenyl)carboxamide;
[0743]
N-(2R-carboxymethyl-5-(2-fluorobiphen-4-yl)pentanoyl)-L-leucine-N'--
(4-(methoxycarbonyl)phenyl)carboxamide;
[0744]
N-(2R-carboxymethyl-4-((biphen-4-yl)thio)butanoyl)-L-t-leucine-N'-(-
pyridin-4-yl)carboxamide;
[0745]
N-(2R-carboxymethyl-5-(4-(2-aminopyridin-5-yl)phenyl)pentanoyl)-L-t-
hreonine-N'-(4S-(methylsulfinyl)phenyl)carboxamide;
[0746]
N-(2R-carboxymethyl-5-(2-hydroxybiphen-4-yl)pentanoyl)-L-threonine--
N'-(4S-(methylsulfinyl)phenyl)carboxamide;
[0747]
N-(2R-carboxymethyl-5-(4'-cyanobiphen-4-yl)pentanoyl)-L-(trans-4-hy-
droxycyclohexyl)glycine-N'-(4S-(methylsulfinyl)phenyl)carboxamide;
[0748]
N-(2R-carboxymethyl-5-(biphen-4-yl)pentanoyl)-L-(4-hydroxytetrahydr-
opyran-4-yl)glycine-N'-(4S-(methylsulfinyl)phenyl)carboxamide;
and
[0749]
N-(2R-carboxymethyl-5-(2R/S-hydroxy-3,3,3-trifluoropropyl)phenyl)pe-
ntanoyl)-L-(cyclohexyl)glycine-N'-(4S-(4-((2-hydroxyethyl)aminosulfonyl)ph-
enyl)carboxamide.
Example 33
[0750] 33A. Formula (Ie) Where R.sup.2 is Biphenyl (and X is
Propanyl), R.sup.3 is t-Butyl, and R.sup.7 is 4-Pyridine (in Place
of the Illustrated Phenyl Group)
[0751] To a solution of 127.4 mg (0.200 mmol) of
N-(2R-carboxymethyl-5-(bi-
phen-4-yl)pentanoyl)-L-t-leucine-N'-(pyridin-4-yl)carboxamide and
40 .mu.L of N-methyl morpholine in 1.0 mL of DMF at room
temperature was added 115 mg (0.26 mmol) of
benzotriazol-1-yl-tris-(dimethylamino)phosphonium
hexafluorophosphate. After 15 min, 42 mg (0.60 mmol) of
hydroxylamine hydrochloride was added in one portion, followed by
addition of an additional 70 .mu.L of N-methyl morpholine. The
mixture was stirred for 24 h at room temperature, then partitoned
between 30 mL of ethyl acetate and 25 mL of 0.5 M aq. sodium
bicarbonate. The organic layer was washed with additional aq.
sodium bicarbonate and with brine/pH 7 buffer, dried over sodium
sulfate, and concentrated. Recrystallization from ethyl acetate
provided 42.2 mg of N-(2R-(N-hydroxycarbamoyl)methyl-5-(biphen-4--
yl)pentanoyl-L-t-leucine-N'-(pyrid-4-yl)carboxamide. Concentration
of the filtrate and purification by radial chromatography (1 mm
plate, 5% to 10% ethanol:dichloromethane) provided, after
recrystallization from 2:1 ethyl acetate:hexane, an additional 21.0
mg of product. Total yield was 63.1 mg (61%) of
N-(2R-(N-hydroxycarbamoyl)methyl-5-(biphen-4-yl)pentanoyl-L-t-le-
ucine-N'-(pyrid-4-yl)carboxamide as a white powder: .sup.1H NMR
(300 MHz, DMSO-d6) .delta. 10.45 (s, 1H), 10.34 (s, 1H), 8.68 (s,
1H), 8.38 (d, 2H, J=7 Hz), 8.04 (d, 1 H, J=9 Hz), 7.25-7.60 (m,
9H), 7.12 (d, 2H, J=7 Hz), 4.39 (d, 1H, J=9 Hz), 2.86-2.97 (m, 1H),
2.36-2.60 (m, 2H, obscured by DMSO-d5 resonance), 2.14 (dd, 1H,
J=15 and 7 Hz), 2.02 (dd, 1H, J=15 and 8 Hz), 1.30-1.53 (m, 4H),
0.94 (s, 9H).
[0752] El. Anal. Calc. for C.sub.30H.sub.36N.sub.4O.sub.4.0.25
H.sub.2O: C, 69.14; H, 7.06, N, 10.75.
[0753] Found: C, 69.15; H, 7.23; N, 10.56.
[0754] 33B. Formula (Ie) Varying R.sup.2, R.sup.3, and R.sup.7
[0755] By following the procedure of part A and substituting
N-(2R-carboxymethyl-5-(biphen-4-yl)pentanoyl)-L-t-leucine-N'-(pyridin-4-y-
l)carboxamide with the following:
[0756]
N-(2R-(carboxymethyl-5-(biphen-4-yl)pentanoyl)-L-threonine-N'-(4S-(-
methylsulfinyl)phenyl)carboxamide;
[0757]
N-(2R-(carboxymethyl-5-(4-(pyridin-4-yl)phenyl)pentanoyl)-L-(.beta.-
-hydroxy)valine-N'-(4S-methylsulfinyl)phenylcarboxamide; and
[0758]
N-(2R-(carboxymethyl-4-methylpentanoyl)-L-t-leucine-N'-(pyridin-4-y-
l)carboxamide;
[0759]
N-(2R-(carboxymethyl-(5-(4-(2S-hydroxypropyl)phenyl)pentanoyl)-L-(t-
rans-4-hydroxycyclohexyl)glycine-N'-(pyridin-4-yl)carboxamide;
[0760]
N-(2R-(carboxymethyl-5-(4-(2-methylthiazol-4yl)phenyl)pentanoyl)-
L-(.beta.-hydroxy)valine-N'-(pyridin-4yl)carboxamide; and
[0761]
N-(2R-(carboxymethyl-5-(4-(2R/S-hydroxy-3,3,3-trifluoropropyl)pheny-
l)pentanoyl)-L-(.beta.-hydroxy)valine-N'(pyridin-4-yl)carboxamide,
there are obtained:
[0762]
N-(2R-(N-hydroxycarbamoyl)methyl-5-(biphen-4-yl)pentanoyl)-L-threon-
ine-N'-(4S-(methylsulfinyl)phenyl)carboxamide;
[0763]
N-(2R-(N-hydroxycarbamoyl)methyl-5-(4-(pyridin-4-yl)phenyl)pentanoy-
l)-L-(.beta.-hydroxy)valine-N'-(4S-methylsulfinyl)phenylcarboxamide;
[0764]
N-(2R-(N-hydroxycarbamoyl)methyl-4-methylpentanoyl)-L-t-leucine-N'--
(pyridin-4-yl)carboxamide;
[0765]
N-(2R-(N-hydroxycarbamoyl)methyl-(5-(4-(2S-hydroxypropyl)phenyl)pen-
tanoyl)L-(trans-4-hydroxycyclohexyl)glycine-N'-(pyridin-4-yl)carboxamide;
[0766]
N-(2R-(N-hydroxycarbamoyl)methyl-5-(4-(2-methylthiazol-4-yl)phenyl)-
pentanoyl)-L-(.beta.-hydroxy)valine-N'-(pyridin-4-yl)carboxamide;
[0767]
N-(2R-(N-hydroxycarbamoyl)methyl-5-(4-(2R/S-hydroxy-3,3,3-trifluoro-
propyl)phenyl)pentanoyl)-L-(.beta.-hydroxy)valine-N'(pyridin-4-yl)carboxam-
ide.
[0768]
N-(2R-(N-hydroxycarbamoyl)methyl-(5-(4-(2S-hydroxypropyl)phenyl)pen-
tanoyl)-L-(trans-4-hydroxycyclohexyl)glycine-N'-(pyridin-4-yl)carboxamide;
[0769]
N-(2R-(N-hydroxycarbamoyl)methyl-5-(4-(2-methylthiazol-4yl)phenyl)p-
entanoyl)-L-(.beta.-hydroxy)valine-N'-(pyridin-4yl)carboxamide;
and
[0770]
N-(2R-(N-hydroxycarbamoyl)methyl-5-(4-(2R/S-hydroxy-3,3,3-trifluoro-
propyl)phenyl)pentanoyl)-L-(.beta.-hydroxy)valine-N'(pyridin-4-yl)carboxam-
ide.
Example 34
[0771] 34A. Formula (C) Where R.sup.3 is t-Butyl, and R.sup.4 and
R.sup.5 are H
[0772] A solution of 2.00 g (6.32 mmol) of the N-hydroxysuccinimide
ester of N-(t-butoxycarbonyl)-L-t-leucine in 9 mL of distilled
aniline was stirred and heated at 100.degree. C. for 30 min. The
mixture was allowed to cool to room temperature, and diluted with
40 mL of ethyl acetate. The solution was washed with 4.times.50 mL
of 1 N aq. sodium bisulfate, and the combined aqueous layers were
extracted with 25 mL of ethyl acetate. The combined organic layers
were washed with brine, dried over sodium sulfate, and
concentrated. The residue was purified by chromatography on silica
gel (2% to 10% ethyl acetate in dichloromethane) to give 1.36 g
(74%) of N-(t-butoxycarbonyl)-L-t-leucine-N'-phenylcarboxamide as a
white solid: .sup.1H NMR (300 MHz, CDCl.sub.3) .delta. 7.49. (d,
2H, J=8 Hz), 7.31 (t, 2H, J=8 Hz), 7.11 (t, 1H, J=7 Hz), 5.30-5.36
(m, 1H), 3.95 (d, 1H, J=9 Hz), 1.44 (s, 9H), 1.07 (s, 9H).
[0773] 34B. Formula (C) Where R.sup.3 is t-Butyl, R.sup.4 is
4-Methylthio, and R.sup.5 is H
[0774] In a manner analogous to that of part A was prepared
N-(t-butoxycarbonyl)-L-t-leucine-N'-(4-(methylthio)phenyl)carboxamide:
.sup.1H NMR (300 MHz, CDCl.sub.3) .delta. 7.65 (br s, 1H), 7.42 (d,
2H, J=8.5 Hz), 7.21 (d, 2H, J=8.5 Hz), 5.32 (br d, 1H), 3.95 (d,
1H, J=8.5 Hz), 2.45 (s, 3H), 1.44 (s, 9H) , 1.06 (s, 9H)
[0775] El. Anal. Calc. for C.sub.18H.sub.23N.sub.2O.sub.3S: C,
61.33; H, 8.01, N, 7.95, S, 9.09.
[0776] Found: C, 61.34; H, 8.06; N, 8.00, S, 9.18.
Example 35
[0777] Formula (C) Having A Trifluoroacetyl Protecting Group, Where
R.sup.3 is t-Butyl, and R.sup.4 and R.sup.5 are H
[0778] To a solution of 1.36 g (4.4 mmol) of
N-(t-butoxycarbonyl)-L-t-leuc- ine-N'-phenylcarboxamide in 10 mL of
dichloromethane was added 5 mL of tifluoroacetic acid. After 45 min
at room temperature, the solution was diluted with toluene and
concentrated. The residue was twice more concentrated from toluene
to remove excess triflouroacetic acid, then dried under vacuum (ca.
1 mm Hg). The residue was then dissolved in 15 mL of
dichloromethane and treated successively with pyridine (0.90 mL, 11
mmol) and trifluoroacetic anhydride (0.70 mL, 4.84 mmol). After 30
min, the mixture was partitioned between dichloromethane (25 mL)
and 1 N aq. sodium bisulfate. The organic layer was washed with
additional aq. sodium bisulfate, brine, dried over sodium sulfate,
and concentrated to give 1.22 g (91%) of
N-(trifluoroacetyl)-L-t-leucine-N'-phenylcarboxamide as a white
solid: mp 201-203.degree. C., .sup.1H NMR (300 MHz, CDCl.sub.3)
.delta. 7.50 (d, 2H, J=8 Hz), 7.36 (t, 2H, J=8 Hz), 7.17 (t, 1H,
J=7 Hz), 4.43(d, 1H, J=9 Hz), 1.10 (s, 9H).
[0779] El. Anal. Calc. for C.sub.14H.sub.17F.sub.3N.sub.2O.sub.2:
C, 55.62; H, 5.67, N, 9.27. Found: C, 55.57; H, 5.60; N, 9.18.
Example 36
[0780] Formula (C) Having A Trifluoroacetyl Protecting Group, Where
R.sup.3 is t-Butyl, R.sup.4 is 4-(2-Hydroxyethyl)aminosulfonyl, and
R.sup.5 is H
[0781] To a solution of 250 mg (0.83 mmol) of
N-(trifluoroacetyl)-L-t-leuc- ine-N'-phenylcarboxamide in 5 mL of
chloroform was added 0.4 mL (6 mmol) of chlorosulfonic acid. The
mixture was heated to reflux for 35 min and then cooled to
0.degree. C. and diluted with ethyl acetate. Ethanolamine (1.5 mL)
was added and the mixture was stirred at 0.degree. C. for 15 min.
The mixture was partitioned between water and ethyl acetate, and
the organic layer was washed with 1 N aq. sodium bisulfate, dried
over sodium sulfate, and concentrated to provide 140 mg (40%) of
N-(trifluoroacetyl)-L-t-leucine-N'-(4-((2-hydroxyethyl)aminosulfonyl)phen-
yl)carboxamide: .sup.1H NMR (300 MHz, CDCl.sub.3) .delta. 7.93 (s,
1H), 7.84 (d, 2H, J=9 Hz), 7.67 (d, 2H, J=9 Hz), 7.15 (br d, 1H),
4.90 (br t, 1H), 4.49 (d, 1H, J=9 Hz), 3.70 (t, 2H, J=5 Hz), 3.11
(q, 2H, J=5 Hz), 1.12 (s, 9H).
Example 37
[0782] Formula (D) Where R.sup.3 is t-Butyl, R.sup.4 is
4-(2-Hydroxyethyl)aminosulfonyl, and R.sup.5 is H
[0783] To a solution of 257 mg (0.6 mmol) of
N-(trifluoroacetyl)-L-t-leuci-
ne-N'-(4-((2-hydroxyethyl)aminosulfonyl)phenyl)carboxamide in 8 mL
of ethanol was added 227 mg (6 mmol) of sodium borohydride. The
mixture was heated to 55.degree. C. for 15 min, allowed to cool to
room temperature, and quenched with 10% ammonium hydroxide in
methanol. After 20 h at room temperature, the mixture was filtered
and the filtrate was absorbed onto silica gel. Chromatography
(dichloromethane to 90:9:1 dichloro-methane:methanol:ammonium
hydroxide) gave 110 mg (56%) of L-t-leucine-N'-(4-((2-hydroxyethyl)
aminosulfonyl)phenyl)carboxamide, as an oil: .sup.1H NMR (300 MHz,
CDCl.sub.3) .delta. 9.48 (br s, 1H), 7.81 (d, 2H, J=9 Hz), 7.72 (d,
2H, J=9 Hz), 3.67 (t, 2H, J=5 Hz), 3.30 (s, 1H), 3.08 (q, 2H, J=5
Hz), 1.06 (s, 9H).
[0784] FAB-MS (M+H.sup.+): expected 330.1488 observed 330.1480
Example 38
[0785] Formula (Ib) Where R.sup.2 is Biphenyl (X is Propanyl),
R.sup.3 is t-Butyl, R.sup.4 is 4R/S-Methylsulfinyl, and R.sup.5 is
H
[0786] To a solution of 60.3 mg (0.100 mmol) of
N-(2R-(t-butoxycarbonyl)-m-
ethyl-5-(biphen-4-yl)pentanoyl)-L-t-leucine-N'-(4-(methylthio)phenyl)-carb-
oxamide in 2 mL of dichloromethane at -78.degree. C. was added a
solution of 26 mg (0.15 mmol) of m-chloroperbenzoic acid in 1 mL of
dichloromethane. The reaction was stirred at -78.degree. C. for 50
min, and then 0.2 mL of dimethyl sulfide was added. The mixture was
allowed to warm to room temperature, and then partitioned between
dichloromethane and sat. aq. sodium bicarbonate. The organic layer
was washed with brine, dried over sodium sulfate, and concentrated
to give 59.6 mg (96%) of
N-(2R-(t-butoxycarbonyl)methyl-5-(biphen-4-yl)pentanoyl)-L-t-leucine-N'-(-
4R/S-(methylsulfinyl)phenyl)carboxamide, as a white solid: .sup.1H
NMR (300 MHz, CDCl.sub.3) .delta. 8.46 (br s, 1H), 7.32-7.68 (m, 11
H), 7.14 (d, 2H, J=8 Hz), 6.63 (d, 1H, J=9 Hz), 4.42 (d, 1H, J=9
Hz), 2.36-2.72 (m, 8H), 1.42-1.76 (s overlapping m, 13 H), 1.09 (s,
9H).
Example 39
[0787] Formula (Fe)
[0788] To a solution of 2.779 g (7.50 mmol) of
N-(2R-(t-butoxycarbonyl)-me-
thyl-4-pentenoyl)-4S-phenylmethyl-2-oxazolidinone in 30 mL of THF
at 0.degree. C. was added 2.55 mL (22.5 mmol) of 30% aq. hydrogen
peroxide, followed by the addition of 7.5 mL (15 mmol) of 2.0 N aq.
lithium hydroxide. The mixture was stirred for 2 h at 0.degree. C.
and for 0.5 h at room temperature. After the mixture was recooled
to 0.degree. C. , 15 mL of 2 M aq. sodium sulfite and 23 mL of sat.
aq. sodium bicarbonate were added. The mixture was stirred an
additional 30 min at 0.degree. C. , and then most of the THF was
removed by concentration in vacuo. The residue was partitioned
between CH.sub.2Cl.sub.2 and H.sub.2, and the aq. layer was
extracted with additional CH.sub.2Cl.sub.2. The combined organic
layers were extracted with aq. sodium bicarbonate, and then the
combined aq. layers were acidified to pH 2 with sodium bisulfate.
The resulting mixture was extracted twice with ethyl acetate, and
the combined organic layers were washed with sat. aq. NaCl, diluted
with 0.25 volume of hexane, dried over Na.sub.2SO.sub.4, and
concentrated to provide 1.47 g (92%) of
2R-(t-Butoxycarbonyl)methyl-4-pentenoic acid as a colorless oil:
.sup.1H NMR (300 MHz, CDCl.sub.3) .delta. 5.68-5.82 (m, 1H),
5.07-5.15 (m, 2H), 2.86-2.96 (m, 1H), 2.60 (dd, 1H, J=18 and 10
Hz), 2.25-2.52 (m, 3H), 1.43 (s, 9H).
Example 40
[0789] Formula (D'-1) Where R.sup.3 is t-Butyl and R.sup.7 is
4-(methoxycarbonyl)phenyl
[0790] In a manner analogous to Example 31, substituting
2R-(t-butoxycarbonyl)methyl-(5-(biphen-4-yl)pentanoic acid with
2R-(t-Butoxycarbonyl)methyl-4-pentenoic acid, and substituting
L-t-leucine-N'-(4-(pyrid-4-yl)carboxamide with
L-leucine-N'-(4-(methoxyca- rbonyl)phenyl)-carboxamide, there was
prepared N-(2R-(t-butoxycarbonyl)met-
hyl-4-pentenoyl)-L-leucine-N'-(4-(methoxycarbonyl)-phenyl)carboxamide:
mp 118-119.degree. C. (cyclohexane); .sup.1H NMR (300 MHz,
CDCl.sub.3) .delta. 8.96 (s, 1H), 7.95 (d, 2H, J=9 Hz), 7.57 (d,
2H, J=9 Hz), 6.32 (d, 1H, J=7 Hz), 5.63-5.77 (m, 1H) 4.95-5.08 (m,
2H), 4.52-4.60 (m, 1H), 3.88(s, 3H), 2.36-2.77 (m, 4H), 2.14-2.27
(m, 1H), 1.60-1.87 (m, 3H), 1.45 (s, 9H), 0.97 (d, 3H, J=7 Hz),
0.91 (d, 3H, J=7 Hz).
[0791] El. Anal. Calc. for C.sub.25H.sub.36N.sub.2O.sub.6: C,
65.20; H, 7.88, N, 6.08. Found: C, 65.04; H, 7.80; N, 6.06.
Example 41
[0792] 41A. Formula (D'-2) Where R.sup.2 is Fluoren-2-yl (X is
Propanyl), R.sup.3 is t-Butyl, and R.sup.7 is
4-(methoxycarbonyl)phenyl
[0793] To a solution of 167 mg (0.36 mmol) of
N-(2R-(t-butoxycarbonyl)-met-
hyl-4-pentenoyl)-L-leucine-N'-(4-(methoxycarbonyl)phenyl)carboxamide,
108 mg (0.44 mmol) of 2-bromofluorene, 21 mg (0.07 mmol) of
tri-o-tolylphosphine, and 69 .mu.L (50 mg, 0.50 mmol) of
triethylamine in 1.0 mL of DMF under argon was added 8.0 mg (0.035
mmol) of palladium diacetate. The solution was heated at
100.degree. C. for 2 h, cooled to room temperature, and then
partitioned between 3:1 ethyl acetate:hexane and water. The organic
layer was washed with 1N aq. sodium bisulfate and with brine/pH 7
buffer, dried over sodium sulfate, and concentrated. Purification
by flash chromatography (20 g silica, 5% to 10% t-butyl methyl
ether in dichloromethane) gave 185 mg (82%) of the product as a
solid containing trace impurities by TLC. Recrystallization from
t-butyl methyl ether/isooctane provided 115 mg (51%) of
N-(2R-(t-butoxycarbonyl)m-
ethyl-5-(fluoren-2-yl)-4E-pentenoyl)-L-leucine-N'-(4-(methoxycarbonyl)phen-
yl)-carboxamide as fine white needles: mp 189-192.degree. C. ;
.sup.1H NMR (300 MHz, CDCl.sub.3) .delta. 8.32 (s, 1H), 7.79 (d,
2H, J=8 Hz), 7.72 (d, 1H, J=8 Hz), 7.57 (d, 1H, J=8 Hz), 7.45-7.53
(m, 3 H), 7.36 (t, 1H, J=7.5 Hz), 7.25-7.32 (m, 2H), 7.17 (d, 1H,
J=7 Hz), 4.52-4.60 (m, 1H), 3.75 (s, 5H), 2.33-2.87 (m, 5H),
1.60-1.90 (m, 3H), 1.45 (s, 9H), 0.92 (apparent t, 6H).
[0794] El. Anal. Calc. for C.sub.38H.sub.44N.sub.2O.sub.6.0.5
H.sub.2O: C, 72.01; H, 7.16, N, 4.42.
[0795] Found: C, 71.87; H, 7.07; N, 4.32.
[0796] 41B. Formula (D'-2) Where R.sup.2 is
7-(N-(benzyloxycarbonyl)glycyl- )-aminofluoren-2-yl (X is
Propanyl), R.sup.3 is t-Butyl, and R.sup.7 is
4-(methoxycarbonyl)phenyl
[0797] To a solution of 600 mg (2.31 mmol) of
2-amino-7-bromofluorene and 483 mg (2.31 mmol) of
N-(benzyloxycarbonyl)glycine in 10 mL of anhydrous pyridine was
added 442 mg (2.31 mmol) of EDC hydrochloride. The reaction was
heated at 60.degree. C. for 4 days, and then the solution was
concentrated. The residue was partitioned between ethyl acetate and
1 N aq. hydrochloric acid, and the organic layer was washhed with
sat. aq. sodium bicarbonate and with brine, dried over magnesium
sulfate, and concentrated to give 813 mg (78%) of
N-(benzyloxycarbonyl)glycine-N'-(7-b- romofluoren-2-yl)carboxamide
as a tan solid: mp 194-195.degree. C.
[0798] By following the procedure of part A and substituting
2-bromofluorene with
N-(benzyloxycarbonyl)glycine-N'-(7-bromofluoren-2-yl- )carboxamide,
there is obtained N-(2R-(t-butoxycarbonyl)methyl-5-(7-(N-(be-
nzyloxy-carbonyl)glycyl)aminofluoren-2-yl)-4E-pentenoyl)-L-leucine-N'-(4-(-
methoxycarbonyl)phenyl)carboxamide: mp 213-214.degree. C.
[0799] FAB-MS (M+Cs).sup.+calculated for
C.sub.48H.sub.54N.sub.4O.sup.9.Cs- : 963.2945; observed:
963.2960.
[0800] El. Anal. Calc. for C.sub.47H.sub.54N.sub.4O.sub.9: C,
69.40; H, 6.51, N, 6.75. Found: C, 69.47; H, 6.51; N, 6.70.
[0801] 41C. Formula (D'-2) Where R.sup.2 is 4-(pyrid-4-yl)phenyl (X
is Propanyl), R.sup.3 is t-Butyl, and R.sup.7 is
4-(methoxycarbonyl)phenyl
[0802] Aq. 2 M sodium carbonate (3 ml)was added to a suspension of
400 mg (2.0 mmol) of 4-bromopyridine in 2 mL of benzene to give 2
clear phases, and argon was bubbled through the mixture for a few
minutes before added 115 mg (0.10 mmol) of palladium
tetrakis(triphenylphosphine). To the resulting mixture was added a
solution of 200 mg (1.00 mmol) of 4-bromophenylboronic acid in 1 mL
of ethanol, and the mixture was heated at reflux for 4 h. After
cooling to room temperature, the mixture was partitioned between
ethyl acetate (25 mL) and water (25 ml). The organic layer was
dried over sodium sulfate and concentrated. Purification of the
residue by silica gel chromatography, eluting with 25% to 50% ethyl
acetate in hexane, gave 193 mg (82%) of 4-(4-bromophenyl)pyridine
as a white solid:
[0803] mp 124-126.degree. C.
[0804] By following the procedure of part A and substituting
2-bromofluorene with 4-(4-bromophenyl)pyridine, there is obtained
N-(2R-(t-butoxy-carbonyl)methyl-5-(4-(pyrid-4-yl)phenyl)-4E-pentenoyl)-L--
leucine-N'-(4-(methoxycarbonyl)phenyl)carboxamide: .sup.1H NMR (300
MHz, CDCl.sub.3) .delta. 8.80 (s, 1H), 8.64 (d, 2H, J=6 Hz), 7.85
(d, 2H, J=8.5 Hz), 7.49 (d, 2H, J=9 Hz), 7.45 (d, 2H, J=6 Hz), 7.19
(d, 2H, J=8 Hz), 6.40 (d, 1H, J=16 Hz), 6.33 (d, 1H, J=8 Hz),
6.09-6.17 (m, 1H), 4.54-4.57 (m, 1H), 3.80 (s, 3H), 2.38-2.81 (m,
5H), 1.48-1.84 (m, 3H), 1.44 (s, 9H), 0.90 and 0.94 (2 d, 6H, J=7
Hz).
Example 42
[0805] 42A. Formula (Ib') Where R.sup.2 is Fluoren-2-yl (X is
Propanyl), R.sup.3 is t-Butyl, and R.sup.7 in
4-(methoxycarbonyl)phenyl
[0806] A solution of 111 mg (0.177 mmol) of
N-(2R-(t-butoxycarbonyl)methyl-
-5-(fluoren-2-yl)-4-pentenoyl)-L-leucine-N'-(4-(methoxycarbonyl)phenyl)-ca-
rboxamide in 7 mL of 4:3 ethyl acetate:ethanol was hydrogenated at
1 atm hydrogen pressure over 30 mg of 10% palladium on carbon for 3
h. The catalyst was removed by filtration through Celite, and the
filtrate was concentrated. Trituration with t-butyl methyl ether
gave 110 mg (99%) of
N-(2R-(t-butoxycarbonyl)methyl-5-(fluoren-2-yl)pentanoyl)-L-leucine-N'-(4-
-(methoxycarbonyl)phenyl)carboxamide as a white solid: mp
166-167.degree. C. (softening at 161.degree. C. ).
[0807] El. Anal. Calc. for C.sub.17H.sub.46N.sub.2O.sub.6: C,
72.29; H, 7.54, N, 4.56. Found: C, 72.32; H, 7.54; N, 4.62.
[0808] 42B. Formula (Ib') Varying R.sup.2
[0809] By following the procedure of part A and substituting
N-(2R-(t-butoxycarbonyl)methyl-5-(fluoren-2-yl)-4-pentenoyl)-L-leucine-N'-
-(4-(methoxycarbonyl)phenyl)carboxamide with:
[0810]
N-(2R-(t-butoxycarbonyl)methyl-5-(7-(N-(benzyloxy-carbonyl)glycyl)a-
minofluoren-2-yl)-4E-pentenoyl)-L-leucine-N'-(4-(methoxycarbonyl)-phenyl)c-
arboxamide; and
[0811]
N-(2R-(t-butoxy-carbonyl)methyl-5-(4-(pyrid-4-yl)phenyl)-4E-penteno-
yl)-L-leucine-N'-(4-(methoxycarbonyl)phenyl)carboxamide, there are
obtained:
[0812]
N-(2R-(t-butoxycarbonyl)methyl-5-(7-(glycyl)aminofluoren-2-yl)penta-
noyl)-L-leucine-N'-(4-(methoxycarbonyl)phenyl)carboxamide
[0813] FAB-MS (M+H).sup.+calculated for
C.sub.40H.sub.51N.sub.4O.sub.7: 699.3758; observed: 699.3770;
and
[0814]
N-(2R-(t-butoxycarbonyl)methyl-5-(4-(pyrid-4-yl)phenyl)pentanoyl)-L-
-leucine-N'-(4-(methoxycarbonyl)phenyl)carboxamide: mp
174-176.degree. C.;
[0815] .sup.1H NMR (300 MHz, CDCl.sub.3) .delta. 8.85 (s, 1H), 8.63
(d, 2H, J=5 Hz), 7.95 (d, 2H, J=8 Hz), 7.57 (d, 2H, J=9 Hz),
7.41-7.47 (m, 4H), 7.07 (d, 2H, J=8 Hz), 6.19 (d, 1H, J=7 Hz),
4.53-4.56 (m, 1H), 3.85 (s, 3H), 2.37-2.66 (m, 5H), 1.45-1.83 (m,
7H), 1.42 (s, 9H), 0.91 and 0.95 (2 d, 6H, J=7 Hz).
Example 43
[0816] Formula (C-1) Where R.sup.4 and R.sup.5 are H
[0817] To a stirred suspension of 4.18 g (20.0 mmol) of
N-(benzyloxy-carbonyl)glycine, 2.73 mL (2.79 g, 30 mmol) of
aniline, and 110 mg (1.0 mmol) of 4-dimethylaminopyridine in 55 mL
of dichlormethane at 0.degree. C. was added 6.53 g (22 mmol) of EDC
methiodide in one portion. The mixture was stirred for 18 h at room
temperature, and then partitioned between 200 mL of 3:1 ethyl
acetate:hexane and water. The organic layer was washed with 1 N aq.
sodium bisulfate, sat. aq. sodium bicarbonate, and finally with
brine/pH 7 phosphate buffer, dried over sodium sulfate, and
concentrated. Recrystallization from 1:1 ethyl acetate:isooctane
gave 3.69 g (65%) of N-(benzyloxycarbonyl)glycine-N'-ph-
enylcarboxamide: mp 143-144.degree. C.
Example 44
[0818] Formula (C-2) Where R.sup.4 and R.sup.5 are H
[0819] To a stirred solution of 1.42 g (5.00 mmol) of
N-(benzyloxycarbonyl)-glycine-N'-phenylcarboxamide in 35 mL of dry
THF at -5.degree. C. was added by syringe 6.15 mL (16.0 mmol) of
2.6 M n-butyllithium in hexane at a rate to maintain the reaction
temperature below 10.degree. C. After ca. 2/3 of the n-butyllithium
had been added, a yellow color began to persist, and the addition
was stopped for ca. 10 min and then resumed in a dropwise fashion
so as to maintain the reaction temperature at about 0.degree. C.
After the addition was complete, the orange solution was stirred at
0.degree. C. for 45 min, and then cooled to -70.degree. C. Acetone
(1.10 mL, 15 mmol) was added in one portion by syringe. After 10
min, the reaction was partitioned between 1 M pH 7 phosphate buffer
and 3:1 ethyl acetate:hexane. The organic layer was washed with
brine, dried over sodium sulfate, and concentrated. The residue was
purified by chromatography on 75 g of silica, eluting with 40%
ethyl acetate:hexane. First to elute was pure products fractions
(pool #1), followed by fractions containing product and the
starting glycinanilide (pool #2). The residue from pool #2 was
recrystallized from ethyl acetate:isooctane to give nearly pure
starting material as a solid, and mother liquors containing mostly
product. The residue from the concentration of the mother liquors
was purified by radial chromatography (4 mm plate, 30% ethyl
acetate:hexane), and the product fractions were combined with pool
#1 to give, after trituration of the gummy residue with
hexane/t-butyl methyl ether, 423 mg (25%) of
N-(benzyloxycarbonyl)-D-
L-.beta.-hydroxyvaline-N'-(phenyl)carboxamide as a pale yellow
solid: mp 128-129.degree. C.; .sup.1H NMR (300 MHz, CDCl.sub.3)
.delta. 8.30 (br s, 1H), 7.39 (d, 2H, J=8 Hz), 7.18-7.28. (m, 7H),
7.06 (t, 1H, J=7 Hz), 5.83 (br d, 1H), 5.05 (s, 2H), 4.04 (d, 1H,
J=9 Hz), 3.73 (s, 1H), 1.33 (s, 3H), 1.16 (s, 3H).
[0820] El. Anal. Calc. for C.sub.19H.sub.22N.sub.2O.sub.4: C,
66.65; H, 6.48, N, 8.18.
[0821] Found: C, 66.66; H, 6.57; N, 8.14.
Example 45
[0822] Formula (C-2) Where R.sup.4 and R.sup.5 are H
[0823] A solution of 400 mg (1.17 mmol) of
N-(benzyloxycarbonyl)-DL-.beta.-
-hydroxyvaline-N'-phenylcarboxamide in 10 mL of ethyl acetate was
hydrogenated over 50 mg of 10% palladium on carbon at 1 atm of
hydrogen pressure for 1.5 h. The catalyst was removed by filtration
through Celite, and the filtrate was concentrated to give 259 mg
(>100%) of DL-.beta.-hydroxyvaline-N'-(phenyl)carboxamide, which
was used without further purification: mp 97-99.degree. C.
[0824] El. Anal. Calc. for C.sub.11H.sub.16N.sub.2O.sub.2: C,
63.44; H, 7.74, N, 13.45.
[0825] Found: C, 63.52; H, 7.79; N, 13.40.
Example 46
[0826] Formula (Ib) Where R.sup.2 is Biphenyl, R.sup.3 is
Hydroxy-t-butyl, and R.sup.4 and R.sup.5 are H
[0827] To a solution of 203 mg (0.55 mmol) of
2R-(t-butoxycarbonyl)methyl-- (5-(biphen-4-yl)pentanoic acid, 104
mg (0.50 mmol) of DL-.beta.-hydroxyvaline-N'-(phenyl)carboxamide,
and 90 .mu.L (0.65 mmol) of triethylamine in 2.5 mL of DMF was
added 265 mg (0.60 mmol) of
benzotriazol-1-yl-tris-(dimethylamino)-phosphonium
hexafluorophosphate. After 24 h, the reaction was partitioned
between 3:1 ethyl acetate:hexane and ca. 0.2 N aq. sodium
bicarbonate. The organic layer was washed with 1N aq. sodium
bisulfate and with brine/pH 7 buffer, dried over sodium sulfate,
and concentrated. The residue was purified by radial chromatography
(4 mm plate), eluting with 25% to 30% ethyl acetate in hexane.
First to elute was 121 mg (43%) of N-(2R-(t-butoxycarbonyl)methyl-
-5-(biphen-4-yl)pentanoyl)-D-.beta.-hydroxyvaline-N'-(phenyl)carboxamide
(diastereomer), followed by 140 mg (50%) of
N-(2R-(t-butoxycarbonyl)methy-
l-5-(biphen-4-yl)pentanoyl)-L-.beta.-hydroxyvaline-N'-(phenyl)carboxamide
as a gummy semi-solid containing, according to NMR analysis, about
1 mole-equivalent of isooctane (the solvent from which the final
sample was concentrated): .sup.1H NMR (300 MHz, CDCl.sub.3) .delta.
8.77 (s, 1H), 7.24-7.56 (m, 11 H), 7.03-7.14 (m, 3H), 6.89 (d, 1H,
J=8.5 Hz), 4.46 (d, 1H, J=8.5 Hz), 4.17 (s, 1H), 2.52-2.70 (m, 4H),
2.36 (br d, 1H, J=12.5 Hz), 1.50-1.70 (m, 4H), 1.43 (B, 3H), 1.40
(s, 9H), 1.25 (s, 3H).
Example 47
[0828] Formula (P-1)
[0829] To a solution of 510 mg (1.97 mmol) of
N-(4-pentenoyl)-4S-phenylmet- hyl-2-oxazolidinone in 8 mL of
dichloromethane at 0.degree. C. was added 2.2 mL (2.2 mmol) of 1 M
titanium tetrachloride in dichloromethane. After 15 min, 0.42 mL
(2.4 mmol) of diisopropylethylamine was added to the thick slurry
to give a deep red solution. After 1 h at 0.degree. C., 216 mg (2.4
mmol) of s-trioxane in 2 mL of dichloromethane was added via
cannula, followed by an additional 2.2 mL of 1M titanium
tetrachloride in dichloromethane. After 4 h at 0.degree. C., the
solution was partitioned between aq. ammonium chloride and
dichloromethane. The organic layer washed with 1 N aq. HCl, with
brine containing pH 7 phosphate buffer, dried over sodium sulfate,
and concentrated. The residue was purified by chromatography on 20
g silica, eluting with 30% to 40% ethyl acetate in hexane.
Recrystallization of the purified product from t-butyl methyl
ether/isooctane provided 404 mg (71%) of
N-(2R-hydroxymethyl-4-pentenoyl)-
-4S-phenylmethyl-2-oxazolidinone.:
[0830] mp 71-72.degree. C. ; .sup.1H NMR (300 MHz, CDCl.sub.3)
.delta. 7.22-7.37 (m, 5H), 5.78 (dddd, J=10, 7, 4, and 3 Hz),
5.03-5.15 (m, 2H), 4.69 (dddd, 1H, J=10, 6, 4, and 3 Hz), 4.17-4.24
(m, 2H), 4.02-4.10 (m, 1H), 3.85-3.91 (m, 2H), 3.29 (dd, 1H, J=4
and 3 Hz), 2.82 (dd, 1H, J=14 and 10 Hz), 2.44 (dt, 1H, J=14 and 7
Hz), 2.31 (dd, 1H, J=14 and 7 Hz), 2.17 (br s, 1H).
Example 48
[0831] Formula (P-2)
[0832] To a suspension of 4.0 g (25.1 mmol) of
O-benzylhydroxylamine hydrochloride in 50 mL of THF at 0.degree. C.
under argon was added 11.4 mL (22.8 mmol) of 2M trimethylaluminum
in toluene. After the addition was complete, the solution was
allowed to warm to room temperature. After 15 min, this solution
was added via cannula to a solution of 2.40 g (8.30 mmol) of
N-(2R-hydroxymethyl-4-pentenoyl)-4S-phenylmethyl-2-oxazolidinone in
100 mL of THF at 0.degree. C. under argon. The reaction was stirred
for 6 h at 0.degree. C. , and then partitioned between 1N HCl/brine
and ethyl acetate/diethyl ether. The organic layer was with 1M pH 7
phosphate buffer and with brine, dried over sodium sulfate, and
concentrated. The residue was purified by chromatography on silica,
eluting with 35% to 45% ethyl acetate in hexane, to give, after
elution of 4S-phenylmethyl-2-oxazolidinone, 2.01 g of the product.
Recrystallization from ethyl acetate: isooctane provided 1.90 g
(97%) of N-benzyloxy-2R-hydroxymethyl-4-pentenamide as a white
powder:
[0833] mp 58-59.degree. C.; .sup.1H NMR (300 MHz, CDCl.sub.3)
.delta. 8.40 (s, 1H), 7.38 (m, 5H), 5.71 (m, 1H), 5.03-5.06 (m,
1H), 4.91 (dd, 1H, J=16 and 12 Hz), 3.73 (m, 1H), 2.20 (m, 2H).
[0834] El. Anal. Calc. for C.sub.13H.sub.17NO.sub.3: C, 66.36; H,
7.28, N, 5.95.
[0835] Found: C, 66.15; H, 7.32; N, 5.99.
Example 49
[0836] Formula (P-3)
[0837] To a solution of 1.92 g (8.17 mmol) of
N-benzyloxy-2R-hydroxymethyl- -4-pentenamide in 10 mL of anhydrous
pyridine at 0.degree. C. was added 1.24 mL (16.3 mmol) of mesyl
chloride. After 3 h, the reaction was poured onto ice, and the
mixture was partitioned between ethyl acetate and 1 N aq. sodium
bisulfate. The organic layer was washed with additional sodium
bisulfate, and the combined aqueous layers were extracted with
ethyl acetate. The combined organic layers were dried over sodium
sulfate and concentrated. The residual oil was dissolved in 30 mL
of acetone and 3.38 g of powdered potassium carbonate was added.
The mixture was heated at reflux for 3 h and then cooled to room
temperature. The precipitate was removed by filtration through
Celite, and the filter cake was washed well with ethyl acetate. The
filtrate was concentrated, and the residue was purified by
chromatography on silica, eluting with 25% ethyl acetate in hexane,
to provide 1.64 g (93%) of N-benzyloxy-3R-(2-propen-1-yl)-2-azeti-
dinone as a slightly orange oil: .sup.1H NMR (300 MHz, CDCl.sub.3)
.delta. 7.37-7.42 (m, 5H), 5.65-5.75 (m, 1H), 5.00-5.06 (m, 1H),
4.93 (s, 2H), 3.32 (ddd, 1H, J=5, 4, and 2 Hz), 2.95 (m, 2H),
2.40-2.47 (m, 1H), 2.20-2.28 (m, 1H).
[0838] El. Anal. Calc. for C.sub.13H.sub.15NO.sub.2: C, 71.86; H,
6.96, N, 6.45.
[0839] Found: C, 71.59; H, 6.88; N, 6.37.
Example 50
[0840] Formula (P-4) Where R.sup.2 is Biphenyl
[0841] A solution of 434 mg (2.00 mmol) of
N-benzyloxy-3R-(2-propen-1-yl)-- 2 azetidinone, 583 mg (2.5 mmol)
of 4-bromobiphenyl, 0.34 mL (2.5 mmol) of triethylamine, 35 mg
(0.11 mmol) of tri(o-tolyl)phosphine, and 14 mg (0.06 mmol) of
palladium diacetate in 7 mL of DMF was heated at 100.degree. C. for
18 h. The reaction solution was cooled to room temperature and
partitioned between ethyl acetate and water. The organic layer was
washed with water, dried over sodium sulfate, and concentrated. The
residue was chromatographed on silica, eluting with 25% ethyl
acetate in hexane, to give slightly impure product, which was
recrystallized from ethyl acetate/isooctane to give 315 mg (43%) of
N-benzyloxy-3R-(3-(biphen- -4-yl)-2-propen-1-yl)-2-azetidinone as
small white flakes: mp 109-110.degree. C. ; .sup.1H NMR (300 MHz,
CDCl.sub.3) .delta. 7.31-7.61 (m, 14H), 6.43 (d, 1H, J=15 Hz), 6.18
(ddd, 1H, J=15, 9, and 7 Hz), 4.94 (s, 2H), 3.36 (dd, 1H, J=10 and
5 Hz), 3.00-3.04 (m, 2H), 2.60-2.75 (m, 1H), 2.20-2.50 (m, 1H).
[0842] El. Anal. Calc. for C.sub.25H.sub.23NO.sub.2; C, 81.27; H,
6.28, N, 3.79.
[0843] Found: C, 81.09; H, 6.31; N, 3.71.
Example 51
[0844] Formula (P-5) Where R.sup.2 is Biphenyl
[0845] To a stirred solution of 62.0 mg (0.168 mmol) of
N-benzyloxy-3R-(3-(biphen-4-yl)-2-propen-1-yl)-2-azetidinone in 5
ml of 4:1 THF:ethanol was added 2 ml of 1N aq. lithium hydroxide.
The mixture was stirred vigorously for 1 h at room temperature, and
the diluted with 10 mL of 0.5 M pH 4 citrate buffer. The mixture
was partitoned between 20 mL of t-butyl methyl ether and brine, and
the organic layer was dried, after dilution with ca. 5 mL hexane,
over sodium sulfate, and concentrated to a residual glass. This
residue was immediately dissolved in 5 mL of dichloromethane,
cooled to 0.degree. C., and 0.10 ml of pyridine was added, followed
by 1.2 mL of a solution of formic anhydride in dichloromethane,
which was prepared by allowing 297 mg (1.00 mmol) of EDC methiodide
and 80 .mu.L (2.00 mmol) of formic acid in 5 mL of dichloromethane
to react at 0.degree. C. for 15 min. After 30 min, the reaction was
partitioned between dichloromethane and 0.5 M pH 3 citrate buffer.
The organic layer was dried over sodium sulfate and concentrated.
Chromatography of the residue on 5 g of silica, eluting with a
gradient of 5% to 10% ethanol in dichloromethane, gave 58 mg (83%)
of
N-((N"-formyl-N"-benzyloxyamino)methyl-5-(biphen-4-yl)-4-pentenoic
acid as a glass. .sup.1H NMR spectrum at room temperature in
CDCl.sub.3 showed broad peaks of amide rotamers.
Example 52
[0846] Formula (P-6) Where R.sup.2 is Biphenyl, R.sup.3 is t-Butyl,
R.sup.7 is 4-Pyridinyl and p is Zero
[0847] To a solution of 99.6 mg (0.24 mmol) of
N-((N"-formyl-N"-benzyloxya- mino)methyl-5-(biphen-4-yl)pentanoic
acid, 125 mg (0.288 mmol) of
L-t-leucine-N'-(pyridin-4-yl)carboxamide bis(trifluroacetate), and
0.125 mL (0.90 mmol) of triethylamine in 4 mL of DMF was added 133
mg (0.30 mmol) of benzotriazol-1-yl-tris-(dimethylamino)phosphonium
hexafluorophosphate. After 16 h at room temperature, the reaction
was partitioned between ethyl acetate and ca. 0.5 M aq. sodium
bicarbonate. The organic layer was washed with 1 M pH 7 phosphate
buffer and with brine, dried over sodium sulfate, and concentrated.
Purification of the residue by chromatography, eluting with 40% to
75% ethyl acetate in hexane, followed by recrystallization from
ethyl acetate/isooctane gave 97.4 mg (67%) of
N-((N"-formyl-N"-benzyloxyamino)methyl-5-(biphen-4-yl)-4-
-pentenoyl)-L-t-leucine-N'-(pyridin-4-yl)carboxamide: mp:
215-216.degree. C.
Example 53
[0848] 53A. Formula (Ih) Where R.sup.2 is Biphenyl, R.sup.3 is
t-Butyl, R.sup.7 is 4-Pyridinyl and p is Zero
[0849] A solution of 87.1 mg (0.143 mmol) of
N-((N"-formyl-N"-benzyloxyami-
no)methyl-5-(biphen-4-yl)pentanoyl)-L-t-leucine-N'-(pyridin-4-yl)carboxami-
de in 5 mL of 3:2 ethyl acetate:ethanol was hydrogenated over 25 mg
of 10% palladium on carbon at 1 atm of hydrogen pressure for 6 h.
The catalyst was removed by filtration through Celite, and the
filtrate was concentrated. Recrystallation of the residue from
ethyl acetate gave 59.0 mg (80%) of
N-((N"-formyl-N"-hydroxyamino)methyl-5-(biphen-4-yl)pentanoyl-
)-L-t-leucine-N'-(pyridin-4-yl)carboxamide as a white powder: mp
190-191.degree. C.;
[0850] .sup.1H NMR (300 MHz, DMSO-d6) .delta. 10.52 (br a, 1H),
9.97 (br s, 1H, 9.53 (br s, 1H), 8.38 (d, 2H, J=7 Hz), 7.20 (br s,
1H), 7.14 (br d, 1H, J=8 Hz), 7.23-7.60 (m, 9H), 7.12 (d, 2H, J=7
Hz), 4.41 (d, 1H, J=9 Hz) 3.40-3.62 (m, 2H), 2.90-3.10 (m, 1H),
2.4-2.6 (m, 2H, partially obscured by DMSO-d5 resonance), 1.28-1.52
(m, 4H), 0.94 (s, 9H).
[0851] El. Anal. Calc. for C.sub.30H.sub.36N.sub.4O.sub.4.0.5
H.sub.2O: C, 68.55; H, 7.10, N. 10.66.
[0852] Found: C, 68.48; H, 7.04; N, 10.63.
[0853] 53B. Formula (Ih) Varying R.sup.2, R.sup.3, and R.sup.7
[0854] By following procedures analogous to Examples 50-52 there
are obtained the following compounds of formula (P-6):
[0855]
N-(2R-(N"-formyl-N"-benzyloxyamino)methyl-5-(4-(2RS-hydroxy-3,3,3-t-
rifluoropropyl)phenyl)-4-pentenoyl)-L-t-leucine-N'-(pyridin-4-yl)carboxami-
de;
[0856]
N-(2R-(N"-formyl-N"-benzyloxyamino)methyl-5-(4-(imidaz-4-yl)phenyl)-
-4-pentenoyl)-L-threonine-N'-((4S-methylsulfinyl)phenyl)carboxamide;
[0857]
N-(2R-(N"-formyl-N"-benzyloxyamino)methyl-5-(4-(2RS-hydroxy-3,3,3-t-
rifluoropropyl)phenyl)-4-pentenoyl)-L-t-leucine-N'-(pyridin-4-yl)carboxami-
de;
[0858]
N-(2R-(N"-formyl-N"-benzyloxyamino)methyl-5-(4-(imidaz-4-yl)phenyl)-
-4-pentenoyl)-L-threonine-N'-((4S-methylsulfinyl)phenyl)carboxamide;
[0859]
N-(2R-(N"-formyl-N"-benzyloxyamino)methyl-5-(4-(pyridin-4-yl)phenyl-
)-4-pentenoyl)-L-t-leucine-N'-(4-((2-hydroxyethyl)-aminosulfonyl)phenyl)ca-
rboxamide;
[0860]
N-(2R-(N"-formyl-N"-benzyloxyamino)methyl-5-(4-(pyridin-4-yl)phenyl-
)-4-pentenoyl)-L-(.beta.-hydroxy)valine-N'-(4S-methylsulfinyl)phenyl)carbo-
xamide;
[0861]
N-(2R,S-(N"-formyl-N"-benzyloxyamino)methyl-(4-(methyl)-4-pentenoyl-
)-L-leucine-N'-(4-(methoxycarbonyl)phenyl)carboxamide; and
[0862]
N-(2R-(N"-formyl-N"-benzyloxyamino)methyl-5-(biphen-4-yl)pentanoyl)-
-L-cyclohexylglycine-N'-(4-((2-(dimethylamino)ethyl)aminosulfonyl)phenyl)c-
arboxamide,
[0863] which, when substituted for
N-((N"-formyl-N"-benzyloxyamino)methyl--
5-(biphen-4-yl)pentanoyl)-L-t-leucine-N'-(pyridin-4-yl)carboxamide
in the procedure of Example 53A, give the following respective
compounds:
[0864]
N-(2R-(N"-formyl-N"-hydroxyamino)methyl-5-(4-(2RS-hydroxy-3,3,3-tri-
fluoropropyl)phenyl)pentanoyl)-L-t-leucine-N'-(pyridin-4-yl)carboxamide;
[0865]
N-(2R-(N"-formyl-N"-hydroxyamino)methyl-5-(4-(imidaz-4-yl)phenyl)-p-
entanoyl)-L-threonine-N'-((4S-methylsulfinyl)phenyl)carboxamide;
[0866]
N-(2R-(N"-formyl-N-hydroxyamino)methyl-5-(4-(pyridin-4-yl)phenyl)pe-
ntanoyl)-L-t-leucine-N'-(4-((2-hydroxyethyl)-aminosulfonyl)phenyl)carboxam-
ide;
[0867]
N-(2R-(N"-formyl-N"-hydroxyamino)methyl-5-(4-(pyridin-4-yl)phenyl)p-
entanoyl)-L-(P-hydroxy)valine-N'-(4S-methylsulfinyl)phenyl)carboxamide;
[0868]
N-(2R,S-(N"-formyl-N"-hydroxyamino)methyl-(4-(methyl)pentanoyl)-L-l-
eucine-N'-(4-(methoxycarbonyl)phenyl)carboxamide: MS (M-H).sup.-:
434.2;(M-CO, H.sub.2O) : 388; and
[0869]
N-(2R-(N"-formyl-N"-hydroxyamino)methyl-5-(biphen-4-yl)pentanoyl)-L-
-cyclohexylglycine-N'-(4-((2-(dimethylamino)ethyl)aminosulfonyl)phenyl)car-
boxamide.
Examples 54-59
[0870] These examples illustrate the preparation of a
representative pharmaceutical formulation containing an active
compound of formula (I), e.g.,
N-(2R-carboxymethyl-5-(biphen-4-yl)pentanoyl)-L-.beta.-hydroxyvalin-
e-N'-(phenyl)carboxamide, or a pharmaceutically acceptable salt
thereof. Other compounds of formula (I) can be used as the active
compound in preparation of the formulations of these examples.
Example 54
[0871] This example illustrates the preparation of representative
pharmaceutical formulations for oral administration.
1 A. Ingredients % wt./wt. Compound of formula (I) 20.0% Lactose
79.5% Magnesium stearate 0.5%
[0872] The above ingredients are mixed and dispensed into
hard-shell gelatin capsules containing 100 mg each, one capsule
would approximate a total daily dosage.
2 B. Ingredients % wt./wt. Compound of formula (I) 20.0% Magnesium
stearate 0.9% Starch 8.6% Lactose 79.6% PVP (polyvinylpyrrolidine)
0.9%
[0873] The above ingredients with the exception of the magnesium
stearate are combined and granulated using water as a granulating
liquid. The formulation is then dried, mixed with the magnesium
stearate and formed into tablets with an appropriate tableting
machine.
3 C. Ingredients Compound of formula (I) 0.1 g Propylene glycol
20.0 g Polyethylene glycol 400 20.0 g Polysorbate 80 1.0 g Water
q.s. 100 mL
[0874] The compound of formula (I) is dissolved in propylene
glycol, polyethylene glycol 400 and polysorbate 80. A sufficient
quantity of water is then added with stirring to provide 100 mL of
the solution which is filtered and bottled.
4 D. Ingredients % wt./wt. Compound of formula (I) 20.0% Peanut Oil
78.0% Span 60 2.0%
[0875] The above ingredients are melted, mixed and filled into soft
elastic capsules.
Example 55
[0876] This example illustrates the preparation of a representative
pharmaceutical formulation for parenteral administration.
5 Ingredients Compound of formula (I) 0.02 g Propylene glycol 20.0
g Polyethylene glycol 400 20.0 g Polysorbate 80 1.0 g 0.9% Saline
solution q.s. 100 mL
[0877] The compound of formula (I) is dissolved in propylene
glycol, polyethylene glycol 400 and polysorbate 80. A sufficient
quantity of 0.9% saline solution is then added with stirring to
provide 100 mL of the I.V. solution which is filtered through a
0.2.mu. membrane filter and packaged under sterile conditions.
Example 56
[0878] This example illustrates the preparation of a representative
pharmaceutical composition in suppository form.
6 Ingredients % wt./wt. Compound of formula (I) 1.0% Polyethylene
glycol 1000 74.5% Polyethylene glycol 4000 24.5%
[0879] The ingredients are melted together and mixed on a steam
bath, and poured into molds containing 2.5 g total weight.
Example 57
[0880] This example illustrates the preparation of a representative
pharmaceutical formulation for insufflation.
7 Ingredients % wt./wt. Micronized compound of formula (I) 1.0%
Micronized lactose 99.0%
[0881] The ingredients are milled, mixed, and packaged in an
insufflator equipped with a dosing pump.
Example 58
[0882] This example illustrates the preparation of a representative
pharmaceutical formulation in nebulized form.
8 Ingredients % wt./wt. Compound of formula (I) 0.005% Water
89.995% Ethanol 10.000%
[0883] The compound of formula (I) is dissolved in ethanol and
blended with water. The formulation is then packaged in a nebulizer
equipped with a dosing pump.
Example 59
[0884] This example illustrates the preparation of a representative
pharmaceutical formulation in aerosol form.
9 Ingredients % wt./wt. Compound of formula (I) 0.10% Propellant
11/12 98.90% Oleic acid 1.00%
[0885] The compound of formula (I) is dispersed in oleic acid and
the propellants. The resulting mixture is then poured into an
aerosol container fitted with a metering valve.
Example 60
[0886] In vitro Matrilysin Assay
[0887] Matrilysin was purified from cloned mammalian cell culture
by Blue-Sepharose and zinc-chelating sepharose column followed by
fast protein liquid chromatography over a MONO S column. The enzyme
was activated by incubation with 1 mmol APMA for 1 hr at
35-37.degree. C.
[0888] Compounds of formula (I) were dissolved in DMSO and added to
a cuvette containing 0.4 .mu.g matrilysin in 1 ml TC buffer (20 mM
Tris, 5 mM CaCl.sub.2, pH 7.5) (2% DMSO final concentration). The
concentrations of the compounds of formula (I) were chosen such
that there was at least one data point for every 20% change in
activity. Enzyme and compounds were permitted to preincubate 3 min
at 37.degree. C. To initiate the reaction,
N-(7-dimethylamino-4-methyl-3-coumarinyl)maleimide ("DACM") (Sigma)
and thiopeptide (Ac-Pro-Leu-Gly-S-"Leu"-Leu-Gly-OEt, Bachem
Bioscience Inc.) were added to 20 .mu.M each. The fluorescence
increase was recorded with excitation and emission wavelengths of
395 and 485 nm, respectively. Each data point is the average of
duplicate experiments. At least six data points, expressed as
change in fluorescence per minute versus compound concentration
were analyzed using the IC.sub.50 fit in the program,
Enzfitter.
[0889] Compounds of formula (I) exhibited the ability to inhibit
matrilysin when tested in this assay.
Example 61
[0890] In vitro Assay
[0891] This assay determines if the compounds of formula (I)
inhibit the release of .sup.35S-labelled glycosaminoglycans (GAG's)
from cartilage explants.
[0892] Small cartilage explants (3 mm diameter) were prepared from
freshly sacrificed bovine knee joints and labeled with
.sup.35SO.sub.4. .sup.35S-labelled glycosaminoglycans (GAG's) are
released into the culture medium in response to the addition of
rhIL-1-alpha, which induces the expression of chondrocyte matrix
metalloproteases (MMP's), including stromelysin and collagenase.
The percent inhibition of labeled GAG's was corrected for
spontaneous release in the absence of rhIL-1-alpha. Results for
each group represent the mean .+-. the S.E.M. for five
explants.
[0893] Compounds of formula (I), when tested in this assay,
displayed the ability to inhibit the release of .sup.35S-labelled
GAG's from cartilage explants.
Example 62
[0894] In vitro Assay
[0895] An in vitro fetal rat long bone model was used to study the
anti-bone resorptive effect of the compounds of formula (I). Bovine
PTH was used to induce bone resorption in vitro. The bone
resorptive effects were expressed by the amounts of .sup.45Ca
released from the .sup.45Ca pre-labelled fetal rat long bones into
the culture medium. The inhibitory effect of the compounds of
formula (I) against bovine PTH induced bone resorption was
expressed as mean percent inhibition.+-.sem.
[0896] .sup.45Ca-prelabelled fetal rat long bones (from forearms)
were dissected and cultured in Linbro dishes at 37.degree. C.
overnight BGJb medium, supplemented with 1 mg/ml BSA. There were
five pairs of bones in each group. The compounds of formula (I)
were dissolved in ethanol first, then diluted to various
concentrations and added simultaneously with Bovine PTH (1-34) at
1.times.10.sup.8M on Day 1. The ethanol concentrations in the
compound solutions were less than 0.05% which did not interfere
with the assay. The assay was terminated on Day 6 with one media
change on Day 3.
[0897] At the end of each medium change, the .sup.45Ca present in
the culture medium was counted. The remaining bones were digested
with 0.1N HCl and the .sup.45Ca presented in the bone digest was
also counted. The results are expressed as % of the total .sup.45Ca
released from each pair of bones. Bovine PTH at 1.times.10.sup.-8M
induces bone resorption to the maximum level which is set as 100%
and this concentration was used as standard. The level of base line
bone resorption in the presence of medium only was set as 0%. All
compound-treated groups were compared with bovine PTH (1-34) at
1.times.10.sup.-8M. The concentration at which a compound inhibited
bone resorption by 50% was defined as IC.sub.50.
[0898] Compounds of formula (I) exhibited the ability to inhibit
bovine PTH-induced bone resorption in this assay.
Example 63
[0899] In vitro Stromelysin Assay
[0900] 63A. Stromelysin enzymatic activity was measured by a
resonance energy transfer fluorogenic assay using the MCA peptide
substrate:
7-methoxycoumarin-4-yl-acetyl-pro-leu-gly-leu-3-(2,4-dinitrophenyl)-L-2,3-
-diaminoproprionyl-ala-arg-NH.sub.2. Cleavage of the substrate at
the gly-leu bond results in the loss of resonance energy transfer
to the 2,4-dinitrophenyl group and an increase in fluorescence of
the MCA (7-methoxycoumarin-4-yl-acetyl) group.
[0901] The assay was performed at 37.degree. C. in buffer
containing 50 mM Tricine, pH 7.5, 10 mM CaCl.sub.2, 200 mM NaCl, 1%
DMSO and 1.4 nM stromelysin. The concentration of MCA substrate was
10 or 20 .mu.M in a final volume of 1.6 ml. In the absence of
compounds to be tested for inhibitory activity, or in the presence
of non-slowbinding inhibitors, fluorescence was measured with
Perkin-Elmer LS-5B and LS-50B spectrofluorimeters with
.lambda..sub.excitation=328 nm and .lambda..sub.emission=393 nm
over a 3 to 5 minute time period and data were fitted to a straight
line. For slow-binding inhibitors, inhibition data were collected
for 45 minutes to 1 hour. Steady-state rates of fluorescence change
were calculated by fitting the curve to an equation for a single
exponential decay containing a linear phase, and taking the fitted
value of the linear phase as the steady-state rate.
[0902] Compounds of formula (I) were tested and found to be active
as inhibitors of MMP activity in this assay. 63B. The MCA assay can
also be used with other matrix metalloproteinases, such as
matrilysin or gelatinase A, by substituting 0.063 nM matrilysin or
0.030 nM gelatinase A for stromelysin.
[0903] While the present invention has been described with
reference to the specific embodiments thereof, it should be
understood by those skilled in the art that various changes may be
made and equivalents may be substituted without departing from the
true spirit and scope of the invention. In addition, many
modifications may be made to adapt a particular situation,
material, composition of matter, process, process step or steps, to
the objective, spirit and scope of the present invention. All such
modifications are intended to be within the scope of the claims
appended hereto. All patents and publications cited above are
hereby incorporated by reference.
* * * * *