U.S. patent application number 09/735968 was filed with the patent office on 2001-09-13 for procollagen c-proteinase inhibitors.
Invention is credited to Bailey, Simon, Billotte, Stephane, Derrick, Andrew Michael, Fish, Paul Vincent, James, Kim, Thomson, Nicholas Murray.
Application Number | 20010021718 09/735968 |
Document ID | / |
Family ID | 27269831 |
Filed Date | 2001-09-13 |
United States Patent
Application |
20010021718 |
Kind Code |
A1 |
Bailey, Simon ; et
al. |
September 13, 2001 |
Procollagen C-proteinase inhibitors
Abstract
1 and their salts, solvates, prodrugs, etc., wherein the
substituents have the values mentioned herein, are Procollagen
C-Proteinase (PCP) inhibitors and have utility in conditions
mediated by PCP.
Inventors: |
Bailey, Simon; (County of
Kent, GB) ; Billotte, Stephane; (County of Kent,
GB) ; Derrick, Andrew Michael; (County of Kent,
GB) ; Fish, Paul Vincent; (County of Kent, GB)
; James, Kim; (County of James, GB) ; Thomson,
Nicholas Murray; (County of Kent, GB) |
Correspondence
Address: |
Paul H. Ginsburg
Pfizer Inc
235 East 42nd Street, 20th Floor
New York
NY
10017-5755
US
|
Family ID: |
27269831 |
Appl. No.: |
09/735968 |
Filed: |
December 13, 2000 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
|
60180527 |
Feb 7, 2000 |
|
|
|
Current U.S.
Class: |
514/364 ;
514/374; 548/131; 548/234 |
Current CPC
Class: |
C07K 5/06139 20130101;
A61K 38/00 20130101; C07D 263/32 20130101; C07C 317/44 20130101;
C07C 259/14 20130101; C07D 271/06 20130101; C07C 2601/14 20170501;
C07K 5/0222 20130101; C07C 233/48 20130101; C07F 9/4006
20130101 |
Class at
Publication: |
514/364 ;
514/374; 548/131; 548/234 |
International
Class: |
A61K 031/42; A61K
031/4245 |
Foreign Application Data
Date |
Code |
Application Number |
Dec 23, 1999 |
GB |
9930570.8 |
Claims
1. A compound of formula (I): 202wherein: X is C.sub.1-6 alkylene
or C.sub.2-6 alkenylene, each of which is optionally substituted by
one or more fluorine atoms; R is aryl or C.sub.3-8 cycloalkyl
optionally substituted by one or more fluorine atoms; W is N or CZ;
Y and Z are each independently H, C.sub.1-4 alkyl (optionally
substituted by one or more substituents independently selected from
halogen, S(O).sub.pR.sup.6, OR.sup.5, CONR.sup.1R.sup.2,
CO.sub.2R.sup.7 and aryl), C.sub.1-4 alkanoyl optionally
substituted by one or more halogen, C.sub.1-4 alkoxycarbonyl
optionally substituted by one or more halogen, or
CONR.sup.1R.sup.2; R.sup.1 and R.sup.2 are each independently
selected from H, C.sub.3-8 cycloalkyl, C.sub.1-4 alkyl (optionally
substituted by C.sub.3-8 cycloalkyl, aryl, CO.sub.2H,
CO.sub.2R.sup.5 and/or NR.sup.3R.sup.4), or R.sup.1 and R.sup.2 can
be taken together with the nitrogen to which they are attached to
represent a 4-to 6-membered heterocyclic ring optionally containing
one or two further hetero atoms in the ring independently selected
from N, O and S, which heterocyclic ring is optionally benzo- or
pyrido-fused, and which heterocyclic ring is optionally substituted
by C.sub.1-4 alkyl, CO.sub.2H, CO.sub.2R.sup.5, aryl and/or
NR.sup.3R.sup.4; R.sup.3 and R.sup.4 are each independently
selected from H, C.sub.1-C.sub.4 alkyl or C.sub.1-4 alkoxycarbonyl
optionally substituted by one or more halogen, or R.sup.3 and
R.sup.4 can be taken together with the nitrogen atom to which they
are attached to represent a morpholine, piperidine, azetidine or
piperazine (optionally N-substituted by C.sub.1-4 alkyl) moiety;
R.sup.5 is C.sub.1-4 alkyl optionally substituted by
CO.sub.2R.sup.7 or CONR.sup.3R.sup.4, or R.sup.5 is aryl; R.sup.6
is C.sub.1-4 alkyl optionally substituted by one or more halogen,
or aryl; R.sup.7 is H or Rr; p is 0 1 or 2; "Aryl"is a mono- or
bicyclic aromatic carbocyclic or heterocyclic system comprising
from 5 to 10 ring atoms, including up to 3 hetero-atoms selected
from N, O and S, where, if there is a N atom in the ring, it can be
present as the N-oxide, which ring system is optionally substituted
by up to 3 substituents independently selected from halogen,
C.sub.1-4 alkyl optionally substituted by one or more halogen,
C.sub.1-4 alkoxy optionally substituted by one or more halogen,
phenyl, pyridyl, CO.sub.2H, CONR.sup.3R.sup.4, CO.sub.2(C.sub.1-4
alkyl), NR.sup.3R.sup.4, OH and OC(O)(C.sub.1-4 alkyl); and the
pharmaceutically acceptable salts, solvates (including hydrates)
and prodrugs thereof.
2. A compound, salt, solvate or prodrug according to claim 1
wherein the compound of formula (I) has the stereochemistry of
formula (IA): 203
3. A compound, salt, solvate or prodrug according to any previous
claim wherein W is N.
4. A compound, salt, solvate or prodrug according to claim 1 or 2
where W is CZ, and at least one of Y and Z is H or C.sub.1-4 alkyl
(optionally substituted by one or more halogen).
5. A compound, salt, solvate or prodrug according to claim 4
wherein W is CZ and Z is H or C.sub.1-4 alkyl optionally
substituted by one or more halogen atoms.
6. A compound, salt, solvate or prodrug according to claim 5
wherein Z is H or methyl optionally substituted by one or more
fluorine atoms.
7. A compound, salt, solvate or prodrug according to claim 6
wherein Z is H or methyl.
8. A compound, salt, solvate or prodrug according to any previous
claim where X is a linear C.sub.2-4 alkylene moiety optionally
substituted by one or more fluorine atoms.
9. A compound, salt, solvate or prodrug according to any previous
claim where R is C.sub.3-8 cycloalkyl optionally substituted by one
or more fluorine atoms.
10. A compound, salt, solvate or prodrug according to any previous
claim where Y is C.sub.1-4 alkyl (optionally substituted by one or
more substituents independently selected from halogen, S(O)
.sub.pR.sup.6, OR.sup.5, CONR.sup.1R.sup.2, CO.sub.2R.sup.7 and
aryl), C.sub.1-4 alkoxycarbonyl, or CONR.sup.1R.sup.2.
11. A compound, salt, solvate or prodrug according to any previous
claim wherein X is propylene.
12. A compound, salt, solvate or prodrug according to any previous
claim wherein R is cyclobutyl or cyclohexyl optionally substituted
by one or more fluorine atoms.
13. A compound, salt, solvate or prodrug according to any previous
claim wherein Y is methyl, isopropyl, methoxymethyl,
2-methoxyethyl, (pyrrolidino)COCH.sub.2, phenylsulphonylmethyl,
4-chlorophenoxymethyl, (pyridin-2-yl)methyl, (pyridin-3-yl)methyl,
(pyridin4-yl)methyl, (imidazol-2-yl)methyl, CO.sub.2(C.sub.1-2
alkyl), CONH.sub.2, CONH(C.sub.1-4 alkyl (optionally substituted by
C.sub.3-8 cycloalkyl, aryl, CO.sub.2H or CO.sub.2R.sup.5)),
CON(C.sub.1-4 alkyl)(C.sub.1-4 alkyl(optionally substituted by
C.sub.3-8 cycloalkyl, aryl, CO.sub.2H or CO.sub.2R.sup.5)), or
CONR.sup.1R.sup.2 where R.sup.1 and R.sup.2 are taken together with
the nitrogen to which they are attached to represent a 4- to
6-membered heterocyclic ring optionally containing one or two
further hetero atoms in the ring independently selected from N, O
and S, and which heterocyclic ring is optionally benzo- or
pyrido-fused, and which heterocyclic ring is optionally substituted
by C.sub.1-4 alkyl, CO.sub.2H, CO.sub.2R.sup.5, aryl or
NR.sup.3R.sup.4.
14. A compound, salt, solvate or prodrug according to any previous
claim wherein R is cyclobutyl or cyclohexyl.
15. A compound, salt, solvate or prodrug according to any previous
claim wherein Y is CO.sub.2C.sub.2H.sub.5, CONH.sub.2,
CONHCH.sub.3, CONH(n-C.sub.3H.sub.7), CONH(i-C.sub.3H.sub.7),
(cyclopropyl)CH.sub.2NHCO- , (cyclobutyl)CH.sub.2NHCO,
(2-methoxyphenyl)CH.sub.2NHCO, (4-methoxyphenyl)CH.sub.2NHCO,
(pyridin-2-yl)CH.sub.2NHCO, CONHCH.sub.2CO.sub.2H,
CON(CH.sub.3)CH.sub.2CO.sub.2CH.sub.3, CON(CH.sub.3).sub.2,
(4-dimethylaminopiperidinyl)CO, (3-morpholinoazetidinyl)CO,
(4-(pyridin4-yl)piperidino)CO, (pyridin-2-yl)CH.sub.2N(CH.sub.3)CO,
CON(CH.sub.3)CH.sub.2CO.sub.2H, (3-methoxycarbonylazetidinyl)CO,
(3-carboxyazetidinyl)CO, methyl, isopropyl, methoxymethyl,
2-methoxyethyl, (pyrrolidino)COCH.sub.2, phenylsulphonylmethyl,
4-chlorophenoxymethyl, (pyridin-2-yl)methyl, (pyridin-3-yl)methyl,
(pyridin4-yl)methyl, (imidazol-2-yl)methyl, benzylaminocarbonyl,
piperidinocarbonyl, (2,3-dihydroisoindol-2-yl)CO,
(1,2,3,4-tetrahydroisoquinolin-2-yl)CO, morpholinocarbonyl,
4-methylpiperazinocarbonyl,
(5-aza-1,2,3,4-tetrahydroisoquinolin-2-yl)CO or
N-methylbenzylaminocarbonyl.
16. A compound, salt, solvate or prodrug according to any previous
claim wherein R is cyclohexyl.
17. A compound, salt, solvate or prodrug according to any previous
claim wherein Y is CONH.sub.2, CONHCH.sub.3 or
CON(CH.sub.3).sub.2.
18. A compound, or salt, solvate or prodrug thereof, of formula (I)
according to claim 1 wherein the substituents are as specified in
the compounds of the Examples described herein.
19. A compound according to claim 1 which is selected from:
5-{(1R)4-cyclohexyl-1-[2-(hydroxyamino)-2-oxoethyl]butyl}-1,2,4-oxadiazol-
e-3-carboxamide;
5-{(1R)-4-cyclohexyl-1-[2-(hydroxyamino)-2-oxoethyl]butyl-
}-N-methyl-1,2,4-oxadiazole-3-carboxamide; and
5-{(1R)-4-cyclohexyl-1-[2-(-
hydroxyaminoy2-oxoethyl]butyl}-N,N-dimethyl-1,2,4-oxadiazole-3-carboxamide-
, and the salts and solvates thereof.
20. A PCP inhibitor which is selective against MMP-1 and/or MMP-2
and/or MMP-9 and/or MMP-14.
21. A substance according to any previous claim for use in
medicine.
22. The use of a substance according to any of claims 1, 2, 3, 4,
5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or 20 in the
manufacture of a medicament for the treatment of a condition
mediated by PCP.
23. A pharmaceutical composition comprising a substance according
to any one of claims 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14,
15, 16, 17, 18, 19 or 20 and a pharmaceutically acceptable diluent,
carrier or adjuvant.
24. A method of treatment of a condition mediated by PCP comprising
administration of a therapeutically-effective amount of a substance
according to any one of claims 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11,
12, 13, 14, 15, 16, 17, 18, 19 or 20.
25. A process to make a compound of formula (I) as defined in claim
(I) which comprises deprotection of a compound of formula (III) or
(VIII), 204as appropriate, where P is an O-protecting group and the
other substituents are as defined in claim 1.
26. A process to make a compound of formula (I) as defined in claim
1 which comprises reaction of a compound of formula (II) or (IX)
205as appropriate, wherein L is a suitable leaving group and the
other substituents are as defined in claim 1, with hydroxylamine or
a salt thereof, if necessary with a base.
27. A process for making a compound of formula (II) or (IX) as
defined in claim 26 which comprises reaction of a compound of
formula (IV) or (X) as appropriate 206with a suitable activating
reagent to introduce the "L"moiety.
28. A process to make a compound of formula (IV) or (X) as defined
in claim 27 which comprises deprotecting a compound of formula (V)
or (XI) as appropriate 207where P is an O-protecting group.
29. A process for making a compound of formula (V) as defined in
claim 28 which comprises internal condensation of a compound of
formula (VI) 208wherein P is an O-protecting group.
30. A process for making a compound of formula (XI) as defined in
claim 28 which comprises dehydrogenation of a compound of formula
(XII) 209wherein P is an O-protecting group.
31. A process for making a compound of formula (VI) as defined in
claim 29 which comprises reaction of a compound of formula (VII)
210with a compound of formula YC(.dbd.NOH)NH.sub.2.
32. A process for making a compound of formula (XII) as defined in
claim 30 which comprises internal condensation of a compound of
formula (XIII): 211
33. A process for making a compound of formula (XIII) as defined in
claim 32 which comprises condensation of a compound of formula
(VII) as defined in claim 31 with a compound of formula
NH.sub.2CH(Y)CH(Z)OH.
34. A compound of formula (II) as defined in claim 26.
35. A compound of formula (III) as defined in claim 25.
36. A compound of formula (IV) as defined in claim 27.
37. A compound of formula (V) as defined in claim 28.
38. A compound of formula (VI) as defined in claim 29.
39. A compound of formula (VII) as defined in claim 31.
40. A compound of formula (VIII) as defined in claim 25.
41. A compound of formula (IX) as defined in claim 26.
42. A compound of formula (X) as defined in claim 27.
43. A compound of formula (XI) as defined in claim 28.
44. A compound of formula (XII) as defined in claim 30.
45. A compound of formula (XIII) as defined in claim 32.
46. (2R)-2-[2-(tert-butoxy)-2-oxoethyl]-5-cyclohexylpentanoic
acid.
47. (4S)-4-benzyl-3-(5-cyclohexylpentanoyl)-1,3-oxazo
idin-2-one.
48. tert-butyl
3-{[(4S)4-benzyl-2-oxo-1,3-oxazolidin-3-yl]carbonyl}-6-cycl-
ohexylhexanoate.
49. 2-[2-(tert-butoxy)-2-oxoethyl]-5-cyclohexylpentanoic acid.
50. 3-(diethoxyphosphoryl)succinic acid 1-tert-butyl ester.
51. (E)-2-[2-(tert-butoxy)-2-oxoethyl]-5phenyl-2-pentenoic
acid.
52. (R)-2-[2-(tert-butoxy)-2-oxoethyl]-5-phenylpentanoic acid.
53. (R)-2-[2-(tert-butoxyy-2-oxoethyl]-5-cyclohexyipentanoic acid
cyclohexylamine salt.
Description
[0001] This invention relates to a certain class of compounds, and
the pharmaceutically acceptable salts, solvates and prodrugs
thereof, which inhibit Procollagen C-proteinase ("PCP"). They are
therefore useful in the treatment of mammals having conditions
alleviable by inhibition of PCP. Especially of interest is an
antiscarring treatment for wounds.
[0002] Fibrotic tissues, including dermal scars, are characterised
by excessive accumulation of extracellular matrix, mainly collagen
type I. It is thought that inhibition of collagen deposition will
reduce formation of scar tissue. Collagen is secreted as the
precursor, procollagen, which is transformed into the insoluble
collagen by cleavage of the C-terminal propeptide by PCP. PCP is a
zinc-dependent metalloprotease which is secreted from
TGF-.beta.-activated fibroblasts belonging to the subfamily of
astacin-like proteases and able to cleave the C-terminal peptide of
types I, II and III procollagens. Furthermore, data suggest that
PCP activates lysyl oxidase, an enzyme essential for the formation
of covalent cross-links which stabilise the fibrous form of
collagen. Therefore, inhibition of PCP may not only reduce collagen
deposition but may also make collagen more accessible for
degradation.
[0003] Collagen is integral to, among other things, the proper
formation of connective tissue. Thus, the over- or under-production
of collagen or the production of abnormal collagen (including
incorrectly processed collagen) has been linked with numerous
connective tissue diseases and disorders. Mounting evidence
suggests that PCP is an essential key enzyme for the proper
maturation of collagen (see for example International Patent
Application publication number WO 97/05865).
[0004] The present invention relates to substances capable of
inhibiting PCP activity in order to regulate, modulate and/or
reduce collagen formation and deposition. More specifically, the
invention relates to the use of compounds and pharmaceutical
compositions thereof for the treatment of various conditions
relating to production of collagen.
[0005] At present more than nineteen types of collagens have been
identified. These collagens, including fibrillar collagen types I,
II, III are synthesized as procollagen precursor molecules which
contain amino- and carboxy-terminal peptide extensions. These
peptide extensions, referred to as "pro-regions," are designated as
N- and C- propepfides, respectively. The pro-regions are typically
cleaved upon secretion of the procollagen triple helical precursor
molecule from the cell to yield a mature triple helical collagen
molecule. Upon cleavage, the "mature" collagen molecule is capable
of association, for example, into highly structured collagen
fibers. See e.g., Fessler and Fessler, 1978, Annu. Rev. Biochem.
47:129-162; Bornstein and Traub, 1979, in: The Proteins (eds.
Neurath, H. and Hill, R. H.), Academic Press, New York, pp.
412-632; Kivirikko et al., 1984, in: Extracellur Matrix
Biochemistry (eds. Piez, K. A. and Reddi. A. H.), Elsevier Science
Publishing Co., Inc., New York, pp. 83-118; Prockop and Kivirikko,
1984, N. Engl, J. Med. 311:376-383; Kuhn, 1987, in: Structure and
Function of Collagen Types (eds. Mayne, R. and Burgeson, R. E.),
Academic Press, Inc., Orlando, Fla., pp. 142.
[0006] An array of conditions has been associated with the
inappropriate or unregulated production of collagen, including
pathological fibrosis or scarring, including endocardial sclerosis,
idiopathic interstitial fibrosis, interstitial pulmonary fibrosis,
perimuscular fibrosis, Symmers' fibrosis, pericentral fibrosis,
hepatitis, dermatofibroma, cirrhosis such as binary cirrhosis and
alcoholic cirrhosis, acute pulmonary fibrosis, idiopathic pulmonary
fibrosis, acute respiratory distress syndrome, kidney
fibrosis/glomerulonephritis, kidney fibrosis/diabetic nephropathy,
scleroderma/systemic, scierodermaalocal, keloids, hypertrophic
scars, severe joint adhesions/arthritis, myelofibrosis, corneal
scarring, cystic fibrosis, muscular dystrophy (duchenne's), cardiac
fibrosis, muscular fibrosis/retinal separation, esophageal
stricture and Pyronie's disease. Further fibrotic disorders may be
induced or initiated by surgery, including scar revision/plastic
surgeries, glaucoma, cataract fibrosis, corneal scarring, joint
adhesions, graft vs. host disease, tendon surgery, nerve
entrapment, dupuytren's contracture, OB/GYN adhesions/fibrosis,
pelvic adhesions, peridural fibrosis, restenosis. Other conditions
where collagen plays a key role include burns. Fibrosis of lung
tissue is also observed in patients suffering from chronic
obstructive airways disease (COAD) and asthma. One strategy for the
treatment of these diseases and conditions is to inhibit the
overproduction and/or deposition and/or unregulation of collagen.
Thus, identification and isolation of molecules which control,
inhibit and/or modulate the production and deposition of collagen
are of major medical interest.
[0007] Recent evidence suggests that PCP is the essential key
enzyme that catalyzes the cleavage of the Procollagen C-propeptide.
This has been demonstrated in fibrillar collagens, including type
I, type II, and type III collagen.
[0008] PCP was first observed in the culture media of human and
mouse fibroblasts (Goldberg et al., 1975, Cell 4:45-50; Kessler and
Goldberg, 1978, Anal. Biochem. 86:463469), and chick tendon
fibroblasts (Duskin et al., 1978, Arch. Biochem. Biophys.
185:326-332; Leung et al., 1979, J. Biol, Chem. 254:224-232). An
acidic proteinase which removes the C-terminal propeptides from
type I procollagen has also been identified (Davidson et al., 1979,
Eur. J. Biochem. 100:551).
[0009] A partially purified protein having PCP activity was
obtained from chick calvaria in 1982. Njieha et al., 1982,
Biochemistry 23:757-764. In 1985, chicken PCP was isolated,
purified and characterized from conditioned media of chick embryo
tendons. Hojima et al., 1985, J. Biol. Chem. 260:15996-16003.
Murine PCP has been subsequently purified from media of cultured
mouse fibroblasts. Kessler et al., 1986, Collagen Relat. Res.
6:249-266; Kessler and Adar, 1989, Eur. J. Biochem. 186:115-121.
Finally, the cDNA encoding human PCP has been identified, as set
forth in the above-referenced articles and references disclosed
therein.
[0010] Experiments conducted with these purified forms of chick and
mouse PCP have indicated that the enzyme is instrumental in the
formation of functional collagen fibers. Fertala et al., 1994, J.
Biol. Chem. 269:11584.
[0011] As a consequence of the enzyme's apparent importance to
collagen production, scientists have identified a number of PCP
inhibitors. See e.g., Hojima et al., supra. For example, several
metal chelators have demonstrated activity as PCP inhibitors.
Likewise, chymostatin and pepstatin A were found to be relatively
strong inhibitors of PCP. Additionally,
.alpha..sub.2-Macroglobuline, ovostatin, and fetal bovine serum
appear to at least partially inhibit PCP activity.
[0012] Dithiothreitol, SDS, concanavalin A, Zn.sup.2+, Cu.sup.2+,
and Cd.sup.2+ are similarly reported to be inhibitory at low
concentrations. Likewise, some reducing agents, several amino
acids, phosphate, and ammonium sulfate were inhibitory at
concentrations of 1-10 mM. Further, the enzyme was shown to be
inhibited by the basic amino acids lysine and arginine (Leung et
al., supra; Ryhnen et al., 1982, Arch. Biochem. Biophys.
215:230-235). Finally, high concentrations of NaCl or Tris-HCl
buffer were found to inhibit PCP's activity. For example, it is
reported that, with 0.2, 0.3, and 0.5M NaCl, the activity of PCP
was reduced 66, 38, and 25%, respectively, of that observed with
the standard assay concentration of 0.1 5M. Tris-HCl buffer in a
concentration of 0.2-0.5M markedly inhibited activity (Hojima et
al., supra). PCP activity and its inhibition have been determined
using a wide array of assays. See e.g., Kessler and Goldberg, 1978,
Anal. Biochem. 86:463; Njieha et al., 1982, Biochemistry
21:757-764. As articulated in numerous publications, the enzyme is
difficult to isolate by conventional biochemical means and the
identity of the cDNA sequence encoding such enzyme was not known
until reported in the above referenced and related patent
applications.
[0013] In view of its essential role in the formation and
maturation of collagen PCP appears to be an ideal target for the
treatment of disorders associated with the inappropriate or
unregulated production and maturation of collagen. However, none of
the inhibitors so far disclosed has proven to be an effective
therapeutic for the treatment of collagen-related diseases and
conditions.
[0014] The identification of effective compounds which specifically
inhibit the activity of PCP to regulate and modulate abnormal or
inappropriate collagen production is therefore desirable and the
object of this invention.
[0015] Matrix metalloproteases (MMPs) constitute a family of
structurally similar zinc-containing metalloproteases, which are
involved in the remodelling, repair and degradation of
extracellular matrix proteins, both as part of normal physiological
processes and in pathological conditions.
[0016] Another important function of certain MMPs is to activate
other enzymes, including other MMPs, by cleaving the pro-domain
from their protease domain. Thus, certain MMPs act to regulate the
activities of other MMPs, so that over-production in one MMP may
lead to excessive proteolysis of extracellular matrix by another,
e.g. MMP-14 activates pro-MMP-2
[0017] During the healing of normal and chronic wounds, MMP-1 is
expressed by migrating keratinocytes at the wound edges (U. K.
Saarialho-Kere, S. O. Kovacs, A. P. Pentland, J. Clin. Invest.
1993, 92, 2858-66). There is evidence which suggests MMP-1 is
required for keratinocyte migration on a collagen type I matrix in
vitro, and is completely inhibited by the presence of the
non-selective MMP inhibitor SC44463
((N4-hydroxy)-Nl-[(1S)-2-(4-methoxyphenyl)methyl-1-((1R)-methylamino)carb-
onyl)]-(2R)-2-(2-methylpropyl)butanediamide) (B. K. Pilcher, J. A.
Dumin, B. D. Sudbeck, S. M. Krane, H. G. Welgus, W. C. Parks, J.
Cell Biol., 1997, 137, 1-13). Keratinocyte migration in vivo is
essential for effective wound healing to occur.
[0018] MMP-2 and MMP-9 appear to play important roles in wound
healing during the extended remodelling phase and the onset of
re-epithelialisation, respectively (M. S. Agren, Brit. J.
Dermatology, 1994, 131, 63440; T. Salo, M. Makanen, M. Kylmniemi,
Lab. Invest., 1994, 70, 176-82). The potent, non-selective MMP
inhibitor BB94
((2S,3R)-5-methyl-3{-[(1S)-1-(methylcarbamoyl)-2-phenylethyl]carbamoyl}-2-
-[(2-thienylthio)methyl]hexanohydroxamic acid, batimastat),
inhibits endothelial cell invasion of basement membrane, thereby
inhibiting anglogenesis (G. Tarboletti, A. Garofalo, D. Belotti, T.
Drudis, P. Borsotti, E. Scanziani, P. D. Brown, R. Giavazzi, J.
Natl. Cancer Inst., 1995, 87, 293-8). There is evidence that this
process requires active MMP-2 and/or 9.
[0019] Thus PCP inhibitors which significantly inhibit MMPs 1
and/or 2 and/or 9 would be expected to impair wound healing. MMP-14
is responsible for the activation of MMP-2, and thus inhibition of
MMP-14 might also result in impaired wound healing.
[0020] For recent reviews of MMPs, see Zask et al, Current
Pharmaceutical Design, 1996, 2, 624-661; Beckett, Exp. Opin. Ther.
Patents, 1996, 6, 1305-1315; and Beckett et al, Drug Discovery
Today, vol 1(no.1), 1996, 16-26.
[0021] Alternative names for various MMPs and substrates acted on
by these are shown in the table below (Zask et al, supra).
1 Enzyme Other names Preferred substrates MMP-1 Collagenase-1,
interstitial Collagens I, II, III, VII, X, collagenase gelatins
MMP-2 Gelatinase A, 72 kDa Gelatins, collagens IV, V, gelatinase
VII, X, elastin, fibronectin; activates pro-MMP-13 MMP-3
Stromelysin-1 Proteoglycans, laminin, fibronectin, gelatins. MMP-7
Pump, Matrilysin Proteoglycans, laminin, fibronectin, gelatins,
collagen IV, elastin, activates pro MMP-1 and -2. MMP-8
Collagenase-2, neutrophil Collagens I, II, III collagenase MMP-9
Gelatinase B, 92 kDa Gelatins, collagens, IV, V, gelatinase elastin
MMP-12 Macrophage metalloelastase Elastin, collagen IV,
fibronectin, activates pro-MMP-2 & 3. MMP-13 Collagensase-3
Collagens I, II, III, gelatins MMP-14 MT-MMP-1 Activates pro-MMP-2
& 13, gelatins MMP-15 MT-MMP-2 unknown MMP-16 MT-MMP-3
Activates pro-MMP-2 MMP-17 MT-MMP-4 unknown
[0022] According to one aspect of the present invention, there are
provided compounds of formula (I): 2
[0023] wherein:
[0024] X is C.sub.1-6alkylene or C.sub.2-6alkenylene, each of which
is optionally substituted by one or more fluorine atoms;
[0025] R is aryl or C.sub.3-8cycloalkyl optionally substituted by
one or more fluorine atoms;
[0026] W is N or CZ;
[0027] Y and Z are each independently H, C.sub.1-4 alkyl
(optionally substituted by one or more substituents independently
selected from halogen, S(O).sub.pR.sup.6, OR.sup.5,
CONR.sup.1R.sup.2, CO.sub.2R.sup.7 and aryl), C.sub.1-4alkanoyl
optionally substituted by one or more halogen,
C.sub.1-4alkoxycarbonyl optionally substituted by one or more
halogen, or CONR.sup.1R.sup.2;
[0028] R.sup.1 and R.sup.2 are each independently selected from H,
C.sub.3-8cycloalkyl, C.sub.1-4alkyl (optionally substituted by
C.sub.3-8cycloalkyl, aryl, CO.sub.2H, CO.sub.2R.sup.5 and/or
NR.sup.3R.sup.4), or R.sup.1 and R.sup.2 can be taken together with
the nitrogen to which they are attached to represent a 4- to
6-membered heterocyclic ring optionally containing one or two
further hetero atoms in the ring independently selected from N, O
and S, which heterocyclic ring is optionally benzo- or
pyrido-fused, and which heterocyclic ring is optionally substituted
by C.sub.1-4alkyl, CO.sub.2H, CO.sub.2R.sup.5, aryl and/or
NR.sup.3R.sup.4;
[0029] R.sup.3 and R.sup.4 are each independently selected from H,
C.sub.1-C.sub.4alkyl or C.sub.1-4alkoxycarbonyl optionally
substituted by one or more halogen, or R.sup.3 and R.sup.4 can be
taken together with the nitrogen atom to which they are attached to
represent a morpholine, piperidine, azetidine or piperazine
(optionally N-substituted by C.sub.1-4alkyl) moiety;
[0030] R.sup.5 is C.sub.1-4alkyl optionally substituted by
CO.sub.2R.sup.7 or CONR.sup.3R.sup.4, or R.sup.5 is aryl;
[0031] R.sup.6 is C.sub.1-4 alkyl optionally substituted by one or
more halogen, or aryl;
[0032] R.sup.7 is H or R.sup.6;
[0033] p is 0, 1 or2;
[0034] "Aryl" is a mono- or bicyclic aromatic carbocyclic or
heterocyclic system comprising from 5 to 10 ring atoms, including
up to 3 hetero-atoms selected from N, O and S, where, if there is a
N atom in the ring, it can be present as the N-oxide, which ring
system is optionally substituted by up to 3 substituents
independently selected from halogen, C.sub.1-4alkyl optionally
substituted by one or more halogen, C.sub.14 alkoxy optionally
substituted by one or more halogen, phenyl, pyridyl, CO.sub.2H,
CONR.sup.3R.sup.4, CO.sub.2(C.sub.1-4alkyl), NR.sup.3R.sup.4, OH
and OC(O)(C.sub.1-4alkyl);
[0035] and the pharmaceutically acceptable salts, solvates
(including hydrates) and prodrugs thereof.
[0036] "Alkyl", "alkylene", "alkoxy", "alkanoyl", and "alkenylene"
groups, including in groups incorporating said moieties, may be
straight chain or branched where the number of carbon atoms
allows.
[0037] Halogen is taken to mean fluorine, chlorine, bromine or
iodine.
[0038] Pharmaceutically-acceptable salts are well known to those
skilled in the art, and for example include those mentioned in the
art cited above, and by Berge et al, in J.Pharm.Sci., 66, 1-19
(1977). Suitable acid addition salts are formed from acids which
form non-toxic salts and indude the hydrochloride, hydrobromide,
hydroiodide, nitrate, sulphate, bisulphate, phosphate,
hydrogenphosphate, acetate, trifluoroacetate, gluconate, lactate,
salicylate, citrate, tartrate, ascorbate, succinate, maleate,
fumarate, gluconate, formate, benzoate, methanesulphonate,
ethanesulphonate, benzenesulphonate, pamoate, camsylate, and
p-toluenesulphonate salts.
[0039] Pharmaceutically acceptable base addition salts are well
known to those skilled in the art, and for example include those
mentioned in the art cited above, and can be formed from bases
which form non-toxic salts and include the aluminium, calcium,
lithium, magnesium, potassium, sodium and zinc salts, and salts of
non-toxic amines such as diethanolamine.
[0040] Certain of the compounds of formula (I) may exist in one or
more zwitterionic forms. It is to be understood that
pharmaceutically acceptable salts includes all such
zwitterions.
[0041] Certain of the compounds of formula (I), their salts,
solvates, prodrugs, etc. may exist in one or more polymorphic
forms. It is to be understood that the invention includes all such
polymorphs.
[0042] The compounds of formula (I), their salts, hydrates,
prodrugs etc. can exhibit isotopic variation, e.g. forms with
enriched .sup.2H, .sup.3H, .sup.13C, .sup.14C, .sup.15N, .sup.18O,
etc. may be prepared, for example by suitable variation of the
synthetic methods described herein using methods and reagents known
in the art or routine modification thereof. All such isotopic
variants are included in the scope of the invention.
[0043] Prodrug moieties are well-known to those skilled in the art
(see for example the article by H Feres, in Drugs of Today, vol 19,
no.9 (1983) pp.499-538, especially section A1), and for example
include those specifically mentioned in A. A. Sinkula's article in
Annual Reports in Medicinal Chemistry, vol 10, chapter 31,
pp.306-326, herein incorporated by reference, and the references
therein. Specific prodrug moieties which may be specifically
mentioned are aliphatic-aromatic, carbonate, phosphate and
carboxylic esters, carbamates, peptides, glycoside, acetals and
ketals, tetrahydropyranyl and silyl ethers. Such prodrug moieties
can be cleaved in situ, e.g. are hydrolysable in physiological
conditions, to give compounds of formula (I).
[0044] Certain of the compounds of the formula (I) may exist as
geometric isomers. The compounds of the formula (I) may possess one
or more asymmetric centres, apart from the specified centres in
formula (I), and so exist in two or more stereoisomeric forms. The
present invention includes all the individual stereoisomers and
geometric isomers of the compounds of formula (I) and mixtures
thereof.
[0045] Preferably the compounds of formula (I) have the following
stereochemistry (IA): 3
[0046] Preferably, for compounds of formula (I) where W is CZ, at
least one of Y and Z is H or C.sub.1-4alkyl (optionally substituted
by one or more halogen).
[0047] Preferably X is a linear C.sub.2-4alkylene moiety optionally
substituted by one or more fluorine atoms. Most preferably X is
propylene.
[0048] Preferably R is C.sub.3-8 cycloalkyl optionally substituted
by one or more fluorine atoms.
[0049] More preferably R is cyclobutyl or cyclohexyl optionally
substituted by one or more fluorine atoms. Yet more preferably R is
cyclobutyl or cyclohexyl. Most preferably R is cyclohexyl.
[0050] Preferably W is N, CH or CCH.sub.3. Most preferably W is
N.
[0051] Preferably Z is H or C.sub.1-4alkyl optionally substituted
by one or more halogen atoms. More preferably Z is H or methyl
optionally substituted by one or more fluorine atoms. Most
preferably Z is H or methyl.
[0052] Preferably Y is C.sub.1-4 alkyl (optionally substituted by
one or more substituents independently selected from halogen,
S(O).sub.pR.sup.6, OR.sup.5, CONR.sup.1R.sup.2, CO.sub.2R.sup.7 and
aryl), C.sub.1-4alkoxycarbonyl, or CONR.sup.1R.sup.2. More
preferably Y is methyl, isopropyl, methoxymethyl, 2-methoxyethyl,
(pyrrolidino)COCH.sub.2- , phenylsulphonylmethyl,
4-chlorophenoxymethyl, (pyridin-2-yl)methyl, (pyridin-3-yl)methyl,
(pyridin-4-yl)methyl, (imidazol-2-yl)methyl, CO.sub.2(C.sub.1-2
alkyl), CONH.sub.2, CONH(C.sub.1-4alkyl (optionally substituted by
C.sub.3-8cycloalkyl, aryl, CO.sub.2H or CO.sub.2R.sup.5)),
CON(C.sub.1-4alkyl)(C.sub.1-4alkyl(optionally substituted by
C.sub.3-8cycloalkyl, aryl, CO.sub.2H or CO.sub.2R.sup.5)), or
CONR.sup.1R.sup.2 where R.sup.1 and R.sup.2 are taken together with
the nitrogen to which they are attached to represent a 4- to
6-membered heterocyclic ring optionally containing one or two
further hetero atoms in the ring independently selected from N, O
and S, and which heterocyclic ring is optionally benzo- or
pyrido-fused, and which heterocyclic ring is optionally substituted
by C.sub.1-4alkyl, CO.sub.2H, CO.sub.2R.sup.5, aryl or
NR.sup.3R.sup.4. Yet more preferably Y is CO.sub.2C.sub.2H.sub.5,
CONH.sub.2, CONHCH.sub.3, CONH(n-C.sub.3H.sub.7),
CONH(i-C.sub.3H.sub.7), (cyclopropyl)CH.sub.2NHCO,
(cyclobutyl)CH.sub.2NHCO, (2-methoxyphenyl)CH.sub.2NHCO,
(4-methoxyphenyl)CH.sub.2NHCO, (pyridin-2-yl)CH.sub.2NHCO,
CONHCH.sub.2CO.sub.2H, CON(CH.sub.3)CH.sub.2CO.sub.2CH.sub.3,
CON(CH.sub.3).sub.2, (4-dimethylaminopiperidinyl)CO,
(3-morpholinoazetidinyl)CO, (4-(pyridin-4-yl)piperidino)CO,
(pyridin-2-yl)CH.sub.2N(CH.sub.3)CO,
CON(CH.sub.3)CH.sub.2CO.sub.2H, (3-methoxycarbonylazetidinyl)CO,
(3-carboxyazetidinyl)CO, methyl, isopropyl, methoxymethyl,
2-methoxyethyl, (pyrrolidino)COCH.sub.2, phenylsulphonylmethyl,
4-chlorophenoxymethyl, (pyridin-2-yl)methyl, (pyridin-3-yl)methyl,
(pyridin-4-yl)methyl, (imidazol-2-yl)methyl, benzylaminocarbonyl,
piperidinocarbonyl, (2,3-dihydroisoindol-2-yl)CO,
(1,2,3,4-tetrahydroisoquinolin-2-yl)CO, morpholinocarbonyl,
4-methylpiperazinocarbonyl,
(5-aza-1,2,3,4-tetrahydroisoquinolin-2-yl)CO or
N-methylbenzylaminocarbonyl. Most preferably Y is CONH.sub.2,
CONHCH.sub.3 or CON(CH.sub.3).sub.2.
[0053] A preferred group of compounds is that in which each
substituent is as specified in the Examples below.
[0054] Another preferred group are the compounds are those of the
Examples below (especially Examples 2,3 and 12) and the salts,
solvates and prodrugs thereof.
[0055] A further aspect of the invention is a PCP inhibitor which
is selective against MMP-1 and/or MMP-2 and/or MMP-9 and/or MMP-14.
A further aspect of the invention is the use of a PCP inhibitor
which is selective against MMP-1 and/or MMP-2 and/or MMP-9 and/or
MMP-14 in medicine. Further related to this aspect of the invention
is the use of a PCP inhibitor which is selective against MMP-1
and/or MMP-2 and/or MMP-9 and/or MMP-14 in the manufacture of an
antiscarring medicament. Further related to this aspect of the
invention is a method of treating a condition mediated by PCP and
in which MMP-1 and/or MMP-2 and/or MMP-9 and/or MMP-14 have a
beneficial effect, with an effective amount of PCP inhibitor which
is selective against MMP-1 and/or MMP-2 and/or MMP-9 and/or MMP-14,
an example of such a condition being a wound. Preferably the PCP
inhibitor mentioned in this aspect of the invention is selective
against at least MMP-1, MMP-2 and MMP-9. Most preferably the said
PCP inhibitor is selective against MMP-1, MMP-2, MMP-9, and MMP-14.
Preferably the said selective PCP inhibitor has an IC.sub.50 vs.
PCP of 0.5.mu.M or lower, and selectivities vs. MMP-2 and MMP-9 of
at least 30-fold, in the tests described herein. Preferably the
selective PCP inhibitor has an IC.sub.50 vs. PCP of 0.1 .mu.M or
lower, and selectivities vs. MMP-1, MMP-2, MMP-9 and MMP-14 of at
least 300-fold, in the tests described herein.
[0056] Another aspect of the invention is the use of the substances
of formula (I) described herein, including the salts, solvates and
prodrugs thereof, in medicine.
[0057] Another aspect of the invention is the use of the substances
of formula (I) described herein, including the salts, solvates and
prodrugs thereof, in the manufacture of an antiscarring
medicament.
[0058] Another aspect of the invention is a pharmaceutical
composition comprising a PCP inhibitor which is selective vs.
MMP-1, MMP-2, MMP-9 and MMP-14, and a pharmaceutically acceptable
diluent, carrier or adjuvant.
[0059] Another aspect of the invention is a pharmaceutical
composition comprising a compound of formula (I), salts thereof,
solvates thereof and/or prodrugs thereof, and a pharmaceutically
acceptable diluent, carrier or adjuvant.
[0060] Another aspect of the invention is the combination of a PCP
inhibitor, preferably a compound of formula (I), or a salt, solvate
or prodrug thereof, with another material useful in treating
wounds, such as:
[0061] (i) a growth factor such as TGF-.beta.-3 ( Renovo), IGF-1
(Genentech), IGF-1 complex (Celtrix), KGF-2 or FGF-10 (Sumitomo),
DWP-401/EGF (Daewoong) or SNK-863 (Sanwa Kagaku Kenkyusho);
[0062] (ii) a growth factor agonist such as Noggin (Regeneron);
[0063] (iii) a growth factor antibody/antisense material, such as
those to: TGF-.beta.-1 or 2 (Renovo, CaT), PDGF (II Yang) or CTGF
(Fibrogen);
[0064] (iv) a hormone such as DHEAS (Pharmadigm), ConXn/Relaxin
(Connetics);
[0065] (v) an antibody to adhesion compounds such as ICAM-1
(Boehringer);
[0066] (vi) a MMP such as Collagenase ABC (BioSpecifics);
[0067] (vii) a barrier such as ADCON (Gliatech);
[0068] (viii) skin products such as artificial skin systems such as
those based on DermaGraft (Advanced Tissue Sciences Inc.), INTEGRA
Artificial Skin (Integra Life Sciences Holding Corp.), cell
cultures such as Apligraf/Graftskin (Novartis), those developed by
Cell Genesys Inc., AlloDerm (LifeCell) or matrix formulation
products such as Argidene gel (Telios Pharmaceuticals Inc.);
[0069] (ix) a uPA inhibitor such as those disclosed in WO
99/01451;
[0070] (x) a MMP-3 inhibitor such as those disclosed in WO
99/35124, WO 99/29667;
[0071] A further aspect of the invention is the use of a substance
according to the above definitions for the manufacture of a
medicament for the treatment of a condition mediated by PCP.
[0072] Yet another aspect of the invention is a method of treatment
of a condition mediated by PCP comprising administration of a
therapeutically-effective amount of a substance according to the
above definitions.
[0073] It is to be appreciated that reference to treatment includes
prophylaxis as well as the alleviation of established symptoms of
PCP-mediated conditions and diseases.
[0074] The invention further provides Methods for the production of
compounds of the invention, which are described below and in the
Examples and Preparations. The skilled man will appreciate that the
compounds of the invention could be made by methods other than
those specifically described herein, by adaptation of the methods
herein described in the sections below and/or adaptation thereof,
for example by methods known in the art. Suitable guides to
synthesis, functional group transformations, use of protecting
groups, etc. are, for example, "Comprehensive Organic
Transformations" by R C Larock, VCH Publishers Inc. (1989),
"Advanced Organic Chemistry" by J March, Wiley lnterscience (1985),
"Designing Organic Synthesis" by S Warren, Wiley Interscience
(1978), "Organic Synthesis--The Disconnection Approach" by S
Warren, Wiley lnterscience (1982), "Guidebook to Organic Synthesis"
by R K Mackie and D M Smith, Longman (1982), "Protective Groups in
Organic Synthesis" by T W Greene and P G M Wuts, John Wiley and
Sons Inc. (1999), and P J Kocienski, in "Protecting Groups", Georg
Thieme Verlag (1994), and any updated versions of said standard
works.
[0075] In the Methods below, unless otherwise specified, the
substituents are as defined above with reference to the compounds
of formula (I) above.
[0076] The compounds of formula (I), where W is N, can be prepared
according to the chemistry outlined in the scheme below: 4
[0077] The hydroxamic acid compounds of formula (I) where W is N
can be made by reaction of the corresponding activated acid
derivative of formula (II), where L is a suitable leaving group,
with hydroxylamine.
[0078] Suitable leaving groups are generally those which would
leave in a more efficient manner than the hydroxide of the parent
acid (IV), in a nucleophilic substitution reaction, such as a
halide, C.sub.1-4 alkoxide optionally substituted by halogen, an
alkylsulphonate such as methylsulphonate or mesylsulphonate, an
arylsulphonate such as p-tosylsulphonate. Other suitable leaving
groups are familiar to those working in the field of amino acid
coupling.
[0079] Such compounds of formula (II) may be made via standard
chemistry from the corresponding acid (IV). Compounds of formula
(II) where L is a leaving group such as Cl, Br, I,
OCO(C.sub.1-4alkyl optionally substituted by one or more halogen),
mesylate, tosylate, and the like, can be made from the
corresponding compound of formula (II) where L is OH by
conventional methods, including methods typified in e.g. Examples
2, etc.
[0080] The hydroxylamine used in this reaction is suitably
generated in situ by treatment of a hydroxylamine salt such as the
hydrochloride salt with a suitable base such as triethylamine.
Suitably the reaction is carried out in a polar solvent such as
DMF. This reaction, leaving groups, solvents, reagents, etc. are
exemplified below in Examples 1-4, 12-16, 18, 20-28, 30, 3340 and
41.
[0081] Alternatively the compounds of formula (I) may be made from
a NHO-protected compound of formula (III), where P is a suitable
O-protecting group, by suitable deprotection. Suitable O-protecting
groups can be found in the text by Greene and Wuts, supra, and
include trialkylsilyl (such as trimethylsilyl), benzyl, etc.
Compounds of formula (III) can be made in an analogous manner to
the compounds of formula (I) from the compounds of formula (II),
using a protected hydroxylamine PONH.sub.2 or a suitable salt
thereof in place of hydroxylamine itself or the hydroxylamine salt.
The deprotection method is determined by the protective group used,
as is well known in the art. E.g. benzyl groups may be
hydrogenated, suitably using a catalytic transfer hydrogenation
method. The reagents and conditions for reaction (III).gtoreq.(I)
are typified in Examples 29, 31, and 32 below, and also in the
other Examples where a protected hydroxylamine reagent (e.g.
O-trimethylsilylhydroxylami- ne) was used (e.g. Examples 2, etc.),
where conveniently the deprotection is carried out in the same
vessel as the previous step.
[0082] Other methods of making hydroxamic acids (I) are known and
may be used, e.g. those mentioned in the text by J.March, supra,
chapters 0-54, 0-57 and 6-4, and relevant references therein.
[0083] Acids of formula (IV) may be made by deprotection of the
O-protected species of formula (V). Suitable O-protecting groups
can be found in the chapter on O-protection in the book by Greene
and Wuts, supra, and include C.sub.1-4alkoxy such as t-butoxy (as
typified in Preparation 4), benzyloxy, trialkylsilyloxy such as
trimethylsilyloxy, etc. The deprotection method is determined by
the protective group used, as is well known in the art (see Greene
and Wuts, supra). E.g. benzyl groups may be removed by
hydrogenation, suitably using a catalytic transfer hydrogenation
method, t-butyl groups may be removed by treatment with an acid
such as trifluoroacetic acid, etc.
[0084] Compounds of formula (V), e.g. where P is a t-butoxy can be
made for example by condensation reaction of a corresponding
compound of formula (VI), for example by heating to elevated
temperature in an inert solvent such as in xylene at about
130.degree. C., this reaction being typified by Preparation 3
below.
[0085] Compounds of formula (VI) can be made for example by
coupling an acid of formula (VII) with a reagent of formula
C(NH.sub.2)(Y).dbd.NOH, which is available via literature methods
or adaptation thereof in a conventional manner, such as typified in
methods described herein (e.g. see Preparation 2, etc.). Typically
the condensation is carried out by adding a solution of the acid
(VII) in a suitable inert solvent such as 1,4-dioxane to a suitable
agent such as 1-hydroxybenzotriazole hydrate, followed by addition
of a suitable coupling agent such as a carbodiimide coupling agent,
e.g. N,N'-dicyclohexylcarbodiimide, then treatment with the reagent
C(NH.sub.2)(Y)=NOH. Suitably the coupling is carried out at ambient
temperature.
[0086] Compounds of formula (VII) can be made by hydrogenation of
the corresonding itaconate derivative, which in turn can be made by
conventional methods such as the Stobbe condensation. Certain
aspects of these preparations related to stereoselective
preparation of certain intermediates, such as are disclosed in
Preparation 1--Route C, are novel and inventive and constitute a
further aspect to this invention.
[0087] The compounds of formula (I), where W is CZ, can be prepared
according to the chemistry outlined in the scheme below: 5
[0088] The hydroxamic acid compounds of formula (I) where W is CZ
can be made by reaction of the corresponding activated acid
derivative of formula (IX), where L.sup.- is a suitable leaving
group, with hydroxylamine. The leaving groups, reagents, etc. are
the same as those mentioned above in relation to the corresponding
compounds of formula (I) where W is N.
[0089] Such compounds of formula (IX) may be made via standard
chemistry from the corresponding acdd (X) using the same or similar
chemistry to that outlined above in relation to the corresponding
compounds of formula (II) where W is N (supra).
[0090] Alternatively the compounds of formula (I) may be made from
a NHO-protected compound of formula (VIII), where P is a suitable
O-protecting group, by suitable deprotection. S Suitable
O-protecting groups can be found in the text by Greene and Wuts,
supra, and include trialkylsilyl (such as trimethylsilyl), benzyl,
etc.
[0091] Compounds of formula (VIII) can be made in an analogous
manner to the compounds of formula (II) from the compounds of
formula (II), using a protected hydroxylamine PONH.sub.2 or a
suitable salt thereof in place of hydroxylamine itself or the
hydroxylamine salt. The deprotection method is determined by the
protective group used, as is well known in the art. E.g. benzyl
groups may be hydrogenated, suitably using a catalytic transfer
hydrogenation method. The reagents and conditions for reaction
(VIII).fwdarw.(I) are typified in Examples 29, 31, and 32 below,
and also in the other Examples where a protected hydroxylamine
reagent (e.g. O-trimethylsilylhydroxylamine) was used (e.g.
Examples 2, etc.), where conveniently the deprotection is carried
out in the same vessel as the previous step.
[0092] Other methods of making hydroxamic acids (I) are known and
may be used, e.g. those mentioned in the text by J.March, supra,
chapters 0-54, 0-57 and 64, and relevant references therein.
[0093] Acids of formula (X) may be made by deprotection of the
O-protected species of formula (XI). Suitable O-protecting groups
can be found in the chapter on O-protection in the book by Greene
and Wuts, supra, and include C.sub.1-4alkoxy such as t-butoxy (as
typified in Preparation 4), benzyloxy, trialkylsilyloxy such as
trimethylsilyloxy, etc. The deprotection method is determined by
the protective group used, as is well known in the art (see Greene
and Wuts, supra). E.g. benzyl groups may be removed by
hydrogenation, suitably using a catalytic transfer hydrogenation
method, t-butyl groups may be removed by treatment with an acid
such as trifluoroacetic acid, etc.
[0094] Compounds of formula (XI), e.g. where P is a t-butoxy group
can be made for example by oxidation of a compound of formula
(XII). Suitably the oxidation is carried out using copper (II)
bromide with hexamethylenetetramine and a base such as DBU. The
reagents, conditions, etc. are typified in Preparation 62
below.
[0095] Compounds of formula (XII) may be made by condensation of
compounds of formula (XIII), for example by treatment of the
compound of formula (XIII) with s suitable agent such as Burgess
Reagent, in an anhydrous solvent such as THF. This reaction is
typified in Preparation 61 below.
[0096] Compounds of formula (XIII) may be made by condensation of
the acid of formula (II) above with an agent of formula
NH.sub.2CH(Y)CH(Z)OH, as typified in Preparation 60 below.
Compounds of formula NH.sub.2CH(Y)CH(Z)OH are available
commercially, from the literature or by routine modification
thereof.
[0097] Certain compounds of formula (I) may be interconverted into
other compounds of formula (I)--for Example where Y is an acid,
this can be converted to an ester and vice versa--typified in
Examples 17 and 19 below.
[0098] It will be apparent to those skilled in the art that other
protection and subsequent deprotection regimes during synthesis of
a compound of the invention may be achieved by conventional
techniques, for example as described in the volumes by Greene and
Wuts, and Kocienski, supra.
[0099] Where desired or necessary the compound of formula (I) is
converted into a pharmaceutically acceptable salt thereof. A
pharmaceutically acceptable salt of a compound of formula (I) may
be conveniently be prepared by mixing together solutions of a
compound of formula (I) and the desired acid or base, as
appropriate. The salt may be precipitated from solution and
collected by filtration, or may be collected by other means such as
by evaporation of the solvent.
[0100] Certain compounds of the invention may be interconverted
into certain other compounds of the invention by methods mentioned
in the Examples and Preparations, and well-known methods from the
literature.
[0101] Compounds of the invention are available by either the
methods described herein in the Methods, Examples and Preparations
or suitable adaptation thereof using methods known in the art. It
is to be understood that the synthetic transformation methods
mentioned herein may be carried out in various different sequences
in order that the desired compounds can be efficiently assembled.
The skilled chemist will exercise his judgement and skill as to the
most efficient sequence of reactions for synthesis of a given
target compound.
[0102] The compounds, salts, solvates and prodrugs of the invention
may be separated and purified by conventional methods.
[0103] Separation of diastereomers may be achieved by conventional
techniques, e.g. by fractional crystallisation, chromatography or
H.P.L.C. of a stereoisomeric mixture of a compound of formula (I)
or a suitable salt or derivative thereof. An individual enantiomer
of a compound of formula (I) may also be prepared from a
corresponding optically pure intermediate or by resolution, such as
by H.P.L.C. of the corresponding racemate using a suitable chiral
support or by fractional crystallisation of the diastereomeric
salts formed by reaction of the corresponding racemate with a
suitably optically active acid or base. In certain cases
preferential crystallisation of one of the enantiomers can occur
from a solution of a mixture of enantiomers, thus enriching the
remaining solution in the other enantiomer.
[0104] For human use, the compounds of formula (I) or their salts
can be administered alone, but will generally be administered in
admixture with a pharmaceutically acceptable diluent or carrier
selected with regard to the intended route of administration and
standard pharmaceutical practice. For example, they can be
administered orally, including sublingually, in the form of tablets
containing such excipients as starch or lactose, or in capsules or
ovules either alone or in admixture with excipients, or in the form
of elixirs, solutions or suspensions containing flavouring or
colouring agents. The compound or salt could be incorporated into
capsules or tablets for targetting the colon or duodenum via
delayed dissolution of said capsules or tablets for a particular
time following oral administration. Dissolution could be controlled
by susceptibility of the formulation to bacteria found in the
duodenum or colon, so that no substantial dissolution takes places
before reaching the target area of the gastrointestinal tract. The
compounds or salts can be injected parenterally, for example,
intravenously, intramuscularly or subcutaneously. For parenteral
administration, they are best used in the form of a sterile aqueous
solution or suspension which may contain other substances, for
example, enough salt or glucose to make the solution isotonic with
blood. They can be administered topically, or transdermally, in the
form of sterile creams, gels, suspensions, lotions, ointments,
dusting powders, sprays, foams, mousses, drug-incorporated
dressings, skin patches, ointments such as petrolatum or white soft
paraffin based ointments or via a skin patch or other device. They
could be administered directly onto a wound. They could be
incorporated into a coated suture. For example they can be
incorporated into a lotion or cream consisting of an aqueous or
oily emulsion of mineral oils; sorbitan monostearate; polysorbate
60; cetyl esters wax; cetearyl alcohol; 2-octyidodecanol; benzyl
alcohol; water; polyethylene glycols and/or liquid paraffin, or
they can be incorporated into a suitable ointment consisting of one
or more of the following--mineral oil; liquid petrolatum; white
petrolatum; propylene glycol; polyoxyethylene polyoxypropylene
compound; emulsifying wax and water, or as hydrogel with cellulose
or polyacrylate derivatives or other viscosity modifiers, or as a
dry powder or liquid spray or aerosol with butane/propane, HFA,
CFC, CO.sub.2 or other suitable propellant, optionally also
including a lubricant such as sorbitan trioleate, or as a
drug-incorporated dressing either as a tulle dressing, with white
soft paraffin or polyethylene glycols impregnated gauze dressings
or with hydrogel, hydrocolloid, alginate or film dressings. The
compound or salt could also be administered intraocularly for
ophthalmic use e.g. in a lens implant or as an eye drop with
appropriate buffers, viscosity modifiers (e.g. cellulose or
polyacrylate derivatives), preservatives (e.g. benzalkonium
chloride (BZK)) and agents to adjust tonicity (e.g. sodium
chloride). Such formulation techniques are well-known in the
art.
[0105] For certain uses, vaginal, rectal and nasal (e.g. by
inhalation of a dry powder or aerosol) administration would be
suitable.
[0106] All such formulations may also contain appropriate
stabilisers and preservatives.
[0107] For oral and parenteral administration to human patients,
the daily dosage level of the compounds of formula (I) or their
salts will be from 0.001 to 20, preferably from 0.01 to 20, more
preferably from 0.1 to 10, and most preferably from 0.5 to 5 mg/kg
(in single or divided doses). Thus tablets or capsules of the
compounds will contain from 0.1 to 500, preferably from 50 to 200,
mg of active compound for administration singly or two or more at a
time as appropriate.
[0108] For topical administration to human patients with
acute/surgical wounds or scars, the daily dosage level of the
compounds, in suspension or other formulation, could be from 0.01
to 50 mg/ml, preferably from 0.3 to 30 mg/ml.
[0109] The dosage will vary with the size of the wound, whether or
not the wound is open or closed or partially closed, and whether or
not the skin is intact.
[0110] The physician in any event will determine the actual dosage
which will be most suitable for a an individual patient and it will
vary with the age, weight and response of the particular patient.
The above dosages are exemplary of the average case; there can of
course be individual instances where higher or lower dosage ranges
are merited, and such are within the scope of this invention.
Biological Test Methods
[0111] PCP Inhibition
[0112] In order to determine potency of PCP inhibitors a
fluorogenic PCP cleavage assay was used. This assay is based on the
template of Beekman et al. (FEBS Letters (1996), 390: 221-225)
using a fluorogenic substrate. The substrate
(Dabcyl-Arg-Tyr-Tyr-Arg-Ala-Asp-Asp-Ala-Asn-Val-Glu(EDANS)--
-NH.sub.2) contains the cleavage site of human PCP (Hojima et al.,
J Biol Chem (1985), 260: 15996-16003). Human PCP has been purified
from supernatant of stable transfected CHO cells using hydrophobic
interaction column followed by Superdex 200 gel filtration. 4 .mu.g
total protein of this enzyme preparation was incubated with various
concentrations of the substance to be tested and 3.times.10.sup.-6
M substrate in assay buffer (50 mM Tris-Base, pH 7.6 containing 150
mM NaCl, 5 mM CaCl.sub.2, 1 .mu.M ZnCl.sub.2 and 0.01 % Brij 35).
The assay was performed in 96-well black fluorimeter plates and
fluorescence was read continuously in a fluorimeter over 2.5 hours
(.lambda..sub.ex=340 nm, .lambda..sub.em=485 nm) at a constant
37.degree. C. with shaking. Release of the fluorogenic signal was
in linear correlation to PCP activity. Reading of the mean velocity
from 30 min after start of experiment until 2.5 hours was
calculated by the Biolise software. IC.sub.50 values were
calculated by plotting % inhibition values against compound
concentration using Tessela add in for Excel spreadsheet.
[0113] MMP Inhibition
[0114] The ability of compounds to inhibit the cleavage of
fluorogenic peptides by MMPs 1, 2, 9, and 14 is described
below.
[0115] The assays for MMPs 2, 9, and 14 are based upon the original
protocol described by Knight et al. (Fed.Euro.Biochem.Soc., 296
(3), 263-266; 1992) with the slight modifications given below.
[0116] Inhibition of MMP-1
[0117] (i) Enzyme Preparation
[0118] Catalytic domain MMP-1 was prepared at Pfizer Central
Research. A stock solution of MMP-1 (1 .mu.M) was activated by the
addition of aminophenylmercuric acetate (APMA), at a final
concentration of 1 mM, for 20 minutes at 37.degree. C. MMP-1 was
then diluted in Tris-HCl assay buffer (50 mM Tris, 200 mM NaCl, 5
mM CaCl.sub.2, 20 .mu.M ZnSO.sub.4, 0.05% Brij 35) pH 7.5 to a
concentration of 10 nM. The final concentration of enzyme used in
the assay was 1 nM.
[0119] (ii) Substrate
[0120] The fluorogenic substrate used in this assay was Dnp-Pro-
-cyclohexyl-Ala-Gly-Cys(Me)-His-Ala-Lys(N-Me-Ala)-NH.sub.2as
originally described by Bickeft et al (Anal. Biochem, 212, 58-64,
1993). The final substrate concentration used in the assay was 10
.mu.M.
[0121] (iii) Determination of Enzyme Inhibition
[0122] Test compounds were dissolved in dimethyl sulphoxide and
diluted with assay buffer so that no more than 1% dimethyl
sulphoxide was present. Test compound and enzyme were added to each
well of a 96 well plate and allowed to equilibrate for 15 minutes
at 37.degree. C. in an orbital shaker prior to the addition of
substrate. Plates were then incubated for 1 hour at 37.degree. C.
prior to determination of fluorescence (substrate cleavage) using a
fluorimeter (Fluostar; BMG LabTechnologies, Aylesbury, UK) at an
excitation wavelength of 355 nm and emission wavelength of 440 nm.
The potency of inhibitors was measured from the amount of substrate
cleavage obtained using a range of test compound concentrations,
and, from the resulting dose-response curve, an IC.sub.50 value
(the concentration of inhibitor required to inhibit 50% of the
enzyme activity) was calculated.
[0123] Inhibition of MMP-2 and MMP-9
[0124] (i) Enzyme Preparation
[0125] Catalytic domain MMP-2 and MMP-9 were prepared at Pfizer
Central Research. A stock solution of MMP-2/MMP-9 (1 M) was
activated by the addition of aminophenylmercuric acetate (APMA).
For MMP-2 and MMP-9, a final concentration of 1 mM APMA was added,
followed by incubation for 1 hour at 37.degree. C. The enzymes were
then diluted in Tris-HCl assay buffer (100 mM Tris, 100 mM NaCl, 10
mM CaCl.sub.2 and 0.16% Brij 35, pH 7.5), to a concentration of 10
nM. The final concentration of enzyme used in the assays was 1
nM.
[0126] (ii) Substrate
[0127] The fluorogenic substrate used in this screen was
Mca-Arg-Pro-Lys-Pro-Tyr-Ala-Nva-Trp-Met-Lys(Dnp)NH.sub.2 (Bachem
Ltd, Essex, UK) as originally described by Nagase et al
(J.Biol.Chem., 269(33), 20952-20957, 1994). This substrate was
selected because it has a balanced hydrolysis rate against MMPs 2
and 9 (k.sub.cat/k.sub.m of 54,000, 59,400 and 55,300 s.sup.-1
M.sup.-1 respectively). The final substrate concentration used in
the assay was 5.mu.M.
[0128] (iii) Determination of Enzyme Inhibition
[0129] Test compounds were dissolved in dimethyl sulphoxide and
diluted with test buffer solution (as above) so that no more than
1% dimethyl sulphoxide was present. Test compound and enzyme were
added to each well of a 96 well plate and allowed to equilibrate
for 15 minutes at 37.degree. C. in an orbital shaker prior to the
addition of substrate. Plates were then incubated for 1 hour at
37.degree. C. prior to determination of fluorescence using a
fluorimeter (Fluostar; BMG LabTechnologies, Aylesbury, UK) at an
excitation wavelength of 328 nm and emission wavelength of 393 nm.
The potency of inhibitors was measured from the amount of substrate
cleavage obtained using a range of test compound concentrations,
and, from the resulting dose-response curve, an IC.sub.50 value
(the concentration of inhibitor required to inhibit 50% of the
enzyme activity) was calculated.
[0130] Inhibition of MMP-14
[0131] (i) Enzyme Preparation
[0132] Catalytic domain MMP-14 was purchased from Prof. Tschesche,
Department of Biochemistry, Faculty of Chemistry, University of
Bielefeld, Germany. A 10 M enzyme stock solution was activated for
20 minutes at 25.degree. C. following the addition of 5 g/ml of
trypsin (Sigma, Dorset, UK). The trypsin activity was then
neutralised by the addition of 50 g/ml of soyabean trypsin
inhibitor (Sigma, Dorset, UK), prior to dilution of this enzyme
stock solution in Tris-HCl assay buffer (100 mM Tris, 100 mM NaCl,
10 mM CaCl.sub.2 and 0.16% Brij 35, pH 7.5) to a concentration of
10 nM. The final concentration of enzyme used in the assay was 1
nM.
[0133] (ii) Substrate
[0134] The fluorogenic substrate used in this screen was
Mca-Pro-Leu-Gly-Leu-Dpa-Ala-Arg-NH.sub.2 (Bachem Ltd, Essex, UK) as
described by Will et al (J.Biol.Chem., 271(29), 17119-17123, 1996).
The final substrate concentration used in the assay was 10
.mu.M.
[0135] Determination of enzyme inhibition by test compounds was
performed in the same manner as described for MMPs-2 and -9
above.
[0136] The compounds of Examples 1-40 and 42-58 had PCP IC.sub.50
values of 0.5 .mu.M and below, and the compounds of Examples 140,
43, 44 and 46 had selectivities vs MMP-2 of more than 100-fold.
[0137] All references mentioned herein in this text are
incorporated by reference in their entirety.
EXAMPLES AND PREPARATIONS
[0138] Melting points were determined using open glass capillary
tubes and a Galtenkamp melting point apparatus and are uncorrected.
Nuclear magnetic resonance (NMR) data were obtained using Varian
Unity Inova-400, Varian Unity Inova-300 or Bruker AC300
spectrometers and are quoted in parts per million from
tetramethylsilane. Mass spectral (MS) data were obtained on a
Finnigan Mat. TSQ 7000 or a Fisons Instruments Trio 1000. The
calculated and observed ions quoted refer to the isotopic
composition of lowest mass. Infra red (IR) spectra were measured
using a Nicolet Magna 550 Fourier transform infra-red spectrometer.
Flash chromatography refers to column chromatography on silica gel
(Kieselgel 60, 230-400 mesh, from E. Merck, Darrnstadt. Kieselgel
60 F.sub.254 plates from E. Merck were used for TLC, and compounds
were visualised using UV light, 5% aqueous potassium permanagate or
Dragendorff's reagent (oversprayed with aqueous sodium nitrite).
Thermal analyses by Differential Scanning Calorimetry (DSC) and
ThermoGravimetric Analysis (TGA) were obtained using Perkin Elmer
DSC7 and TGA7. Moisture sorption characteristics were recorded
using Surface Measurement Systems Ltd. Automated Water Sorption
Analyser DVS 1. Water content was determined on a Mitsubishi CA100
(Coulometric Karl Fisher Titrator). Powder X-ray diffraction (PXRD)
pattern was determined using a Siemens D5000 powder X-ray
diffractometer fitted with an automatic sample changer, a
theta-theta goniometer, automatic beam divergence slits, a
secondary monochromator and a scintillation counter. Other
measurements were taken using standard equipment. Hexane refers to
a mixture of hexanes (hplc grade) b.p. 65-70.degree. C. Ether
refers to diethyl ether. Acetic acid refers to glacial acetic acid.
1-Hydroxy-7-aza-1H-1,2,3-benzotriazole (HOAt),
N-[(dimethylamino)-1H-1,2,3-triazolo[4,5-b]pyridin-1-ylmethylene]-N-methy-
lmethaninium hexafluorophosphate N-oxide (HATU) and
7-azabenzotriazol-1-yloxytris(pyrrolidino)phosphonium
hexafluorophosphate (PyAOP) were purchased from PerSeptive
Biosystems U.K. Ltd. "DIPE" refers to diisopropyl ether.
Reverse-phase silica gel for flash chromatography was obtained from
Fluka (Fluka 100, C.sub.18, 40-63.mu.). Pentane refers to High
Performance Liquid Chromatography (HPLC) grade n-pentane
(b.pt.35-37.degree. C.). Nomenclature has been allocated using a
program available from IUPAC. Standard abbreviations are used
throughout, e.g. "Me" is methyl, "Et" is ethyl, "Pr" is propyl,
"Ph" is phenyl, etc. It was noticed that during certain repetitions
of the methods disclosed in the Examples and Preparations that some
racemisation appeared to have taken place. It was found in some
cases that specific desired enantiomers can be separated from
mixtures thereof by routine methods such as by differential
crystallisation.
[0139] .sup.aHPLC autopurification performed using 2
columns--Phenomonex LUNA C8 150.times.21.2 mm, 10 .mu.m and
Phenomonex MAGELLEN C18 150.times.21.2 mm, 5 .mu.m, eluting with a
gradient system of organic solvent [ammonium acetate (aq) 100
mM:acetonitrile (1:9)]:aqueous solvent [ammonium acetate (aq) 100
mM: acetonitrile (9:1)]
[0140] .sup.bHPLC autopurification performed using 2
columns--Phenomonex LUNA C8 150.times.21.2 mm, 10 .mu.m and
Phenomonex MAGELLEN C18 150.times.21.2 mm, 5 .mu.m, eluting with a
gradient system of organic solvent ( acetonitrile) : aqueous
solvent (0.1% aqueous trifluoroacetic acid)
Example 1
Ethyl
5-{(1R)-4-cyclohexyl-1-[2-(hydroxyamino)-2-oxoethyobutyl}-1,2,4-oxad-
iazole-3-carboxylate
[0141] 6
[0142] A solution of
(3R)-6-cyclohexyl-3-[3-(ethoxycarbonyl)-1,2,4-oxadiaz-
ol-5-yl]hexanoic acid (Preparation 4) (140 mg, 0.41 mmol) and
N,N-diisopropylethylamine (320 .mu.l, 2.07mmol) in
N,N-dimethylformamide (5 ml) was treated with
O-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethylu- ronium
hexafluorophosphate (235 mg, 0.61 mmol) and the resulting solution
was stirred at room temperature under a nitrogen atmosphere for 30
minutes. Hydroxylamine hydrochloride (113 mg, 0.61 mmol) was then
added and the mixture was stirred at room temperature for 20 hours.
The mixture was poured into hydrochloric acid (1 M, 20 ml) and
extracted with ethyl acetate (.times.3). The combined organic
layers were washed sequentially with a saturated aqueous solution
of sodium hydrogen carbonate and brine, dried over anhydrous sodium
sulphate, filtered and the solvent removed under reduced pressure.
The residue was purified by column chromatography on silica gel
eluting with a gradient system of dichloromethane:methanol
(99.5:0.5) gradually changing to dichloromethane:methanol (97:3) to
afford the title compound as a yellow oil (62 mg).
[0143] MS:354 (MH.sup.+)
[0144] .sup.1H-NMR (CDCl.sub.3) .delta.: 4.52 (2H, m), 3.70 (1 H,
br m), 2.94-2.52 (2H, br m), 1.96-1.00 (18H, m), 0.85 (2H, m).
Example 2 (a)
5-{(1
R)-4-Cyclohexyl-1-[2-(hydroxyamino)-2-oxoethyl]butyl}-1,2,4-oxadiazo-
le-3-carboxamide
[0145] 7
[0146] A solution of
(3R)-3-[3-(aminocarbonyl)-1,2,4-oxadiazol-5-yl]-6-cyc-
lohexylhexanoic acid (Preparation 6) (6.00 g, 19.42 mmol) in
anhydrous tetrahydrofuran (200 ml) was cooled to 0.degree. C. then
treated with N-methylmorpholine (2.40 ml, 22.0 mmol) and isobutyl
chloroformate (2.8 ml, 22.0 mmol) and stirred under a nitrogen
atmosphere for 1 hour. O-(Trimethylsilyl)hydroxylamine (7.0 ml,
60.0 mmol) was added and the mixture was stirred for 18 hours,
being allowed to warm to room temperature over this time. The
mixture was then treated with methanol (50 ml) and stirred for a
further 30 minutes. This mixture was partitioned between ethyl
acetate and water. The combined organic layers were washed with
brine, dried over anhydrous sodium sulphate, filtered and the
solvent removed under reduced pressure. The solid was
recrystallised from isopropyl acetate (500 ml) to afford the title
compound as a white solid (4.46 g). Chiral HPLC analysis indicated
this material to be 87.6% ee. A sample of this material (2.9 g) was
dissolved in ethyl acetate (450 ml) at reflux then allowed to cool.
The precipitate was filtered off and the solvent was removed from
the filtrate under reduced pressure to afford a white solid, which
was then recrystallised from isopropyl acetate (120 ml) to afford
the title compound (1.42 g). Chiral HPLC analysis showed this
material to be 98.3% ee.
[0147] MS: 323 (MH.sup.-)
[0148] .sup.1H-NMR (DMSO-d.sub.6) .delta.: 10.50 (1H, br s), 8.81
(1 H, br s), 8.26 (1H, br s), 8.05 (1H, br s), 3.47 (1H, m),
2.56-2.39 (2H, m), 1.73-1.46 (7H, m), 1.27-0.99 (8H, m), 0.80 (2H,
m).
[0149] Analysis: Found C, 52.76; H, 7.64; N, 16.34%;
C.sub.15H.sub.24N.sub.4O.sub.4.H.sub.2O requires C, 52.62; H, 7.65;
N, 16.36%
[0150] MPt.: 136-138.degree. C.
Example 2(b)
5-{(1R)4-Cyclohexyl-1-[2-(hydroxyamino)-2-oxoethyl]butyl}-1,2,4-oxadiazole-
-3-carboxamide monohydrate
[0151] 8
[0152] A solution of
5-{(1R)-4-cyclohexyl-1-[2-(hydroxyamino)-2-oxoethyl]b-
utyl}-1,2,4-oxadiazole-3-carboxamide (1.58 Kg, 4.86 mol) in
tetrahydrofuran (15 litres) was heated to 40.degree. C. and
demineralised water (26 litres) was added to give a hazy solution.
The mixture was allowed to cool to ambient temperature where it was
stirred for 24 hours. The mixture was cooled to 5.degree. C. and
stirred for 1 hour. The precipitate was collected by filtration and
dried in vacuo (38.degree. C., 100 mbar) to afford the title
compound as a colourless solid (1.37 Kg).
[0153] .sup.1H-NMR (DMSO-d6) .delta.: 10.50 (1H, br s), 8.81 (1H,
br s), 8.26 (1H, br s), 3.47 (1H, m), 2.56-2.39 (2H, m), 1.73-1.46
(7H, m), 1.27-0.99 (8H, m), 0.80 (2H, m)
[0154] Water Content (Coulometric Karl Fisher): 5.0%
[0155] Dynamic Vapour Sorption: 5.17% w/w @20% RH (30.degree. C.),
4.84% w/w @2% RH (30.degree. C.), significant dehydration below 1%
RH (30.degree. C.).
2 TGA (Weight Loss): 24.degree. C.-117.degree. C. = 4.93%
117.degree. C.-160.degree. C. = 0.26%
Example 2(c)
5-{(1R)-4-Cyclohexyl-1-[2-(hydroxyamino)-2-oxoethyl]butyl}-1,2,4-oxadiazol-
e-3-carboxamide (anhydrous)
[0156] 9
[0157]
5-{(1R)-4-Cyclohexyl-1-[2-(hydroxyamino)-2-oxoethyl]butyl}-1,2,4-ox-
adiazole-3-carboxamide (3.20 g, 9.86 mmol) was dissolved in ethyl
acetate (80 ml) at reflux. The mixture was allowed to cool to
ambient temperature, stirred for 3 hours. The precipitate was
collected by filtration and dried in vacuo (40-45.degree. C., 48
hours) to afford the title compound as a colourless solid (2.40
g).
[0158] .sup.1H-NMR (DMSO-d.sub.6) .delta.: 10.50 (1H, br s), 8.81
(IH, br s), 8.26 (1H, br s), 3.47 (1H, m), 2.56-2.39 (2H, m),
1.73-1.46 (7H, m), 1.27-0.99 (8H, m), 0.80 (2H, m)
[0159] Dynamic Vapour Sorption: 0.18% w/w @15% RH (30.degree. C.),
0.23% w/w @30% RH (30.degree. C.), 0.34% w/w @45% RH (30.degree.
C.). Title compound hydrates slowly at 60% RH (30.degree. C.)
converting to the monohydrate (XRD used to confirm), eventually
equilibrates at 5.3% w/w at 90% RH (30.degree. C.).
3 TGA (Weight Loss): 25.degree. C.-91.degree. C. = 0.22% 91.degree.
C.-138.degree. C. = 0.33%
Example 3
5-{(1R)-4-Cyclohexyl-1-[2-(hydroxyamino)-2-oxoethyl]butyl}-N-methyl-1,2,4--
oxadiazole-3-carboxamide
[0160] 10
[0161] A solution of
(3R)-6-cyclohexyl-3-{3-[(methylamino)carbonyl]-1,2,4--
oxadiazol-5-yl}hexanoic acid (Preparation 8) (285 mg, 0.88 mmol)
and N-methylmorpholine (110 .mu.l, 1.00 mmol) in anhydrous
tetrahydrofuran (10 ml) was cooled to 0.degree. C., treated with
isobutyl chloroformate (130 .mu.l, 1.00 mmol) and stirred under a
nitrogen atmosphere for 1 hour. O-(Trimethylsilyl)hydroxylamine
(130 .mu.l, 1.06 mmol) was added and the mixture was stirred for 18
hours, being allowed to warm to room temperature over this time.
The mixture was then treated with aqueous citric acid solution (10%
w/v, 10 ml) and stirred for 2 hours. This mixture was diluted with
water and extracted with ethyl acetate (.times.3). The combined
organic layers were washed sequentially with water and brine, dried
over anhydrous sodium sulphate, filtered and the solvent removed
under reduced pressure. The solid was then recrystallised from
hexane: ethyl acetate to afford the title compound as a white solid
(130 mg).
[0162] .sup.1H-NMR (DMSO-d.sub.6) .delta.: 10.48 (1H, br s), 8.87
(1H, m), 8.77 (1H, br s), 3.48 (1H, m), 3.19 (2H, m), 2.16 (1H, m),
1.87 (1H, m), 1.57 (9H, m), 1.14 (8H, m), 0.80 (5H, m).
Example 4
5{-(1R)-4-Cyclohexyl-1-[2-(hydroxyamino)-2-oxoethyl]butyl}-N-propyl-1,2,4--
oxadiazole-3-carboxamide
[0163] 11
[0164] A solution of
(3R)-6-cyclohexyl-3-{3-[(propylamino)carbonyl]-1,2,4--
oxadiazol-5-yl}hexanoic acid (Preparation 9) (66 mg, 0.18 mmol) and
N,N-diisopropylethylamine (31 .mu.l, 0.18 mmol) in
N-methyl-2-pyrrolidinone (3 ml) was treated with
O-(7-azabenzotriazol-1-y- l)-N,N,N',N'-tetramethyluronium
hexafluorophosphate (102 mg, 0.27 mmol) and the resulting mixture
was stirred at room temperature under a nitrogen atmosphere for 1
hour. N,N-diisopropylethylamine (93 .mu.l, 0.54 mmol) was then
added, followed by hydroxylamine hydrochloride (37.5 mg, 0.54 mmol)
and the mixture was stirred at room temperature for 18 hours. The
mixture was partitioned between ethyl acetate and pH 7 aqueous
buffer. The combined organic layers were washed sequentially with
aqueous citric acid solution (5% w/v) and brine, dried over
anhydrous sodium sulphate, filtered and the solvent removed under
reduced pressure. The residue was purified by column chromatography
on silica gel eluting with a gradient system of
dichloromethane:ethyl acetate (90:10) gradually changing to
dichloromethane:ethyl acetate (0:100) then to
dichloromethane:methanol (95:5) to afford the title compound as an
oil (31 mg).
[0165] MS: 367(MH.sup.+)
[0166] .sup.1H-NMR (DMSO-d.sub.6).delta.: 10.48(1H, brs),8.87(1H,
br t),8.76 (1H, brs), 3.47 (1H, m), 3.19 (2H, m), 2.16 (1H, m),
1.88 (1H, m), 1.76-1.43 (9H, m), 1.30-1.00 (8H, m), 0.90-70 (5H,
m).
Example 5
5-{(1R)-4-Cyclohexyl-1-[2-(hydroxyamino)-2-oxoethyl]butyl}N-isopropyl-1,2,-
4-oxadiazole-3-carboxamide
[0167] 12
[0168] A solution of
(3R)-6-cyclohexyl-3-[3-(ethoxycarbonyl)-1,2,4-oxadiaz-
ol-5-yl]hexanoic acid (Preparation 4) (500 mg, 1.47 mmol) in
ethanol (13 ml) was treated with a solution of isopropylamine (434
mg, 7.35 mmol) in ethanol (2 ml) and the resulting mixture was
heated at 80.degree. C. under a nitrogen atmosphere for 10 hours.
The solvent was removed under reduced pressure and the residue was
dissolved in hydrochloric acid (1M, 10 ml) then extracted with
ethyl acetate (.times.2). The combined organic layers were dried
over anhydrous magnesium sulphate, filtered and the solvent removed
under reduced pressure. This residue (531 mg) was then dissolved in
dichloromethane (15 ml), cooled to 0.degree. C. and treated
sequentially with N-methylmorpholine (180 .mu.l, 1.63 mmol) and
isobutyl chloroformate (210 .mu.l, 1.62 mmol). This mixture was
stirred at 0.degree. C. under a nitrogen atmosphere for 1 hour,
then treated with O-(trimethylsilyl)hydroxylamine (220 .mu.l, 1.80
mmol) and stirring continued for 10 minutes at 0.degree. then for
17 hours at room temperature. The mixture was then treated with
trifluoroacetic acid:water (5 ml, 9:1) and the solution stirred for
30 minutes. The solvent was removed under reduced pressure and the
residue azeotroped from toluene (.times.2). The residue was
purified by reverse phase HPLC.sup.a to afford the title compound
as a foam (43 mg).
[0169] MS: 367(MH.sup.+), 389 (MNa.sup.+)
[0170] .sup.1H-NMR (CDCl.sub.3) .delta.: 9.06 (1H, br s), 7.63 (1H,
br s), 6.74 (1H, br d),4.27 (1H, m), 3.70 (1H, m), 2.83-2.60 (2H,
m), 1.78-1.52 (7H, m), 1.35-1.07 (14H, m), 0.83 (2H, m).
Example 6
5-{(1R)-4-Cyclohexyl-1-[2-(hydroxyamino)-2-oxoethyl]butyl}-N-(cyclopropylm-
ethyl)-1,2,4-oxadiazole-3-carboxamide
[0171] 13
[0172] A solution of
(3R)-6-cyclohexyl-3-[3-(ethoxycarbonyl)-1,2,4-oxadiaz-
ol-5-yl]hexanoic acid (Preparation 4) (500 mg, 1.47 mmol) in
ethanol (13ml) was treated with a solution of
cyclopropylmethylamine (522 mg, 7.35 mmol) in ethanol (2 ml) and
the resulting solution was heated at 80.degree. C. under a nitrogen
atmosphere for 10 hours. The solvent was removed under reduced
pressure and the residue was dissolved in hydrochloric acid (1M, 10
ml) then extracted with ethyl acetate (.times.2). The combined
organic layers were dried over anhydrous magnesium sulphate,
filtered and the solvent removed under reduced pressure. This
residue (349 mg) was then dissolved in dichloromethane (15 ml),
cooled to 0.degree. C. and treated sequentially with
N-methylmorpholine (120 .mu.l, 1.09 mmol) and isobutyl
chloroformate (140 .mu.l, 1.09 mmol). This mixture was stirred at
0.degree. C. under a nitrogen atmosphere for 1 hour, then treated
with O-(trimethylsilyl)hydro- xylamine (140 .mu.l, 1.15 mmol) and
stirring continued for 10 minutes at 0.degree. then for 17 hours at
room temperature. The mixture was then treated with trifluoroacetic
acid:water (5 ml, 9:1) and the solution stirred for 30 minutes. The
solvent was removed under reduced pressure and the residue
azeotroped from toluene (.times.2). The residue was purified by
HPLC.sup.a to afford the title compound as a foam (88 mg).
[0173] MS: 379 (MH.sup.+), 401 (MNa.sup.+)
[0174] .sup.1H-NMR (CDCl.sub.3) .delta.: 9.20 (1H, br s), 7.05 (1H,
br s), 3.72 (1H, m), 3.33 (2H, m), 2.87- 2.60 (2H, m), 1.88-1.52
(7H, m), 1.40-1.00 (8H, m), 0.83 (3H, m), 0.59 (2H, m), 0.30 (2H,
m).
Example 7
N-Cyclobutyl-5-{1R)-4-cyclohexyl-1-[2-(hydroxyamino)-2-oxoethyl]butyl}-1,2-
,4-oxadiazole-3-carboxamide
[0175] 14
[0176] A solution of
(3R)-6-cyclohexyl-3-[3-(ethoxycarbonyl)-1,2,4-oxadiaz-
ol-5-yl]hexanoic acid (Preparation 4) (500 mg, 1.47 mmol) in
ethanol (13 ml) was treated with a solution of cyclobutylamine (522
mg, 7.35 mmol) in ethanol (2 ml) and the resulting solution was
heated at 80.degree. C. under a nitrogen atmosphere for 10 hours.
The solvent was removed under reduced pressure and the residue was
dissolved in hydrochloric acid (1M, 10 ml) then extracted with
ethyl acetate (.times.2). The organic layers were combined, dried
over anhydrous magnesium sulphate, filtered and the solvent removed
under reduced pressure. This residue (462 mg) was then dissolved in
dichloromethane (15 ml), cooled to 0.degree. C. and treated
sequentially with N-methylmorpholine (150 .mu.l, 1.36 mmol)and
isobutyl chloroformate (180 .mu.l, 1.39 mmol). This mixture was
stirred at 0.degree. C. under a nitrogen atmosphere for 1 hour,
then treated with O-(trimethylsilyl)hydroxylamine (190 .mu.l, 1.55
mmol) and stirring continued for 10 minutes at 0.degree. then for
17 hours at room temperature. The mixture was then treated with
trifluoroacetic acid:water (5 ml, 9:1) and the solution stirred for
30 minutes. The solvent was removed under reduced pressure and the
residue azeotroped from toluene (.times.2). The residue was
purified by HPLCA to afford a residue (63 mg) which was dissolved
in dichloromethane (3 ml), cooled to 0.degree. C. and treated
sequentially with N-methylmorpholine (20 .mu.l, 0.18 mmol)and
isobutyl chloroformate (23 .mu.l, 0.18 mmol). This mixture was
stirred at 0.degree. C. under a nitrogen atmosphere for 1 hour,
then treated with O-(trimethylsilyl)hydroxylamine(24 1.mu.l, 0.20
mmol) and stirring continued for 3 hours. Further
O-(trimethylsilyl)hydroxylamine(30 .mu.l, 0.25 mmol) was added and
stirring continued for 17 hours. The reaction was quenched with
methanol and the solvent removed under reduced pressure. The
residue was dissolved in ethyl acetate, washed with water, dried
over anhydrous sodium sulphate, filtered and the solvent removed
under reduced pressure. The residue was then dissolved in methanol
(5 ml), treated with potassium carbonate (110 mg) and stirred at
room temperature for 17 hours. The mixture was treated with a few
drops of acetic acid and the solvent removed under reduced
pressure. The residue was partitioned between ethyl acetate and
water. The organic phase was separated, dried over anhydrous sodium
sulphate, filtered and the solvent removed under reduced pressure
to afford the title compound as a foam (24 mg)
[0177] MS :379 (MH.sup.+), 401 (MNa.sup.+)
[0178] .sup.1H-NMR (CDCl.sub.3) .delta.: 8.80 (1H, br s), 7.06 (1H,
br d), 4.57 (1H, m), 3.70 (1H, m), 2.85-2.54 (2H, m), 2.43 (1H, m),
2.05 (1H, m), 1.87-1.41 (11 H, m), 1.38-0.98 (8H, m), 0.84 (2H,
m).
Example 8
5-{(1R)-4-Cyclohexyl-1-[2-(hydroxyamino)-2-oxoethyl]butyl}-N-(2-methoxyben-
zyl)-1,2,4-oxadiazole-3-carboxamide
[0179] 15
[0180] A solution of
(3R)-6-cyclohexyl-3-[3-(ethoxycarbonyl)-1,2,4-oxadiaz-
ol-5-yl]hexanoic acid (Preparation 4) (500 mg, 1.47 mmol) in
ethanol (13ml) was treated with a solution of 2-methoxybenzylamine
(1.00 g, 7.35 mmol) in ethanol (2 ml) and the resulting solution
was heated at 80.degree. C. under a nitrogen atmosphere for 10
hours. The solvent was removed under reduced pressure and the
residue was dissolved in hydrochloric acid (1M, 10 ml) then
extracted with ethyl acetate (.times.2). The combined organic
layers were dried over anhydrous magnesium sulphate, filtered and
the solvent removed under reduced pressure. This residue (672 mg)
was then dissolved in dichloromethane (15 ml), cooled to 0.degree.
C. and treated sequentially with N-methylmorpholine (180 .mu.l,
1.64 mmol) and isobutyl chloroformate (210 g, 1.63 mmol). This
mixture was stirred at 0.degree. C. under a nitrogen atmosphere for
1 hour, then treated with O-(trimethylsilyl)hydroxylamine (220
.mu.l, 1.80 mmol) and stirring continued for 10 minutes at
0.degree. then for 17 hours at room temperature. The mixture was
then treated with trifluoroacetic acid:water (5 ml, 9:1) and the
solution stirred for 30 minutes. The solvent was removed under
reduced pressure and the residue azeotroped from toluene
(.times.2). The residue was purified by HPLC.sup.a to afford the
title compound as a foam (140 mg).
[0181] MS: 445 (MH.sup.+), 462 (MNH.sub.4.sup.+)
[0182] .sup.1H-NMR (CDCl.sub.3) .delta.: 7.44 (1H, br s), 7.29 (2H,
m), 6.91 (2H, m), 4.63 (2H, d, J=5Hz), 3.89 (3H, s), 3.71 (1H, m),
2.86-2.54 (2H, m), 1.90-1.48 (7H, m), 1.39-1.00 (8H, m), 0.83 (2H,
m).
Example 9
5-{(1R)4-Cyclohexyl-1-[2-(hydroxyamino)-2-oxoethyl]butyl}-N-(4-methoxybenz-
yl)-1,2,4-oxadiazole-3-carboxamide
[0183] 16
[0184] A solution of (3R)-6-cyclohexyl-3-[3-(ethoxycarbonyl)-1
,2,4-oxadiazol-5-yl]hexanoic acid (Preparation 4) (500 mg, 1.47
mmol) in ethanol (13 ml) was treated with a solution of
4-methoxybenzylamine (1.00 g, 7.35 mmol) in ethanol (2 ml) and the
resulting solution was heated at 80.degree. C. under a nitrogen
atmosphere for 10 hours. The solvent was removed under reduced
pressure and the residue was dissolved in hydrochloric acid (1M, 10
ml) then extracted with ethyl acetate (.times.2). The combined
organic layers were dried over an hydrous magnesium sulphate,
filtered and the solvent removed under reduced pressure. This
residue (578 mg) was then dissolved in dichloromethane (15 ml),
cooled to 0.degree. C. and treated sequentially with
N-methylmorpholine (160 .mu.l, 1.45 mmol) and isobutyl
chioroformate (190 .mu.l, 1.47 mmol). This mixture was stirred at
0.degree. under a nitrogen atmosphere for 1 hour, then treated with
O-(trimethylsilyl)hydroxylamine (195 .mu.l, 1.60 mmol) and stirring
continued for 10 minutes at 0.degree. then for 17 hours at room
temperature. The mixture was then treated with trifluoroacetic
acid:water (ml, 9:1) and the solution stirred for 30 minutes. The
solvent was removed under reduced pressure and the residue
azeotroped from toluene (.times.2). The residue was purified by
HPLC.sup.a to afford the title compound as a foam (74 mg).
[0185] MS: 445 (MH.sup.+), 462 (MNH.sub.4.sup.+)
[0186] .sup.1H-NMR (CD.sub.3OD) .delta.: 7.37 (2H, d, J=8Hz), 6.89
(2H, d, J=8Hz), 4.50 (2H, s), 3.76 (3H, s), 3.60 (1H, m), 2.73-2.50
(2H, m), 1.86-1.66 (7H, m), 1.40-1.09 (8H, m), 0.85 (2H, m).
Example 10
5-{(1R)-4-Cyclohexyl-1-[2-(hydroxyamino)-2-oxoethyl]butyl}-N-(2-pyridinylm-
ethyl)-1,2,4-oxadiazole-3-carboxamide
[0187] 17
[0188] A solution of (3R)-6-cyclohexyl-3-[3-(ethoxycarbonyl)-1
,2,4-oxadiazol-5-yl]hexanoic acid (Preparation 4) (500 mg, 1.47
mmol) in ethanol (13 ml) was treated with a solution of
2-aminomethylpyridine (794 mg, 7.35 mmol) in ethanol (2 ml) and the
resulting solution was heated at 80.degree. C. under a nitrogen
atmosphere for 10 hours. The solvent was removed under reduced
pressure. The residue was dissolved in hydrochloric acid (1M, 10
ml), neutral ised to pH 4 with a saturated aqueous solution of
ammonium hydroxide then extracted with ethyl acetate (.times.2).
The combined organic layers were dried over anhydrous magnesium
sulphate, filtered and the solvent removed under reduced pressure.
This residue (467 mg) was then dissolved in dichloromethane (15
ml), cooled to 0.degree. C. and treated sequentially with
N-methylmorpholine (140 .mu.l, 1.27 mmol) and isobutyl
chloroformate (165 .mu.l , 1.28 mmol). This mixture was stirred at
0.degree. C. under a nitrogen atmosphere for I hour, then treated
with O-(trimethylsilyl)hydroxylamine (170 .mu.l, 1.39 mmol) and
stirring continued for 10 minutes at 0.degree. then for 17 hours at
room temperature. The mixture was then treated with trifluoroacetic
acid:water (5 ml, 9:1) and the solution stirred for 30 minutes. The
solvent was removed under reduced pressure and the residue
azeotroped from toluene (.times.2). The residue was purified by
reverse phase HPLC.sup.b to afford a solid (130 mg) which was
dissolved in dichloromethane (5 ml), cooled to 0.degree. C. and
treated sequentially with N-methylmorpholine (33 .mu.l, 0.30
mmol)and isobutyl chloroformate (38 .mu.l, 0.30 mmol). This mixture
was stirred at 0.degree. C. under a nitrogen atmosphere for 1 hour,
then treated with O-(trimethylsilyl)hydro- xylamine (40 .mu.l, 0.32
mmol) and stirring continued for 3 hours. The reaction was quenched
with methanol and the solvent removed under reduced pressure. The
residue was dissolved in ethyl acetate, washed with water, dried
over anhydrous sodium sulphate, filtered and the solvent removed
under reduced pressure. The residue was dissolved in methanol (5
ml), treated with potassium carbonate (110 mg, 0.80 mmol) and
stirred at room temperature for 17 hours. The mixture was treated
with a few drops of acetic acid and the solvent removed under
reduced pressure. The residue was partitioned between ethyl acetate
and water. The organic phase was separated, dried over anhydrous
sodium sulphate, filtered and the solvent removed under reduced
pressure. The residue was purified by HPLCI to afford the title
compound as a solid (34 mg).
[0189] MPt.:128-130.degree. C.
[0190] MS: 416 (MH.sup.+)
[0191] .sup.1H-NMR (CDCl.sub.3) .delta.: 8.50 (2H, m), 7.72 (1H,
dd, J=7, 7Hz), 7.40-7.18 (2H, m), 4.74 (2H, d, J=5Hz), 3.67 (1H,
m), 2.83-2.50 (2H, m), 1.86-1.42 (7H, m), 1.40-0.97 (8H, m), 0.80
(2H, m).
Example 11
2-{[(5-{(1R)-4-Cyclohexyl-1-[2-(hydroxyamino)-2-oxoethyl]butyl}-1,2,4-oxad-
iazol-3-yl)carbonyl]amino}acetic acid
[0192] 18
[0193] A solution of
(3R)-6-cyclohexyl-3-[3-(ethoxycarbonyl)-1,2,4-oxadiaz-
ol-5-yl]hexanoic acid (Preparation 4) (527 mg, 1.56 mmol) in
ethanol (20 ml) was treated with triethylamine (508 mg, 5.02 mmol)
and glycine t-butyl ester hydrochloride (743 mg, 4.43 mmol) and the
resulting solution was heated at 80.degree. C. under a nitrogen
atmosphere for 30 hours. The solvent was removed under reduced
pressure and the residue was dissolved in hydrochloric acid (1M, 20
ml) then extracted with ethyl acetate (.times.2). The combined
organic layers were dried over anhydrous sodium sulphate, filtered
and the solvent removed under reduced pressure. The residue was
purified by column chromatography on silica gel eluting with
dichloromethane: ethyl acetate (1:1) then dichloromethane: methanol
(19:1). This residue (430 mg, 1.02 mmol) was dissolved in
dichloromethane (10 ml), cooled to 0.degree. C. and treated
sequentially with N-methylmorpholine (130 .mu.l, 1.20 mmol) and
isobutyl chloroformate (150 .mu.l, 1.20 mmol). This mixture was
stirred at 0.degree. C. under a nitrogen atmosphere for 1 hour,
then treated with O-(trimethylsilyl)hydro- xylamine (160 .mu.l,
1.30 mmol). The mixture was stirred for 17 hours being allowed to
warm to room temperature over this time. The mixture was diluted
with dichloromethane, washed with aqueous citric acid solution (10%
w/v), dried over anhydrous magnesium sulphate, filtered and the
solvent removed under reduced pressure. The residue was dissolved
in dichloromethane (10 ml), treated with trifluoroacetic acid (10
ml) and the solution was stirred for 2.5 hours at room temperature.
The solvent was removed under reduced pressure and the residue
azeotroped from toluene (.times.2). The residue was purified by
HPLC.sup.b to afford the title compound as a white solid (350
mg).
[0194] MPt.:141-142.degree. C.
[0195] MS: 383 (MH.sup.+), 405 (MNa.sup.+)
[0196] Analysis: Found C, 50.80; H, 7.00; N, 13.90%;
C,.sub.7H.sub.26N.sub.40.sub.6.H.sub.20 requires C, 50.99; H, 7.05;
N, 13.99%
[0197] .sup.1H-NMR (CD.sub.3OD) .delta.:4.10 (2H, s), 3.63 (1H, m),
2.72-2.51 (2H, m), 1.85-1.60 (7H, m), 1.38-1.10 (8H, m), 0.86 (2H,
m).
Example 12
5-{(1R)-4-Cyclohexyl-1-[2-(hydroxyamino)-2-oxoethyl]butyl}-N,N-dimethyl-1,-
2,4-oxadiazole-3-carboxamide
[0198] 19
[0199] A solution of
(3R)-6-cyclohexyl-3-{3-[(dimethylamino)carbonyl]-1,2,-
4-oxadiazol-5-yl}hexanoic acid (Preparation 11) (60 mg, 0.18 mmol)
and N,N-diisopropylethylamine (30 .mu.l, 0.18 mmol) in
N-N-dimethylformamide (3 ml) was treated with
O-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethylu- ronium
hexafluorophosphate (100 mg, 0.27 mmol) and the resulting mixture
was stirred at room temperature under a nitrogen atmosphere for 1
hour. Further N,N-diisopropylethylamine (90 .mu.l, 0.52 mmol) was
then added, followed by hydroxylamine hydrochloride (35 mg, 0.52
mmol) and the mixture was stirred at room temperature for 18 hours.
The mixture was partitioned between ethyl acetate and pH 7 aqueous
buffer. The layers were separated and the aqueous phase extracted
with ethyl acetate (.times.2). The combined organic layers were
washed sequentially with aqueous citric acid solution (5% w/v) and
brine, dried over anhydrous sodium sulphate, filtered and the
solvent removed under reduced pressure. The residue was purified by
column chromatography on silica gel eluting with a gradient system
of dichloromethane:ethyl acetate (90:10 ) gradually changing to
dichloromethane:ethyl acetate (50:50) then to dichloromethane:
methanol (95:5). The residue was further purified by column
chromatography on silica gel eluting with a gradient system of
dichloromethane: methanol (99:1) gradually changing to
dichloromethane:methanol (95:5) to afford the title compound as an
oil which crystallised on standing (35 mg).
[0200] MS: 353 (MH.sup.+)
[0201] .sup.1H-NMR (DMSO-d.sub.6) .delta.: 10.50 (1H, br s), 8.81
(1H, br s), 3.48 (1H, m), 3.03 (3H, s), 2.93 (3H, s), 2.50 (2H, m),
1.76-1.52 (7H, m), 1.32-1.03 (8H, m), 0.80 (2H, m).
Example 13
(3R)-6-Cyclohexyl-3-(3-{[4-(dimethylamino)-1-piperidinyl]carbonyl}-1,2,4-o-
xadiazol-5-yl)N-hydroxyhexanamide
[0202] 20
[0203] A solution of (3R)-6-cyclohexyl-3-(3-{[4-(dimethylaminoy1
-piperidinyl]carbonyl}-1,2,4-oxadiazol-5-yl)hexanoic acid
trifluoroacetate (Preparation 26) (702 mg, 1.31 mmol) and
N-methylmorpholine (159 .mu.l, 1.45 mmol) in N,N-dimethylformamide
(15 ml) was cooled to 0.degree. C. and then treated with dropwise
with isobutyl chloroformate (188 .mu.l, 1.45 mmol). The resulting
solution was stirred at 0.degree. C. under a nitrogen atmosphere
for 45 minutes. Hydroxylamine hydrochloride (274 mg, 3.94 mmol) was
then added followed by further N-methylmorpholine (433 .mu.l, 3.94
mmol) and the mixture was stirred for 18 hours being allowed to
warm to room temperature over this time. The solvent was removed
under reduced pressure and the residue was dissolved in a saturated
aqueous solution of sodium carbonate. The pH of the solution was
adjusted to 9 by dropwise addition of acetic acid and the product
was then extracted with ethyl acetate (.times.2). The combined
organic phases were washed with brine, dried over anhydrous sodium
sulphate, filtered and the solvent removed under reduced pressure.
The residue was purified by column chromatography on silica gel
eluting with a gradient system of dichloromethane:methanol:0.88
ammonia (90:10:1) then dichloromethane:methanol:0.88 ammonia
(80:20:2) to afford the title compound as an oil (23 mg)
[0204] MS: 436 (MH.sup.+)
[0205] .sup.1H-NMR (CD.sub.3OD) .delta.:4.68 (1H, m), 3.95 (1H, m),
3.48 (1H, m), 3.18 (1H, m), 2.91 (1H, m), 2.72-2.54 (3H, m), 2.36
(6H, s), 2.04 (1H, m), 1.96 (1H, m), 1.84-1.60 (7H, m), 1.50 (2H,
m), 1.35-1.10 (8H, m), 0.88 (2H, m).
Example 14
(3R)-6-Cyclohexyl-N-hydroxy-3-(3-{[3-(4-morpholinyl)-1
-azetidinyl]carbonyl}-1,2,4-oxadiazol-5-yl)hexanamide
[0206] 21
[0207] A solution of
(3R)-6-cyclohexyl-3-(3-{[3-(4-morpholinyl)-1-azetidin-
yl]carbonyl)1,2,4-oxadiazol-5-yl)hexanoic acid trifluoroacetate
(Preparation 28) (1173 mg, 2.70 mmol) and N-methylmorpholine (327
.mu.l, 2.97 mmol) in N,N-dimethylformamide (25 ml) was cooled to
0.degree. C., treated dropwise with isobutyl chloroformate (386
.mu.l, 2.97 mmol) and stirred under a nitrogen atmosphere at
0.degree. C. for 45 minutes. Hydroxylamine hydrochloride (564 mg,
8.11 mmol) was then added followed by further N-methylmorpholine
(891 .mu.l, 8.11 mmol) and the mixture was stirred for 18 hours
being allowed to warm to room temperature over this time. The
solvent was removed under reduced pressure and the residue
dissolved in a saturated aqueous solution of sodium carbonate. The
pH of the solution was adjusted to 9 by dropwise addition of acetic
acid. Water was added (75 ml) and the product was then extracted
with ethyl acetate (.times.2). The combined organic phases were
washed with brine, dried over anhydrous sodium sulphate, filtered
and the solvent removed under reduced pressure. The residue was
purified by column chromatography on silica gel eluting with a
gradient system of dichloromethane:methanol:0.8- 8 ammonia
(90:10:1) then dichloromethane:methanol:0.88 ammonia (80:20:2). The
product was further purified by HPLC to afford the title compound
as a white solid (151 mg)
[0208] MS: 450 (MH.sup.+)
[0209] Analysis: Found C, 56.19; H, 7.61; N, 14.81%;
C.sub.22H.sub.35N.sub.5O.sub.5.H.sub.2O requires C, 56.51; H, 7.96;
N, 14.98%
[0210] .sup.1H-NMR (CD.sub.3OD) .delta.: 4.63 (1H, m), 4.42 (1H,
m), 4.44 (1H, m), 4.03 (1H, m), 3.75 (4H, m), 3.41 (1H, m), 3.30
(1H, m), 2.72-2.53 (2H, m), 2.45 (4H, m), 1.83-1.56 (7H, m),
1.36-1.07 (8H, m), 0.87 (2H, m).
[0211] MPt.: 52-63.degree. C.
Example 15
(3R)-6-Cyclohexyl-N-hydroxy-3-(3-([4-(4-pyridinyl)-1-piperidinyl]carbonyl}-
-1,2,4-oxadiazol-5-yl)hexanamide
[0212] 22
[0213] A solution of
(3R)-6-cyclohexyl-3-(3-{[4-(4-pyridinyl)-1-piperidiny-
l]carbonyl}-1,2,4-oxadiazol-5-yl)hexanoic acid (Preparation 32)
(311 mg, 0.68 mmol) and N-methylmorpholine (83 .mu.l, 0.75 mmol) in
N,N-dimethylformamide (15 ml) was cooled to 0.degree. C., treated
dropwise with isobutyl chloroformate (98 .mu.l, 0.75 mmol) and the
resulting solution was stirred at 0.degree. C. for 45 minutes.
Hydroxylamine hydrochloride (142 mg, 2.05 mmol) was then added
followed by further N-methylmorpholine (226 .mu.l, 2.05 mmol) and
the mixture was stirred for 18 hours being allowed to warm to room
temperature over this time. The solvent was removed under reduced
pressure and the residue partitioned between ethyl acetate (50 ml)
and water (50 ml). The layers were separated and the aqueous phase
extracted with ethyl acetate (.times.2). The combined organic
phases were washed with brine, dried over anhydrous sodium
sulphate, filtered and the solvent removed under reduced pressure.
The residue was purified by HPLC to afford a residue that was
partitioned between ethyl acetate and water. The organic phase was
separated, dried over anhydrous sodium sulphate, filtered and the
solvent removed under reduced pressure. The residue was triturated
with diethyl ether to yield the title compound as a white solid (94
mg).
[0214] MS: 470 (MH.sup.+)
[0215] .sup.1H-NMR (CD.sub.3OD) .delta.: 8.45 (2H, d, J=4Hz), 7.37
(2H, d, J=4Hz), 4.79 (1H, m), 4.00 (1H, m), 3.60 (1H, m), 3.32 (1H,
m), 3.00 (2H, m), 2.70-2.52 (2H, m), 2.06-1.85 (2H, m), 1.83-1.60
(9H, m), 1.39-1.12 (8H, m), 0.86 (2H, m).
Example 16
5{(1R)-4-Cyclohexyl-1-[2-(hydroxyamino)-2-oxoethyl]butyl}-N-methyl-N-(2-py-
ridinylmethyl)-1,2,4-oxadiazole-3-carboxamide
[0216] 23
[0217] A solution of
(3R)-6-cyclohexyl-3-(3-{([methyl(2-pyridinylmethyl)am-
ino]carbonyl}-1,2,4-oxadiazol-5-yl)hexanoic acid (Preparation 36)
(453 mg, 1.09 mmol)and N-methylmorpholine (132 .mu.l, 1.20 mmol) in
anhydrous tetrahydrofuran (10 ml) was cooled to 0.degree. C.,
treated with isobutyl chloroformate (155 .mu.l, 1.20 mmol) and
stirred under a nitrogen atmosphere for 2 hours.
O-(trimethylsilyl)hydroxylamine (160 .mu.l, 1.31 mmol) was added
and the mixture was stirred for 18 hours, being allowed to warm to
room temperature over this time. The mixture was then treated with
saturated aqueous ammonium chloride solution (10 ml) and stirred
for 1 hour. This mixture was then diluted with water and extracted
with ethyl acetate (.times.3). The combined organic layers were
sequentially washed with water and brine, dried over anhydrous
sodium sulphate, filtered and the solvent removed under reduced
pressure. The residue was purified by column chromatography on
silica gel eluting with ethyl acetate: isopropanol (95:5) then a
gradient system of dichloromethane:methanol (90:10) to
dichloromethane: methanol (80:20) to afford a residue (260 mg, 0.31
mmol) which was dissolved in methanol (5 ml) and treated with
potassium carbonate (138 mg, 1.00 mmol) and the mixture was stirred
at room temperature for 17 hours. The solvent was removed under
reduced pressure and the residue partitioned between ethyl acetate
and a saturated aqueous solution of ammonium chloride. The layers
were separated and the aqueous layer was extracted with ethyl
acetate (.times.4). The combined organic layers were washed with
brine, dried over anhydrous sodium sulphate, filtered and the
solvent removed under reduced pressure. The residue was purified by
HPLC.sup.a to afford the title compound as a white solid (84
mg)
[0218] MS: 430 (MH.sup.+)
[0219] .sup.1H-NMR (CDCl.sub.3) (mixture of rotamers) .delta.: 8.55
(1H, m), 7.72 (1H, m), 7.42-7.20 (2H, m), 4.84 (2H, m), 3.64 (1H,
m), 3.24 (1.2H, s), 3.18 (1.8H, s), 2.84-2.53 (2H, m), 1.86-1.45
(7H, m), 1.39-0.88 (8H, m), 0.80 (2H, m).
Example 17
2-[[(5-{(1R)-4-Cyclohexyl-1-[2-(hydroxyamino)-2-oxoethyl]butyl}-1,2,4-oxad-
iazol-3-yl)carbonyl](methyl)amino]acetic acid
[0220] 24
[0221] A solution of methyl 2-[[(5-(1R)4-cyclohexyl-1
-[2-(hydroxyamino)-2-oxoethyl]butyl)-1,2,4-oxadiazol-3-yl)carbonyl](methy-
l)amino]acetate (Preparation 40) (180 mg, 0.44 mmol) in 1,4-dioxane
(4 ml) was cooled to 0.degree. C. and treated with lithium
hydroxide monohydrate (42 mg, 1 mmol). The mixture was stirred for
1 hour being allowed to warm to room temperature over this time.
The mixture was then treated with hydrochloric acid (2M), diluted
with water and extracted with ethyl acetate (.times.2). The
combined organic layers were washed with brine, dried over
anhydrous sodium sulphate, filtered and the solvent removed under
reduced pressure. The residue was purified by HPLC.sup.b to afford
the title compound as a white foam (95 mg).
[0222] MS: 397(MH.sup.+), 419 (MNa.sup.+)
[0223] .sup.1H-NMR (CD.sub.3OD) (mixture of rotamers) .delta.: 4.39
(1H, s), 4.27 (1H, s), 3.60 (1H, m), 3.23 (1.5H, s), 3.16 (1.5H,
s), 2.73-2.50 (2H, m), 1.86-1.58 (7H, m), 1.39-1.08 (8H, m), 0.87
(2H, m).
Example 18
Methyl
1-[(5{(1R)-4-cyclohexyl-1-[2-(hydroxyamino)-2-oxoethyl]butyl}-1,2,4-
-oxadiazol-3-yl)carbonyl]-3-azetidinecarboxylate
[0224] 25
[0225] A solution of
(3R)-6-cyclohexyl-3-(3-[3-(methoxycarbonyl)-1-azetidi-
nyl]carbonyl)1,2,4-oxadiazol-5-yl)hexanoic acid (Preparation 43)
(200 mg, 0.49 mmol) and N-methylmorpholine (59 .mu.l, 0.54 mmol) in
anhydrous dichloromethane (7ml) was cooled to 0.degree. C., treated
with isobutyl chloroformate (70 .mu.l, 0.54 mmol) and stirred under
a nitrogen atmosphere for 1 hour. O-(Trimethylsilyl)hydroxylamine
(175 .mu.l, 1.47 mmol) was added and the mixture was for 18 hours,
being allowed to warm to room temperature over this time. The
mixture was then treated with aqueous citric acid solution (10%
w/v, 10 ml) and stirred for 2 hours. This mixture was then diluted
with water and extracted with ethyl acetate (.times.3). The
combined organic layers were washed with water and brine, dried
over anhydrous sodium sulphate, filtered and the solvent removed
under reduced pressure. The residue was purified by HPLC to afford
the title compound as a foam (150 mg).
[0226] MS: 423 (MH.sup.+), 440 (MNH.sub.4.sup.+)
[0227] .sup.1H-NMR (CD.sub.3OD) .delta.: 4.804.62 (2H, m), 4.464.25
(2H, m), 3.77 (3H, s), 3.63 (2H, m), 2.75-2.50 (2H, m), 1.83-1.57
(7H, m), 1.39-1.08 (8H, m), 0.86 (2H, m).
Example 19
1-[(5-{(1R)4-Cyclohexyl-1-[2-(hydroxyamino)-2-oxoethyl]butyl}-1,2,4-oxadia-
zol-3-yl)carbonyl]-3-azetidinecarboxylic acid
[0228] 26
[0229] A solution of methyl 1-[(5-{(1R)-4-cyclohexyl-1
-[2-(hydroxyamino)-2-oxoethyl]butyl}-1,2,4-oxadiazol-3-yl)carbonyl]-3-aze-
tidinecarboxylate (Example 18) (130 mg, 0.31 mmol) in 1,4-dioxane
(33 ml) was cooled to 0.degree. C. and treated with lithium
hydroxide monohydrate (25 mg, 0.60 mmol). The mixture was stirred
for 1 hour being allowed to warm to room temperature over this
time. Further lithium hydroxide monohydrate (25 mg, 0.60 mmol) was
added and the mixture stirred for 3 hours. The mixture was then
treated with hydrochloric acid (2M), diluted with water and
extracted with ethyl acetate (.times.2). The combined organic
layers were washed with brine, dried over anhydrous sodium
sulphate, filtered and the solvent removed under reduced pressure.
The residue was purified by HPLC.sup.b to afford the title compound
as a foam (110 mg).
[0230] MS: 409 (MH.sup.+), 431 (MNa.sup.+)
[0231] .sup.1H-NMR (CDCl.sub.3) .delta.: nmr-needs redoing 4.71
(2H, m), 4.35 (2H, m), 3.69-3.43 (2H, m), 2.86-2.56 (2H, m),
1.90-1.52 (7H, m), 1.40-0.99 (8H, m), 0.83 (2H, m).
Example 20
(3R)-6-Cyclohexyl-N-hydroxy-3-(3-methyl-1,2,4-oxadiazol-5-yl)hexanamide
[0232] 27
[0233] A solution of
(3R)-6-cyclohexyl-3-(3-methyl-1,2,4-oxadiazol-5-yl)he- xanoic acid
(Preparation 45) (170 mg, 0.61 mmol) in anhydrous tetrahydrofuran
(2 ml) was treated with 1,1'-carbonyldiimidazole (98 mg, 0.61 mmol)
and the mixture stirred at room temperature under a nitrogen
atmosphere for 20 minutes. Hydroxylamine hydrochloride (42 mg, 0.61
mmol) was then added and the mixture stirred for 18 hours. The
solvent was removed under reduced pressure and the residue
partitioned between hydrochloric acid (0.5M) and ethyl acetate. The
layers were separated and the organic phase was washed with water,
dried over anhydrous sodium sulphate, filtered and solvent removed
under reduced pressure. The residue was purified by column
chromatography on silica gel eluting with dichloromethane:methanol
(98:2) to afford the title compound as an oil (82 mg).
[0234] MS: 296 (MH.sup.+)
[0235] .sup.1H-NMR (CDCl.sub.3) .delta.: 3.56 (1H, m), 2.70 (1H,
m), 2.58 (1H, m), 2.38 (3H, s), 1.88-1.42 (7H, m), 1.39-1.03 (8H,
m), 0.83 (2H, m)
Example 21
(3R)6-Cyclohexyl-N-hydroxy-3-(3-isopropyl-1,2,4-oxadiazol-5-yl)hexanamide
[0236] 28
[0237] A solution of
(3R)-6-cyclohexyl-3-(3-isopropyl-1,2,4-oxadiazol-5-yl- )hexanoic
acid (Preparation 47) (175 mg, 0.57 mmol) in anhydrous
tetrahydrofuran (10 ml) was treated with 1,1'-carbonyldiimidazole
(93 mg, 0.57 mmol) and the mixture stirred at room temperature
under a nitrogen atmosphere for 1 hour. Hydroxylamine hydrochloride
(40 mg, 0.57 mmol) was then added and the mixture stirred for 18
hours. The mixture was filtered and the solvent was removed from
the filtrate under reduced pressure. The residue was purified by
column chromatography on silica gel eluting with
dichloromethane:methanol:0.88 ammonia (95:5:0.5) to afford the
title compound as an oil (44 mg).
[0238] MS: 324 (MH.sup.+)
[0239] .sup.1H-NMR (CDCl.sub.3) .delta.: 3.54 (1H, m), 3.06 (1H,
m), 2.75-2.48 (2H, m), 1.88-1.58 (7H, m), 1.39-1.05 (14H, m), 0.83
(2H, m)
Example 22
(3R)-6-Cyclohexyl-N-hydroxy-3-[3-(methoxymethyl)-1,2,4-oxadiazol-5-yl]hexa-
namide
[0240] 29
[0241] A solution of
(3R)-6-cyclohexyl-3-[3-(methoxymethyl)-1,2,4-oxadiazo-
l-5-yl]hexanoic acid (Preparation 49) (250 mg, 0.81 mmol) in
anhydrous tetrahydrofuran (10 ml) was treated with
1,1'-carbonyldiimidazole (130 mg, 0.81 mmol) and the mixture
stirred at room temperature under a nitrogen atmosphere for 2
hours. Hydroxylamine hydrochloride (56 mg, 0.81 mmol) was then
added and the mixture stirred for 18 hours. The solvent was removed
under reduced pressure and the residue was purified by column
chromatography on silica gel eluting with
dichloromethane:methanol:0.88 ammonia (95:5:0.5) to afford the
title compound as an oil (75 mg).
[0242] MS: 326 (MH.sup.+), 348 (MNa.sup.+)
[0243] .sup.1H-NMR (CDCl.sub.3) .delta.: 4.57 (2H, s), 3.62 (1H,
m), 3.46 (3H, s), 2.80-2.52 (2H, m), 1.88-1.42 (7H, m), 1.39-1.03
(8H, m), 0.83 (2H, m)
Example 23
(3R)-6-Cyclohexyl-N-hydroxy-3-[3-(2-methoxyethyl)-1,2,4-oxadiazol-5-yl]hex-
anamide
[0244] 30
[0245] A solution of
((3R)-6-cyclohexyl-3-[3-(2-methoxyethyl)-1,2,4-oxadia-
zol-5-yl]hexanoic acid (Preparation 51) (245 mg, 0.76 mmol) in
anhydrous tetrahydrofuran (10 ml) was treated with
1,1'-carbonyldiimidazole (123 mg, 0.76 mmol) and the mixture
stirred at room temperature under a nitrogen atmosphere for 1 hour.
Hydroxylamine hydrochloride (53 mg, 0.76 mmol) was then added and
the mixture stirred for 18 hours. The solvent was removed under
reduced pressure and the residue was purified by column
chromatography on silica gel eluting with
dichloromethane:methanol:0.88 ammonia (95:5:0.5). The residue was
dissolved in ethyl acetate, washed with hydrochloric acid (1M),
dried over anhydrous magnesium sulphate, filtered and the solvent
removed under reduced pressure to afford the title compound as an
oil (100 mg)
[0246] MS: 340 (MH.sup.+), 362 (MNa.sup.+)
[0247] .sup.1H-NMR (CDCl.sub.3) .delta.: 3.77 (2H, m), 3.55 (1H,
m), 3.36 (3H, s), 3.00 (2H, t, J=6Hz), 2.80-2.52 (2H, m), 1.88-1.52
(7H, m), 1.39-1.05 (8H, m), 0.83 (2H, m)
Example 24
(3R)-6-Cyclohexyl-N-hydroxy-3-{3-[2-oxo-2-(1
-pyrrolidinyl)ethyl]-1,2,4-ox- adiazol-5-yl}hexanamide
[0248] 31
[0249] A solution of (3R)-6-cyclohexyl-3-{3-[2-oxo-2-(1
-pyrrolidinyl)ethyl]-1,2,4-oxadiazol-5-yl}hexanoic acid
(Preparation 53) (200 mg, 0.53 mmol) in anhydrous tetrahydrofuran
(10 ml) was treated with 1,1'-carbonyldiimidazole (86 mg, 0.53
mmol) and the mixture stirred at room temperature under a nitrogen
atmosphere for 2 hours. Hydroxylamine hydrochloride (37 mg, 0.53
mmol) was then added and the mixture stirred for 18 hours. The
solvent was removed under reduced pressure and the residue was
purified by column chromatography on silica gel eluting with
dichloromethane:methanol:0.88 ammonia (90:10:1) to afford the title
compound as a colourless oil (50 mg).
[0250] .sup.1H-NMR (CDCl.sub.3) .delta.: 3.80 (2H, m), 3.67-3.40
(5H, m), 2.80-2.52 (2H, m), 2.13-0.98 (19H, m), 0.83 (2H, m)
Example 25
(3R)-6-Cyclohexyl-N-hydroxy-3-{3-[(phenylsulfonyl)methyl]-1,2,4-oxadiazol--
5-yl}hexanamide
[0251] 32
[0252] A solution of
(3R)-6-cyclohexyl-3-{3-[(phenylsulfonyl)methyl]-1,2,4-
-oxadiazol-5-yl)hexanoic acid (Preparation 55) (60 mg, 0.14 mmol)
in anhydrous tetrahydrofuran (4 ml) was treated with
1,1'-carbonyldiimidazol- e (25 mg, 0.15 mmol) and the mixture
stirred at room temperature under a nitrogen atmosphere for 2
hours. Hydroxylamine hydrochloride (10 mg, 0.14 mmol) was then
added and the mixture was stirred for 18 hours. The solvent was
removed under reduced pressure and the residue was purified by
column chromatography on silica gel eluting with
dichloromethane:methanol:0.88 ammonia (95:5:0.5) to afford the
title compound as an oil (15 mg).
[0253] MS: 436 (MH.sup.+), 453 (MNH.sub.4+)
[0254] .sup.1H-NMR (CDCl.sub.3) .delta.: 8.93 (1H, brs), 8.01-7.40
(5H, m), 4.53 (2H, s), 3.60 (1H, m), 2.85-2.52 (2H, m), 1.84-1.43
(7H, m), 1.39-1.00 (8H, m), 0.85 (2H, m)
Example 26
(3R)-3-{3-[(4-Chlorophenoxy)methyl]-1,2,4-oxadiazol-5-yl}-6-cyclohexyl-N-h-
ydroxyhexanamide
[0255] 33
[0256] A solution of
(3R)-3-3-[(4-chlorophenoxy)methyl]-1,2,4-oxadiazol-5--
yl}6-cyclohexylhexanoic acid (Preparation 57) (180 mg, 0.44 mmol)
in anhydrous tetrahydrofuran (10 ml) was treated with
1,1'-carbonyidiimidazole (71 mg, 0.44 mmol) and the mixture stirred
at room temperature under a nitrogen atmosphere for 2 hours.
Hydroxylamine hydrochloride (30 mg, 0.44 mmol) was then added and
the mixture stirred for 18 hours. The solvent was removed under
reduced pressure and the residue was partitioned between ethyl
acetate and water. The layers were separated and the organic phase
was washed with brine, dried over anhydrous magnesium sulphate,
filtered and the solvent removed under reduced pressure. The
residue was purified by column chromatography on silica gel eluting
with dichloromethane:methanol:0.88 ammonia (95:5:0.5) to afford the
title compound as a glass (53 mg).
[0257] .sup.1H-NMR (CDCl.sub.3) .delta.: 7.24 (2H, m), 6.94 (2H, d,
J=8Hz), 5.13 (2H, s), 3.70-3.44 (1H, m), 3.00-2.50 (2H, m),
1.85-1.53 (7H, m), 1.37-1.02 (8H, m), 0.82 (2H, m)
Example 27
(3R)-6-Cyclohexyl-N-hydroxy-3-[3-(2-pyridinylmethyly1,2,4-oxadiazol-5-yl]h-
exanamide
[0258] 34
[0259] A solution of
(3R)-6-cyclohexyl-3-[3-(2-pyridinylmethyl)-1,2,4-oxad-
iazol-5-yl]hexanoic acid trifluoroacetate (Preparation 59) (214 mg,
0.45 mmol) in anhydrous tetrahydrofuran (5 ml) was treated with
1,1'-carbonyidiimidazole (194 mg, 1.20 mmol) and the mixture
stirred at room temperature under a nitrogen atmosphere for 2
hours. Hydroxylamine hydrochloride (83 mg, 1.20 mmol) was then
added and the mixture stirred for 18 hours. The solvent was removed
under reduced pressure and the residue was partitioned between
ethyl acetate and water. The layers were separated and the organic
phase was washed with water then brine, dried over anhydrous
magnesium sulphate, filtered and the solvent removed under reduced
pressure. The residue was purified by column chromatography on
silica gel eluting with dichloromethane:methanol: 0.88 ammonia
(95:5:0.5) to afford the title compound as a colourless oil (120
mg).
[0260] MS: 373 (MH.sup.+), 395 (MNa.sup.+)
[0261] .sup.1H-NMR (DMSO-d.sub.6) .delta.: 8.79 (1H, br s), 8.45
(1H, d, J=5Hz), 7.74 (1H, t, J=5Hz), 7.38-7.20 (2H, m), 4.20 (2H,
s), 3.40 (1H, m), 2.48-2.31 (2H, m), 1.89-1.43 (7H, m), 1.29-0.95
(8H, m), 0.76 (2H, m)
Example 28
Ethyl
2-{(1R)-4-cyclohexyl-1-[2-(hydroxyamino)-2-oxoethyl]butyl}-1,3-oxazo-
le-4-carboxylate
[0262] 35
[0263] A solution of
(3R)-6-cyclohexyl-3-[4-(ethoxycarbonyl)-1,3-oxazol-2-- yl]hexanoic
acid (Preparation 63) (130 mg, 0.39 mmol) and
N,N-diisopropylethylamine (67 .mu.l, 0.39 mmol) in
N,N-dimethylformamide (6 ml) was treated with
O-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethylu- ronium
hexafluorophosphate (220 mg, 0.58 mmol) and the resulting mixture
was stirred at room temperature under a nitrogen atmosphere for 45
minutes. Further N,N-diisopropylethylamine (270 .mu.l, 1.54 mmol)
was then added, followed by hydroxylamine hydrochloride (80 mg,
1.16 mmol) and the mixture was stirred at room temperature for 18
hours. The solvent was removed under reduced pressure and the
residue partitioned between ethyl acetate and pH 7 aqueous buffer.
The organic layer was separated, washed sequentially with water and
brine, dried over anhydrous magnesium sulphate, filtered and the
solvent removed under reduced pressure. The residue was purified by
column chromatography on silica gel eluting with a gradient system
of dichloromethane:methanol (99:1) gradually changing to
dichloromethane:methanol (98:2) to afford the title compound (20
mg).
[0264] MS: 353 (MH.sup.+)
[0265] .sup.1H-NMR (CDCl.sub.3) .delta.: 8.14 (1H, s), 4.37 (2H, q,
J=7Hz), 3.50 (1H, m), 2.76 (1H, m), 2.57 (1H, m), 1.88-1.52 (7H,
m), 1.37 (3H, t, J=7Hz), 1.32-1.02 (8H, m), 0.83 (2H, m).
Example 29
2-{(1R)-4-Cyclohexyl-1-[2-(hydroxyamino)-2-oxoethyl]butyl}-N,N-dimethyl-1,-
3-oxazole-4-carboxamide
[0266] 36
[0267] A solution of 2-((1R)-1
-{2-[(benzyloxy)amino]-2-oxoethyl}-4-cycloh-
exylbutyl)-N,N-dimethyl-1,3-oxazole-4-carboxamide (Preparation 66)
(100 mg, 0.23 mmol) in ethanol (5 ml) was treated with 5% palladium
on barium sulphate (50 mg), pressurised to 1 atm with hydrogen in a
sealed vessel and the mixture was stirred at room temperature for 2
hours. The solution was filtered through Arbocel.RTM. and the
solvent removed from the filtrate under reduced pressure. The
residue was azeotroped from dichloromethane (.times.3) then
dichloromethane:ether (1 ml:4 ml) to afford the title compound as a
foam (75 mg).
[0268] MS: 351 (M.sup.+)
[0269] .sup.1H-NMR (CDCl.sub.3) .delta.: 7.94 (1H, s), 3.52 (1H,
m), 3.38-2.94 (6H, br d), 2.73-2.45 (2H, m), 1.85-1.47 (7H, m),
1.41-0.98 (8H, m), 0.83 (2H, m).
Example 30
(3R)-6-Cyclohexyl-N-hydroxy-3-[3-(1-pyrrolidinylcarbonyl)-1,2,4-oxadiazol--
5-yl]hexanamide
[0270] 37
[0271] A solution of
(3R)-6-cyclohexyl-3-[3-(1-pyrrolidinylcarbonyl)-1,2,4-
-oxadiazol-5-yl]hexanoic acid (Preparation 13) (288 mg, 0.79 mmol)
and N,N-diisopropylethylamine (138 .mu.l, 0.79 mmol) in
N,N-dimethylformamide (6ml) was treated with
O-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethylur- onium
hexafluorophosphate (452 mg, 1.19 mmol) and the resulting mixture
was stirred at room temperature under a nitrogen atmosphere for 1
hour. Further N,N-diisopropylethylamine (552 .mu.l, 3.16 mmol) was
then added, followed by the hydroxylamine hydrochloride (165 mg,
2.38 mmol) and the mixture stirred at room temperature for 18
hours. The solvent was removed under reduced pressure and the
residue wasa partitioned between ethyl acetate and pH 7 aqueous
buffer. The organic layer was separated, washed sequentially with
water and brine, dried over anhydrous magnesium sulphate, filtered
and the solvent removed under reduced pressure. The residue was
purified by column chromatography on silica gel eluting with
dichloromethane:methanol (98:2) to afford the title compound as a
foam (126 mg).
[0272] MS: 379 (MH.sup.+), 396 (MNH.sub.4.sup.+)
[0273] .sup.1H-NMR(CD.sub.3OD) .delta.: 3.75 (2H, m), 3.60 (3H, m),
2.66 (1H, dd, J=13, 8Hz), 2.57 (1H, dd, J=13, 3Hz), 1.98 (4H, m),
1.86-1.58 (7H, m), 1.40-1.07 (8H, m), 0.87 (2H, m).
Example 31
2-{(1R)-4-Cyclohexyl-1-[2-(hydroxyamino)-2-oxoethyl]butyl}-N,N,5-trimethyl-
-1,3-oxazole-4-carboxamide
[0274] 38
[0275] A solution of
2-((1R)-1-{2-[(benzyloxy)amino]-2-oxoethyl}-4-cyclohe-
xylbutyl)-N,N,5-trimethyl-1,3-oxazole-4-carboxamide (Preparation
73) (105 mg, 0.23 mmol) in ethanol (5 ml) was treated with 5%
palladium on barium sulphate (45 mg) and ammonium formate (73 mg,
1.15 mmol) and the mixture was heated at 45.degree. C. for 17
hours. Further palladium on barium sulphate (30 mg) and ammonium
formate (60 mg, mmol) were added and the heating was continued for
2 hours. The solution was filtered through Arbocel.RTM. and the
solvent removed from the filtrate under reduced pressure. The
residue was azeotroped from dichloromethane (.times.3) then
purified by column chromatography on silica gel eluting with
dichloromethane:methanol:0.88 ammonia (98:2:0.2 to 90:10:1) to
afford the title compound as a foam (25 mg).
[0276] MS: 366(MH.sup.+), 388 (MNa.sup.+)
[0277] Analysis: Found C, 61.77; H, 8.62; N, 11.22%;
C.sub.19H.sub.31N.sub.3O.sub.4.0.1H.sub.2O requires C, 62.14; H,
8.56; N, 11.44%
[0278] .sup.1H-NMR(CD.sub.3).delta.: 10.37 (1H, brs), 8.66 (1H,
brs),3.25(1H, m), 3.14 (3H, brs), 2.91 (3H, brs), 2.43-2.21 (5H,
m), 1.66-1.50 (7H, m), 1.24-1.00 (8H, m), 0.79 (2H, m).
Example 32
2-{(1R)-4-Cyclohexyl-1-[2-(hydroxyamino)-2-oxoethyl]butyl}-5-methyl-1,3-ox-
azole-4-carboxamide
[0279] 39
[0280] A solution of 2-((1R)-1
-2-[(benzyloxy)amino]-2-oxoethyl)-4-cyclohe-
xylbutyl)-5-methyl-1,3-oxazole-4-carboxamide (Preparation 74) (133
mg, 0.31 mmol) in ethanol (6 ml) was treated with 5% palladium on
barium sulphate (80 mg) and ammonium formate (196 mg, 3.10 mmol)
and the mixture was heated at 45.degree. C. for 2.5 hours. The
solution was filtered through Arboce.RTM. and the solvent removed
from the filtrate under reduced pressure. The residue was
azeotroped from dichloromethane (.times.3) then purified by column
chromatography on silica gel eluting with
dichloromethane:methanol:0.88 ammonia (98:2:0.2 to 90:10:1) to
afford the title compound as a white solid (56 mg).
[0281] MS: 338 (MH.sup.+), 360 (MNa.sup.+)
[0282] Analysis: Found C, 61.77; H, 8.62; N, 11.22%;
C.sub.19H.sub.31N.sub.3O.sub.4.0.1H.sub.2O requires C, 62.14; H,
8.56; N, 11.44%
[0283] .sup.1H-NMR (DMSO-d.sub.6) .delta.: 7.12 (2H, br s), 3.52
(1H, m), 2.56-2.21 (2H, m), 1.67-1.49 (7H, m), 1.23-1.00 (8H, m),
0.79 (2H, m).
[0284] Other compounds prepared by the same general method, using
appropriate starting materials (see Preparations section below),
are listed in Table 1 below.
4TABLE 1 40 Example R' Elemental Analysis LRMS .sup.1H,.delta. 33
NHCH.sub.2Ph Found C, 62.93;H, 7.37;N, 13.29%; 415(MH.sup.+)
(CD.sub.3OD):7.40-7.21(5H, m), 4.57(2H, s), 3.60
C.sub.22H.sub.30N.sub.4O.sub.4, 0.25H.sub.2O requires C, 63.06, H,
432(MNH.sub.4.sup.+) (1H, m), 2.72-2.53(2H, m), 1.86-1.59(7H, m),
7.34;N, 13.37% 1.40-1.10(8H, m), 0.86(2H, m) 34 41 393(MH.sup.+)
410(MNH.sub.4.sup.+) (CD.sub.3OD):3.73(2H, m), 3.60(1H, m), 3.47
(2H, m), 2.73--2.50(2H, m), 1.91-1.53(13H, m), 1.43-1.05(8H, m),
0.86(2H, m). 35 42 428(M2H.sup.+) (CD.sub.3OD):7.42-7.27(4H, m),
5.20(2H, s), 4.98 (2H, s), 3.64(1H, m), 2.70(1H, dd, J=13, 8Hz),
2.50(1H, dd, J=13, 4Hz), 1.89-1.58(7H, m), 1.40-1.10(8H, m),
0.88(2H, m). 36 43 Found C, 64.53;H, 7.43;N, 12.58%;
C.sub.24H.sub.32N.sub.4O.sub.4, 0.25H.sub.2O requires C, 64.77, H,
7.36;N, 12.59% 441(MH.sup.+), 463(MNa.sup.+) (CD.sub.3OD):(mixture
of rofamers)7.32-7.04(4H, m), 4.83(1.2H, s), 4.73(0.8H, s),
4.00(0.8H, m), 3.83(1.2H, m), 3.64(1H, m), 3.00(2H, m),
7.77-2.54(2H, m), 1.90-1.53(7H, m), 1.44-1.02 (8H, m), 0.88(2H, m)
37 44 395(MH.sup.+), 412(MNH.sub.4.sup.+) (CD.sub.3OD):3.75(4H, m),
3.71-3.53(5H, m), 2.62(2H, m), 1.87-1.58(7H, m), 1.40-1.06(6H, m),
0.86(2H, m) 38 45 408(MH.sup.+) (CD.sub.3OD:3.80(2H, m), 2.60(3H,
m), 2.72-2.43(6H, m), 2.33(3H, s), 1.82-1.58(7H, m), 1.40-1.09(8H,
m), 0.87(2H, m) 39 46 442(MH.sup.+) (CD.sub.3OD):(mixture of
rofamers[1:2])8.39(1H, m), 7.73(0.67H, d, J=7Hz), 7.62(0.33H, d,
J=7Hz), 7.32(1H, m), 4.94(1.34H, m), 4.85 (0.66H, m), 4.12(0.66H,
t, J=4Hz), 3.95 (1.34H, t, J=Hz), 3.63(1H, m), 3.10(2H, t, J=Hz),
2.75-2.54(2H, m), 1.86-1.59(7H, m), 1.40-1.07(8H, m), 0.89(2H, m)
40 47 429(MH+) (CD.sub.3OD) (mixture of rofamers[1:1])7.40-7.23
(5H, m), 4.80(1H, s), 4.64(1H, s), 3.62(1H, m), 3.03(1.5H, s),
2.99(1.5H, s), 2.72-2.51 (2H, m), 1.86-1.55(7H, m), 1.38-1.03(8H,
m), 0.84(2H, m).
Example 41/Preparation 40
Methyl
2-[[(5-{(1R)-4-cyclohexyl-1-[2-(hydroxyamino)-2-oxoethyl]butyl}-1,2-
,4-oxadiazol-3-yl)carbonyl](methyl)amino]acetate
[0285] 48
[0286] A solution of
(3R)-6-cyclohexyl-3-{(3-[(2-methoxy-2-oxoethyl)(methy-
l)amino]carbonyl}-1,2,4-oxadiazol-5-yl)hexanoic acid (Preparation
39) (273 mg, 0.70 mmol) and N-methylmorpholine (85 .mu.l, 0.77
mmol) in anhydrous dichloromethane (10 ml) was cooled to 0.degree.
C., treated with isobutyl chloroformate (100 .mu.l, 0.77 mmol) and
the resulting mixture was stirred under a nitrogen atmosphere for
30 minutes. O-(Trimethylsilyl)hydroxylamine (250 .mu.l, 2.10 mmol)
was then added and the mixture stirred under a nitrogen atmosphere
for 1 hour, being allowed to warm to room temperature over this
time. The mixture was then quenched with methanol (10 ml) and
stirred for 10 minutes. The solvent was removed under reduced
pressure and the residue partitioned between ethyl acetate and
water. The layers were separated and the organic layer was
sequentially washed with water and brine, dried over anhydrous
sodium sulphate, filtered and the solvent removed under reduced
pressure. The residue was purified by HPLC.sup.a to afford the
title compound as a colourless oil (187 mg).
[0287] MS: 411 (MH.sup.+), 433 (MNa.sup.+)
[0288] .sup.1H-NMR (CDCl.sub.3) .delta.: (mixture of rotamers)
4.50-4.21 (2H, m), 3.84-3.60 (4H, m), 3.32 (1.8H, s), 3.21 (1.2H,
s), 2.81-2.56 (2H, m), 1.90-1.50 (7H, m), 1.40-1.03 (8H, m), 0.82
(2H, m).
Example 42
(3R)-6-Cyclohexyl-3-(3-{[3-(dimethylamino)-1-azetidinyl]carbonyl}-1,2,4-ox-
adiazol-5-yl)N-hydroxyhexanamide
[0289] 49
[0290] A solution of
(3R)-6-cyclohexyl-3-(3-{[3-(dimethylamino)-1-azetidin-
yl]carbonyl}-1,2,4-oxadiazol-5-yl)hexanoic acid (Preparation 76)
(200 mg, 0.51 mmol) and N-methylmorpholine (112 .mu.l, 1.00 mmol)
in anhydrous tetrahydrofuran (15 ml) was cooled to 0.degree. C.,
treated with isobutyl chloroformate (79 .mu.l, 0.69 mmol) and
stirred under a nitrogen atmosphere for 2 hours.
O-(Trimethylsilyl)hydroxylamine (85 .mu.l, 0.61 mmol) was added and
the mixture was stirred for 18 hours, being allowed to warm to room
temperature over this time. The mixture was then treated with
methanol and stirred at room temperature for 2 hours. The solvent
was removed under reduced pressure. The residue was dissolved in
ethyl acetate (100 ml) and washed with water (2.times.25 ml) and
brine (25 ml), dried over anhydrous sodium sulphate, filtered and
the solvent removed under reduced pressure to afford the title
compound as a foam (180 mg).
[0291] MS: 408 (MH.sup.+)
[0292] .sup.1H-NMR (CD.sub.3OD) .delta.:4.60 (1H, m), 4.38 (1H, m),
4.22 (1H, m), 3.99 (1H, m), 3.60 (1H, m), 3.24 (1H, m), 2.64 (1H,
dd), 2.57 (1H, dd), 2.20 (6H, s), 1.58-1.80 (6H, m), 1.08-1.36 (7H,
m), 0.85 (2H, m).
Example 43
tert-Butyl
3-{[(5-{(1R)4-cyclohexyl-1-[2-(hydroxyamino)-2-oxoethyl]butyl}--
1,2,4-oxadiazol-3-yl)carbonyl]amino}-1-azetidinecarboxylate
[0293] 50
[0294] A solution of
(3R)-3-[3-({[1-(tert-butoxycarbonyl)-3-azetidinyl]ami-
no}carbonyl)-1,2,4-oxadiazol-5-yl]-6-cyclohexylhexanoic acid
(Preparation 79) (350 mg, 0.75 mmol) and N-methylmorpholine (163
.mu.l, 1.48 mmol) in anhydrous tetrahydrofuran (15 ml) was cooled
to 0.degree. C., treated with isobutyl chloroformate (116 .mu.l,
0.89 mmol) and stirred under a nitrogen atmosphere for 2 hours.
O-(Trimethylsilyl)hydroxylamine (272 .mu.l, 2.22 mmol) was added
and the mixture was stirred for 18 hours, being allowed to warm to
room temperature over this time. The mixture was then treated with
methanol and stirred at room temperature for 2 hours. The solvent
was removed under reduced pressure. The residue was dissolved in
ethyl acetate (100 ml) and washed with water (2.times.25 ml) and
brine (25 ml), dried over anhydrous sodium sulphate, filtered and
the solvent removed under reduced pressure. The solid was purified
by column chromatography on silica gel eluting with a gradient
system of 98.75:1.25:0.13 (dichloromethane:methanol:ammonia)
gradually changing to 90:10:1 (dichloromethane:methanol:ammonia) to
afford the title compound as a glass.
[0295] MS: 497 (MNH.sub.4.sup.+)
[0296] .sup.1H-NMR (CD.sub.3OD) .delta.:4.23 (2H, t), 3.98 (2H, t),
3.59 (1H, m), 2.64 (1H, dd), 2.57 (1H, dd), 1.58-1.83 (7H, m), 1.41
(9H, s), 1.08-1.37 (8H, m), 0.84 (2H, m).
Example 44
N-(3-Azetidinyl)-5-{(1R)-4-cyclohexyl-1-[2-(hydroxyamino)-2-oxoethyl]butyl-
)1,2,4-oxadiazole-3-carboxamide
[0297] 51
[0298] A solution of tert-butyl 3-{[(5-{(1R)-4-cyclohexyl-1
-[2-(hydroxyamino)-2-oxoethyl]butyl}-1,2,4-oxadiazol-3-yl)carbonyl]amino}-
-1-azetidinecarboxylate (Example 43) (160 mg, 0.33 mmol) in
dichloromethane (10 ml) was cooled to 0.degree. C. and treated with
trifluoroacetic acid (7 ml) and the resulting mixture was stirred
at 0.degree. C. under a nitrogen atmosphere for 45 minutes. The
solvent was removed under reduced pressure and the residue
azeotroped from dichloromethane (.times.2).
[0299] The residue was triturated with diethylether, filtered and
dried to afford the title compound as a white solid (123 mg).
[0300] MS: 380 (MH.sup.+)
[0301] .sup.1H-NMR (CD.sub.3OD) 8 :4.83 (1H, m), 4.30 (4H, d), 3.58
(1H, m), 2.50-2.70 (2H, m), 1.77 (2H, m), 1.62 (5H, m), 1.08-1.39
(8H, m), 0.83 (2H, m).
Example 45
Di(tert-butyl)
1-[(5-{(1R)-4-cyclohexyl-1-[2-(hydroxyamino)-2-oxoethyl]but-
yl}-1,2,4-oxadiazol-3-yl)carbonyl]-3-azetidinylimidodicarbonate
[0302] 52
[0303] A solution of
(.sup.3R)-3-[3-({3-[bis(tert-butoxycarbonyl)amino]-1--
azetidinyl}carbonyl)-1,2,4-oxadiazol-5-yl]-6-cyclohexylhexanoic
acid (Preparation 80) (430 mg, 0.76 mmol) and N-methylmorpholine
(167 .mu.l, 1.52 mmol) in anhydrous tetrahydrofuran (20 ml) was
cooled to 0.degree. C., treated with isobutyl chloroformate (120
.mu.l, 0.93 mmol) and stirred under a nitrogen atmosphere for 2
hours. O-(Trimethylsilyl)hydrox- ylamine (280 .mu.l, 2.29 mmol) was
added and the mixture was stirred for 18 hours, being allowed to
warm to room temperature over this time. The mixture was then
treated with methanol and stirred at room temperature for 2 hours.
The solvent was removed under reduced pressure. The residue was
dissolved in ethyl acetate (100 ml) and washed with water
(2.times.25 ml) and brine (25 ml), dried over anhydrous sodium
sulphate, filtered and the solvent removed under reduced pressure.
The solid was purified by column chromatography on silica gel
eluting with a gradient system of 98.75:1.25:0.13
(dichloromethane:methanol:ammonia) gradually changing to 90:10:1
(dichloromethane:methanol:ammonia) to afford the title compound as
a sticky foam.
[0304] MS: 578 (M-H.sup.-)
[0305] .sup.1H-NMR (CD.sub.3OD) .delta.: 4.88 (1H, m), 4.62 (1H,m),
4.40 (1H, t), 4.24 (1H,m), 3.60 (1H. m), 2.65 (1H, dd), 2.56 (1H,
dd), 1.58-1.80 (6H, m), 1.50 (18H, s), 1.08-1.35 (9H, m), 0.83 (2H,
m).
Example 46
(3R)-3-{3-[(3-Amino-1-azetidinyl)carbonyl]-1,2,4-oxadiazol-5-yl}-6-cyclohe-
xyl-N-hydroxyhexanamide
[0306] 53
[0307] A solution of di(tert-butyl) 1-[(5-{(1R)-4-cyclohexyl-1
-[2-(hydroxyamino)-2-oxoethyl]butyl)-1,2,4-oxadiazol-3-yl)carbonyl]-3-aze-
tidinylimidodicarbonate (Example 45) (250 mg, 0.43 mmol) in
dichloromethane (10 ml) was cooled to 0.degree. C. and treated with
trifluoroacetic acid (7 ml) and the resulting mixture was stirred
at 0.degree. C. under a nitrogen atmosphere for 45 minutes. The
solvent was removed under reduced pressure and the residue
azeotroped from dichloromethane (.times.2). The residue was
triturated with diethylether, filtered and dried to afford the
title compound as a white solid.
[0308] MS: 380 (MH.sup.+)
[0309] .sup.1H-NMR (CD.sub.3OD) .delta.:4.90 (1H, m), 4.53 (2H, m),
4.19 (2H, m), 3.58 (1H, m), 2.58-2.65 (2H, m), 1.76 (2H, m), 1.66
(5H, m), 1.06-1.38 (8H, m), 0.84 (2H,m).
Example 47
5-{(1R)-4-Cyclohexyl-1-[2-(hydroxyamino)-2-oxoethyl]butyl}-N-[2-(dimethyla-
mino)ethyl]-N-methyl-1,2,4-oxadiazole-3-carboxamide
[0310] 54
[0311] A solution
(3R)-6-cyclohexyl-3-(3-{[[2-(dimethylamino)ethyl](methyl-
)amino]carbonyl}-1,2,4-oxadiazol-5-yl)hexanoic acid (Preparation
82) (450 mg, 0.72 mmol) and N-methylmorpholine (284 .mu.l, 2.53
mmol) in dichloromethane (10 ml) was cooled to 0.degree. C.,
treated with isobutyl chloroformate (112 .mu.l, 0.87 mmol) and
stirred under an argon atmosphere for 1 hour.
O-(Trimethylsilyl)hydroxylamine (355 .mu.l, 2.90 mmol) was added
and the mixture was stirred for 3.5 hours, being allowed to warm to
room temperature over this time. The mixture was then treated with
methanol (2.5 ml) and stirred at room temperature for 20 minutes.
The solvent was removed under reduced pressure. The residue was
dissolved in ethyl acetate (100 ml) and washed with water (20 ml),
dried over anhydrous magnesium sulphate, filtered and the solvent
removed under reduced pressure. The solid was purified by column
chromatography on silica gel eluting with a gradient system of
97:3:0.3 (dichloromethane:methanol:ammonia) gradually changing to
90:10:1 (dichloromethane:methanol:ammonia) to afford the title
compound as a gum (123 mg).
[0312] MS: 410 (MH.sup.+)
[0313] Analysis: Found, C, 57.02; H, 8.50; N, 16.43%;
C.sub.20H.sub.35N.sub.50.sub.4. 0.25H.sub.20. 0.1CH.sub.2Cl.sub.2
requires C, 58.35; H, 8.72; N, 16.14%
[0314] .sup.1H-NMR (CDCl.sub.3) .delta.: 3.36-3.76 (3H, br d), 3.11
(3H, s), 2.45-2.74 (3H, m), 2.30 (1H, s), 2.25-2.35 (7H, m),
1.60-1.85 (7H, m), 1.05-1.40 (8H, m), 0.83 (2H, m).
Example 48
5-{(1R)-4-Cyclohexyl-1-[2-(hydroxyamino)-2-oxoethyl]butyl}-N-[3-(dimethyla-
mino)propyl]-N-methyl-1,2,4-oxadiazole-3-carboxamide
[0315] 55
[0316] A solution
(3R)-6-cyclohexyl-3-(3-[[3-(dimethylamino)propyl](methyl-
)amino]carbonyl}-1,2,4-oxadiazol-5-yl)hexanoic acid (Preparation
84) (566 mg, 0.88 mmol) and N-methylmorpholine (390 .mu.l, 3.52
mmol) in dichloromethane (10 ml) was cooled to 0.degree. C, treated
with isobutyl chloroformate (340 .mu.l, 2.64 mmol) and stirred
under an argon atmosphere for 1 hour.
O-(Trimethylsilyl)hydroxylamine (540 .mu.l, 4.40 mmol) was added
and the mixture was stirred for 4.5 hours, being allowed to warm to
room temperature over this time. The mixture was then treated with
methanol (2.5 ml) and stirred at room temperature for 20 minutes.
The solvent was removed under reduced pressure. The residue was
dissolved in ethyl acetate (100 ml) and washed with water (20 ml),
dried over anhydrous magnesium sulphate, filtered and the solvent
removed under reduced pressure. The solid was purified by column
chromatography on silica gel eluting with a gradient system of
97:3:0.3 (dichloromethane:methanol:ammonia) gradually changing to
90:10:1 (dichloromethane:methanol:ammonia) to afford the title
compound (150 mg).
[0317] MS: 424 (MH.sup.+)
[0318] Analysis: Found, C, 58.30; H, 8.64; N, 15.33%;
C.sub.21H.sub.37N.sub.5O.sub.4. 0.2 H.sub.2O. 0.08 CH.sub.2Cl.sub.2
requires C, 57.14; H, 8.50; N, 16.57%
[0319] .sup.1H-NMR (CDCl.sub.3) .delta.: 3.68 (2H, m), 3.30 (2H,
m), 3.18 (3H, s), 2.28-2.40 (3H, m), 2.23 (6H, s), 1.58-1.88 (9H,
m), 1.08-1.40 (8H, m), 0.88 (2H, m).
Example 49
Ethyl
[(5{(1R)-4-cyclohexyl-1-[2-(hydroxyamino)-2-oxoethyl]butyl}-1,2,4-ox-
adiazol-3-yl)methoxy]acetate
[0320] 56
[0321] A solution of
(3R)-6-cyclohexyl-3-3-[(2-ethoxy-2-oxoethoxy)methyl]--
1,2,4-oxadiazol-5-yl}hexanoic acid (Preparation 87) (330 mg, 0.86
mmol) and N-methylmorpholine (160 .mu.l, 1.46 mmol) in anhydrous
tetrahydrofuran (14 ml) was cooled to 0.degree. C., treated with
isobutyl chloroformate (120 .mu.l, 0.93 mmol) and stirred under a
nitrogen atmosphere for 2.5 hours. O-(Trimethylsilyl)hydroxylamine
(350 .mu.l, 2.86 mmol) was added and the mixture was stirred for 18
hours, being allowed to warm to room temperature over this time.
The mixture was then treated with methanol and stirred at room
temperature for 1 hour. The solvent was removed under reduced
pressure. The residue was dissolved in ethyl acetate and washed
with water and brine, dried over anhydrous sodium sulphate,
filtered and the solvent removed under reduced pressure. The oil
was purified by column chromatography on silica gel eluting with a
gradient system of 98:2 (dichloromethane:methanol) gradually
changing to 90:10 (dichloromethane:methanol) to afford the title
compound as a colourless oil (272 mg).
[0322] MS: 420 (MNa.sup.+)
[0323] .sup.1H-NMR (CD.sub.3OD) .delta.:4.70 (2H, s), 4.20 (4H, m),
3.54 (1H, m), 2.61 (1H, dd), 2.50 (1H, dd), 1.60-1.80 (7H, m),
1.10-1.30 (11H, m), 0.82 (2H, m).
Example 50
[(5-{(1R)-4-Cyclohexyl-1-[2-(hydroxyamino)-2-oxoethyl]butyl}-1,2,4-oxadiaz-
ol-3-yl)methoxy]acetic acid
[0324] 57
[0325] A solution of ethyl
[(5-{(1R)-4-cyclohexyl-1-[2-(hydroxyamino)-2-ox-
oethyl]butyl)1,2,4-oxadiazol-3-yl)methoxy]acetate (Example 49) (158
mg, 0.40 mmol) in 1,4-dioxan (4 ml) and water (2 ml) was treated
with lithium hydroxide monohydrate (2 mg, 0.48 mmol) and stirred at
room temperature for 2 hours. The reaction mixture was diluted with
water and washed with diethylether (.times.2). The aqueous layer
was acidified to pH 1 with hydrochloric acid (2M) and washed with
ethyl acetate (.times.3). The combined organic layers were washed
with brine, dried over anhydrous sodium sulphate, filtered and the
solvent was removed under reduced pressure to afford the title
compound as a colourless oil (141 mg).
[0326] MS: 370 (MH.sup.+)
[0327] .sup.1H-NMR (CD.sub.3OD) .delta.: 4.70 (2H, s), 4.20 (2H,
s), 3.55 (1H, m), 2.60 (1H, dd), 2.50 (1H, dd), 1.60-1.80 (7H, m),
1.10-1.30 (8H, m), 0.84 (2H, m).
Example 51
Ethyl
2-[(5-{(1R)-4-cyclohexyl-1-[2-(hydroxyamino)-2-oxoethyl]butyl}-1,2,4-
-oxadiazol-3-yl)methoxy]propanoate
[0328] 58
[0329] A solution of (3R)-6-cyclohexyl-3-{3-[(2-ethoxy-1
-methyl-2-oxoethoxy)methyl]-1,2,4-oxadiazol-5-yl}hexanoic acid
(Preparation 89 ) (370 mg, 0.93 mmol) and N-methylmorpholine (170
.mu.l, 1.54 mmol) in anhydrous tetrahydrofuran (15 ml) was cooled
to 0.degree. C., treated with isobutyl chloroformate (130 .mu.l,
100 mmol) and stirred under a nitrogen atmosphere for 2.5 hours.
O-(Trimethylsilyl)hydroxylamin- e (380 .mu.l, 3.10 mmol) was added
and the mixture was stirred for 18 hours, being allowed to warm to
room temperature over this time. The mixture was then treated with
methanol (4 ml) and stirred at room temperature for 1 hour. The
solvent was removed under reduced pressure. The residue was
dissolved in ethyl acetate and washed with water and brine, dried
over anhydrous sodium sulphate, filtered and the solvent removed
under reduced pressure. The oil was purified by column
chromatography on silica gel eluting with a gradient system of
100:0 (dichloromethane:methanol) gradually changing to 90:10
(dichloromethane methanol) to afford the title compound as a
colourless oil (330 mg).
[0330] MS: 434 (MNa.sup.+)
[0331] Analysis: Found, C, 57.55; H, 8.14; N, 9.81%;
C.sub.20H.sub.33N.sub.3O.sub.6. 0.1 H.sub.2O. 0.05 DCM requires C,
57.68; H, 8.04; N, 10.06%
[0332] .sup.1H-NMR (CD.sub.3OD) .delta.:4.75 (1H, d), 4.59 (1H, d),
4.20 (3H, m), 3.55 (1H, m), 6.61 (1H, dd), (2.50 (1H, dd),
1.69-1.80 (7H, m), 1.38 (3H, d), 1.10-1.30 (I1H, m), 0.82 (2H,
m).
Example 52
2-[(5-{(1R)-4-Cyclohexyl-1-[2-(hydroxyamino)-2-oxoethyl]butyl}-1,2,4-oxadi-
azol-3-yl)methoxy]propanoic acid
[0333] 59
[0334] A solution of ethyl 2-[(5-{(1R)-4-cyclohexyl-1
-[2-(hydroxyamino)-2-oxoethyl]butyl}-1,2,4-oxadiazol-3-yl)methoxy]propano-
ate (Example 51) (184 mg, 0.45 mmol) in 1,4-dioxan (4 ml) and water
(2 ml) was cooled to 0.degree. C. and treated with lithium
hydroxide monohydrate (2 mg, 0.48 mmol) and stirred at 0.degree. C.
for 2 hours. The solvent was partially removed under reduced
pressure and diluted with water and washed with diethylether
(.times.2). The aqueous layer was acidified to pH 1 with
hydrochloric acid (2M) and washed with ethyl acetate (.times.2).
The combined organic layers were washed with brine, dries over
anhydrous sodium sulphate, filtered and the solvent was removed
under reduced pressure to afford the title compound as a colourless
oil (160 mg).
[0335] MS: 384 (MH.sup.+)
[0336] .sup.1H-NMR (CD.sub.3OD) .delta.: 4.77 (1H, d), 4.59 (1H,
d), 4.17 (3H, q), 3.45 (1H, m), 2.61 (1H, dd), 2.50 (1H, dd),
1.60-1.80 (7H, m), 1.39 (3H, d), 1.07-1.30 (8H, m), 0.83 (2H,
m).
Example 53
(3R)-3-{3-[(2-Amino-2-oxoethoxy)methyl]-1,2,4-oxadiazol-5-yl}-6-cyclohexyl-
-N-hydroxyhexanamide
[0337] 60
[0338] A solution of
(3R)-3-{3-[(2-amino-2-oxoethoxy)methyl]-1,2,4-oxadiaz-
ol-5-yl}6-cyclohexylhexanoic acid (Preparation 91) (178 mg, 0.50
mmol) and N-methylmorpholine (1001 .mu.l, 0.91 mmol) in anhydrous
tetrahydrofuran (8 ml) was cooled to 0.degree. C., treated with
isobutyl chloroformate (70 .mu.l, 0.54 mmol) and stirred under a
nitrogen atmosphere for 2.5 hours. O-(Trimethylsilyl)hydroxylamine
(200 .mu.l, 1.63 mmol) was added and the mixture was stirred for 18
hours, being allowed to warm to room temperature over this time.
The mixture was then treated with methanol and stirred at room
temperature for 1 hour. The solvent was removed under reduced
pressure. The residue was dissolved in ethyl acetate and washed
with water and brine, dried over anhydrous sodium sulphate,
filtered and the solvent removed under reduced pressure. The oil
was purified by column chromatography on silica gel eluting with a
gradient system of 98:2 (dichloromethane:methanol) gradually
changing to 90:10 (dichloromethane:methanol) and then neat methanol
to afford the title compound as a colourless oil (60 mg).
[0339] MS: 391 (MH.sup.+)
[0340] Analysis: Found, C, 53.67; H, 7.55; N, 14.19%;
C.sub.17H.sub.28N.sub.4O.sub.5. 0.2 CH.sub.2Cl.sub.2 requires C,
53.60; H, 7.43; N, 14.54%
[0341] .sup.1H-NMR (CD.sub.3OD) .delta.:4.70 (2H, s), 4.08 (2H, s),
3.55 (1H, m), 2.45-2.70 (2H, m), 1.60-1.80 (7H, m), 1.10-1.30 (8H,
m), 0.82 (2H, m).
Example 54
Ethyl
3-(5-{(1R)-4-cyclohexyl-1-[2-(hydroxyamino)-2-oxoethyl]butyl}-1,2,4--
oxadiazol-3-yl)propanoate
[0342] 61
[0343] A solution of
3R)-6-cyclohexyl-3-[3-(3-ethoxy-3-oxopropyl)-1,2,4-ox-
adiazol-5-yl]hexanoic acid (Preparation 94) (110 mg, 0.30 mmol) and
N-methylmorpholine (60 .mu.l, 0.54 mmol) in anhydrous
tetrahydrofuran (6 ml) was cooled to 0.degree. C., treated with
isobutyl chloroformate (42 .mu.l, 0.32 mmol) and stirred under a
nitrogen atmosphere for 2.5 hours. O-(Trimethylsilyl)hydroxylamine
(120 .mu.l, 1.00 mmol) was added and the mixture was stirred for 18
hours, being allowed to warm to room temperature over this time.
The mixture was then treated with methanol and stirred at room
temperature for 3 hours. The solvent was removed under reduced
pressure. The residue was dissolved in ethyl acetate and washed
with water and brine, dried over anhydrous sodium sulphate,
filtered and the solvent removed under reduced pressure. The oil
was purified by column chromatography on silica gel eluting with a
gradient system of 100:0 (dichloromethane:methanol) gradually
changing to 90:10 (dichloromethane:methanol) to afford the title
compound as a yellow oil (94 mg).
[0344] MS :404 (MNa.sup.+)
[0345] .sup.1H-NMR (CDCl.sub.3) .delta.:4.15 (2H, q), 3.52 (1H, m),
3.03 (2H, t), 2.78 (4H, m), 2.58 (1H, dd), 1.60-1.80 (7H, m),
1.10-1.35 (11 H, m), 0.82 (2H, m).
Example 55
3-(5-{(1R)-4-Cyclohexyl-1-[2-(hydroxyamino)-2-oxoethyl]butyl}1,2,4-oxadiaz-
ol-3-yl)propanoic acid
[0346] 62
[0347] A solution of ethyl 3-(5-{(1R)-4-cyclohexyl-1
-[2-(hydroxyamino)-2-oxoethyl]butyl}-1,2,4-oxadiazol-3-
yl)propanoate (Example 54) (58 mg, 0.15 mmol) in 1,4-dioxan (2 ml)
and water (1 ml) was treated with lithium hydroxide monohydrate (13
mg, 0.31 mmol) and stirred at room temperature for 2 hours. The
reaction mixture was diluted with water and washed with
diethylether (.times.2). The aqueous layer was acidified with
hydrochloric acid (2M) (2 ml) and washed with ethyl acetate
(.times.2). The combined organic layers were washed with brine,
dried over anhydrous sodium sulphate, filtered and the solvent was
removed under reduced pressure. The oil was purified by column
chromatography on silica gel eluting with a gradient system of
100:0:0 (dichloromethane:methanol:acetic acid) gradually changing
to 90:10:1 (dichloromethane:methanol:acetic acid) to afford the
title compound as an orange gum (36 mg).
[0348] MS: 376 (MNa.sup.+)
[0349] Analysis: Found, C, 55.98; H, 7.61; N, 11.12%;
C.sub.17H.sub.27N.sub.3O.sub.5. 0.2 CH.sub.2Cl.sub.2 requires C,
55.77; H, 7.46; N, 11.34%
[0350] .sup.1H-NMR (CD.sub.3OD) .delta.:3.50 (1H, m), 2.98 (2H, t),
2.76 (2H, t), 2.59 (1H, dd), 2.48 (1H, dd), 1.60-1.80 (7H, m),
1.07-1.35 (8H, m), 0.81 (2H, m).
Example 56
(3R)-6-Cyclohexyl-N-hydroxy-3-{3-[(propylsulfonyl)methyl]-1,2,4-oxadiazol--
5-yl}hexanamide
[0351] 63
[0352] A solution of
(3R)-6-cyclohexyl-3-{3-[(propylsulfonyl)methyl]-1,2,4-
-oxadiazol-5-yl}hexanoic acid (Preparation 97) (298 mg, 0.77 mmol)
and 2,6-lutidine (135 .mu.l, 1.16 mmol) in dichloromethane (10 ml)
was cooled to 0.degree. C., treated with isobutyl chloroformate
(100 .mu.l, 0.77 mmol) and stirred under a nitrogen atmosphere for
1 hour. O-(Trimethylsilyl)hydroxylamine (310 .mu.l, 2.31 mmol) was
added and the mixture was stirred for 4 hours, being allowed to
warm to room temperature over this time. The mixture was then
treated with methanol (5 ml) and stirred at room temperature for 1
hour. The solvent was removed under reduced pressure. The residue
was dissolved in ethyl acetate and washed with hydrochloric acid
(1M) and brine, dried over anhydrous magnesium sulphate, filtered
and the solvent removed under reduced pressure. The oil was
purified by column chromatography on silica gel eluting with a
gradient system of 100:0 (dichloromethane:methanol) gradually
changing to 95:5 (dichloromethane:methanol) to afford the title
compound as a white solid (140 mg).
[0353] MS: 424 (MNa.sup.+)
[0354] Analysis: Found, C, 53.65; H, 7.80; N, 10.26%;
C.sub.18H.sub.31N.sub.3O.sub.5S requires C, 53.84; H, 7.78; N,
10.47%
[0355] .sup.1H-NMR (DMSO) .delta.: 10.39 (1H, br s), 8.65 (1H. br
s), 4.70 (2H, s), 3.45 (1H, m), 3.20 (2H, obs), 2.50 (2H, obs),
1.76 (1H, m), 1.50-1.70 (7H, m), 1.08-1.25 (8H, m), 0.98 (3H, t),
0.80 (2H, m).
Example 57
2-{(1R)-4-Cyclohexyl-1-[2-(hydroxyamino)-2-oxoethyl]butyl)N-[2-(dimethylam-
ino)ethyl]-5-methyl-1,3-oxazole-4-carboxamide
[0356] 64
[0357] A solution 2-((1R)-1
-{2-[(benzyloxy)amino]-2-oxoethyl}4-cyclohexyl-
butyl)-N-[2-(dimethylamino)ethyl]-5-methyl-1,3-oxazole-4-carboxamide
(Preparation 98) (193 mg, 0.39 mmol) in ethanol (10 ml) was treated
with ammonium formate (244 mg, 3.90 mmol) and 5% palladium on
barium sulphate (100 mg) and heated at 43.degree. C., under a
nitrogen atmosphere for 2 hours. The reaction mixture was filtered
and the solvent removed under reduced pressure. The residue was
purified by column chromatography on silica gel eluting with a
gradient system of 98:2:0.2 (dichloromethane:methanol:ammonia)
gradually changing to 90:10:1 (dichloromethane:methanol:ammonia) to
afford the title compound as a white solid (80 mg).
[0358] MS: 409 (MH.sup.+)
[0359] Analysis: Found, C, 60.93; H, 9.06; N, 13.54%;
C.sub.21H.sub.36N.sub.4O.sub.4. 0.2 H.sub.2O requires C, 61.20; H,
8.90; N, 13.59%
[0360] .sup.1H-NMR (d.sub.6-DMSO) .delta.: 8.61 (1H, br s), 7.59
(1H, brs), 3.28 (2H, q), 3.18 (1H, obs), 2.45 (2H, obs), 2.35 (2H,
t), 2.17 (6H, s), 1.50-1.65 (7H, m), 1.10-1.20 (7H, m), 0.80 (2H,
m).
Example 58
{[(2-{(1R)-4-Cyclohexyl-1-[2-(hydroxyamino)-2-oxoethyl]butyl}-5-methyl-1,3-
-oxazol4-yl)carbonyl]amino}acetic acid
[0361] 65
[0362] A solution ({[2-((1R)-1
-2-[(benzyloxy)amino]-2-oxoethyl}4-cyclohex-
ylbutyl)-5-methyl-1,3-oxazol-4-yl]carbonyl}amino)acetic acid
(Preparation 100) (60 mg, 0.12 mmol) in ethanol (3 ml) was treated
with ammonium formate (78 mg, 1.23 mmol) and palladium hydroxide
(20 mg) and heated at 43.degree. C., under a nitrogen atmosphere
for 4 hours. The reaction mixture was filtered and the solvent
removed under reduced pressure. The residue was purified by column
chromatography on silica gel eluting with a gradient system of
99:1:0.1 (dichloromethane:methanol:acetic acid) gradually changing
to 90:10:1 (dichloromethane:methanol:acetic acid) to afford the
title compound as a white solid (17 mg).
[0363] MS : 394 (M-H).sup.-
[0364] .sup.1H-NMR (CDCl.sub.3) .delta.: 7.68 (1H, brs), 4.95-4.10
(2H, m), 3.34 (1H, m), 2.40-2.60 (5H, m), 1.50-1.80 (7H, m),
1.05-1.35 (8H, m), 0.81 (2H, m).
Example 59
(3R)6-Cyclohexyl-N-hydroxy-3-[3-(1H-imidazol-2-ylmethyl)-1,2,4-oxadiazol-5-
-yl]hexanamide
[0365] 66
[0366] A solution of
(3R)-6-cyclohexyl-3-[3-(1H-imidazol-2-ylmethyl)-1,2,4-
-oxadiazol-5-yl]hexanoic acid (Preparation 103) (200 mg, 0.43 mmol)
in anhydrous tetrahydrofuran (5 ml) was treated with
1,1'-carbonyidiimidazol- e (77 mg, 0.48 mmol) and stirred under a
nitrogen atmosphere for 30 minutes. O-(Trimethylsilyl)hydroxylamine
(160 .mu.l, 1.30 mmol) was added and the mixture was stirred for 18
hours. The mixture was then treated with methanol (4 ml) and
stirred at room temperature for 1 hour. The solvent was removed
under reduced pressure. The residue was purified by column
chromatography on silica gel eluting with 90:10:1
(dichloromethane:methanol:ammonia) to afford a colourless oil which
began to crystallise once a little methanol and dichloromethane
were added. The solid was triturated with dichloromethane and dried
under reduced pressure to afford the title compound as a solid (49
mg).
[0367] MS : 362 (MH.sup.+)
[0368] Analysis: Found, C, 59.24; H, 7.42; N, 19.13%;
C.sub.18H.sub.27N.sub.5O.sub.3. 0.2H.sub.20 requires C, 59.22; H,
7.57; N, 19.18%
[0369] .sup.1H-NMR (d.sub.6-DMSO) .delta.: 10.38 (1H, s), 8.65 (1H,
s), 7.70 (1H, s), 7.17 (1H, s), 6.88 (1H, s), 5.39 (2H, s), 3.42
(1H, m), 2.45 (2H, m), 1.5-1.70 (7H, m), 1.00-1.15 (8H, m), 0.78
(2H, m).
Example 60
(3R)-6-cyclohexyl-N-hydroxy-3-[3-(4-pyridinylmethyl)-1,2,4-oxadiazol-5-yl]-
hexanamide
[0370] 67
[0371] A solution of
(3R)-6-cyclohexyl-3-[3-(4-pyridinylmethyl)-1,2,4-oxad-
iazol-5-yl]hexanoic acid (Preparation 106) (380 mg, 0.96 mmol) in
dichloromethane (10 ml) was treated with 1,1'-carbonyldiimidazole
(156 mg, 0.96 mmol) and stirred under a nitrogen atmosphere for 1
hour. O-(Trimethylsilyl)hydroxylamine (388 .mu.l, 2.89 mmol) was
added and the mixture was stirred for 3.5 hours. The mixture was
then treated with methanol (5 ml) and stirred at room temperature
for 2 hours. The solvent was removed under reduced pressure. The
residue was dissolved in ethyl acetate and washed with water and
brine, dried over anhydrous magnesium sulphate, filtered and the
solvent removed under reduced pressure. The residue was purified by
column chromatography on silica gel eluting with 95:5
(dichloromethane:methanol) to afford a white solid. Analysis
suggests that 10% imidazole remains. The solid was dissolved in
ethyl acetate and washed with water, dried over anhydrous magnesium
sulphate, filtered and the solvent was removed under reduced
pressure to afford the title compound as a white solid (128
mg).
[0372] MS: 373 (MH.sup.+)
[0373] Analysis: Found, C, 64.13; H, 7.59; N, 14.86%;
C.sub.20H.sub.28N.sub.4O.sub.3 requires C, 64.49; H, 7.58; N,
15.04%
[0374] .sup.1H-NMR (d.sub.6-DMSO) .delta.: 10.38 (1H, s), 8.65 (1H,
s), 8.49 (2H, d), 7.25 (2H, d), 4.10 (2H, s), 3.39 (1H, m), 2.20
(2H, m), 1.50-1.65 (7H, m), 1.00-1.20 (8H, m), 0.87 (2H, m).
Example 61
(3R)-6-Cyclohexyl-N-hydroxy-3-[3-(3-pyridinylmethyl)-1,2,4-oxadiazol-5-yl]-
hexanamide
[0375] 68
[0376] A solution of
(3R)-6-cyclohexyl-3-[3-(3-pyridinylmethyl)-1,2,4-oxad-
iazol-5-yl]hexanoic acid (Preparation 109) (410 mg, 1.04 mmol) in
dichloromethane (10 ml) was treated with 1,1'-carbonyidiimidazole
(169 mg, 1.04 mmol) and stirred under a nitrogen atmosphere for 1
hour. O-(Trimethylsilyl)hydroxylamine (418 .mu.l, 3.12 mmol) was
added and the reation mixture was stirred for 18 hours. The
reaction mixture was then treated with methanol (5 ml) and stirred
at room temperature for 3 hours. The solvent was removed under
reduced pressure. The residue was dissolved in ethyl acetate and
washed with water (3.times.30 ml) and brine, dried over anhydrous
magnesium sulphate, filtered and the solvent removed under reduced
pressure. The residue was purified by column chromatography on
silica gel eluting with a solvent gradient of 100:0
(dichloromethane:methanol) gradually changing to 95:5
(dichloromethane:methanol) to afford a white waxy solid. The solid
was recrystallised from ethyl acetate to afford the title compound
as a white solid (128 mg).
[0377] MS: 373 (MH.sup.+)
[0378] Analysis: Found, C, 64.53; H, 7.63; N, 14.53%;
C.sub.20H.sub.28N.sub.4O.sub.3.0.15 EtOAc requires C, 64.15; H,
7.63; N, 14.53%
[0379] .sup.1H-NMR (d.sub.6-DMSO) .delta.: 10.36 (1H, s), 8.62 (1H,
s), 8.54 (1H, s), 8.43 (1H, d), 7.65 (1H, d), 7.30 (1H, m), 4.10
(2H, s), 3.40 (1H, m), 2.70 (1H, m), 2.40 (1H, m), 1.50-1.70 (7H,
m), 1.00-1.20 (8H, m), 0.76 (2H, m).
Example 62
2-{(1R)-4-Cyclobutyl-1-[2-(hydroxyamino)-2-oxoethyl]butyl}-N,N-dimethyl-1,-
3-oxazole-4-carboxamide
[0380] 69
[0381] A solution of
(3R)-6-cyclobutyl-3-{4-[(dimethylamino)carbonyl]-1,3--
oxazol-2-yl}hexanoic acid (Preparation 121) (155 mg, 0.50 mmol) and
N-methylmorpholine (90 .mu.l, 0.82 mmol) in anhydrous
tetrahydrofuran (8 ml) was cooled to 0.degree. C., treated with
isobutyl chloroformate (90 .mu.l, 0.70 mmol) and stirred under a
nitrogen atmosphere for 2 hours. O-(Trimethylsilyl)hydroxylamine
(200 .mu.l, 1.63 mmol) was added and the mixture was stirred for 18
hours, being allowed to warm to room temperature over this time.
The mixture was then treated with methanol (2 ml) and stirred at
room temperature for 10 minutes. The solvent was removed under
reduced pressure. The residue was dissolved in ethyl acetate and
washed with water, dried over anhydrous sodium sulphate, filtered
and the solvent removed under reduced pressure. The oil was
purified by column chromatography on silica gel eluting with a
gradient system of 98:2 (dichloromethane:methanol) gradually
changing to 90:10 (dichloromethane:methanol). The solid was
triturated with diethyl ether, filtered and dried under reduced
pressure to afford the title compound as a white solid (64 mg).
[0382] MS: 346 (MNa.sup.+)
[0383] Analysis: Found, C, 59.03; H, 7.81; N, 12.92%;
C.sub.16H.sub.25N.sub.3O.sub.4. 0.1 H.sub.2O requires C, 59.10; H,
7.81; N, 12.92%
[0384] .sup.1H-NMR (d.sub.6-DMSO) .delta.: 10.45 (1H, s), 8.78 (1H,
s), 8.38 (1H, s), 3.25 (1H, t), 3.28 (3H, s), 2.90 (3H, s), 2.40
(1H, dd), 2.28 (1H, dd), 2.12 (1H, m), 1.91 (2H, m), 1.70 (2H, m),
1.57 (2H, m), 1.45 (2H, m), 1.27 (2H, m), 1.02 (2H, m).
Preparation 1
(2R)-2-[2-(tert-Butoxy)-2-oxoethyl]-5-cyclohexylpentanoic acid
[0385] Route A
[0386] (2R)-2-[2-(tert-butoxy)-2-oxoethyl]-5-cyclohexylpentanoic
acid 70
[0387] A solution of
(2R)-2-[2-(tert-butoxy)-2-oxoethyl]-5-phenylpentanoic acid (Syn.
Lett.; 1998; 637-639) (10.00 g, 34.2 mmol) in acetic acid (120 ml)
was treated with 5% Rhodium on alumina catalyst, pressurised to 60
psi with hydrogen in a sealed vessel and stirred at room
temperature for 17 hours. The mixture was filtered through a pad of
Arbocel.RTM. and the solvent was removed from the filtrate under
reduced pressure. The residue was azeotroped from toluene to afford
the title compound (7.53 g).
[0388] MS: 299 (MH.sup.+)
[0389] .sup.1H-NMR (CDCl.sub.3) .delta.: 2.80 (1H, m), 2.61 (1H,
m), 2.38 (1H, m), 1.75-1.56 (7H, m), 1.55-1.04 (17H, m), 0.84 (2H,
m).
[0390] Route B
[0391] (4S)-4-Benzyl-3-(5-cyclohexylpentanoyl)-1,3-oxazolidin-2-one
71
[0392] A solution of 5-cyclohexylpentanoic acid (63.50 g, 345 mmol)
in N,N-dimethylformamide (0.5 ml) and dichloromethane (350 ml) was
cooled to 5.degree. C. and treated dropwise with oxalyl chloride
(31.6 ml, 362 mmol) over 30 minutes. The mixture was stirred at
0.degree. C. for 3 hours then the solvent was removed under reduced
pressure to afford 5-cyclohexylpentanoyl chloride as a pale yellow
solid (70.0 g).
[0393] A solution of n-butyllithium (100 ml, 250 mmol, 2.5M in
hexanes) was added via a cannula to a solution of
(4S)-4-benzyl-1,3-oxazolidin-2-o- ne (44.30 g, 250 mmol) in
anhydrous tetrahydrofuran (400 ml) at -78.degree. C. The yellow
solution was then stirred for 45 minutes. A solution of
5-cyclohexylpentanoyl chloride (55.5 g, 275 mmol) in
tetrahydrofuran (100 ml) was then added over 1 hour. The mixture
was stirred at -78.degree. C. for 30 minutes then warmed to room
temperature over 1 hour. The mixture was quenched with an aqueous
solution of ammonium chloride (20% w/v, 400 ml) and extracted with
ethyl acetate. The layers were separated and the organic layer was
washed with brine, dried over anhydrous sodium sulphate and the
solvent removed under reduced pressure. The solid was
recrystallised from hexane (500 ml) to afford the title compound as
a white solid (81.0 g).
[0394] MS: 344 (MH.sup.+)
[0395] .sup.1H-NMR (CDCl.sub.3) .delta.: 7.41-7.13 (5H, m), 4.68
(I1H, m), 4.27-402 (2H, m), 3.31 (1H, dd, J=16, 4Hz), 3.06-2.70
(3H, m), 1.81-1.53 (7H, m), 1.49-1.04 (8H, m), 0.88 (2H, m)
[0396] tert-Butyl
3-{[(4S)-benzyl-2-oxo-1,3-oxazolidin-3-yl]carbonyl}-6-cy-
clohexylhexanoate 72
[0397] A solution of
(4S)-4-benzyl-3-(5-cyclohexylpentanoyl)-1,3-oxazolidi- n-2-one
(70.0 g, 204 mmol) in anhydrous tetrahydrofuran (650 ml) was cooled
to -70.degree. C. and treated dropwise with sodium
hexamethyldisilazide (1M in tetrahydrofuran, 224 ml, 224 mmol) over
45 minutes. The mixture was stirred for a further 45 minutes before
being treated with t-butylbromoacetate (31.6 ml, 214 mmol). This
mixture was stirred at -70.degree. C. for 30 minutes then warmed to
-30.degree. C. and quenched with an aqueous solution of ammonium
chloride (20% w/v, 400 ml) and warmed to room temperature. The
mixture was extracted with ethyl acetate and the combined organic
extracts were washed with brine, dried over anhydrous sodium
sulphate and the solvent removed under reduced pressure. The solid
was recrystallised from hexane to afford the title compound as a
white solid (71.4 g).
[0398] MS: 458(MH.sup.+)
[0399] .sup.1H-NMR (CDCl.sub.3) .delta.: 7.41-7.13 (5H, m), 4.66
(1H, m), 4.23-4.03 (3H, m), 3.35 (1H, dd, J=16, 4Hz), 2.95-2.68
(3H, m), 2.47 (1H, m), 1.80-1.07 (24H, m), 0.85 (2H, m)
[0400] 2-[2-(tert-Butoxy)-2-oxoethyl]-5-cyclohexylpentanoic acid
73
[0401] A solution of tert-butyl
3-{[(4S)-4-benzyl-2-oxo-1,3-oxazolidin-3-y-
l]carbonyl}-6-cyclohexylhexanoate (64.0 g, 139.9 mmol) in
tetrahydrofuran:water (3:1, 800 ml) was cooled to 5.degree. C. then
treated sequentially with hydrogen peroxide (30% w/v water, 87 ml,
769 mmol) then lithium hydroxide hydrate (10.0 g, 238 mmol). The
reaction was stirred for 1 hour then quenched by dropwise addition
of an aqueous solution of sodium thiosulphate (500 ml) keeping the
temperature below 20.degree. C. The mixture was extracted with
ethyl acetate (discarded) and the aqueous phase was acidifed to pH
2 with solid citric acid and extracted with ethyl acetate. The
combined organic phases were washed with brine, dried over
anhydrous sodium sulphate and the solvent removed under reduced
pressure. The residue was purified by column chromatography on
silica gel eluting with a gradient system of hexane : ethyl acetate
(2:1) gradually changing to hexane:ethyl acetate (1:1) to afford
the title compound (40.7 g)
[0402] Route C
[0403] 3-(Diethoxyphosphoryl)succinic acid 1 -tert-butyl ester
74
[0404] Triethylphosphonoacetate (102 g, 0.45 mol) was added
dropwise over 11 min to a stirred solution of potassium
tert-butoxide (60 g, 0.54 mol) in THF (500 ml), at 0.degree. C.,
under nitrogen. The mixture was stirred for 1 hour at 0.degree. C.
and then dichloromethane (300 ml) was added and the reaction
mixture was warmed to 25-30.degree. C. The mixture was stirred at
25-30.degree. C. for 1 hour and then added dropwise over 33 minutes
to a solution of tert-butyl bromoacetate (96 g, 0.49 mol) in THF
(500 ml), at 0.degree. C., under nitrogen. The mixture was stirred
at 0-5.degree. C. for 2 hours and then a solution of citric acid
(174 g, 0.91 mol) in demineralised water (250 ml) was added. The
mixture was concentrated in vacuo to remove most of the THF and
then toluene (750 ml) was added. The organic phase was separated,
washed with brine (2.times.150 ml) and concentrated in vacuo to
leave a colourless oil. The oil was taken up in ethanol and a
solution of potassium hydroxide (36. g, 0.64 mol) in demineralised
water (150 ml) was added dropwise over 15 mins. The mixture was
stirred at 0.degree. C. for 4 hours and then a solution of citric
acid (158 g, 0.82 mol) in demineralised water (600 ml), and toluene
(600 ml), were added. The organic phase was separated and the
aqueous phase was re-extracted with toluene (600 ml). The combined
organic phases were washed with demineralised water (2.times.150
ml) and concentrated in vacuo to leave a white solid. Toluene (150
ml) was added and the slurry was re-concentrated in vacuo to leave
a white solid. The product was purified by crystallisation from
tert-butylmethyl ether (300 ml) and cyclohexane (600 ml) to give
the title compound as a solid (79 g).
[0405] .sup.1H-NMR (CDCl.sub.3) .delta.: 4.20-4.10 (4H, m),
3.49-3.36 (1H, m), 3.00-2.85 (1H, m), 2.72-2.60 (1H, m), 1.20 (9H,
s), 1.37-1.27 (6H, m)
[0406] Alternative Preparation
[0407] Triethylphosphonoacetate (12.0 Kg, 53.5 mol) was added over
30 minutes to a stirred solution of potassium tert-butoxide (7.20
Kg, 64.2 mol) in THF (118 litres), between 0 and 5.degree. C.,
under nitrogen. The mixture was warmed to 25-30.degree. C. where it
was stirred for 1 hour and then added over 45 minutes to a solution
of tert-butyl bromoacetate (11.5 Kg, 59.0 mol) in THF (28 litres),
between 0 and 5.degree. C., under nitrogen. The mixture was stirred
at 0-5.degree. C. for 1 hour and then demineralised water (6.1
litres) and ethanol (30 litres) were added. A solution of potassium
hydroxide (4.2 Kg, 75.0 mol) in demineralised water (84 litres) was
then added over 2 hours, between -5 and 0.degree. C. The mixture
was stirred at -10.degree. C. for 16 hours and then a solution of
citric acid (16.5 Kg, 85.8 mol) in demineralised water (32 litres)
was added. The mixture was concentrated in vacuo to a volume of 180
litres and then ethyl acetate (90 litres) was added. The organic
phase was separated and the aqueous phase was re-extracted with
ethyl acetate (30 litres). The combined organic phases were washed
with water (30 litres) and then stripped and replaced with
cyclohexane by distillation at atmospheric pressure, at a constant
volume of 72 litres. tert-Butylmethyl ether (18 litres) was added
and the mixture was stirred at ambient temperature for 12 hours and
then filtered. The residue was washed with a mixture of cyclohexane
(16 litres) and tert-butylmethyl ether (3.6 litres) then dried in
vacuo for 16 hours to give the title compound as a colourless solid
(10.0 Kg, 60%).
[0408] .sup.1H-NMR (CDCl.sub.3) .delta.: 4.20-4.10 (4H, m),
3.49-3.36 (1H, m), 3.00-2.85 (1H, m), 2.72-2.60 (1H, m), 1.20 (9H,
s), 1.37-1.27 (6H, m)
[0409] (E)-.sup.2-[2-(tert-Butoxy)-2-oxoethyl]-5-phenyl-2-pentenoic
acid 75
[0410] A solution of 3-(diethoxyphosphoryl)succinic acid
1-tert-butyl ester (100 g, 0.32 mol) in THF (300 ml) was added
dropwise over 15 min to a stirred solution of potassium
tert-butoxide (110 g, 0.98 mol) in THF (300 ml), between -10 and
-5.degree. C., under nitrogen. The mixture was stirred at
-10.degree. C. for 15 min and then a solution of
hydrocinnamaidehyde (46.8 g, 0.35 mmol) in THF (100 ml) was added
dropwise over 15 min, between -13 and -8.degree. C. The mixture was
stirred at -10.degree. C. for 30 min and then a solution of citric
acid (111 g, 0.58 mol) in demineralised water (500 ml), and ethyl
acetate (500 ml), were added. The pH was adjusted to pH 4 with
aqueous sodium hydroxide solution (50%) and the phases were
separated. The aqueous fraction was washed with ethyl acetate (500
ml) and the combined organic fractions were washed with saturated
sodium bicarbonate solution (500 ml), citric acid solution (10%,
500 ml) and demineralised water (500 ml) and then concentrated in
vacuo. The resulting solid was slurried in cyclohexane (470 ml) for
1 hour and then the mixture was filtered. The residue was washed
with cyclohexane (2.times.50 ml) and dried in vacuo to leave the
title compound as a colourless solid (76 g, 81%).
[0411] MS: 289 [(M-H)].sup.-
[0412] .sup.1H-NMR (CDCl.sub.3) .delta.: 7.33-7.16 (5H, m), 7.05
(1H, br t), 3.20 (2H, s), 2.89 (2H, br t), 2.50 (2H, br dd), 1.41
(9H, s)
[0413] (R)-2-[2-(tert-Butoxy)-2-oxoethyl]-5-phenylpentanoic acid
76
[0414] A stirred solution of
(E)-2-[2-(tert-butoxy)-2-oxoethyl]-5-phenyl-2- -pentenoic acid (100
g, 0.34mol), cyclohexylamine (39 ml, 0.34 mol) and
[(S)-2,2'-bis(diphenylphosphino-1,1'-binaphthyl]chloro(p-cymene)ruthenium
chloride (0.64 g, 0.69 mmol) in methanol (1000 ml) was heated to
60.degree. C., under hydrogen (60 p.s.i.), for 42 hours and then
allowed to cool to room temperature. The mixture was filtered
through celite and then concentrated in vacuo to a yellow solid
which was purified by re-crystallisation from acetone (850 ml). The
resulting solid was partitioned between ethyl acetate (1200 ml) and
citric acid solution (10%, 1200 ml) and the organic phase was
separated, washed with demineralised water (1200 ml) and
concentrated in vacuo to leave the title compound as an oil (80
g).
[0415] H-NMR (CDCl.sub.3) .delta.: 7.30-7.17 (5H, m), 2.85-2.78
(1H, m), 2.66-2.58 (3H, m), 2.37 (1H, br dd), 1.75-1.51 (4H, m),
1.40 (9H, s)
[0416] Preparation of cyclohexylamine salt 77
[0417] A stirred solution of cyclohexylamine (266 ml, 2.32 mol),
(E)-2-[2-(tert-butoxy)-2-oxoethyl]-5-phenyl-2-pentenoic acid (688
g, 2.37 mol) and
[(S)-2,2'-bis(diphenylphosphino-1,1'-binaphthyl]chloro(p-cymene)-
ruthenium chloride (4.4 g, 4.7 mmol) in methanol (6.9 litres) was
heated to 60.degree. C., under hydrogen (60 p.s.i.), for 47 hours
and then allowed to cool to room temperature (enantiomeric
excess=88%). The mixture was filtered through celite and then the
solvent was stripped and replaced with acetone by distillation at
atmospheric pressure, at a constant volume of 4.2 litres. The
resulting suspension was cooled to room temperature where it was
stirred for 4 hours and then filtered. The residue was washed with
acetone (2.times.1 litre) and then dried in vacuo at 45.degree. C.
for 16 hours to leave the title compound as a colourless solid (590
g, 64%, enantiomeric excess=98.9%).
[0418] .sup.1H-NMR (CD.sub.3OD) .delta.: 7.23-7.09 (5H, m),
3.05-2.98 (1H, m), 2.64-2.56 (3H, m), 2.53 (1H, dd, J 15.2, 7.2Hz),
2.23 (1H, dd, J 15.2, 7.2Hz), 2.00-1.97, (2H, m), 1.85-1.81 (2H,
m), 1.72-1.20 (10 H, m), 1.40 (9H, s)
(R)-2-[2-(tert-Butoxy)-2-oxoethyl]-5-cyclohexylpentanoic acid
cyclohexylamine salt
[0419] 78
[0420] (R)-2-[2-(tert-Butoxy)-2-oxoethyl]-5-phenylpentanoic acid
cyclohexylamine salt (691 g, 1.77 mol) and ethyl acetate (7.0
litres) were added to an aqueous solution of citric acid (10%, 6.3
litres) and the organic phase was separated, washed with water (7.0
litres) and concentrated in vacuo to a yellow oil. A solution of
the oil and 5% rhodium on carbon (51.6 g) in methanol (7.0 litres)
was stirred at ambient temperature, under hydrogen (150 p.s.i.) for
48 hours and then filtered through celite. To the filtrate was
added cyclohexylamine (202 ml, 1.77 mol) and the methanol solution
was stripped and replaced with methylethyl ketone by distillation
at atmospheric pressure, to a volume of 5.5 litres. The mixture was
allowed to cool to ambient temperature where it was stirred for 48
hours and then filtered. The residue was washed with methylethyl
ketone (2.times.500 ml) and then dried in vacuo at 45.degree. C.
for 4 hours to leave the title compound as a colourless solid (495
g, 71%).
[0421] .sup.1H-NMR (CD.sub.3OD) .delta.: 3.06-2.99 (1H, m),
2.63-2.56 (1H, m), 2.53 (1H, dd, J 15.2, 7.2Hz), 2.23 (1H, dd, J
15.2, 7.2Hz), 2.02-1.97 (2H, m), 1.77-1.15 (21 H, m), 1.43 (9H, s),
0.93-0.82 (2H, m)
[0422] (R)-2-[2-(tert-Butoxy)-2-oxoethyl]-5-cyclohexylpentanoic
acid 79
[0423] A solution of
(R)-2-[2-(tert-butoxy)-2-oxoethyl]-5-phenylpentanoic acid (2.2 g,
7.5 mmol) and 5% Rh/C (0.22 g) in methanol (220 ml) was stirred at
room temperature, under hydrogen (150 p.s.i.) for 24 hours and then
filtered through celite. The filtrate was concentrated in vacuo to
leave the title compound as an oil (2.0 g).
[0424] .sup.1H-NMR (CDCl.sub.3) .delta.: 2.82-2.76 (1H, m), 2.60
(1H, br dd), 2.37 (1H, br dd), 1.70-1.60 (6H, m), 1.51-1.30 (3H,
m), 1.42 (9H, s), 1.23-1.11 (6H, m), 0.96-0.80 (2H, m)
Preparation 2
tert-Butyl
(3R)-3-[({[(Z)-1-amino-2-ethoxy-2-oxoethylidene]amino}oxy)carbo-
nyl]-6-cyclohexylhexanoate
[0425] 80
[0426] A solution of
(2R)-2-[2-(tert-butoxy)-2-oxoethyl]-5-cyclohexylpenta- noic acid
(Preparation 1) (7.53 g, 25.2 mmol) in 1,4-dioxane (175 ml) was
treated with 1 -hydroxybenzotriazole hydrate (3.75 g, 27.8 mmol)
and the mixture cooled to 0.degree. C.
N,N'-Dicyclohexylcarbodiimide (5.47 g, 26.5 mmol) was then added
and the mixture was stirred for 3 hours being allowed to warm to
room temperature over this time. The mixture was then filtered and
washed with 1,4-dioxane (2.times.50 ml). The filtrate was then
treated with sodium carbonate (4.01 g, 37.8 mmol) and ethyl
2-amino-2-(hydroxyimino)acetate (J.Org.Chem.;23; 1958; 1794) (3.33
g, 25.2 mmol). The resulting mixture was stirred at room
temperature for 17 hours. The solvent was removed under reduced
pressure and the residue was partitioned between ethyl acetate and
water. The layers were separated and the aqueous phase was
extracted with ethyl acetate (.times.2). The combined organic
layers were dried over anhydrous magnesium sulphate, filtered and
the solvent removed under reduced pressure. The residue was
purified by column chromatography on silica gel eluting with a
gradient system of ethyl acetate:pentane (30:70) gradually changing
to ethyl acetate: pentane (50:50) to afford the title compound as a
white solid (6.50 g).
[0427] MS: 413 (MH.sup.+)
[0428] .sup.1H-NMR (CDCl.sub.3) .delta.: 5.71 (2H, br s), 4.39 (2H,
q), 2.92 (1H, m), 2.67 (1H, dd), 2.44 (1H, dd), 1.75-1.32 (22H, m),
1.26-1.04 (5H, m), 0.84 (2H, m).
Preparation 3
Ethyl
5-{(1R)-1-[2-(tert-butoxy)-2-oxoethyl]-4-cyclohexylbutyl)-1,2,4-oxad-
iazole-3-carboxylate
[0429] 81
[0430] A solution of tert-Butyl
(3R)-3-[({[(Z)-1-amino-2-ethoxy-2-oxoethyl-
idene]amino}oxy)carbonyl]-6-cyclohexylhexanoate (Preparation 2)
(21.0 g, 50.82 mmol) in xylene (400 ml) was heated at 130.degree.
C. for 17 hours, then allowed to cool to room temperature. The
residue was purified by column chromatography on silica gel eluting
with a gradient system of ethyl acetate : pentane (5:95) gradually
changing to ethyl acetate: pentane (20:80) to afford the title
compound as a colourless oil (20.0 g).
[0431] MS : 395 (MH.sup.+), 412 (MNH.sub.4.sup.+)
[0432] .sup.1H-NMR (CDCl.sub.3) .delta.: 4.51 (2H, m), 3.54 (1H,
m), 2.86 (1H, dd), 2.65 (1H, dd), 1.86-1.57 (7H, m), 1.50-1.33
(12H, m), 1.30-1.03 (8H, m), 0.82 (2H, m).
Preparation 4
(3R)-6-Cyclohexyl-3-[3-(ethoxycarbonyl)-1,2,4-oxadiazol-5-yl]hexanoic
acid
[0433] 82
[0434] A solution of ethyl
5-{(1R)-1-[2-(tert-butoxy)-2-oxoethyl]-4-cycloh-
exylbutyl}-1,2,4-oxadiazole-3-carboxylate (Preparation 3) (820 mg,
2.08 mmol) in dichloromethane (10 ml) was cooled to 0.degree. C.
and treated with trifluoroacetic acid (5 ml). The mixture was
stirred for 2.5 hours being allowed to warm to room temperature
over this time. The solvent was removed under reduced pressure and
the residue azeotroped with toluene (.times.2). The residue was
then dissolved in ethyl acetate, washed sequentially with an
aqueous solution of sodium dihydrogen citrate and brine, dried over
anhydrous sodium sulphate, filtered and the solvent removed under
reduced pressure to afford the title compound as an oil (740
mg).
[0435] MS: 339 (MH.sup.+)
[0436] .sup.1H-NMR (CDCl.sub.3) .delta.:4.49 (2H, q J=7Hz), 3.57
(1H, m), 3.05 (1H, dd J=17, 8Hz), 2.81 (1H, dd J=17, 4Hz),
1.92-1.55 (7H, m), 1.45 (3H, t J=7Hz), 1.35-1.02 (8H, m), 0.84 (2H,
m)
Preparation 5
tert-Butyl
(3R)-3-[3-(aminocarbonyl)-1,2,4-oxadiazol-5-yl]-6-phenylhexanoa-
te
[0437] 83
[0438] A solution of ethyl
5-{(1R)-1-[2-(tert-butoxy)-2-oxoethyl]-4-cycloh-
exylbutyl}-1,2,4-oxadiazole-3-carboxylate (Preparation 3) (400 mg,
1.01 mmol) in ethanol saturated with ammonia gas (20 ml) was
stirred at room temperature for 18 hours. The solvent was removed
under reduced pressure and the residue was purified by column
chromatography on silica gel eluting with a gradient system of
hexane:ethyl acetate (90:10) gradually changing to hexane: ethyl
acetate (60:40) to afford the title compound as a white solid ( 260
mg).
[0439] MPt: 77-79.degree. C.
[0440] MS: 366 (MH.sup.+), 383 (MNa.sup.+)
[0441] Analysis: Found C, 62.42; H, 8.59; N, 11.48%;
C.sub.19H.sub.31N.sub.3O.sub.4 requires C, 62.44; H, 8.55; N,
11.50%
[0442] .sup.1H-NMR (CDCl.sub.3) .delta.: 6.80 (1H, br s), 5.90 (1H,
br s), 3.53 (1H, m), 2.87 (1H, dd, J=17, 9Hz), 2.66 (1H, dd, J=17,
5Hz), 1.90-1.50 (7H, m), 1.46-1.02 (17H, m), 0.83 (2H, m).
[0443] Alternative preparation of tert-butyl
(3R)-3-[3-(aminocarbonyl)-1,2-
,4-oxadiazol-5-yl]-6-cyclohexylhexanoate 84
[0444]
tert-Butyl(3R)-3-[({[(Z)-1,2-diamino-2-oxoethylidene]amino}oxy)carb-
onyl]-6-cyclohexylhexanoate (Preparation 101) (4.10 g, 10.7 mmol)
in mixed xylenes (25 ml) was heated to reflux for 48 hours. The
solvent was removed under reduced pressure and the residue was
purified by column chromatography on silica gel eluting with
isocratic system of n-hexane:ethyl acetate (75:25) to afford a
yellow oil. The oil was crystallised from cyclohexane to afford the
title compound as a colourless solid (0.60 g).
[0445] .sup.1H-NMR (CDCl.sub.3) .delta.: 6.80 (1H, br s), 5.90 (1H,
br s), 3.53 (1H, m), 2.87 (1H, dd, J=17, 9 Hz), 2.66 (1H, dd, J=17,
5 Hz), 1.90-1.50 (7H, m), 1.46-1.02 (17H, m), 0.93-0.82 (2H,
m).
Preparation 6
(3R)-3-[3-(Aminocarbonyl)-1,2,4-oxadiazol-5-yl]-6-cyclohexylhexanoic
acid
[0446] 85
[0447] A solution of tert-butyl
(3R)-3-[3-(aminocarbonyl)-1,2,4-oxadiazol-- 5-yl]-6-phenylhexanoate
(Preparation 5) (250 mg, 0.68 mmol) in dichloromethane (10 ml) was
cooled to 0.degree. C. and treated with trifluoroacetic acid (5
ml). The mixture was stirred for 2 hours, being allowed to warm to
room temperature over this time. The solvent was removed under
reduced pressure and the residue was azeotroped with toluene
(.times.2) then hexane to afford the title compound as a white
solid (204 mg).
[0448] MPt.: 172-174.degree. C.
[0449] Analysis: Found C, 58.03; H, 7.48; N, 13.38%;
C.sub.15H.sub.23N.sub.30.sub.4 requires C, 58.24; H, 7.49; N,
13.19%
[0450] .sup.1H-NMR (CD.sub.3OD) .delta.: 3.55 (1H, m), 2.93 (1H,
dd, J=17, 9Hz), 2.80 (1H, dd, J=17, 4Hz), 1.84-1.59 (7H, m),
1.40-1.08 (8H, m), 0.86 (2H, m).
Preparation 7
tert-Butyl
(3R)-6-cyclohexyl-3-{3-[(methylamino)carbonyl]-1,2,4-oxadiazol--
5-yl)hexanoate
[0451] 86
[0452] A solution of ethyl
5-{1R)-1-[2-(tert-butoxy)-2-oxoethyl]-4-cyclohe-
xylbutyl)1,2,4-oxadiazole-3-carboxylate (Preparation 3) (4.70 g,
11.9 mmol) in ethanol (80 ml) was treated with methylamine (33% w/v
in ethanol, 12.0 ml, 96.0 mmol) and the solution was stirred at
room temperature for 18 hours. The solvent was removed under
reduced pressure and the residue was purified by column
chromatography on silica gel eluting with a gradient system of
dichloromethane:ethyl acetate (9:1) gradually changing to
dichloromethane:ethyl acetate (8:2) to afford the title compound as
a pale yellow oil which crystallised on standing (4.23 g).
[0453] MS: 380 (MH.sup.+)
[0454] .sup.1H-NMR (CDCl.sub.3) .delta.: 6.97 (1H, br m), 3.48 (1H,
m), 3.04 (3H, d), 2.84 (1H, dd, J=1 7, 9Hz), 2.66 (1H, dd, J=17,
4Hz), 1.84-1.55 (7H, m), 1.39 (9H, s), 1.33-1.02 (8H, m), 0.83 (2H,
m).
Preparation 8
(3R)-6-Cyclohexyl-3-3-[(methylamino)carbonyl]-1,2,4-oxadiazol-5-yl}hexanoi-
c acid
[0455] 87
[0456] A solution of tert-butyl
(3R)-6-cyclohexyl-3-{3-[(methylamino)carbo-
nyl]-1,2,4-oxadiazol-5-yl}hexanoate (Preparation 7, 380 mg, 1.00
mmol) in dichloromethane (10 ml) was treated with trifluoroacetic
acid (5 ml) and the mixture was stirred at room temperature for 3.5
hours. The solvent was removed under reduced pressure and the
residue azeotroped with toluene (.times.2). The residue was
dissolved in ethyl acetate and washed sequentially with a saturated
aqueous solution of sodium citrate then brine. The organic layer
was dried over anhydrous sodium sulphate, filtered and the solvent
removed under reduced pressure. The residue was crystallised from
hexane to afford the title compound as a white solid (310 mg).
[0457] MPt.:83-86.degree. C.
[0458] MS: 341 (MNH.sub.4.sup.+)
[0459] Analysis: Found C, 59.24; H, 7.75; N, 12.77%;
C.sub.16H.sub.25N.sub.3O.sub.4 requires C, 59.43; H, 7.79; N,
12.79
[0460] .sup.1H-NMR (CDCl.sub.3) .delta.:6.94 (1H, brm), 3.55 (1H,
m), 3.01 (4H, m), 2.79 (1H, dd, J=14, 3), 1.86-1.53 (7H, m),
1.35-1.06 (8H, m), 0.83 (2H, m).
Preparation 9
(3R)-6-Cyclohexyl-3{-3-[(propylamino)carbonyl]-1,2,4-oxadiazol-5-yl}hexano-
ic acid
[0461] 88
[0462] A solution of
(3R)-6-cyclohexyl-3-[3-(ethoxycarbonyl)-1,2,4-oxadiaz-
ol-5-yl]hexanoic acid (Preparation 4) (107 mg, 0.31 mmol) in
toluene (2 ml) was treated with n-propylamine (250 .mu.l, 3.10
mmol) and the mixture was heated at 125.degree. C. in a sealed
vessel for 2 hours. The mixture was cooled to room temperature,
diluted with ethyl acetate and washed sequentially with aqueous
citric acid solution (5% w/v), water and brine, dried over
anhydrous sodium sulphate, filtered and the solvent removed under
reduced pressure. The residue was purified by column chromatography
on silica gel eluting with a gradient system of
dichloromethane:ethyl acetate (80:20) gradually changing to
dichloromethane:ethyl acetate (60:40) then to
dichloromethane:methanol (90:10) to afford the title compound as an
oil (76 mg).
[0463] MS: 352 (MH.sup.+)
[0464] .sup.1H-NMR (DMSO-d.sub.6) .delta.: 8.90 (1H, m), 3.64-3.00
(3H, m), 2.80 (2H, m), 1.81-1.43 (9H, m), 1.39-1.00 (8H, m), 0.83
(5H, m).
Preparation 10
tert-Butyl
(3R)-6-cyclohexyl-3-{3-[(dimethylamino)carbonyl]-1,2,4-oxadiazo-
l-5-yl}hexanoate
[0465] 89
[0466] A solution of ethyl
5{(1R)-1-[2-(tert-butoxy)-2-oxoethyl]4-cyclohex-
ylbutyl)-1,2,4-oxadiazole-3-carboxylate (Preparation 3) (1.00 g,
2.53 mmol) in ethanol (8 ml) was cooled to 0.degree. C. and treated
dropwise with dimethylamine (5.6M in ethanol, 4.50 ml, 25.3 mmol).
The solution was stirred for 17 hours being allowed to warm up to
room temperature over this time. The solvent was removed under
reduced pressure and the residue was purified by column
chromatography on silica gel eluting with dichloromethane:ethyl
acetate (4:1) to afford the title compound as a yellow oil (0.93
g).
[0467] MS: 394 (MH.sup.+), 411 (MNH.sub.4.sup.+)
[0468] .sup.1H-NMR (CDCl.sub.3) .delta.:3.50 (1H, m), 3.12 (6H, d),
2.85 (iH, dd, J=16, 7Hz), 2.65 (1H, dd, J=16, 5Hz), 1.84-1.57 (7H,
m), 1.39 (9H, s), 1.34-1.05 (8H, m), 0.83 (2H, m).
Preparation 11
(3R)-6-Cyclohexyl-3-{3-[(dimethylamino)carbonyl]-1,2,4-oxadiazol-5-yl}hexa-
noic acid
[0469] 90
[0470] A solution of tert-butyl
(3R)-6-cyclohexyl-3-{3-[(dimethylamino)car-
bonyl]-1,2,4-oxadiazol-5-yl}hexanoate (Preparation 10) (2.35 g,
5.97 mmol) in dichloromethane (10 ml) was treated with
trifluoroacetic acid (2 ml) and the resulting mixture was stirred
at room temperature under a nitrogen atmosphere for 17 hours. The
solvent was removed under reduced pressure and the residue
azeotroped from dichloromethane. The residue was purified by column
chromatography on silica gel eluting with dichloromethane:methanol
(98:2) to afford the title compound (1.27 g).
[0471] MS: 360 (MNa.sup.+), 355 (MNH.sub.4.sup.+)
[0472] Analysis: Found C, 60.63; H, 8.16; N, 12.30%;
C.sub.17H.sub.27N.sub.3O.sub.4 requires C, 60.51; H, 8.07; N,
12.45%
[0473] .sup.1H-NMR (DMSO-d.sub.6) .delta.: 3.45 (1H, m), 2.99 (3H,
s), 3.91 (3H, s), 2.76 (2H, m), 1.73-1.52 (7H, m), 1.30-1.02 (8H,
m), 0.80 (2H, m).
Preparation 12
tert-Butyl
(3R)-6-cyclohexyl-3-[3-(1-pyrrolidinylcarbonyl)-1,2,4-oxadiazol-
-5-yl]hexanoate
[0474] 91
[0475] A solution of ethyl 5-{(1R)-1
-[2-(tert-butoxy)2-oxoethyl]4-cyclohe-
xylbutyl}-1,2,4-oxadiazole-3-carboxylate (Preparation 3) (300 mg,
0.76 mmol) in ethanol (4 ml) was treated dropwise with pyrrolidine
(0.63 ml, 7.60 mmol) and the resulting solution was heated at
60.degree. C. for 9 hours. The solvent was removed under reduced
pressure and the residue was purified by column chromatography on
silica gel eluting with dichloromethane:ethyl acetate (4:1) to
afford the title compound as a pale yellow oil (360 mg).
[0476] MS :420 (MH.sup.+), 437 (MN H.sub.4.sup.+)
[0477] .sup.1H-NMR (CDCl.sub.3) .delta.:3.71 (4H, m), 3.50 (1H, m),
2.86 (1H, dd, J=16, 8Hz), 2.64 (1H, dd, J=16, 3Hz), 1.96 (4H, m),
1.74-1.55 (7H, m), 1.38 (9H, s), 1.33-1.04 (8H, m), 0.82 (2H,
m).
Preparation 13
(3R)-6-Cyclohexyl-3-[3-(1
-pyrrolidinylcarbonyl)-1,2,4-oxadiazol-5-yl]hexa- noic acid
[0478] 92
[0479] A solution of tert-butyl
(3R)-6-cyclohexyl-3-[3-(1-pyrrolidinylcarb-
onyl)-1,2,4-oxadiazol-5-yl]hexanoate (Preparation 12) (356 mg, 0.85
mmol) in dichloromethane (4 ml) was treated with trifluoroacetic
acid (1 ml) and the resulting mixture was stirred at room
temperature under a nitrogen atmosphere for 17 hours. The solvent
was removed under reduced pressure and the residue azeotroped from
toluene to afford the title compound (288 mg).
[0480] MS: 364 (MH.sup.+), 381 (MNH.sub.4.sup.+)
[0481] .sup.1H-NMR (CD.sub.3OD) .delta.: 3.83-3.45 (5H, m), 2.94
(1H, dd, J=16, 8Hz), 2.81 (1H, dd, J=16, 4Hz), 1.98 (4H, m),
1.87-1.54 (7H, m), 1.44-1.06 (8H, m), 0.88 (2H, m).
Preparation 14
tert-Butyl
(3R)-6-cyclohexyl-3-[3-(1-piperidinylcarbonyl)-1,2,4-oxadiazol--
5-yl]hexanoate
[0482] 93
[0483] A solution of ethyl 5-(1R)-1
-[2-(tert-butoxy)-2-oxoethyl]4-cyclohe- xylbutyl}-1
,2,4-oxadiazole-3-carboxylate (Preparation 3) (300 mg, 0.76 mmol)
in ethanol (4 ml) was treated dropwise with piperidine (0.75 ml,
7.60 mmol) and the resulting mixture was heated at 60.degree. C.
under a nitrogen atmosphere for 9 hours. The mixture was cooled and
the solvent removed under reduced pressure. The residue was
purified by column chromatography on silica gel eluting with
dichloromethane:ethyl acetate (80:20) to afford the title compound
as a pale yellow oil (334 mg).
[0484] MS: 434 (MH.sup.+), 451(MNH.sub.4.sup.+)
[0485] .sup.1H-NMR (CDCl.sub.3) .delta.: 3.72 (2H, m), 3.48 (3H,
m), 2.84 (1H, dd, J=14, 8Hz), 2.65 (1H, dd, J=14, 4Hz), 1.86-1.53
(13H, m), 1.39 (9H, s), 1.33-1.05 (8H, m), 0.82 (2H, m).
Preparation 15
(3R)-6-Cyclohexyl-3-[3-(1
-piperidinylcarbonyl)-1,2,4-oxadiazol-5-yl]hexan- oic acid
[0486] 94
[0487] A solution of tert-butyl
(3R)-6-cyclohexyl-3-[3-(1-piperidinylcarbo-
nyl)-1,2,4-oxadiazol-5-yl]hexanoate (Preparation 14) (334 mg, 0.77
mmol) in dichloromethane (4 ml) was treated with trifluoroacetic
acid (1 ml) and the resulting mixture was stirred at room
temperature under a nitrogen atmosphere for 17 hours. The solvent
was removed under reduced pressure and the residue azeotroped from
toluene to afford the title compound as a beige solid (266 mg).
[0488] MS: 378 (MH.sup.+), 395 (MNH.sub.4.sup.+)
[0489] .sup.1H-NMR (CD.sub.3OD) .delta.: 3.73 (2H, m), 3.55 (1H,
m), 3.43 (2H, m), 2.97-2.75 (2H, m), 1.86-1.57 (13H, m), 1.40-1.07
(8H, m), 0.87 (2H, m).
Preparation 16a
tert-Butyl
(3R)-6-cyclohexyl-3-[3-(1,3-dihydro-2H-isoindol-2-ylcarbonyl)-1-
,2,4-oxadiazol-5-yl]hexanoate
Preparation 16b
tert-Butyl
(3R)-3-{3-[(benzylamino)carbonyl]-1,2,4-oxadiazol-5-yl}-6-cyclo-
hexylhexanoate
[0490] 95
[0491] A solution of ethyl
5{(1R)-1-[2-(tert-butoxy)-2-oxoethyl]4-cyclohex-
ylbutyl}-1,2,4-oxadiazole-3-carboxylate (Preparation 3) (300 mg,
0.76 mmol) in ethanol (4 ml) was treated with isoindoline
hydrochloride (0.59 g, 3.80 mmol) (which also contained
benzylamine) and triethylamine (0.74 ml, 5.32 mmol) and the
resulting mixture was heated at 60.degree. C. under a nitrogen
atmosphere for 16 hours. The solvent was removed under reduced
pressure and the residue was purified by column chromatography on
silica gel eluting with dichloromethane:ethyl acetate (90:10). The
residue was further purified by column chromatography on silica gel
eluting with a gradient system of dichloromethane:ethyl acetate
(99:1) to afford the title compound 16a (91 mg).
[0492] MS: 468 (MH.sup.+), 485 (MNH.sub.4.sup.+)
[0493] .sup.1H-NMR (CDCl.sub.3) .delta.: 7.33 (4H, m), 5.19 (2H,
s), 5.04 (2H, s), 3.56 (1H, m), 2.92 (1H, dd, J=15, 7Hz), 2.71 (1H,
dd, J=15, 3Hz), 1.90-1.58 (7H, m), 1.41 (9H, s), 1.38-1.05 (8H, m),
0.83 (2H, m).
[0494] Further elution with dichloromethane:ethyl acetate (95:5)
then gave compound 16b (173 mg)
[0495] MS: 473 (MNH.sub.4.sup.+)
[0496] .sup.1H-NMR (CDCl.sub.3) .delta.: 7.41-7.17 (5H, m), 4.66
(2H, d, J=5Hz), 3.50 (1H, m), 2.84 (1H, dd, J=15, 8Hz), 2.65 (1H,
dd, J=15, 3Hz), 1.83-1.57 (7H, m), 1.39 (9H, s), 1.34-1.04 (8H, m),
0.83 (2H, m).
Preparation 17
(3R)-6-Cyclohexyl-3-[3-(1,3-dihydro-2H-isoindol-2-ylcarbonyl)-1,2,4-oxadia-
zol-5-yl]hexanoic acid
[0497] 96
[0498] A solution of tert-butyl
(3R)-6-cyclohexyl-3-[3-(1,3-dihydro-2H-iso-
indol-2-ylcarbonyl)-1,2,4-oxadiazol-5-yl]hexanoate (Preparation
16a) (91 mg, 0.19 mmol) in dichloromethane (4 ml) was treated with
trifluoroacetic acid (1 ml) and the resulting mixture was stirred
at room temperature under a nitrogen atmosphere for 17 hours. The
solvent was removed under reduced pressure and the residue
azeotroped from toluene to afford the title compound as a beige
solid (82 mg).
[0499] MS: 412 (MH.sup.+), 429 (MNH.sub.4.sup.+)
[0500] .sup.1H-NMR (CD.sub.3OD) .delta.: 7.46-7.25 (4H, m), 5.19
(2H, s), 4.99 (2H, s), 3.62 (1H, m), 2.98 (1H, dd, J=17, 9Hz), 2.84
(1H, dd, J=17, 5Hz), 2.94-2.77 (7H, m), 1.47-1.06 (8H, m), 0.90
(2H, m).
Preparation 18
(3R)-3-{3-[(Benzylamino)carbonyl]-1,2,4-oxadiazol-5-yl}-6-cyclohexylhexano-
ic acid
[0501] 97
[0502] A solution of tert-butyl
(3R)-3-{3-[(benzylamino)carbonyl]-1,2,4-ox-
adiazol-5-yl}6-cyclohexylhexanoate (Preparation 16b) (173 mg, 0.38
mmol) in dichloromethane (4 ml) was treated with trifluoroacetic
acid (1 ml) and the resulting mixture was stirred at room
temperature under a nitrogen atmosphere for 17 hours. The solvent
was removed under reduced pressure and the residue azeotroped from
toluene to afford the title compound as a beige solid (155 mg).
[0503] MS: 400 (MH.sup.+), 417 (MNH.sub.4.sup.+)
[0504] .sup.1H-NMR (CD.sub.3OD) .delta.7.40-7.20 (5H, m), 4.56 (2H,
s), 3.54 (1H, m), 2.93 (1H, dd, J=16, 8Hz), 2.80 (1H, dd, J=16,
3Hz), 1.83-1.67 (7H, m), 1.40-1.06 (8H, m), 0.86 (2H, m).
Preparation 19
tert-Butyl
(3R)-6-cyclohexyl-3-3-[3,4-dihydro-2(1H)-isoquinolinylcarbonyl]-
-1,2,4-oxadiazol-5-yl}hexanoate
[0505] 98
[0506] A solution of ethyl
5-{1R)-1-[2-(tert-butoxy)-2-oxoethyl]-4-cyclohe-
xylbutyl}-1,2,4-oxadiazole-3-carboxylate (Preparation 3) (300 mg,
0.76 mmol) in ethanol (4 ml) was treated with
1,2,3,4-tetrahydroisoquinoline (0.95 ml, 7.60 mmol) and the
resulting mixture was heated at 60.degree. C. under a nitrogen
atmosphere for 9 hours. The mixture was cooled and the solvent
removed under reduced pressure. The residue was purified by column
chromatography on silica gel eluting with dichloromethane:ethyl
acetate (80:20). The residue was further purified by column
chromatography on silica gel eluting with a gradient system of
pentane:ethyl acetate (90:10) gradually changing to pentane:ethyl
acetate (70:30) to afford the title compound (343 mg).
[0507] MS: 482 (MH.sup.+), 499 (MNH.sub.4.sup.+)
[0508] .sup.1H-NMR (CDCl.sub.3) .delta.: (mixture of rotamers)
7.26-6.97 (4H, m), 4.92 (1.2H, s), 4.80 (0.8H, s), 4.00 (0.8H, m),
3.82 (1.2H, m), 3.53 (1H, m), 3.01-2.83 (3H, m), 2.67 (1H, dd,
J=15, 3Hz), 1.87-1.58 (7H, m), 1.40 (9H, s), 1.36-1.08 (8H, m),
0.83 (2H, m)
Preparation 20
(3R)-6-Cyclohexyl-3-3-[3,4-dihydro-2(1H)-isoquinolinylcarbonyl]-1,2,4-oxad-
iazol-5-yl}hexanoic acid
[0509] 99
[0510] A solution of tert-butyl
(3R)-6-cyclohexyl-3-3-[3,4-dihydro-2(1H)-i-
soquinolinylcarbonyl]-1,2,4-oxadiazol-5-yl)hexanoate (Preparation
19) (343 mg, 0.71 mmol) in dichloromethane (4 ml) was treated with
trifluoroacetic acid (1 ml) and the resulting mixture was stirred
at room temperature under a nitrogen atmosphere for 17 hours. The
solvent was removed under reduced pressure and the residue
azeotroped from toluene to afford the title compound (281 mg).
MS: 426 (MH.sup.+), 443 (MNH.sub.4.sup.+)
[0511] .sup.1H-NMR (CD.sub.3OD) .delta.: (mixture of rotamers)
7.28-7.00 (4H, m), 4.87 (1.2H, s), 4.74 (0.8H, s), 3.99 (0.8H, m),
3.78 (1.2H, m), 3.57 (1H, m), 3.05-2.78 (4H, m), 1.86-1.57 (7H, m),
1.45-1.08 (8H, m), 0.87 (2H, m)
Preparation 21
tert-Butyl
(3R)-6-cyclohexyl-3-[3-(4-morpholinylcarbonyl)-1,2,4-oxadiazol--
5-yl]hexanoate
[0512] 100
[0513] A solution of ethyl
5-{(1R)-1-[2-(tert-butoxy)-2-oxoethyl]-4-cycloh-
exylbutyl)-1,2,4-oxadiazole-3-carboxylate (Preparation 3) (300 mg,
0.76 mmol) in ethanol (4 ml) was cooled to 0.degree. C. then
treated with morpholine (0.066 ml, 7.60 mmol). The resulting
mixture was warmed to room temperature and stirred under a nitrogen
atmosphere for 17 hours. Further morpholine (0.53 ml, 6.08 mmol)
was added and the mixture heated to 60.degree. C. for 8 hours. The
mixture was cooled and the solvent was removed under reduced
pressure. The residue was purified by column chromatography on
silica gel eluting with dichloromethane:ethyl acetate (80:20) to
afford title compound as a yellow oil (269 mg)
[0514] MS: 436 (MH.sup.+), 453 (MNH.sub.4.sup.+)
[0515] .sup.1H-NMR (CDCl.sub.3) .delta.: 3.80 (4H, m), 3.67 (4H,
m), 3.49 (1H, m), 2.84 (2H, dd, J=14, 8Hz), 2.65 (1H, dd, J=14,
3Hz), 1.84-1.57 (7H, m), 1.39 (9H, s), 1.33-1.06 (8H, m), 0.81 (2H,
m)
Preparation 22
(3R)-6-Cyclohexyl-3-[3-(4-morpholinylcarbonyl)-1,2,4-oxadiazol-5-yl]hexano-
ic acid
[0516] 101
[0517] A solution of tert-butyl
(3R)-6-cyclohexyl-3-[3-(4-morpholinylcarbo-
nyl)-1,2,4-oxadiazol-5-yl]hexanoate (Preparation 21) (269 mg, 0.62
mmol) in dichloromethane (4 ml) was treated with trifluoroacetic
acid (1 ml) and the resulting mixture was stirred at room
temperature under a nitrogen atmosphere for 17 hours. The solvent
was removed under reduced pressure and the residue azeotroped from
toluene to afford the title compound (219 mg).
[0518] MS: 380 (MH.sup.+), 397 (MNH.sub.4.sup.+)
[0519] .sup.1H-NMR (CDCl.sub.3) .delta.: 3.80 (4H, m), 3.68 (4H,
m), 3.54 (1H. m), 3.00 (1H, dd, J=14, 8Hz), 2.78 (1H, dd, J=14,
3Hz), 1.88-1.57 (7H, m), 1.40-1.04 (8H, m), 0.85 (2H, m)
Preparation 23
tert-Butyl
(3R)-6-cyclohexyl-3-(3-[(4-methyl-1-piperazinyl)carbonyl]-1,2,4-
-oxadiazol-5-yl)hexanoate
[0520] 102
[0521] A solution of ethyl
5-{(1R)-1-[2-(tert-butoxy)-2-oxoethyl]4-cyclohe-
xylbutyl}-1,2,4-oxadiazole-3-carboxylate (Preparation 3) (300 mg,
0.76 mmol) in ethanol (4 ml) was treated with 1-methylpiperazine
(0.84 ml, 7.60 mmol) and the resulting mixture was heated at
60.degree. C. under a nitrogen atmosphere for 16 hours. The mixture
was cooled and the solvent removed under reduced pressure. The
residue was purified by column chromatography on silica gel eluting
with a gradient system of dichloromethane:methanol (98:2) gradually
changing to dichloromethane methanol (95:5) to afford the title
compound (312 mg).
[0522] MS: 449 (MH.sup.+)
[0523] .sup.1H-NMR (CDCl.sub.3) .delta.:3.80 (2H, m), 3.59 (2H, m),
3.47 (1H, m), 2.85 (1H, m), 2.65 (1H, m), 2.54-2.36 (4H, m), 2.30
(3H, s), 1.85-1.54 (7H, m), 1.37 (9H, s), 1.33-1.04 (8H, m), 0.81
(2H, m)
Preparation 24
(3R)-6-Cyclohexyl-3-3-[(4-methyl-1-piperazinyl)carbonyl]-1,2,4-oxadiazol-5-
-yl}hexanoic acid trifluoroacetate
[0524] 103
[0525] A solution of tert-butyl
(3R)-6-cyclohexyl-3-{3-[(4-methyl-1-pipera-
zinyl)carbonyl]-1,2,4-oxadiazol-5-yl}hexanoate (Preparation 23)
(312 mg, 0.70 mmol) in dichloromethane (4 ml) was treated with
trifluoroacetic acid (1 ml) and the resulting mixture was stirred
at room temperature under a nitrogen atmosphere for 17 hours. The
solvent was removed under reduced pressure and the residue
azeotroped from toluene to afford the title compound as a white
foam (320 mg).
[0526] MS: 393 (MH.sup.+)
[0527] .sup.1H-NMR (CD.sub.3OD) .delta.: 4.03 (4H, br m), 3.56 (1H.
m), 3.41 (4H, m), 2.98-2.78 (5H, m), 1.83 (7H, m), 1.40-1.12 (8H,
m), 0.87 (2H, m)
Preparation 25
tert-Butyl
(3R)-6-cyclohexyl-3-(3-[4-(dimethylamino)-1-piperidinyl]carbony-
l}-1,2,4-oxadiazol-5-yl)hexanoate
[0528] 104
[0529] A solution of ethyl
5-{(1R)-1-[2-(tert-butoxy)-2-oxoethyl]4-cyclohe-
xylbutyl}-1,2,4-oxadiazole-3-carboxylate (Preparation 3) (800 mg,
2.03 mmol) in ethanol (10 ml) was treated with
N,N-dimethyl-N-(4-piperidinyl)a- mine (1.23 g, 9.61 mmol) and the
resulting mixture was heated under reflux under a nitrogen
atmosphere for 3 hours. The solvent was removed under reduced
pressure and the residue was partitioned between ethyl acetate and
water. The layers were separated and the aqueous layer was
extracted with ethyl acetate. The combined organic layers were
washed sequentially with water and brine, dried over anhydrous
sodium sulphate, filtered and the solvent was removed under reduced
pressure. The residue was purified by column chromatography on
silica gel eluting with dichloromethane:methanol (90:10) to give a
residue which was further purified by column chromatography on
silica gel eluting with dichloromethane:methanol: 0.88 ammonia
(90:10:0.5) to afford the title compound as a yellow oil (653
mg).
[0530] MS: 477 (MH.sup.+)
[0531] .sup.1H-NMR (CDCl.sub.3) .delta.: 4.77 (1H, m), 4.05 (1H,
m), 3.50 (1H, m), 3.13 (1H, m), 2.92-2.60 (4H, m), 2.40 (6H, s),
2.04-1.50 (9H, m), 1.40 (9H, s), 1.36-1.07 (10H, m), 0.84 (2H,
m)
Preparation 26
(3R)-6-Cyclohexyl-3-(3-{[4-(dimethylamino)-1-piperidinyl]carbonyl)-1,2,4-o-
xadiazol-5-yl)hexanoic acid trifluoroacetate
[0532] 105
[0533] A solution of tert-butyl
(3R)-6-cyclohexyl-3-(3-[4-(dimethylamino)--
1-piperidinyl]carbonyl}-1,2,4-oxadiazol-5-yl)hexanoate (Preparation
25) (652 mg, 1.37 mmol) in dichloromethane (15 ml) was cooled to
0.degree. C. and treated with trifluoroacetic acid (5m1). The
resulting mixture was stirred for 2 hours being allowed to warm to
room temperature over this time. The solvent was removed under
reduced pressure and the residue azeotroped from toluene. The
residue was purified by column chromatography on silica gel eluting
with a gradient system of dichloromethane:methanol:0.88 ammonia
(90:10:1) gradually changing to dichloromethane:methanol:0.88
ammonia (70:30:2) to afford the title compound as a colourless oil
(702 mg).
[0534] MS: 421 (MH.sup.+)
[0535] .sup.1H-NMR (CD.sub.3OD) .delta.: 4.80 (1H, m), 4.09 (1H,
m), 3.55 (1H, m), 3.22 (2H, m), 2.93 (1H, m), 2.83-2.60 (8H, m),
2.20-2.00 (2H, m), 1.82-1.54 (9H, m), 1.41-1.07 (8H, m), 0.87 (2H,
m)
Preparation 27
tert-Butyl
(3R)-6-cyclohexyl-3-(3-{[3-(4-morpholinyl)-1-azetidinyl]carbony-
l}-1,2,4-oxadiazol-5-yl)hexanoate
[0536] 106
[0537] A solution of ethyl 5-{(1R)-1
-[2-(tert-butoxy)-2-oxoethyl]-4-cyclo-
hexylbutyl}-1,2,4-oxadiazole-3-carboxylate (Preparation 3) (1.00 g,
2.54 mmol) in ethanol (10 ml) was treated with
4-(3-azetidinyl)morpholine dihydrochloride (2.72 g, 12.6 mmol) and
triethylamine (2.56 g, 25 mmol) and the resulting mixture was
heated under reflux under a nitrogen atmosphere for 24 hours. The
solvent was removed under reduced pressure and the residue was
partitioned between ethyl acetate and water. The layers were
separated and the aqueous layer was extracted with ethyl acetate.
The combined organic layers were washed sequentially with water and
brine, dried over anhydrous sodium sulphate, filtered and the
solvent removed under reduced pressure. The residue was purified by
column chromatography on silica gel eluting with a gradient system
of dichloromethane:methanol (97:3) gradually changing to
dichloromethane:methanol (95:5) to afford the title compound as a
colourless oil (1.32 g).
[0538] MS: 491 (MH.sup.+), 508 (MNH.sub.4.sup.+)
[0539] .sup.1H-NMR (CD.sub.3OD) .delta.:4.64 (1H, m), 4.43 (1H, m),
4.25 (1H, m), 4.05 (1H, m), 3.73 (4H, m), 3.52 (1H, m), 3.31 (1H.
m), 2.84-2.66 (2H, m), 2.45 (4H, m), 1.85-1.55 (7H, m), 1.46-1.05
(17H, m), 0.86 (2H, m)
Preparation 28
(3R)-6-Cyclohexyl-3-(3-{[3-(4-morpholinyl)-1
-azetidinyl]carbonyl}-1,2,4-o- xadiazol-5-yl)hexanoic acid
trifluoroacetate
[0540] 107
[0541] A solution of tert-butyl
(3R)-6-cyclohexyl-3-(3-{[3-(4-morpholinyl)-
-1-azetidinyl]carbonyl)-1,2,4-oxadiazol-5-yl)hexanoate (Preparation
27) (1.32 g, 2.70 mmol) in dichloromethane (15 ml) was cooled to
0.degree. C. and treated with trifluoroacetic acid (5 ml). The
resulting mixture was stirred for 3 hours being allowed to warm to
room temperature over this time. The solvent was removed under
reduced pressure and the residue azeotroped from toluene (.times.3)
then dichloromethane to afford the title compound as a white foam
(1.31 g).
[0542] MS: 435 (MH.sup.+)
[0543] .sup.1H-NMR (CD.sub.3OD) .delta.:4.86 (1H, m), 4.50 (1H, m),
4.39 (1H, m), 4.14 (1H, m), 3.92 (4H, m), 3.56 (1H, m), 3.37-3.17
(5H, m), 2.93 (1H, dd, J=13, 8Hz), 2.82 (1H, dd, J=13, 3Hz),
1.83-1.59 (7H, m), 1.39-1.09 (8H, m), 0.86 (2H, m)
Preparation 29
tert-Butyl
(3R)-6-cyclohexyl-3-3-[7,8-dihydro[1,6]naphthyridin-6(5H)-ylcar-
bonyl]-1,2,4-oxadiazol-5-yl}hexanoate
[0544] 108
[0545] A solution of ethyl 5-{(1R)-1
-[2-(tert-butoxy)-2-oxoethyl]-4-cyclo-
hexylbutyl}-1,2,4-oxadiazole-3-carboxylate (Preparation 3) (300 mg,
0.76 mmol) in ethanol (4 ml) was treated with
5,6,7,8-tetrahydro[1,6]naphthyri- dine dihydrochloride
(Chem.Pharm.Bull.; 32; 7; 1984; 2522-2529) (0.79 g, 3.80 mmol) and
triethylamine (1.27 ml, 9.13 mmol) and the resulting mixture was
heated at 60.degree. C. under a nitrogen atmosphere for 16 hours.
The mixture was cooled and the solvent removed under reduced
pressure. The residue was purified by column chromatography on
silica gel eluting with a gradient system of dichloromethane:ethyl
acetate (90:10) gradually changing to dichloromethane:ethyl acetate
(50:50) to afford the title compound (281 mg).
[0546] MS: 483 (MH.sup.+), 505 (MNa.sup.+)
[0547] .sup.1H-NMR (CDCl.sub.3) .delta.: (mixture of rotamers) 8.44
(1H, d, J=3Hz), 7.48 (0.67H, d, J=6Hz), 7.33 (0.33H, d, J=6Hz),
7.20-7.10 (1H, m), 4.92 (1.34H, s), 4.83 (0.66H, s), 4.13 (0.66H,
t, J=5Hz), 3.94 (1.34H, t, J=5Hz), 3.52 (1H, m), 3.13 (2H, m), 2.87
(1H, dd, J=14, 7Hz), 2.67 (1H, dd, J=14, 3Hz), 1.85-1.56 (7H, m),
1.39 (9H, d), 1.35-1.04 (8H, m), 0.83 (2H, m)
Preparation 30
(3R)-6-Cyclohexyl-3-{3-[7,8-dihydro[1,6]naphthyridin-6(5H)-ylcarbonyl]-1,2-
,4-oxadiazol-5-yl}hexanoic acid trifluoroacetate
[0548] 109
[0549] A solution of tert-butyl
(3R)-6-cyclohexyl-3-{3-[7,8-dihydro[1,6]na-
phthyridin-6(5H)-ylcarbonyl]-1,2,4-oxadiazol-5-yl}hexanoate
(Preparation 29) (281 mg, 0.58 mmol) in dichloromethane (4 ml) was
treated with trifluoroacetic acid (1 ml) and the resulting mixture
was stirred at room temperature under a nitrogen atmosphere for 17
hours. The solvent was removed under reduced pressure and the
residue azeotroped from toluene to afford the title compound (245
mg).
[0550] MS: 427 (MH.sup.+)
[0551] .sup.1H-NMR (CD.sub.3OD) .delta.: (mixture of rotamers) 8.56
(1H. d, 5Hz), 8.17 (0.67H, d, J=8Hz), 8.01 (0.33H, d, J=8Hz),
7.73-7.56 (1H, m), 5.05 (1.34H, s), 5.00 (0.66H, s), 4.16 (0.66H,
m), 4.01 (1.34H, m), 3.59 (1H, m), 3.24 (2H, m), 3.04-2.76 (2H, m),
1.92-1.55 (7H, m), 1.46-1.06 (8H, m), 0.87 (2H, m)
Preparation 31
tert-Butyl
(3R)-6-cyclohexyl-3-(3-{[4-(4-pyridinyl)-1-piperidinyl]carbonyl-
}-1,2,4-oxadiazol-5-yl)hexanoate
[0552] 110
[0553] A solution of ethyl 5-{(1R)-1
-[2-(tert-butoxy)-2-oxoethyl]4-cycloh-
exylbutyl}-1,2,4-oxadiazole-3-carboxylate (Preparation 3) (0.50 g,
1.27 mmol) in ethanol (10 ml) was treated with
4-(4-pyridinyl)piperidine (Monatsh.Chem.; 3; 1882; 867) (0.41 g,
2.54 mmol) and the resulting mixture was heated under reflux under
a nitrogen atmosphere for 72 hours. Further
4-(4-pyridinyl)piperidine (0.21 g, 1.27 mmol) was added and the
mixture heated under reflux for 24 hours. The solvent was removed
under reduced pressure and the residue was partitioned between
ethyl acetate and water. The layers were separated and the aqueous
layer extracted with ethyl acetate. The combined organic layers
were washed sequentially with water and brine, dried over anhydrous
sodium sulphate, filtered and the solvent removed under reduced
pressure. The residue was purified by column chromatography on
silica gel eluting with a gradient system of
dichloromethane:methanol (99:1) gradually changing to
dichloromethane:methanol (95:5) to afford the title compound as a
pale yellow oil (0.39 g).
[0554] MS: 511 (MH.sup.+)
[0555] .sup.1H-NMR (CD.sub.3OD) .delta.: 8.44 (2H, d, J=4Hz), 7.35
(2H, d, J=4Hz), 4.79 (1H, m), 4.00 (1H, m), 3.53 (1H, m), 3.34 (1H,
m), 3.00 (2H, m), 2.84 (1H, dd, J=14, 8Hz), 2.75 (1H, dd, 14, 5Hz),
2.02 (1H, m), 1.92 (1H, m), 1.83-1.58 (9H, m), 1.44-1.09 (17H, m),
0.86 (2H, m)
Preparation 32
(3R)-6-Cyclohexyl-3-(3-{[4-(4-pyridinyl)-1
-piperidinyl]carbonyl}-1,2,4-ox- adiazol-5-yl)hexanoic acid
[0556] 111
[0557] A solution of tert-butyl
(3R)-6-cyclohexyl-3-(3-{[4-(4-pyridinyl)-1-
-piperidinyl]carbonyl}-1,2,4-oxadiazol-5-yl)hexanoate (Preparation
31) (376 mg, 0.74 mmol) in dichloromethane (15 ml) was cooled to
0.degree. C. and treated with trifluoroacetic acid (5 ml). The
resulting mixture was stirred for 3 hours being allowed to warm to
room temperature over this time. The solvent was removed under
reduced pressure and the residue azeotroped from toluene (.times.3)
then dichloromethane. A saturated solution of sodium carbonate was
added to the residue until a pH of 12 was achieved followed by
dropwise addition of an aqueous citric acid solution solution (10%
w/v) until the pH became 3.5. The solution was then diluted with
water and extracted with ethyl acetate (.times.3). The combined
organic layers were washed with brine, dried over anhydrous sodium
sulphate, filtered and the solvent removed under reduced pressure
to afford the title compound as a white solid (310 mg).
[0558] MS: 455 (MH.sup.+)
[0559] .sup.1H-NMR (CD.sub.3OD) .delta.: 8.44 (2H, d, J=4Hz), 7.36
(2H, d, J=4Hz), 4.78 (1H, m), 3.97 (1H, m), 3.56 (1H, m), 3.34 (1H,
m), 3.06-2.75 (4H, m), 2.01 (1H, m), 1.92 (1H, m), 1.83-1.56 (9H,
m), 1.40-1.04 (8H, m), 0.85 (2H, m)
Preparation 33
tert-Butyl
(3R)-3-(3-{[benzyl(methyl)amino]carbonyl}-1,2,4-oxadiazol-5-yl)-
-6-cyclohexylhexanoate
[0560] 112
[0561] A solution of ethyl
5{(1R)-1-[2-(tert-butoxy)-2-oxoethyl]-4-cyclohe-
xylbutyl}-1,2,4-oxadiazole-3-carboxylate (Preparation 3) (300 mg,
0.76 mmol) in ethanol (4 ml) was treated with
N-benzyl-N-methylamine (0.98 ml, 7.60 mmol) and the resulting
mixture was heated at 60.degree. C. under a nitrogen atmosphere for
16 hours. The mixture was cooled and the solvent removed under
reduced pressure. The residue was purified by column chromatography
on silica gel eluting with a gradient system of
dichloromethane:ethyl acetate (100:0) gradually changing to
dichloromethane:ethyl acetate (95:5) to afford the title compound
(296 mg).
[0562] MS: 470 (MH.sup.+), 487 (MNH.sub.4.sup.+)
[0563] .sup.1H-NMR (CD.sub.3OD) .delta.: (mixture of rotamers)
7.42-7.21 (5H, m), 4.79 (1H, d, J=1 5Hz), 4.63 (1H, d, J=15Hz),
3.52 (1H, m), 3.00 (3H, m), 2.90-2.67 (2H, m), 1.85-1.57 (7H, m),
1.41-1.04 (17H, m), 0.84 (2H, m)
Preparation 34
(3R)-3-(3{[Benzyl(methyl)amino]carbonyl}-1,2,4-oxadiazol-5-yl)-6-cyclohexy-
lhexanoic acid
[0564] 113
[0565] A solution of tert-butyl
(3R)-3-(3-{[benzyl(methyl)amino]carbonyl}--
1,2,4-oxadiazol-5-yl)-6-cyclohexylhexanoate (Preparation 33) (296
mg, 0.63 mmol) in dichloromethane (4 ml) was treated with
trifluoroacetic acid (1 ml) and the resulting mixture was stirred
at room temperature under a nitrogen atmosphere for 17 hours. The
solvent was removed under reduced pressure and the residue
azeotroped from toluene to afford the title compound (226 mg).
[0566] MS: 414 (MH.sup.+), 431 (MNH.sub.4.sup.+)
[0567] .sup.1H-NMR (CD.sub.3OD) .delta.: (mixture of rotamers)
7.42-7.22 (5H, m), 4.79 (1H, d, J=15Hz), 4.58 (1H, d, 15Hz), 3.56
(1H, m), 3.00 (3H, d), 2.97-2.73 (2H, m), 1.84-1.55 (7H, m),
1.41-1.03 (8H, m), 0.83 (2H, m)
Preparation 35
tert-Butyl
(3R)-6-cyclohexyl-3-(3{[methyl(2-pyridinylmethyl)amino]carbonyl-
}1,2,4-oxadiazol-5-yl)hexanoate
[0568] 114
[0569] 1-1'-Azobis(N,N-dimethylformamide) (645 mg, 3.75 mmol) was
added to a cooled solution of tert-butyl
(3R)-6-cyclohexyl-3-{3-[(methylamino)carb-
onyl]-1,2,4-oxadiazol-5-yl}hexanoate (Preparation 7) (1.42 g, 3.75
mmol), tributylphosphine (930 .mu.l, 3.75 mmol) and
2-hydroxymethylpyridine (240 .mu.l, 2.50 mmol) in toluene (10 ml)
and the resulting mixture was stirred at 0.degree. C. under a
nitrogen atmosphere for 15 minutes, then at room temperature for 72
hours. The mixture was filtered and the solvent removed from the
filtrate under reduced pressure. The residue was purified by column
chromatography on silica gel eluting with a gradient system of
hexane:ethyl acetate (90:10) gradually changing to hexane:ethyl
acetate (50:50) to afford the title compound as a pale yellow oil
(530 mg).
[0570] MS :472 (MH.sup.+)
[0571] .sup.1H-NMR (CDCl.sub.3) .delta.: (mixture of rotamers) 8.54
(1H, m), 7.68 (1H, m), 7.44-7.15 (2H, m), 4.86 (1H, s), 4.79 (1H,
s), 3.51 (1H, m), 3.16 (1.5H, s), 3.10 (1.5H, s), 2:84 (1H, m),
2.63 (1H, m), 1.89-1.52 (7H, m), 1.36 (9H, d), 1.31-1.00 (8H, m),
0.81 (2H, m)
Preparation 36
(3R)-6-Cyclohexyl-3-(3-{[methyl(2-pyridinylmethyl)amino]carbonyl}-1,2,4-ox-
adiazol-5-yl)hexanoic acid
[0572] 115
[0573] A solution of tert-butyl
(3R)-6-cyclohexyl-3-(3-{[methyl(2-pyridiny-
lmethyl)amino]carbonyl}-1,2,4-oxadiazol-5-yl)hexanoate (Preparation
35) (527 mg, 1.12 mmol) in dichloromethane (20 ml) was treated with
trifluoroacetic acid (10 ml) and the resulting mixture was stirred
at room temperature under a nitrogen atmosphere for 2 hours. The
solvent was removed under reduced pressure and the residue was
azeotroped from toluene. The residue was dissolved in a saturated
aqueous solution of sodium hydrogen carbonate (3 ml) and the pH was
adjusted to pH 4 with aqueous citric acid solution (10% w/v). The
aqueous phase was extracted with ethyl acetate (.times.2) and the
combined organic layers were washed with brine, dried over
anhydrous sodium sulphate, filtered and the solvent removed under
reduced pressure to afford the title compound as a yellow oil (456
mg)
[0574] MS: 414 (M+)
[0575] .sup.1H-NMR (CDCl.sub.3) .delta.: (mixture of rotamers) 8.51
(1H, m), 7.75 (1H, m), 7.47 (0.5H, d, J=6Hz), 7.40 (0.5H, d,
J=6Hz), 7.23 (1H, m), 4.96-4.66 (2H, m), 3.51 (1H, m), 3.10 (3H,
d), 3.05-2.66 (2H, m), 1.89-1.52 (7H, m), 1.40-1.01 (8H, m), 0.82
(2H, m)
Preparation 37
tert-Butyl
(3R)-6-cyclohexyl-3-(3-{[(2-methoxy-2-oxoethyl)amino]carbonyl}--
1,2,4-oxadiazol-5-yl)hexanoate
[0576] 116
[0577] A solution of ethyl 5-{(1R)-1
-[2-(tert-butoxy)-2-oxoethyl]-4-cyclo-
hexylbutyl}-1,2,4-oxadiazole-3-carboxylate (Preparation 3) (1.18 g,
3.00 mmol) and triethylamine (1.51 g, 15.00 mmol) in ethanol (30
ml) was treated with glycine methyl ester hydrochloride (1.88 g,
15.00 mmol) and the resulting mixture was heated at 80.degree. C.
under a nitrogen atmosphere for 16 hours. The mixture was cooled
and the solvent removed under reduced pressure. The residue was
partitioned between water and ethyl acetate. The combined organic
layers were washed with brine, dried over anhydrous sodium
sulphate, filtered and the solvent removed under reduced pressure.
The residue was purified by column chromatography on silica gel
eluting with a gradient system of hexane:ethyl acetate (90:10)
gradually changing to hexane:ethyl acetate (50:50) to afford the
title compound (456 mg).
[0578] MS: 455 (MNH.sub.4.sup.+)
[0579] .sup.1H-NMR (CDCl.sub.3) .delta.: 7.42 (1H, br t), 4.25 (2H,
d, J=4Hz), 3.80 (3H, s), 3.51 (1H, m), 2.84 (1H, dd, J=14, 9Hz),
2.66 (1H, dd, J=14, 3Hz), 1.83-1.55 (7H, m), 1.38 (9H, s),
1.34-1.07 (8H, m), 0.84 (2H, m)
Preparation 38
tert-Butyl
(3R)-6-cyclohexyl-3-(3-{[(2-methoxy-2-oxoethyl)(methyl)amino]ca-
rbonyl)1 ,2,4-oxadiazol-5-yl)hexanoate
[0580] 117
[0581] A solution of tert-butyl
(3R)-6-cyclohexyl-3-(3-{[(2-methoxy-2-oxoe-
thyl)amino]carbonyl}-1,2,4-oxadiazol-5-yl)hexanoate (Preparation
37) (440 mg, 1.00 mmol) in anhydrous dimethylsulphoxide (10 ml) was
treated with iodomethane (310 .mu.l, 5.00 mmol) and cesium
carbonate (975 mg, 3.00 mmol) and the resulting mixture was heated
at 40.degree. under a nitrogen atmosphere for 3 hours then stirred
at room temperature for 17 hours. The mixture was diluted with
water and extracted with diethyl ether (.times.3). The combined
organic extracts were washed with brine, dried over anhydrous
sodium sulphate, filtered and the solvent removed under reduced
pressure. The residue was purified by column chromatography on
silica gel eluting with hexane:ethyl acetate (2:1) to afford the
title compound as a colourless oil (315 mg)
[0582] MS: 452 (MH.sup.+)
[0583] .sup.1H-NMR (CDCl.sub.3) .delta.: (mixture of rotamers) 4.36
(1H, s), 4.30 (1H, s), 3.76 (3H, d), 3.51 (1H, m), 3.25 (1.5H, s),
3.20 (1.5H, s), 2.86 (1H, m), 2.66 (1H. m), 1.85-1.57 (7H, m), 1.39
(9H, s), 1.34-1.04 (8H, m), 0.84 (2H, m)
Preparation 39
(3R)-6-Cyclohexyl-3-(3-{[(2-methoxy-2-oxoethyl)(methyl)amino]carbonyl}-1,2-
,4-oxadiazol-5-yl)hexanoic acid
[0584] 118
[0585] A solution of tert-butyl
(3R)-6-cyclohexyl-3-(3-{[(2-methoxy-2-oxoe-
thyl)(methyl)amino]carbonyl)-1,2,4-oxadiazol-5-yl)hexanoate
(Preparation 38) (315 mg, 0.70 mmol) in dichloromethane (10 ml) was
treated with trifluoroacetic acid (5 ml) and the resulting mixture
was stirred at room temperature under a nitrogen atmosphere for 2
hours. The solvent was removed under reduced pressure and the
residue azeotroped from toluene. The residue was dissolved in ethyl
acetate and washed with a saturated aqueous solution of sodium
citrate and brine, dried over anhydrous sodium sulphate, filtered
and the solvent removed under reduced pressure to afford the title
compound as an oil (273 mg)
[0586] MS: 396 (MH.sup.+), 418 (MNa.sup.+)
[0587] .sup.1H-NMR (CDCl.sub.3) .delta.: (mixture of rotamers) 4.26
(1.6H, m), 4.11 (0.4H, m), 3.75 (3H, d), 3.52 (1H, m), 3.21 (3H,
d), 3.00 (1H, m), 2.78 (1H, m), 1.89-1.41 (7H, m), 1.38-0.95 (8H,
m), 0.80 (2H, m)
Preparation 40/Example 41
Methyl
2-[[(5-{(1R)-4-cyclohexyl-1-[2-(hydroxyamino)-2-oxoethyl]butyl)-1,2-
,4-oxadiazol-3-yl)carbonyl](methyl)amino]acetate
[0588] 119
[0589] A solution of
(3R)-6-cyclohexyl-3-(3-[(2-methoxy-2-oxoethyl)(methyl-
)amino]carbonyl)-1,2,4-oxadiazol-5-yl)hexanoic acid (Preparation
39) (273 mg, 0.70 mmol) and N-methylmorpholine (85 .mu.l, 0.77
mmol) in anhydrous dichloromethane (10 ml) was cooled to 0.degree.
C., treated with isobutyl chloroformate (100 .mu.l, 0.77 mmol) and
the resulting mixture was stirred under a nitrogen atmosphere for
30 minutes. O-(Trimethylsilyl)hydroxylamine (250 .mu.l, 2.10 mmol)
was then added and the mixture stirred under a nitrogen atmosphere
for 1 hour, being allowed to warm to room temperature over this
time. The mixture was then quenched with methanol (10 ml) and
stirred for 10 minutes. The solvent was removed under reduced
pressure and the residue partitioned between ethyl acetate and
water. The layers were separated and the organic layer was
sequentially washed with water and brine, dried over anhydrous
sodium sulphate, filtered and the solvent removed under reduced
pressure. The residue was purified by HPLC.sup.a to afford the
title compound as a colourless oil (187 mg).
[0590] MS: 411 (MH.sup.+), 433 (MNa.sup.+)
[0591] .sup.1H-NMR (CDCl.sub.3) .delta.: (mixture of rotamers)
4.50-4.21 (2H, m), 3.84-3.60 (4H, m), 3.32 (1 .8H, s), 3.21 (1.2H,
s), 2.81-2.56 (2H, m), 1.90-1.50 (7H, m), 1.40-1.03 (8H, m), 0.82
(2H, m)
Preparation 41
1-[(5-{(1R)-1-[2-(tert-Butoxy)-2-oxoethyl]-4-cyclohexylbutyl}1,2,4-oxadiaz-
ol-3-yl)carbonyl]-3-azetidinecarboxylic acid
[0592] 120
[0593] A solution of ethyl 5-{(1R)-1
-[2-(tert-butoxy)-2-oxoethyl]4-cycloh-
exylbutyl}-1,2,4-oxadiazole-3-carboxylate (Preparation 3) (790 mg,
2.00 mmol) in dimethylsulphoxide (25 ml) was treated with
3-azetidine carboxylic acid (505 mg, 5.00 mmol) and potassium
carbonate (690 mg, 5.00 mmol) and the resulting mixture was heated
at 95.degree. C. under a nitrogen atmosphere for 16 hours. The
mixture was cooled and the mixture treated with hydrochloric acid
(1M, 25 ml) then diluted further with water (25 ml) and extracted
with ethyl acetate (.times.3). The combined organic layers were
washed with brine, dried over anhydrous sodium sulphate, filtered
and the solvent removed under reduced pressure. The residue was
purified by column chromatography on silica gel eluting with a
gradient system of dichloromethane:methanol (99:1) gradually
changing to dichloromethane:methanol (90:10) to afford the title
compound as a pale yellow oil (490 mg).
[0594] .sup.1H-NMR (CDCl.sub.3) .delta.: 4.76 (2H, m), 4.45 (2H,
m), 3.64-3.43 (2H, m), 2.89 (1H, dd, J=15, 8Hz), 2.65 (1H, dd,
J=15, 4Hz), 1.84-1.56 (7H, m), 1.38 (9H, s), 1.34-1.03 (8H, m),
0.84 (2H, m)
Preparation 42
Methyl
1-[(5-{(1R)-1-[2-(tert-butoxy)2-oxoethyl]-4-cyclohexylbutyl}-1,2,4--
oxadiazol-3-yl)carbonyl]-3-azetidinecarboxylate
[0595] 121
[0596] A solution of
1-[(5-{(1R)-1-[2-(tert-butoxy)-2-oxoethyl]-4-cyclohex-
ylbutyl}-1,2,4-oxadiazol-3-yl)carbonyl]-3-azetidinecarboxylic acid
(Preparation 41) (480 mg, 1.07 mmol) and N-methylmorpholine (130
.mu.l, 1.17 mmol) in dichloromethane (10 ml) was cooled to
0.degree. C. and then treated with isobutyl chloroformate (150
.mu.l, 1.17 mmol). The mixture was stirred at 0.degree. C. for 30
minutes then allowed to warm to room temperature over 1 hour. The
mixture was quenched with methanol (5 ml) and the solvent removed
under reduced pressure. The residue was partitioned between ethyl
acetate and hydrochloric acid (1M). The combined organic layers
were washed with brine, dried over anhydrous sodium sulphate,
filtered and the solvent removed under reduced pressure. The
residue was purified by column chromatography on silica gel eluting
with a gradient system of dichloromethane:ethyl acetate (95:5)
gradually changing to dichloromethane:ethyl acetate (90:10) to
afford the title compound (230 mg)
[0597] MS: 464 (MH.sup.+)
[0598] .sup.1H-NMR (CDCl.sub.3) .delta.: 4.73 (2H, m), 4.40 (2H,
m), 3.78 (3H, s), 3.52 (2H, m), 2.86 (1H, dd, J=15, 8Hz), 2.64 (1H,
dd, J=15, 3Hz), 1.83-1.58 (7H, m), 1.38 (9H, s), 1.33-1.06 (8H, m),
0.83 (2H, m)
Preparation 43
(3R)-6-Cyclohexyl-3-(3-{[3-(methoxycarbonyl)-1
-azetidinyl]carbonyl}-1,2,4- -oxadiazol-5-yl)hexanoic acid
[0599] 122
[0600] A solution of methyl
1-[(5-{(1R)-1-[2-(tert-butoxy)-2-oxoethyl]-4-c-
yclohexylbutyl}-1,2,4-oxadiazol-3-yl)carbonyl]-3-azetidinecarboxylate
(Preparation 42) (225 mg, 0.48 mmol) in dichloromethane (8 ml) was
treated with trifluoroacetic acid (4 ml) and the resulting mixture
was stirred at room temperature under a nitrogen atmosphere for 5
hours. The solvent was removed under reduced pressure and the
residue azeotroped from toluene (.times.2). The residue was
dissolved in ethyl acetate and washed sequentially with a saturated
aqueous solution of sodium citrate and brine, dried over anhydrous
sodium sulphate, filtered and the solvent removed under reduced
pressure to afford the title compound as an oil (200 mg).
[0601] MS: 408 (MH.sup.+), 430 (MNa.sup.+)
[0602] .sup.1H-NMR (CDCl.sub.3) .delta.:4.73 (2H, m), 4.40 (2H, m),
3.78 (3H, s), 3.53 (2H, m), 3.04 (1H, m), 2.80 (1H, m), 1.87-1.41
(7H, m), 1.36-1.00 (8H, m), 0.83 (2H, m)
Preparation 44
tert-Butyl
(3R)-6-cyclohexyl-3-(3-methyl-1,2,4-oxadiazol-5-yl)hexanoate
[0603] 123
[0604] A solution of
(2R)-2-[2-(tert-butoxy)-2-oxoethyl]-5-cyclohexylpenta- noic acid
(Preparation 1) (596 mg, 2.00 mmol) in dichloromethane (8ml) was
treated with 1,1'-carbonyldiimidazole (364 mg, 2.25 mol) and the
solution was stirred at room temperature for 15 minutes. The
N-hydroxyacetamidine (Chem.Ber.; 17; 1884; 2746) (148 mg, 2.00
mmol) was then added and the mixture was stirred for 1 hour. The
solvent was removed under reduced pressure and the residue was
heated neat under a nitrogen atmosphere for 90 minutes. The crude
product was then purified by column chromatography on silica gel
eluting with dichloromethane to afford the title compound (385
mg).
[0605] MS: 337 (MH.sup.+)
[0606] .sup.1H-NMR (CDCl.sub.3) .delta.: 3.41 (1H, m), 2.76 (1H,
dd, J=14, 8Hz), 2.60 (1H, dd, J=14, 5Hz), 2.36 (3H, s), 1.76-1.56
(7H, m), 1.39 (9H, s), 1.32-1.04 (8H, m), 0.82 (2H, m)
Preparation 45
(3R)-6-Cyclohexyl-3-(3-methyl-1,2,4-oxadiazol-5-yl)hexanoic
acid
[0607] 124
[0608] tert-Butyl
(3R)-6-cyclohexyl-3-(3-methyl-1,2,4-oxadiazol-5-yl)hexan- oate
(Preparation 44) (350 mg, 1.04 mmol) was treated with
trifluoroacetic acid (3 ml) and the resulting mixture was stirred
at room temperature under a nitrogen atmosphere for 45 minutes. The
solvent was removed under reduced pressure and the residue
azeotroped from toluene then dichioromethane to afford the title
compound (185 mg).
[0609] MS: 298 (MNH.sub.4.sup.+)
[0610] .sup.1H-NMR (CDCl.sub.3) .delta.: 3.41 (1H, m), 2.96 (1H,
dd, J=16, 8Hz), 2.75 (1H, dd, J=16, 6Hz), 2.36 (3H, s), 1.80-1.40
(7H, m), 1.35-0.93 (8H, m), 0.82 (2H, m)
Preparation 46
tert-Butyl
(3R)-6-cyclohexyl-3-(3-isopropyl-1,2,4-oxadiazol-5-yl)hexanoate
[0611] 125
[0612] A solution of
(2R)-2-[2-(tert-butoxy)2-oxoethyl]-5-cyclohexylpentan- oic acid
(Preparation 1) (500 mg, 1.70 mmol) in dichloromethane (30 ml) was
treated with 1,1'-carbonyldiimidazole (272 mg, 1.70mol) and the
solution was stirred at room temperature for 1 hour. The
N'-hydroxy-2-methylpropanimidamide (Monatsh.Chem.; 113; 1982;
781-792) (174 mg, 1.70 mmol) was then added and the mixture was
stirred for 30 minutes. The solvent was removed under reduced
pressure and the residue was heated neat at 120.degree. C. under a
nitrogen atmosphere for 18 hours. The crude product was then
purified by column chromatography on silica gel eluting with
dichloromethane to afford the title compound as a colourless oil
(250 mg).
[0613] MS: 365 (MH.sup.+)
[0614] .sup.1H-NMR (CDCl.sub.3) .delta.: 3.42 (1H, m), 3.05 (1H,
m), 2.78 (1H, dd, J=16, 8Hz), 2.60 (1H, dd, J=16, 4Hz), 1.70-1.57
(7H, m), 1.39 (9H, s), 1.34-1.06 (14H, m), 0.81 (2H, m)
Preparation 47
(3R)-6-Cyclohexyl-3-(3-isopropyl-1,2,4-oxadiazol-5-yl)hexanoic
acid
[0615] 126
[0616] tert-Butyl
(3R)-6-cyclohexyl-3-(3-isopropyl-1,2,4-oxadiazol-5-yl)he- xanoate
(Preparation 46) (250 mg, 0.69 mmol) was treated with
trifluoroacetic acid (5 ml) and the resulting mixture was stirred
at room temperature under a nitrogen atmosphere for 45 minutes. The
solvent was removed under reduced pressure and the residue
azeotroped from toluene then dichloromethane to afford the title
compound as a white solid (220 mg).
[0617] MS: 309 (MH.sup.+)
[0618] .sup.1H-NMR (CDCl.sub.3) .delta.: 3.50 (1H, m), 3.09 (1H,
m), 2.95 (2H, dd, J=16, 8Hz), 2.76 (1H, dd, J=16, 4Hz), 1.84-1.56
(7H, m), 1.40-1.05 (14H, m), 0.81 (2H, m)
Preparation 48
tert-Butyl
(3R)-6-cyclohexyl-3-[3-(methoxymethyl)-1,2,4-oxadiazol-5-yl]hex-
anoate
[0619] 127
[0620] A solution of
(2R)-2-[2-(tert-butoxy)-2-oxoethyl]-5-cyclohexylpenta- noic acid
(Preparation 1) (500 mg, 1.70 mmol) in dichloromethane (30 ml) was
treated with 1,1'-carbonyldiimidazole (272 mg, 1.70 mol) and the
solution was stirred at room temperature for 1 hour. The
N'-hydroxy-2-methoxyethanimidamide (J.Med.Chem,; 40; 8; 1997;
1230-1246) (177 mg, 1.70 mmol) was then added and the mixture was
stirred for 17 hours. The solvent was removed under reduced
pressure and the residue was heated neat at 120.degree. C. under a
nitrogen atmosphere for 2 hours. The crude product was then
purified by column chromatography on silica gel eluting with
dichloromethane:methanol (99:1) to afford the title compound as an
oil (350 mg).
[0621] MS: 367 (MH.sup.+), 389 (MNa.sup.+)
[0622] .sup.1H-NMR (CDCl.sub.3) .delta.: 4.56 (2H, s), 3.49 (4H,
m), 2.84 (1H, dd, J=16, 8Hz), 2.65 (1H, dd, J=16, 5Hz), 1.85-1.52
(7H, m), 1.40 (9H, s), 1.36-1.05 (8H, m), 0.84 (2H, m)
Preparation 49
(3R)-6-Cyclohexyl-3-[3-(methoxymethyl)-1,2,4-oxadiazol-5-yl]hexanoic
acid
[0623] 128
[0624] tert-Butyl
(3R)-6-cyclohexyl-3-[3-(methoxymethyl)-1,2,4-oxadiazol-5-
-yl]hexanoate (Preparation 48) (350 mg, 0.96 mmol) was treated with
trifluoroacetic acid (3 ml) and the resulting mixture was stirred
at room temperature under a nitrogen atmosphere for 2 hours. The
solvent was removed under reduced pressure and the residue
azeotroped from toluene then dichloromethane to afford the title
compound as a colourless oil (250 mg).
[0625] MS: 311 (MH.sup.+), 333 (MNa.sup.+)
[0626] .sup.1H-NMR (CDCl.sub.3) .delta.:4.57 (2H, s), 3.50 (4H, m),
3.00 (1H, m), 2.89 (1H, m), 1.90-1.51 (7H, m), 1.40-1.01 (8H, m),
0.84 (2H, m)
Preparation 50
tert-Butyl
(3R)-6-cyclohexyl-3-[3-(2-methoxyethyl)-1,2,4-oxadiazol-5-yl]he-
xanoate
[0627] 129
[0628] A solution of
(2R)-2-[2-(tert-butoxy)-2-oxoethyl]-5-cyclohexylpenta- noic acid
(Preparation 1) (500 mg, 1.70 mmol) in dichloromethane (30 ml) was
treated with 1,1'-carbonyidiimidazole (272 mg, 1.70 mmol) and the
solution was stirred at room temperature for 1 hour.
N'-hydroxy-3-methoxypropanimidamide (J.Amer.Chem.Soc.; 80; 1958;
3769-3771) (201 mg, 1.70 mmol) was then added and the mixture was
stirred for 1 hour. The solvent was removed under reduced pressure
and the residue was heated neat at 120.degree. C. under a nitrogen
atmosphere for 2 hours. The crude product was then purified by
column chromatography on silica gel eluting with
dichloromethane:methanol (99:1) to afford the title compound as a
colourless oil (410 mg).
[0629] MS: 381 (MH.sup.+), 403 (MNa.sup.+)
[0630] .sup.1H-NMR (CDCl.sub.3) .delta.: 3.73 (2H, t, J=6Hz), 3.42
(1H, m), 3.34 (3H, s), 2.98 (2H, t, J=6Hz), 2.77 (1H, dd, J=16,
9Hz), 2.61 (1H, dd, J=16, 5Hz), 1.79-1.54 (7H, m), 1.38 (9H, s),
1.32-1.03 (8H, m), 0.81 (2H, m)
Preparation 51
(3R)-6-Cyclohexyl-3-[3-(2-methoxyethyl)-1,2,4-oxadiazol-5-yl]hexanoic
acid
[0631] 130
[0632] tert-Butyl
(3R)-6-cyclohexyl-3-[3-(2-methoxyethyl)-1,2,4-oxadiazol--
5-yl]hexanoate (Preparation 50) (410 mg, 1.08 mmol) was treated
with trifluoroacetic acid (3 ml) and the resulting mixture was
stirred at room temperature under a nitrogen atmosphere for 1 hour.
The solvent was removed under reduced pressure and the residue
azeotroped from toluene then dichloromethane to afford the title
compound (250 mg).
[0633] MS: 325 (MH.sup.+)
[0634] .sup.1H-NMR (CDCl.sub.3) .delta.: 7.46 (1H, br s), 2.77 (2H,
t, J=6Hz), 3.48 (1H, m), 3.36 (3H, s), 3.07-2.87 (3H, m), 2.75 (1H,
dd, J=16, 5Hz), 1.86-1.53 (7H, m), 1.38-1.02 (8H, m), 0.83 (2H,
m)
Preparation 52
tert-Butyl (3R)-6-cyclohexyl-3-3-[2-oxo-2-(1
-pyrrolidinyl)ethyl]-1,2,4-ox- adiazol-5-yl}hexanoate
[0635] 131
[0636] A solution of
(2R)-2-[2-(tert-butoxy)-2-oxoethyl]-5-cyclohexylpenta- noic acid
(Preparation 1) (500 mg, 1.70 mmol) in dichloromethane (30 ml) was
treated with 1,1'-carbonyidiimidazole (272 mg, 1.70 mmol) and the
solution was stirred at room temperature for 1 hour.
N'-hydroxy-3-oxo-3-(1-pyrrolidinyl)propanimidamide (Patent FR
73-36858 731016) (291 mg, 1.70 mmol) was then added and the mixture
was stirred for 17 hours. The solvent was removed under reduced
pressure and the residue was heated neat at 110.degree. C. under a
nitrogen atmosphere for 2 hours. The crude product was then
purified by column chromatography on silica gel eluting with
dichloromethane:methanol (99:1) to afford the title compound as a
colourless oil (309 mg).
[0637] MS: 434 (MH.sup.+)
[0638] .sup.1H-NMR (CDCl.sub.3) .delta.: 3.76 (2H, s), 3.56-3.39
(5H, m), 2.80 (1H, dd, J=15, 8Hz), 2.61 (1H, dd, J=15, 4Hz), 1.97
(2H, m), 1.86 (2H, m), 1.81-1.58 (7H, m), 1.41-1.05 (17H, m), 0.83
(2H, m)
Preparation 53
(3R)-6-Cyclohexyl-3-{3-[2-oxo-2-(1-pyrrolidinyl)ethyl]-1,2,4-oxadiazol-5-y-
l}hexanoic acid
[0639] 132
[0640] tert-Butyl
(3R)-6-cyclohexyl-3-{3-(2-oxo-2-(1-pyrrolidinyl)ethyl]-1-
,2,4-oxadiazol-5-yl}hexanoate (Preparation 52) (309 mg, 0.71 mmol)
was treated with trifluoroacetic acid (2 ml) and the resulting
mixture was stirred at room temperature under a nitrogen atmosphere
for 2 hours. The solvent was removed under reduced pressure and the
residue azeotroped from toluene to afford the title compound as a
pale yellow oil (200 mg).
[0641] MS: 378 (MH.sup.+), 400 (MNa.sup.+)
[0642] .sup.1H-NMR (CDCl.sub.3) .delta.:3.83 (2H, s), 3.64-3.40
(5H, m), 2.93 (1H, dd, J=17, 8Hz), 2.76 (1H, dd, J=17, 5Hz),
2.11-1.50 (11 H, m), 1.43-1.02 (8H, m), 0.84 (2H, m)
Preparation 54
tert-Butyl
(3R)-6-cyclohexyl-3-{3-[(phenylsulfonyl)methyl]-1,2,4-oxadiazol-
-5-yl}hexanoate
[0643] 133
[0644] A solution of
(2R)-2-[2-(tert-butoxy)-2-oxoethyl]-5-cyclohexylpenta- noic acid
(Preparation 1) (300 mg, 1.00 mmol) in dichloromethane (15m) was
treated with 1,1'-carbonyidiimidazole (162 mg, 1.00 mmol) and the
solution was stirred at room temperature for 2 hours.
N'-hydroxy-2-(phenylsulfonyl)ethanimidamide (J.Heterocycl.Chem.;
16; 1979; 1197-1200) (214 mg, 1.00 mmol) was then added and the
mixture was stirred for 17 hours. The solvent was removed under
reduced pressure and the residue was heated neat at 130.degree. C.
under a nitrogen atmosphere for 2 hours. The crude product was then
purified by column chromatography on silica gel eluting with
dichloromethane:methanol (99:1) to afford the title compound (78
mg).
[0645] MS: 499 (MNa.sup.+)
[0646] .sup.1H-NMR (CDCl.sub.3) .delta.: 7.83 (2H, d, J=7Hz), 7.69
(1H, dd, J=7,7Hz), 7.54 (2H, dd, J=7,7Hz), 4.52 (2H, s), 3.43 (1H,
m), 2.78 (1H, dd, J=16, 9Hz), 2.60 (1H, dd, J=16, 5Hz), 1.78-1.58
(7H, m), 1.40 (9H, s), 1.30-1.06 (8H, m), 0.85 (2H, m)
Preparation 55
(3R)-6-Cyclohexyl-3-3-[(phenylsulfonyl)methyl]-1,2,4-oxadiazol-5-yl}hexano-
ic acid
[0647] 134
[0648] tert-Butyl
(3R)-6-cyclohexyl-3-{3-[(phenylsulfonyl)methyl]-1,2,4-ox-
adiazol-5-yl}hexanoate (Preparation 54) (78 mg, 0.16 mmol) was
treated with trfluoroacetic acid (2 ml) and the resulting mixture
was stirred at room temperature under a nitrogen atmosphere for 4
hours. The solvent was removed under reduced pressure and the
residue azeotroped from toluene then dichloromethane to afford the
title compound as an oil which crystallised on standing (60
mg).
[0649] MS: 421 (MH.sup.+), 438 (MNH.sub.4.sup.+)
[0650] .sup.1H-NMR (CDCl.sub.3) .delta.: 7.80 (2H, d, J=7Hz), 7.66
(1H, dd, J=7, 7Hz), 7.53 (2H, dd, J=7, 7Hz), 4.53 (2H, s), 3.45
(1H, m), 2.91 (1H, dd, J=16, 9Hz), 2.73 (1H, dd, J=5Hz), 1.80-1.52
(7H, m), 1.39-1.01 (8H, m), 0.84 (2H, m)
Preparation 56
tert-Butyl
(3R)-3-{3-[(4-chlorophenoxy)methyl]-1,2,4-oxadiazol-5-yl}-6-cyc-
lohexylhexanoate
[0651] 135
[0652] A solution of
(2R)-2-[2-(tert-butoxy)-2-oxoethyl]-5-cyclohexylpenta- noic acid
(Preparation 1) (300 mg, 1.00 mmol) in dichloromethane (15 ml) was
treated with 1,1'-carbonyldiimidazole (162 mg, 1.00 mmol) and the
solution was stirred at room temperature for 2 hours.
2-(4-chlorophenoxy)-N'-hydroxyethanimidamide (Patent US 97-815671
970313) (197 mg, 0.98 mmol) was then added and the mixture was
stirred for 17 hours. The solvent was removed under reduced
pressure and the residue was heated neat at 120.degree. C. under a
nitrogen atmosphere for 2 hours. The crude product was then
purified by column chromatography on silica gel eluting with
dichloromethane:methanol (99:1) to afford the title compound as a
colourless oil (220 mg).
[0653] .sup.1H-NMR (CDCl.sub.3) .delta.: 7.25 (2H, d, J=8Hz), 6.93
(2H, d, J=8Hz), 5.13 (2H, s), 3.48 (1H, m), 2.82 (1H, dd, J=17,
9Hz), 2.64 (1H, dd, J=17, 4Hz), 1.81-1.57 (7H, m), 1.36 (9H, s),
1.33-1.03 (8H, m), 0.83 (2H, m)
Preparation 57
(3R)-3-{3-[(4-Chlorophenoxy)methyl]-1,2,4-oxadiazol-5-yl}-6-cyclohexylhexa-
noic acid
[0654] 136
[0655] tert-Butyl
(3R)-3-{3-[(4-chlorophenoxy)methyl]-1,2,4-oxadiazol-5-yl-
}6-cyclohexylhexanoate (Preparation 56) (215 mg, 0.46 mmol) was
treated with trifluoroacetic acid (5 ml) and the resulting mixture
was stirred at room temperature under a nitrogen atmosphere for 4
hours. The solvent was removed under reduced pressure and the
residue azeotroped from toluene to afford the title compound (189
mg).
[0656] .sup.1H-NMR (CDCl.sub.3) .delta.: 7.25 (2H, d, J=9Hz), 6.94
(2H, d, J=9Hz), 5.14 (2H, s), 3.53 (1H, m), 2.99 (1H, dd, J=15,
9Hz), 2.78 (1H, dd, J=15, 4Hz), 1.85-1.56 (7H, m), 1.35-1.04 (8H,
m), 0.84 (2H, m)
Preparation 58
tert-Butyl
(3R)-6-cyclohexyl-3-[3-(2-pyridinylmethyl)-i,2,4-oxadiazol-5-yl-
]hexanoate
[0657] 137
[0658] A solution of
(2R)-2-[2-(tert-butoxy)-2-oxoethyl]-5-cyclohexylpenta- noic acid
(Preparation 1) (300 mg, 1.00 mmol) in dichloromethane (15 ml) was
treated sequentially with
1-[3-(dimethylamino)propyl]-3-ethylcarbodii- mide hydrochloride
(192 mg, 1.00 mmol), 4-(dimethylamino)pyridine (125 mg, 1.02 mmol)
and N-hydroxy-2-(2-pyridinyl)ethanimidamide (Chem.Pharm.Bull.; 21;
10; 1973; 2146-2160) (152 mg, 1.00 mmol). The resulting mixture was
stirred at room temperature for 17 hours. The solvent was removed
under reduced pressure and the residue was heated neat at
120.degree. C. under a nitrogen atmosphere for 2 hours. The crude
product was then purified by column chromatography on silica gel
eluting with dichloromethane:methanol (99:1). The residue was
further purified by column chromatography on silica gel eluting
with ethyl acetate: pentane (30:70) to afford the title compound as
an oil (107 mg).
[0659] MS: 414 (MNa.sup.+), 436 (MNa.sup.+)
[0660] .sup.1H-NMR (CDCl.sub.3) .delta.: 8.55 (1H, d, J=5Hz), 7.62
(1H, dd, J=7, 7Hz), 7.32-7.10 (2H, m), 4.27 (2H, s), 3.45 (1H, m),
2.79 (1H, dd, J=16, 8Hz), 2.60 (1H, dd, J=16, 5Hz), 1.83-1.51 (7H,
m), 1.41-1.00 (17H, m), 0.82 (2H, m)
Preparation 59
(3R)-6-Cyclohexyl-3-[3-(2-pyridinylmethyl)-1,2,4-oxadiazol-5-yl]hexanoic
acid trifluoroacetate
[0661] 138
[0662] tert-Butyl
(3R)-6-cyclohexyl-3-[3-(2-pyridinylmethyl)-1,2,4-oxadiaz-
ol-5-yl]hexanoate (Preparation 58) (247 mg, 0.60 mmol) was treated
with trifluoroacetic acid (7 ml) and the resulting mixture was
stirred at room temperature under a nitrogen atmosphere for 4
hours. The solvent was removed under reduced pressure and the
residue azeotroped from toluene then dichloromethane to afford the
title compound as an oil (262 mg).
[0663] MS: 358 (MH.sup.+)
[0664] .sup.1H-NMR (CDCl.sub.3) .delta.: 8.75 (1H, d, J=5Hz), 8.04
(1H, dd, J=7, 7Hz), 7.55 (2H, m), 4.48 (2H, s), 3.49 (1H, m),
2.99-2.61 (2H, m), 1.84-1.44 (7H, m), 1.42-1.00 (8H, m), 0.84 (2H,
m)
Preparation 60
tert-Butyl
(3R)-6-cyclohexyl-3-({[(1S)-2-ethoxy-1-(hydroxymethyl)-2-oxoeth-
yl]amino}carbonyl)hexanoate
[0665] 139
[0666] A solution of
(2R)-2-[2-(tert-butoxy)-2-oxoethyl]-5-cyclohexylpenta- noic acid
(Preparation 1) (5.00 g, 16.76 mmol) in dichloromethane (75 ml) was
treated sequentially with 1-hydroxybenzotriazole hydrate (2.49 g,
18.43 mmol), serine ethyl ester hydrochloride (3.13 g, 18.43 mmol)
and N,N-diisopropylethylamine (6.13 ml, 35.19 mmol) and the
resulting mixture was stirred at 0.degree. C. under a nitrogen
atmosphere for 15 minutes.
1-[3-(dimethylamino)propyl]-3-ethylcarbodiimide hydrochloride (3.53
g, 18.43 mmol) was then added and the mixture was stirred for 48
hours being allowed to warm to room temperature over this time. The
mixture was diluted with dichloromethane (200 ml), washed
sequentially with water, aqueous citric acid solution (10% w/v), a
saturated aqueous solution of sodium hydrogen carbonate and brine,
dried over anhydrous magnesium sulphate, filtered and the solvent
removed under reduced pressure. The residue was then purified by
column chromatography on silica gel eluting a gradient system of
ethyl acetate:pentane (10:90) to (50:50) to afford the title
compound as a colourless oil (5.41 g).
[0667] MS :413 (M.sup.+)
[0668] Analysis: Found C, 63.20; H, 9.52; N, 3.27%;
C.sub.22H.sub.39NO.sub.6. 0.33 EtOAc requires C, 63.28; H, 9.48; N,
3.16%
[0669] .sup.1H-NMR (CDCl.sub.3) .delta.: 6.50 (1H, br d, J=6Hz),
4.60 (1H, m), 4.26 (2H, q, J=8Hz), 4.09 (1H, m), 3.85 (1H, m), 3.18
(1H, m), 2.70 (1H, dd, J=18, 9Hz), 2.51 (1H, m), 2.37 (1H, dd,
J=18, 3Hz), 1.78-1.52 (7H, m), 1.50-1.02 (20H, m), 0.85 (2H, m)
Preparation 61
Ethyl
(4S)-2-{(1R)-1-[2-(tert-butoxy)-2-oxoethyl]4-cyclohexylbutyl}-4,5-di-
hydro-1,3-oxazole-4-carboxylate
[0670] 140
[0671] A solution of tert-butyl (3R)-6-cyclohexyl-3-({[(1
S)-2-ethoxy-1-(hydroxymethyl)-2-oxoethyl]amino}carbonyl)hexanoate
(Preparation 60) (4.14 g, 10 mmol) in anhydrous tetrahydrofuran (40
ml) was treated with (methoxycarbonylsulfamoyl)triethylammonium
hydroxide, inner salt [Burgess Reagent] (2.62 g, 11 mmol) and the
resulting mixture was heated under reflux under a nitrogen
atmosphere for 1 hour. The solvent was removed under reduced
pressure and the residue purified by column chromatography on
silica gel eluting with a gradient system of pentane:ethyl acetate
(80:20) to (50:50) to afford the title compound as a colourless oil
(3.10 g)
[0672] .sup.1H-NMR (CDCl.sub.3) .delta.: 4.69 (1H, m), 4.52-4.33
(2H, m), 4.22 (2H, m), 2.87 (1H, m), 2.63 (1H, dd, J=16, 7Hz), 2.40
(1H, dd, J=16, 6Hz), 1.76-1.03 (27H, m), 0.85 (2H, m)
Preparation 62
Ethyl 2-{(1R)-1
-[2-(tert-butoxy)-2-oxoethyl]4-cyclohexylbutyl}-1,3-oxazol-
e-4-carboxylate
[0673] 141
[0674] A suspension of copper (II) bromide (2.08 g, 9.31 mmol) and
hexamethylenetetramine (1.30 g, 9.31 mmol) in degassed
dichloromethane (25 ml) was treated with
1,8-diazabicyclo[5.4.0]undec-7-ene (1.39 ml, 9.31 mmol) and then
cooled in a cold water bath and stirred for 5 minutes. This
suspension was then treated dropwise with a solution of ethyl
(4S)-2-(1R)-1
-[2-(tert-butoxy)-2-oxoethyl]-4-cyclohexylbutyl}-4,5--
dihydro-1,3-oxazole-4-carboxylate (Preparation 61) (0.92 g, 2.33
mmol) in dichloromethane (5 ml) and the resulting mixture was
stirred at room temperature under a nitrogen atmosphere for 17
hours. The solvent was removed under reduced pressure and the
residue partitioned between ethyl acetate and a solution of 0.88
ammonia:saturated aqueous solution of ammonium chloride (1:1, 100
mls). The layers were separated and the aqueous layer was extracted
with ethyl acetate (.times.2). The organic layers were combined,
washed sequentially with hydrochloric acid (2M), saturated aqueous
sodium hydrogen carbonate solution and brine, dried over anhydrous
magnesium sulphate, filtered and the solvent removed under reduced
pressure. The residue was purified by column chromatography on
silica gel eluting with ethyl acetate:pentane (10:90) to afford the
title compound as a pale yellow oil (0.59 g).
[0675] MS: 394 (MH.sup.+)
[0676] .sup.1H-NMR (CDCl.sub.3) .delta.: 8.13 (1H, s), 4.39 (2H, q,
J=7Hz), 3.39 (1H, m), 2.80 (1H, dd, J=17, 8Hz), 2.58 (1H, dd, J=17,
6Hz), 1.84-1.53 (7H, m), 1.49-1.02 (20H, m), 0.84 (2H, m)
Preparation 63
(3R)-6-Cyclohexyl-3-[4-(ethoxycarbonyl)-1,3-oxazol-2-yl]hexanoic
acid trifluoroacetate
[0677] 142
[0678] A solution of ethyl 2-{(1R)-1
-[2-(tert-butoxy)-2-oxoethyl]-4-cyclo-
hexylbutyl}-1,3-oxazole-4-carboxylate (Preparation 62) (1.58 g,
4.01 mmol) in anhydrous dichloromethane (25 ml) was treated with
trifluoroacetic acid (7 ml) and the resulting mixture was stirred
at room temperature under a nitrogen atmosphere for 4 hours. The
solvent was removed under reduced pressure and the residue
azeotroped from dichloromethane to afford the title compound (1.66
g).
[0679] MS: 338 (MH.sup.+)
[0680] .sup.1H-NMR (CDCl.sub.3) .delta.: 9.62 (1H, br s), 8.12 (1H,
s), 4.34 (2H, q, J=7Hz), 3.39 (1H, m), 2.93 (1H, dd, J=17, 8Hz),
2.67 (1H, dd, J=17, 5Hz), 1.84-1.47 (7H, m), 1.34 (3H, t, J=7Hz),
1.29-0.98 (8H, m), 0.80 (2H, m)
Preparation 64
Ethyl 2-((1R)-1
-{2-[(benzyloxy)amino]-2-oxoethyl}4-cyclohexylbutyl)-1,3-o-
xazole4-carboxylate
[0681] 143
[0682] A mixture of
(3R)-6-cyclohexyl-3-[4-(ethoxycarbonyl)-1,3-oxazol-2-y- l]hexanoic
acid trifluoroacetate (Preparation 63) (1.66 g, 3.68 mmol),
1,1'-carbonyldiimidazole (0.80 g, 4.93 mmol), O-benzylhydroxylamine
hydrochloride (1.57 g, 9.84 mmol) and N,N-diisopropylethylamine
(1.71 ml, 9.82 mmol) in anhydrous tetrahydrofuran (10 ml) was
stirred at room temperature for 72 hours. The mixture was diluted
with ethyl acetate (50 ml) and washed sequentially with a saturated
solution of sodium hydrogen carbonate, aqueous citric acid solution
(10% w/v) and brine, dried over anhydrous magnesium sulphate,
filtered and the solvent removed under reduced pressure. The
residue was purified by column chromatography on silica gel eluting
with dichloromethane:methanol (98:2) then further purified by
column chromatography on silica gel eluting with ethyl
acetate:pentane (50:50) to afford the title compound as an orange
oil (1.41 g)
[0683] MS: 443 (MH.sup.+), 465 (MNa.sup.+)
[0684] .sup.1H-NMR (CDCl.sub.3) .delta.: 8.07 (1H, s), 7.40-7.22
(5H, m), 4.85 (2H, s), 4.36 (2H, q, J=7Hz), 3.45 (1H, m), 2.89-2.40
(2H, m), 1.82-1.56 (7H, m), 1.36 (3H, t, J=7Hz), 1.32-1.05 (8H, m),
0.84 (2H, m)
Preparation 65
2-((1R)-1-{2-[(Benzyloxy)amino]-2-oxoethyl}-4-cyclohexylbutyl)-1,3-oxazole-
-4-carboxylic acid
[0685] 144
[0686] A solution of ethyl
2-((1R)-1-{2-[(benzyloxy)amino]-2-oxoethyl}4-cy-
clohexylbutyl)-1,3-oxazole4-carboxylate (Preparation 64) (1.31 g,
2.96 mmol) in 1,4-dioxane:water (10 ml:5 ml) was treated with
lithium hydroxide monohydrate (0.19 g, 4.44 mmol) and stirred in a
cold water bath for 3.5 hours. The mixture was diluted with water
(100 ml) and washed with ethyl acetate. The layers were separated
and the aqueous layer was acidified with solid citric acid then
extracted with ethyl acetate. The layers were separated and the
organic layer was dried over anhydrous sodium sulphate, filtered
and the solvent removed under reduced pressure to afford the title
compound as a white solid (0.75 g).
[0687] MS: 415 (MH.sup.+), 437 (MNa.sup.+)
[0688] .sup.1H-NMR (DMSO-d.sub.6) .delta.: 12.87 (1H, br s), 11.03
(1H, br s), 8.58 (1H, s), 7.41-7.20 (5H, m), 4.70 (2H, s), 3.26
(1H, m), 2.50-2.26 (2H, m), 1.67-1.45 (7H, m), 1.22-0.99 (8H, m),
0.79 (2H, m)
Preparation 66
2-((1R)-1-{2-[(Benzyloxy)amino]-2-oxoethyl}-4-cyclohexylbutyl)-N,N-dimethy-
l-1,3-oxazole4-carboxamide
[0689] 145
[0690] A solution of
2-((1R)-1-{2-[(benzyloxy)amino]-2-oxoethyl}-4-cyclohe-
xylbutyl)-1,3-oxazole-4-carboxylic acid (Preparation 65) (200 mg,
0.48 mmol) in dichloromethane (6 ml) was treated sequentially with
1-hydroxybenzotriazole hydrate (72 mg, 0.53 mmol), dimethylamine
hydrochloride (79 mg, 0.96 mmol), N-methylmorpholine (160 .mu.l,
1.45 mmol) and 1-[3-(dimethylamino)propyl]-3-ethylcarbodiimide
hydrochloride (111 mg, 0.58 mmol) and the resulting mixture was
stirred at room temperature under a nitrogen atmosphere for 4
hours. The solvent was removed under reduced pressure and the
residue was dissolved in ethyl acetate and washed sequentially with
aqueous citric acid solution (10% w/v), a saturated aqueous
solution of sodium hydrogen carbonate and brine, dried over
anhydrous magnesium sulphate, filtered and the solvent removed
under reduced pressure. The residue was then purified by column
chromatography on silica gel eluting with
dichloromethane:methanol:0.88 ammonia (95:5:0.5) to afford the
title compound as a colourless oil (200 mg).
[0691] MS: 442 (MH.sup.+), 464 (MNa.sup.+)
[0692] .sup.1H-NMR (CDCl.sub.3) .delta.: 7.99 (1H, s), 7.43-7.20
(5H, m), 4.85 (2H, s), 3.44 (1H, m), 3.39-2.91 (6H, br m),
2.80-2.32 (2H, m), 1.80-1.51 (7H, m), 1.37-1.04 (8H, m), 0.84 (2H,
m)
Preparation 67
tert-Butyl
(3R)-6-cyclohexyl-3-({[2-hydroxy-1-(methoxycarbonyl)propyl]amin-
o}carbonyl) hexanoate
[0693] 146
[0694] An ice-cooled solution of
(2R)-2-[2-(tert-butoxy)-2-oxoethyl]-5-cyc- lohexylpentanoic acid
(Preparation 1) (6.40 g, 21.45 mmol) in dichloromethane (75 ml) was
treated sequentially with 1-hydroxybenzotriazole hydrate (3.19 g,
23.60 mmol), threonine methyl ester hydrochloride (4.00 g, 23.60
mmol), N,N-diisopropylethylamine (7.85 ml, 45.10 mmol) and
1-[3-(dimethylamino)propyl]-3-ethylcarbodiimide hydrochloride (4.52
g, 23.58 mmol) and the mixture was stirred for 17 hours being
allowed to warm to room temperature over this time. The solvent was
removed under reduced pressure and the residue was dissolved in
ethyl acetate then washed sequentially with water, aqueous citric
acid solution (10% w/v), a saturated aqueous sodium hydrogen
carbonate solution and brine, dried over anhydrous magnesium
sulphate, filtered and the solvent removed under reduced pressure.
The residue was dissolved in diethyl ether (100 ml) and treated
with pentane (150 ml) to produce a white precipitate. This was
filtered off and washed with pentane to afford the title compound
as a white powder (6.48 g).
[0695] MS: 436 (MNa.sup.+)
[0696] .sup.1H-NMR (CDCl.sub.3) .delta.: 6.35 (1H, br d), 4.63 (1H,
m), 4.26 (1H, m), 3.76 (3H, s), 2.73-2.53 (2H, m), 2.34 (1H, m),
1.73-1.56 (7H, m), 1.45-1.09 (20H, m), 0.84 (2H, m)
Preparation 68
tert-Butyl (3R)-6-cyclohexyl-3-({[1
-(methoxycarbonyl)-2-oxopropyl]amino}c- arbonyl) hexanoate
[0697] 147
[0698] A solution of tert-butyl
(3R)-6-cyclohexyl-3-({[2-hydroxy-1-(methox-
ycarbonyl)propyl]amino}carbonyl) hexanoate (Preparation 67) (6.48
g, 15.69 mmol) in dichloromethane (60 ml) was treated with
Dess-Martin periodinane
[1,1,1-triacetoxy-1,1-dihydro-1,2-benziodoxol-3(1H)-one] (7.32 g,
17.26 mmol) and the resulting mixture was stirred at room
temperature under a nitrogen atmosphere for 1 hour. A solution of
sodium thiosulphate (6 g in 50 ml water) and a saturated aqueous
sodium hydrogen carbonate solution (50ml) were then added to the
mixture which was stirred for a further 10 minutes. The layers were
separated and the aqueous layer was extracted with dichloromethane.
The combined organic layers were sequentially washed with water and
brine, dried over anhydrous magnesium sulphate, filtered and the
solvent removed under reduced pressure. The residue was purified by
column chromatography on silica gel eluting with a gradient system
of pentane:ethyl acetate (100:0 to 90:10) to afford the title
compound as a colourless oil (4.86 g)
[0699] MS: 412 (MH.sup.+), 434 (MNa.sup.+)
[0700] .sup.1H-NMR (CDCl.sub.3) .delta.: 6.82 (1H, br d), 5.21 (1H,
m), 3.78 (3H, s), 2.72-2.52 (2H, m), 2.40-2.25 (4H, m), 1.72-1.53
(7H, m), 1.45-1.04 (17H, m), 0.83 (2H, m)
Preparation 69
Methyl 2-{(1R)-1
-[2-(tert-butoxy)-2-oxoethyl]-4-cyclohexylbutyl)-5-methyl-
-1,3-oxazole-4-carboxylate
[0701] 148
[0702] A suspension of triphenylphosphine (9.49 g, 36.18 mmol),
iodine (7.98 g, 31.44 mmol) and triethylamine (8.49 ml, 60.75 mmol)
in tetrahydrofuran was cooled to -78.degree. C. then treated with
tert-butyl
(3R)-6-cyclohexyl-3-({[1-(methoxycarbonyl)-2-oxopropyl]amino}carbonyl)
hexanoate (Preparation 68) (4.86 g, 11.80 mmol) over 15 minutes.
The mixture was stirred at -78.degree. C. for 30 minutes then at
0-5.degree. C. for 2 hours. The mixture was diluted with water and
extracted with dichloromethane. The combined organic layers were
dried over anhydrous magnesium sulphate, filtered and the solvent
removed under reduced pressure. The residue was purified by column
chromatography on silica gel eluting with a gradient system of
ethyl acetate:pentane (0:100 to 10:90) to afford the title compound
as a colourless oil (2.92 g).
[0703] MS: 394 (MH.sup.+), 416 (MNa.sup.+)
[0704] .sup.1H-NMR (CDCl.sub.3) .delta.: 3.89 (3H, s), 3.29 (1H,
m), 2.74 (1H, dd, J=14, 6Hz), 2.60-2.49 (4H, m), 1.79-1.54 (7H, m),
1.38 (9H, s), 1.31-1.05 (8H, m), 0.82 (2H, m)
Preparation 70
(3R)-6-Cyclohexyl-3-[4-(methoxycarbonyl)-5-methyl-1,3-oxazol-2-yl]hexanoic
acid
[0705] 149
[0706] A solution of methyl
2-(1R)-1-[2-(tert-butoxy)-2-oxoethyl]4-cyclohe-
xylbutyl}-5-methyl-1,3-oxazole-4-carboxylate (Preparation 69) (2.92
g, 7.43 mmol) in anhydrous dichloromethane (15 ml) was treated with
trifluoroacetic acid (7.5 ml) and the resulting mixture was stirred
at room temperature under a nitrogen atmosphere for 24 hours. The
solvent was removed under reduced pressure and the residue was
azeotroped with dichloromethane (.times.3). The residue was
purified by column chromatography on silica gel eluting with a
gradient system of ethyl acetate:pentane (0:100 to 40:60) to afford
the title compound as a colourless oil (2.50 g).
[0707] MS: 338 (MH.sup.+), 360 (MNa.sup.+)
[0708] Analysis: Found C, 62.90; H. 8.18; N, 3.93%;
C.sub.18H.sub.27NO.sub.5. 0.3 EtOAc requires C, 63.38;H, 8.14; N,
3.85%
[0709] .sup.1H-NMR (CDCl.sub.3) .delta.:3.86 (3H, s), 3.33 (1H, m),
2.92 (1H, dd, J=17, 8Hz), 2.67 (1H, dd, J=17, 5Hz), 2.58 (3H, s),
1.81-1.56 (7H, m), 1.34-1.03 (8H, m), 0.81 (2H, m)
Preparation 71
Methyl
2-((1R)-1-{2-[(benzyloxy)amino]-2-oxoethyl}4-cyclohexylbutyl)-5-met-
hyl-1,3-oxazole-4-carboxylate
[0710] 150
[0711] A solution of
(3R)-6-cyclohexyl-3-[4-(methoxycarbonyl)-5-methyl-1,3-
-oxazol-2-yl]hexanoic acid (Preparation 70) (2.48 g, 7.36 mmol) was
cooled to 0.degree. and treated with 1-hydroxybenzotriazole hydrate
(994 mg, 7.36 mmol),
1-[3-(dimethylamino)propyl]-3-ethylcarbodiimide hydrochloride (2.12
g, 11.06 mmol) and N-methylmorpholine (1.21 ml, 11.04 mmol). The
mixture was stirred for 15 minutes then treated with
O-benzylhydroxyamine (1.17 g, 7.36 mmol) and further
N-methylmorpholine (0.81 ml, 7.36 mmol). The mixture was stirred
for 1 hour being allowed to warm to room temperature over this
time. The solvent was removed under reduced pressure and the
residue was dissolved in ethyl acetate, washed sequentially with
water, a saturated solution of sodium hydrogen carbonate and brine,
dried over anhydrous magnesium sulphate, filtered and the solvent
removed under reduced pressure to afford the title compound as a
colourless oil (3.22 g)
[0712] MS: 443 (MH.sup.+), 465 (MNa.sup.+)
[0713] Analysis: Found C, 66.78; H, 7.77; N, 6.19%;
C.sub.25H34N.sub.2O.sub.5. 0.3 EtOAc requires C, 67.10;H, 7.82; N,
5.97%
[0714] .sup.1H-NMR (CDCl.sub.3) .delta.: 8.62 (1H, br s), 7.33 (5H,
m), 4.84 (2H, s), 3.84 (3H, s), 3.36 (1H, m), 2.70-2.33 (5H, m),
1.78-1.54 (7H, m), 1.30-1.03 (8H, m), 0.82 (2H, m)
Preparation 72
2-((1R)-1-{2-[(Benzyloxy)amino]-2-oxoethyl}-4-cyclohexylbutyl)-5-methyl-1,-
3-oxazole-4-carboxylic acid
[0715] 151
[0716] A solution of methyl
2-((1R)-1-2-[(benzyloxy)amino]-2-oxoethyl)-4-c-
yclohexylbutyl)-5-methyl-1,3-oxazole-4-arboxylate (Preparation 71)
(1.00 g, 2.26 mmol) in 1,4-dioxane (10 ml) was treated with an
aqueous sodium hydroxide solution (1N, 1.5 ml) and stirred at room
temperature for 4 hours. Further aqueous sodium hydroxide (1N, 3.0
ml) was added and the mixture was stirred for 20 hours. The solvent
was removed under reduced pressure and the residue was dissolved in
water and washed with ethyl acetate. The layers were separated and
the aqueous layer was acidified with solid citric acid then
extracted with ethyl acetate. The combined organic layers were
dried over anhydrous magnesium sulphate, filtered anid the solvent
was removed under reduced pressure to afford the title compound as
a white foam (0.83 g).
[0717] MS: 427 (MH.sup.-)
[0718] .sup.1H-NMR (CDCl.sub.3) .delta.: 7.34 (5H, m), 4.85 (2H,
s), 3.40 (1H, m), 2.71-2.33 (5H, m), 1.76-1.51 (7H, m), 1.32-1.03
(8H, m), 0.83 (2H, m)
Preparation 73
2-((1R)-1-{2-[(Benzyloxy)amino]-2-oxoethyl}-4-cyclohexylbutyl)-N,N,5-trime-
thyl-1,3-oxazole-4-carboxamide
[0719] 152
[0720] A solution of methyl
2-((1R)-1-{2-[(benzyloxy)amino]-2-oxoethyl}-4--
cyclohexylbutyl5-methyl-1,3-oxazole-4-carboxylate (Preparation 72)
(152 mg, 0.36 mmol) in dichloromethane (5 ml) was treated
sequentially with 1-hydroxybenzotriazole hydrate (48 mg, 0.36
mmol), N-methylmorpholine (82 .mu.l, 0.75 mmol),
1-[3-(dimethylamino)propyl]-3-ethylcarbodiimide hydrochloride (75
mg, 0.39 mmol) and dimethylamine hydrochloride (29 mg, 0.36 mmol)
and the resulting mixture was stirred at room temperature under a
nitrogen atmosphere for 17 hours. The solvent was removed under
reduced pressure and the residue was dissolved in ethyl acetate and
washed sequentially with water, a saturated aqueous solution of
sodium hydrogen carbonate and brine, dried over anhydrous magnesium
sulphate, filtered and the solvent removed under reduced pressure.
The residue was purified by column chromatography on silica gel
eluting with dichloromethane:methanol:0.88 ammonia (95:5:0.5) to
afford the title compound as a colourless oil which crystallised on
standing (109 mg).
[0721] MS: 456 (MH.sup.+), 478 (MNa.sup.+)
[0722] .sup.1H-NMR (CDCl.sub.3) .delta.: 8.53 (1H, br s), 7.34 (5H,
m), 4.84 (2H, s), 3.36 (1H, m), 3.27-2.86 (6H, br m), 2.64-2.31
(5H, m), 1.74-1.50 (7H, m), 1.31-1.03 (8H, m), 0.82 (2H, m)
Preparation 74
2-((1R)-1-{2-[(Benzyloxy)amino]-2-oxoethyl}-4-cyclohexylbutyl)-5-methyl-1,-
3-oxazole-4-carboxamide
[0723] 153
[0724] A solution of methyl
2-((1R)-1-{2-[(benzyloxy)amino]-2-oxoethyl}4-c-
yclohexylbutyl)-5-methyl-1,3-oxazole4-carboxylate (Preparation 72)
(200 mg, 0.47 mmol) in dichloromethane (10 ml) was treated
sequentially with 1-hydroxybenzotriazole hydrate (63 mg, 0.47
mmol), N-methylmorpholine (77 .mu.l, 0.70 mmol) and
1-[3-(dimethylamino)propyl]-3-ethylcarbodiimide hydrochloride (134
mg, 0.70 mmol) and the resulting mixture was stirred at room
temperature under a nitrogen atmosphere for 10 minutes.
Concentrated ammonia solution (0.88, 50 .mu.l, 1.00 mmol) was then
added and the mixture was stirred for 17 hours. The mixture was
diluted with dichloromethane and washed sequentially with water, a
saturated aqueous solution of sodium hydrogen carbonate and brine,
dried over anhydrous magnesium sulphate, filtered and the solvent
removed under reduced pressure. The residue was then purified by
column chromatography on silica gel eluting with
dichloromethane:methanol:0.88 ammonia (97:3:0.3) to afford the
title compound as a white solid (140 mg).
[0725] MS: 426(MH.sup.-)
[0726] Analysis: Found C, 67.45; H, 7.85; N, 9.64%;
C.sub.24H33N.sub.3O.sub.4 requires C, 67.42;H, 7.78; N, 9.83%
[0727] .sup.1H-NMR (CDCl.sub.3) .delta.: 7.36 (5H, m), 4.86 (2H,
s), 3.36 (1H, m), 2.72-2.37 (5H, m), 1.75-1.53 (7H, m), 1.42-1.05
(8H, m), 0.84 (2H, m).
Preparation 75
tert-Butyl
(3R)-6-cyclohexyl-3-(3-{[3-(dimethylamino)-l-azetidinyl]carbony-
l}-I,2,4-oxadiazol-5-yl)hexanoate
[0728] 154
[0729] A solution of ethyl
5{(1R)-1-[2-(tert-butoxy)-2-oxoethyl]4-cyclohex-
ylbutyl}-1,2,4-oxadiazole-3-carboxylate (Preparation 3) (360 mg,
0.91 mmol) and triethylamine (370 mg, 3.65 mmol) in ethanol (5 ml)
was treated with N,N-dimethyl-3-azetidinamine (J.Med.Chem.;36; 801;
1993) (300 mg, 0.91 mmol) and the resulting mixture was heated at
80.degree. C. under a nitrogen atmosphere for 16 hours. The mixture
was partitioned between ethyl acetate (150 ml) and saturated
aqueous ammonium chloride solution (150 ml). The organic layer was
washed with saturated ammonium chloride solution (150 ml), water
(150 ml) and brine (150 ml), dried over anhydrous sodium sulphate,
filtered and the solvent removed under reduced pressure. The
residue was purified by column chromatography on silica gel eluting
with a gradient system of pentane:ethyl acetate:diethylamine
(87.5:12.5:0.25) gradually changing to pentane:ethyl acetate:
diethylamine (50:50:1) to afford the title compound as a colourless
oil (355 mg).
[0730] MS: 449(MH.sup.+), 471 (MNa.sup.+)
[0731] .sup.1H-NMR (CDCl.sub.3) .delta.4.56 (1H, dd), 4.38 (1H,
dd), 4.21 (1H, dd), 4.05 (1H,dd), 3.50 (1H, m), 3.17 (1H, m), 2.86
(1H, dd), 2.63 (1H, dd), 2.19 (6H, s), 1.78 (1H, m), 1.65 (6H, br
t), 1.39 (9H, s), 1.08-1.28 (8H, m), 0.82 (2H, m).
Preparation 76
(3R)-6-Cyclohexyl-3-(3-{[3-(dimethylamino)-1
-azetidinyl]carbonyly1,2,4-ox- adiazol-5-yl)hexanoic acid
[0732] 155
[0733] A solution of tert-butyl
(3R)-6-cyclohexyl-3-(3-{[3-(dimethylamino)- -1
-azetidinyl]carbonyl}-1,2,4-oxadiazol-5-yl)hexanoate (Preparation
75) (340 mg, 0.76 mmol) in trifluoroacetic acid (10 ml) was stirred
at room temperature for 2 hours. The solvent was removed under
reduced pressure and the residue azeotroped from dichloromethane.
The residue was dissolved in ethyl acetate (50 ml) and washed with
saturated aqueous ammonium chloride solution (50 ml) and brine (50
ml), dried over anhydrous sodium sulphate, filtered and the solvent
removed under reduced pressure to afford the title compound as a
white solid (30 mg). The combined aqueous washes were extracted
with dichloromethane, containing <10% methanol, (3.times.150
ml). The combined organic layers were dried over anhydrous sodium
sulphate, filtered and the solvent removed under reduced pressure
to afford the title compound as a white solid (180 mg), combined
total mass 210 mg.
[0734] MS: 393 (MH.sup.+), 415 (MNa.sup.+)
[0735] .sup.1H-NMR (CD.sub.3OD) .delta.:4.88 (1H, m), 4.70 (1H, m),
4.49 (1H, dd), 4.31 (1H, dd), 4.13 (1H, m), 3.05 (1H, m), 2.91 (1H,
dd), 2.35 (6H, s), 2.79 (1H, dd), 1.78 (2H, q), 1.58-1.73 (5H, m),
1.09-1.3.5 (8H, m), 0.70-0.93 (2H, br q).
Preparation 77
tert-Butyl 3-(1,3-dioxo-1,3-dihydro-2H-isoindol-2-yl)-1
-azetidinecarboxylate
[0736] 156
[0737] A solution of tert-butyl 3-iodocyclobutanecarboxylate (EP
992493) (5.0 g, 17.7 mmol) in dimethylformamide was treated with
potassium phthalimide (5.0 g, 27.0 mmol) and heated at 100.degree.
C. for 18 hours. The reaction mixture was filtered, to remove the
excess potassium phthalimide, which was washed with
dimethylformamide (10 ml). The filtrate was concentrated under
reduced pressure and the residue azeotroped with xylene (2.times.30
ml). The residue was purified by column chromatography on silica
gel eluting with a gradient system of 95:5 (pentane:ethyl acetate)
gradually changing to 55:45 (pentane:ethyl acetate) to afford the
title compound as a white solid (3.78 g).
[0738] MS: 325 (MNa.sup.+)
[0739] .sup.1H-NMR (CDCl.sub.3) .epsilon.: 7.84 (2H, m), 7.72 (2H,
m), 5.01 (1H m), 4.50 (2H, m), 4.20 (2H, m), 1.40 (9H, s).
Preparation 78
tert-Butyl 3-amino-1-azetidinecarboxylate
[0740] 157
[0741] A solution of tert-butyl
3-(1,3-dioxo-1,3-dihydro-2H-isoindol-2-yl)- -1-azetidinecarboxylate
(Preparation 77) in methylamine in methanol (2M) (10 ml) was
stirred in a sealed tube at 55.degree. C. for 3 hours. On cooling a
precipitate formed and the reaction mixture was concentrated under
reduced pressure. The residue was dissolved in ethyl acetate (100
ml) and washed with water (100 ml, containing (2M) hydrochloric
acid (3 ml)) and water (50 ml, containing (2M) hydrochloric acid (2
ml)). The combined aqueous was basified with (2M) sodium hydroxide
solution (20 ml) and extracted with ethyl acetate (3.times.75ml).
The combined organic layers were dried over anhydrous magnesium
sulphate, filtered and concentrated under reduced pressure. The
residue was purified by column chromatography on silica gel eluting
with a gradient system of 97.5:2.5:0.25
(dichloromethane:methanol:ammonia) gradually changing to 90:10:1
(dichloromethane:methanol:ammonia) to afford the title compound
(935 mg) of approximately 90% purity, which was used in preparation
without further purification.
[0742] MS: 173 (MH.sup.+)
[0743] .sup.1H-NMR (CDCl.sub.3) .delta.: 4.11 (2H, m), 3.76 (1H,
m), 3.57 (2H, m), 1.41 (9H, s).
Preparation 79
(3R)-3-[3-({[1-(tert-Butoxycarbonyl)-3-azetidinyl]amino}carbonyl)-1,2,4-ox-
adiazol-5-yl]-6-cyclohexylhexanoic acid
[0744] 158
[0745] A solution of
(3R)-6-cyclohexyl-3-[3-(ethoxycarbonyl)-1,2,4-oxadiaz-
ol-5-yl]hexanoic acid (Preparation 4) (480 mg, 1.42 mmol) and
triethylamine (1 ml, 7.18 mmol) in ethanol (8 ml) was treated with
tert-butyl 3-amino-1-azetidinecarboxylate (Preparation 78) (300 mg,
1.74 mmol) and the resulting mixture was heated at 80.degree. C.
under a nitrogen atmosphere for 18 hours. The solvent was removed
under reduced pressure. The residue was purified by column
chromatography on silica gel eluting with a gradient system of
dichloromethane:methanol (99:1) gradually changing to
dichloromethane:methanol (90:10) to afford the title compound
contaminated with starting amine. The solid was dissolved in ethyl
acetate and washed with hydrochloric acid (0.5M) (2.times.), water
and brine, dried over magnesium sulphate, filtered and the solvent
was removed under reduced pressure to afford the title compound as
a sticky foam.
[0746] MS: 463 (M-H).sup.-
[0747] .sup.1H-NMR (CDCl.sub.3) .delta.7.64 (1H, d), 4.80 (1H, m),
4.25 (2H, t), 3.88 (2H, dd), 3.97 (1H, dd), 2.79 (1H, dd),
1.57-1.80 (8H, m), 1.38 (9H, s), 1.03-1.20 (8H, m), 0.80 (2H,
m).
Preparation 80
(3R)-3-[3-({3-[bis(tert-Butoxycarbonyl)amino]-1-azetidinyl}carbonyl)-1,2,4-
-oxadiazol-5-yl]-6-cyclohexylhexanoic acid
[0748] 159
[0749] A solution of
(3R)-6-cyclohexyl-3-[3-(ethoxycarbonyl)-1,2,4-oxadiaz-
ol-5-yl]hexanoic acid (Preparation 4) (480 mg, 1.26 mmol) and
triethylamine (350 l, 2.50 mmol) in ethanol (8 ml) was treated with
di(tert-butyl) 3-azetidinylimidodicarbonate (EP 153163, EP 106489)
(1.00 g, 4.56 mmol) and the resulting mixture was heated at
80.degree. C. under a nitrogen atmosphere for 18 hours. The solvent
was removed under reduced pressure. The solid was dissolved in
ethyl acetate and washed with water, to which the minimal amount of
hydrochloric acid (2M) was added to reach pH 2, hydrochloric acid
(0.5M) (2.times.), water and brine, dried over magnesium sulphate,
filtered and the solvent was removed under reduced pressure. The
residue was purified by column chromatography on silica gel eluting
with a gradient system of dichloromethane:methanol (99:1) gradually
changing to dichloromethane:methanol (90:10) to afford the title
compound (440 mg).
[0750] MS: 563 (M-H).sup.-
[0751] .sup.1H-NMR (CDCl.sub.3) .delta.: 4.78 (2H, m), 4.60 (1H,
m), 4.45 (1H, t), 4.26 (1H, dd), 2.98 (1H, dd), 2.78 (1H, dd),
1.60-2.0 (7H, m), 1.49 (9H, s), 1.08-1.35 (8H, m), 0.82 (2H,
m).
Preparation 81
tert-Butyl
(3R)-6-cyclohexyl-3-(3-{[[2-(dimethylamino)ethyl](methyl)amino]-
carbonyl)-1,2,4-oxadiazol-5-yl)hexanoate
[0752] 160
[0753] A solution of ethyl
5-{(1R)-1-[2-(tert-butoxy)-2-oxoethyl]-4-cycloh-
exylbutyl)-1,2,4-oxadiazole-3-carboxylate (Preparation 3) (560 mg,
1.42 mmol) in ethanol (2 ml) was treated with
N,N,N'-trimethylethylenediamine (900 l, 0.91 mmol) and the
resulting mixture was heated at 85.degree. C. in a sealed tube for
4.5 hours. The solvent was removed under reduced pressure and the
residue was purified by column chromatography on silica gel eluting
with a system of 95:5:0.5 (dichloromethane:methanol:ammonia) to
afford the title compound as a colourless oil (632 mg).
[0754] MS: 451 (MH.sup.+)
[0755] .sup.1H-NMR (CDCl.sub.3) .delta.: 3.64 (1H, br s), 3.51 (2H,
m), 3.14 (3H, s), 2.83 (1H, dd), 2.65 (1H, dd), 2.43-2.60 (2H br
d), 2.28 (3H, br s), 2.19 (3H, br s), 1.58-1.85 (7H, m), 1.39 (9H,
s), 1.07-1.36 (8H, m), 0.84 (2H, m).
Preparation 82
(3R)-6-Cyclohexyl-3-(3-[[2-(dimethylamino)ethyl](methyl)amino]carbonyl}-1,-
2,4-oxadiazol-5-yl)hexanoic acid
[0756] 161
[0757] A solution of tert-butyl
(3R)-6-cyclohexyl-3-(3-{[[2-(dimethylamino-
)ethyl](methyl)amino]carbonyl}-1,2,4-oxadiazol-5-yl)hexanoate
(Preparation 81) (630 mg, 1.40 mmol) in dichloromethane (5ml) was
treated with trifluoroacetic acid (5 ml) and stirred at room
temperature for 4 hours. The solvent was removed under reduced
pressure and the residue azeotroped from dichloromethane. The
residue was purified by column chromatography on silica gel eluting
with a gradient system of 100:0 (dichloromethane: methanol)
gradually changing to 90:10 (dichloromethane:methanol) to afford
the title compound as a colourless gum (453 mg).
[0758] MS: 395 (MH.sup.+)
[0759] Analysis: Found, C, 46.50; H. 6.00; N, 9.13%;
C.sub.20H34N.sub.4O.sub.4. 2 CF.sub.3CO.sub.2H requires C, 46.30;
H, 5.83; N, 9.00%
[0760] .sup.1H-NMR (CDCl.sub.3) .delta.: 3.88 (1H, m), 3.76 (1H,
m), 3.52 (1H, m), 3.05-3.40 (5H, m), 2.70-3.00 (7H, m), 1.57-1.90
(6H, m), 1.08-1.40 (7H, m), 0.84 (2H, m).
Preparation 83
tert-Butyl
(3R)-6-cyclohexyl-3-(3-{[[3-(dimethylamino)propyl](methyl)amino-
]carbonyl)-1,2,4-oxadiazol-5-yl)hexanoate
[0761] 162
[0762] A solution of ethyl
5-(1R)-1-[2-(tert-butoxy)-2-oxoethyl]-4-cyclohe-
xylbutyl}-1,2,4-oxadiazole-3-carboxylate (Preparation 3) (550 mg,
1.40 mmol) in ethanol (2 ml) was treated with
N,N,N'-trimethyl-1,3-propanediam- ine (1.02 ml, 7.00 mmol) and the
resulting mixture was heated at 85.degree. C. in a sealed tube for
3 hours. The solvent removed under reduced pressure. The residue
was purified by column chromatography on silica eluting with a
gradient system of 97:3:0.3 (dichloromethane:methan- ol:ammonia)
gradually changing to 90:10:1 (dichloromethane:methanol:ammoni- a)
to afford the title compound as a colourless oil (512 mg).
[0763] MS: 465 (MH.sup.+)
[0764] Analysis: Found, C, 64.44; H, 9.72; N, 12.07%;
C.sub.28H44N.sub.4O.sub.4 requires C, 64.62; H, 9.54; N, 12.06%
[0765] .sup.1H-NMR (CDCl.sub.3) .delta.: 3.58 (1H, t), 3.50 (1H.
t), 3.44 (1H, t), 3.09 (3H, d), 2.83 (1H, m), 2.64 (1H, dd), 2.35
(1H, t), 2.23 (3H, s), 2.20 (1H, m), 2.15 (3H, s), 1.58-1.88 (8H,
m), 1.40 (9H, s), 1.06-1.37 (7H, m), 0.84 (2H, m).
Preparation 84
(3R-6-Cyclohexyl-3-(3-[[3-(dimethylamino)propyl](methyl)amino]carbonyl}1,2-
,4-oxadiazol-5-yl)hexanoic acid
[0766] 163
[0767] A solution of tert-butyl
(3R)-6-cyclohexyl-3-(3-[[3-(dimethylamino)-
propyl](methyl)amino]carbonyl}-1,2,4-oxadiazol-5-yl)hexanoate
(Preparation 83) (500 mg, 1.08 mmol) in dichloromethane (5 ml) was
treated with trifluoroacetic acid (5 ml) and stirred at room
temperature for 4.5 hours. The solvent was removed under reduced
pressure and the residue azeotroped from dichioromethane. The
residue was purified by column chromatography on silica gel eluting
with a gradient system of 100:0 (dichloromethane:methanol)
gradually changing to 80:20 (dichloromethane:methanol) to afford
the title compound as a colourless glass (595 mg).
[0768] MS: 409 (MH.sup.+)
[0769] Analysis: Found, C, 45.48; H, 5.84; N, 8.50%; C.sub.2,
H36N.sub.4O.sub.4. 2 CF.sub.3CO.sub.2H. 2H.sub.2O requires C,
45.87; H, 6.16; N, 8.56%
[0770] .sup.1H-NMR (CDCl.sub.3) .delta.: 3.52 (2H, m), 3.37 (1H,
m), 3.09 (3H, m), 2.99 (2H, t), 2.81 (6H, s), 1.98 (2H, m),
1.58-1.90 (7H, m), 1.05-1.44 (8H, m), 0.88 (2H, m).
Preparation 85
tert-Butyl
(3R)-6-cyclohexyl-3-[3-(hydroxymethyl)-1,2,4-oxadiazol-5-yl]hex-
anoate
[0771] 164
[0772] A solution of ethyl
5-{(1R)-1-[2-(tert-butoxy)-2-oxoethyl]-4-cycloh-
exylbutyl)1,2,4-oxadiazole-3-carboxylate (Preparation 3) (15.2 g,
38.50 mmol) in ethanol (120 ml) was treated with portions of sodium
borohydride (1.46 g, 38.50 mmol) and the resulting mixture was was
stirred at room temperature under a nitrogen atmosphere for 5
hours. Aqueous citric acid (5% w/v solution) was added slowly and
the mixture was stirred at room temperature for a further 30
minutes. The organic solvent was removed under reduced pressure.
The aqueous layer was diluted with water and extracted with ethyl
acetate giving an emulsion. Anhydrous sodium chloride was added to
break up the emulsion. The combined organic layers were washed with
saturated sodium hydrogen carbonate solution and brine, dried over
anhydrous sodium sulphate, filtered and the solvent removed under
reduced pressure to afford the title compound as a colourless oil
(13.4 g).
[0773] MS: 375 (MH.sup.+)
[0774] .sup.1H-NMR (CDCl.sub.3) .delta.: 4.77 (2H, s), 3.46 (1H,
m), 2.80 (1H, dd), 2.62 (1H, dd), 1.58-1.80 (7H, m), 1.39 (9H, s),
1.07-1.33 (8H, m), 0.82 (2H, m).
Preparation 86
tert-Butyl
(3R)-6-cyclohexyl-3-{3-[(2-ethoxy-2-oxoethoxy)methyl]-1,2,4-oxa-
diazol-5-yl}hexanoate
[0775] 165
[0776] A suspension of sodium hydride 60% suspension in mineral oil
(13 mg, 0.33 mmol) in anhydrous tetrahydrofuran (1 ml) was cooled
to 0.degree. C. and treated with a solution of tert-butyl
(3R)-6-cyclohexyl-3-[3-(hydroxymethyl)-1,2,4-oxadiazol-5-yl]hexanoate
(Preparation 85) (105 mg, 0.30 mmol) in anhydrous tetrahydrofuran
(1 ml) and stirred under a nitrogen atmosphere for 1 hour. Ethyl
bromoacetate (37 .mu.l, 0.33 mmol) was added and the mixture was
stirred for 18 hours, being allowed to warm to room temperature
over this time. The reaction mixture was diluted with ethyl acetate
and washed with water and brine, dried over anhydrous sodium
sulphate, filtered and the solvent removed under reduced pressure.
The solid was purified by column chromatography on silica gel
eluting with a gradient system of pentane:ethyl acetate (99:1) to
pentane:ethyl acetate (80:20) to afford the title compound as a
colourless oil (84 mg).
[0777] MS: 461 (MNa.sup.+)
[0778] Analysis: Found, C, 62.87; H, 8.77; N, 6.39%;
C.sub.23H.sub.38N.sub.2O, requires C, 62.99; H, 8.73; N, 6.39%
[0779] .sup.1H-NMR (CDCl.sub.3) .delta.: 4.75 (2H, s), 4.20 (4H,
m), 3.45 (1H, m), 2.80 (1H, dd), 2.60 (1H, dd), 1.6-1.8 (7H, m),
1.40 (9H, s), 1.10-1.30 (11 H, m), 0.94 (2H, m).
Preparation 87
(3R)-6-Cyclohexyl-3-3-[(2-ethoxy-2-oxoethoxy)methyl]-1,2,4-oxadiazol-5-yl}-
hexanoic acid
[0780] 166
[0781] A solution of tert-butyl
(3R)-6-cyclohexyl-3-{3-[(2-ethoxy-2-oxoeth-
oxy)methyl]-1,2,4-oxadiazol-5-yl)hexanoate (Preparation 86) (500
mg, 1.08 mmol) in dichloromethane (7 ml) was treated with
trifluoroacetic acid (3ml) and stirred at room temperature for 5
hours. The solvent was removed under reduced pressure and the
residue azeotroped from toluene and dichloromethane. The oil was
purified by column chromatography on silica gel eluting with a
gradient system of 100:0:0 (dichloromethane:methanol:acetic acid)
gradually changing to 90:10:1 (dichloromethane:methanol:acetic
acid) to afford the title compound as a colourless oil (374
mg).
[0782] MS : 383 (MH.sup.+)
[0783] Analysis: Found, C, 59.67; H. 7.91; N, 7.32%;
C.sub.19H.sub.30N.sub.2O.sub.6. 0.05 H.sub.2O. 0.05
CH.sub.2CI.sub.2 requires C, 59.62; H, 7.90; N, 7.22%
[0784] .sup.1H-NMR (CDCl.sub.3) .delta.: 4.75 (2H, s), 4.20 (4H,
m), 3.51 (1H, m), 2.98 (1H, dd), 2.75 (1H, dd), 1.60-1.80 (7H, m),
1.10-1.30 (11H ,m), 0.82 (2H, m).
Preparation 88
tert-Butyl
(3R)-6-cyclohexyl-3-{3-[(2-ethoxy-1-methyl-2-oxoethoxy)methyl]--
1,2,4-oxadiazol-5-yl}hexanoate
[0785] 167
[0786] A suspension of sodium hydride 60% suspension in mineral oil
(18 mg, 0.45 mmol) in anhydrous tetrahydrofuran (1 ml) was cooled
to 0.degree. C. and treated with a solution of tert-butyl
(3R)-6-cyclohexyl-3-[3-(hydroxymethyl)-1,2,4-oxadiazol-5-yl]hexanoate
(Preparation 85) (142 mg, 0.40 mmol) in anhydrous tetrahydrofuran
(1 ml) and stirred under a nitrogen atmosphere for 0.5 hours. Ethyl
2-bromopropionate (37 .mu.l, 0.33 mmol) was added and the mixture
was stirred for 2 days, being allowed to warm to room temperature
over this time. The reaction mixture was diluted with ethyl acetate
and washed with water and brine, dried over anhydrous sodium
sulphate, filtered and the solvent removed under reduced pressure.
The oil was purified by column chromatography on silica gel eluting
with a gradient system of pentane : ethyl acetate (99:1) to
pentane:ethyl acetate (0:100) to afford the title compound as a
colourless oil (90 mg).
[0787] MS : 475 (MNa.sup.+)
[0788] Analysis: Found, C, 63.57; H, 8.93; N, 6.19%;
C.sub.24H.sub.40N.sub.2O.sub.6 requires C, 63.69; H, 8.91; N,
6.19%
[0789] .sup.1H-NMR (CDCl.sub.3) .delta.: 4.79 (1H, d), 4.58 (1H,
d), 4.20 (3H, m), 3.45 (1H, m), 2.78 (1H, dd), 2.60 (1H, dd),
1.60-1.80 (7H, m), 1.42 (3H, d), 1.35 (9H, s), 1.10-1.30 (11H, m),
0.83 (2H, m).
Preparation 89
(3R)-6-Cyclohexyl-3-{3-[(2-ethoxy-1
-methyl-2-oxoethoxy)methyl]-1,2,4-oxad- iazol-5-yl}hexanoic
acid
[0790] 168
[0791] A solution of tert-butyl
(3R)-6-cyclohexyl-3-{3-[(2-ethoxy-1-methyl-
-2-oxoethoxy)methyl]-1,2,4-oxadiazol-5-yl}hexanoate (Preparation
88) (480 mg, 1.06 mmol) in dichloromethane (8 ml) was treated with
trifluoroacetic acid (3.5 ml) and stirred at room temperature for 5
hours. The solvent was removed under reduced pressure and the
residue azeotroped from toluene and dichloromethane. The oil was
purified by column chromatography on silica gel eluting with a
gradient system of 100:0:0 (dichloromethane:methanol:acetic acid)
gradually changing to 95:5:0.5 (dichloromethane:methanol:acetic
acid) to afford the title compound as a colourless oil (395
mg).
[0792] MS: 419 (MNa.sup.+)
[0793] Analysis: Found, C, 60.03; H, 8.21; N, 6.97%;
C.sub.20H.sub.32N.sub.2O.sub.6. 0.2 CH.sub.2Cl.sub.2 requires C,
60.04; H, 8.16; N, 7.00%
[0794] .sup.1H-NMR (CDCl.sub.3) .delta.: 4.80 (1H, d), 4.58 (1H,
d), 4.10-4.30 (3H, m), 3.50 (1H, m), 2.98 (1H, dd), 2.75 (1H, dd),
1.60-1.80 (7H, m), 1.45 (3H, d), 1.10-1.30 (11 H, m), 0.83 (2H,
m).
Preparation 90
tert-Butyl
(3R)-3-{3-[(2-amino-2-oxoethoxy)methyl]-1,2,4-oxadiazol-5-yl}-6-
-cyclohexylhexanoate
[0795] 169
[0796] A suspension of sodium hydride 60% suspension in mineral oil
(51 mg, 1.28 mmol) in anhydrous tetrahydrofuran (3 ml) was cooled
to 0.degree. C. and treated with a solution of tert-butyl
(3R)-6-cyclohexyl-3-[3-(hydroxymethyl)-1,2,4-oxadiazol-5-yl]hexanoate
(Preparation 85) (105 mg, 0.30 mmol) in anhydrous tetrahydrofuran
(3 ml) and stirred under a nitrogen atmosphere for 30 minutes.
2-Bromoacetamide (235 mg, 1.70 mmol) was added and the mixture was
allowed to warm to room temperature and then heated at 40.degree.
C. for 15 hours. The cooled reaction mixture was diluted with ethyl
acetate and washed with water and brine, dried over anhydrous
sodium sulphate, filtered and the solvent removed under reduced
pressure. The solid was purified by column chromatography on silica
gel eluting with a gradient system of pentane:ethyl acetate (99:1)
to pentane:ethyl acetate (0:100) to afford the title compound as a
colourless oil (330 mg).
[0797] MS : 432 (MNa.sup.+)
[0798] .sup.1H-NMR (CDCl.sub.3) .delta.: 4.68 (2H, s), 4.10 (2H,
s), 3.46 (1H, m), 2.79 (1H, dd), 2.62 (1H, dd), 1.6-1.8 (7H, m),
1.40 (9H, s), 1.10-1.30 (8H, m), 0.84 (2H, m).
Preparation 91
(3R)-3-{3-[(2-Amino-2-oxoethoxy)methyl]-1,2,4-oxadiazol-5-yl}-6-cyclohexyl-
hexanoic acid
[0799] 170
[0800] A solution of tert-butyl
(3R)-3-{3-[(2-amino-2-oxoethoxy)methyl]-1,-
2,4-oxadiazol-5-yl}-6-cyclohexylhexanoate (Preparation 90) (280 mg,
0.86 mmol) in dichloromethane (7 ml) was treated with
trifluoroacetic acid (3 ml) and stirred at room temperature for 4
hours. The solvent was removed under reduced pressure and the
residue azeotroped from toluene and dichloromethane. The oil was
purified by column chromatography on silica gel eluting with a
gradient system of 100:0:0 (dichloromethane:methanol:a- cetic acid)
gradually changing to 90:10:1 (dichloromethane:methanol:acetic
acid) to afford a colourless oil which was triturated with diethyl
ether and filtered to afford the title compound (155 mg).
[0801] MS: 376 (MNa.sup.+)
[0802] .sup.1H-NMR (CDCl.sub.3) .delta.: 6.60-6.80 (2H, br d) 4.70
(2H, s), 4.08 (2H, dd), 3.48 (1H, m), 2.85 (1H, dd), 2.73 (1H, dd),
1.60-1.80 (7H, m), 1.10-1.40 (8H, m), 0.82 (2H, m).
Preparation 92
tert-Butyl
(3R)-6-cyclohexyl-3-[3-({[(4-methylphenyl)sulfonyl]oxy}methyl)1-
,2,4-oxadiazol-5-yl]hexanoate
[0803] 171
[0804] A suspension of sodium hydride 60% suspension in mineral oil
(1.52 g, 38.00 mmol) in anhydrous tetrahydrofuran (30 ml) was
cooled to 0.degree. C. and treated with a solution of tert-butyl
(3R)-6-cyclohexyl-3-[3-(hydroxymethyl)-1,2,4-oxadiazol-5-yl]hexanoate
(Preparation 85) (13.40 g, 38.00 mmol) in anhydrous tetrahydrofuran
(120 ml) and stirred under a nitrogen atmosphere for 30 minutes.
p-Toluene sulphonyl chloride (7.25 g, 38.00 mmol) was added
portionwise and the mixture was allowed to warm to room temperature
over 18 hours. The solvent was removed under reduced pressure The
residue was dissolved in ethyl acetate and washed with water,
saturated sodium hydrogen carbonate solution and brine, dried over
anhydrous sodium sulphate, filtered and the solvent removed under
reduced pressure. The oil was purified by column chromatography on
silica gel eluting with dichloromethane to afford the title
compound (10.75 g).
[0805] MS: 529 (MNa.sup.+)
[0806] .sup.1H-NMR (CDCl.sub.3) .delta.:7.80 (2H, d), 7.33 (2H, d),
5.12 (2H, s), 3.40 (1H, m), 2.72 (1H, dd), 2.58 (1H, dd), 2.43 (3H,
s), 1.56-1.78 (7H, m), 1.35 (9H, s), 1.05-1.30 (8H, m), 0.82 (2H,
m).
Preparation 93
1-tert-Butyl 3-ethyl
2-({5-[(1R)-1-(2-tert-butoxy-2-oxoethyl)-4-cyclohexyl-
butyl]-1,2,4-oxadiazol-3-yl}methyl)malonate
[0807] 172
[0808] A suspension of sodium hydride 60% suspension in mineral oil
(35 mg, 38.00 mmol) in anhydrous tetrahydrofuran (3 ml) was cooled
to 0.degree. C. and treated with tert-butylethyl malonate (13.40 g,
38.00 mmol), stirred under a nitrogen atmosphere for 5 minutes and
then allowed to warm to room temperature. A solution of tert-butyl
(3R)-6-cyclohexyl-3-[3-({[(4-methylphenyl)sulfonyl]oxy}methyl)-1,2,4-oxad-
iazol-5- yl]hexanoate (Preparation 92) (400 mg, 0.78 mmol) in
anhydrous tetrahydrofuran (3 ml) was added and the mixture was
stirred at room temperature for 18 hours. The solvent was removed
under reduced pressure. The residue was dissolved in ethyl acetate
and washed with water and brine, dried over anhydrous sodium
sulphate, filtered and the solvent removed under reduced pressure
to afford the title compound as a colourless oil (518 mg) of
approximately 70% purity which was used without further
purification.
[0809] MS: 546 (MNa.sup.+)
[0810] .sup.1H-NMR (CDCl.sub.3) .delta.:4.20 (2H, m), 3.80 (1H, t),
3.40 (1H, m), 3.25 (2H, d), 2.75 (1H, dd), 2.59 (1H, dd), 1.60-1.75
(7H, m), 1.43 (9H, s), 1.39 (9H, s), 1.10-1.30 (11H, m), 0.83 (2H,
m).
Preparation 94
(3R)-6-Cyclohexyl-3-[3-(3-ethoxy-3-oxopropyl)-1,2,4-oxadiazol-5-yl]hexanoi-
c acid
[0811] 173
[0812] A solution of 1-tert-butyl 3-ethyl
2-({5-[(1R)-1-(2-tert-butoxy-2-o-
xoethyl)-4-cyclohexylbutyl]-1,2,4-oxadiazol-3-yl}methyl)malonate
(Preparation 93) (510 mg, 0.79 mmol) in dichloromethane (10 ml) was
treated with trifluoroacetic acid (5 ml) and stirred at room
temperature for 5 hours. The solvent was removed under reduced
pressure and the residue azeotroped from toluene. The solid was
dissolved in xylene (10 ml) and heated at 140.degree. C. for 7
hours. The solvent was removed under reduced pressure. The oil was
purified by column chromatography on silica gel eluting with a
gradient system of 95:5:0 (dichloromethane:isopropyl alcohol:acetic
acid) gradually changing to 90:10:1 (dichloromethane:isopropyl
alcohol:acetic acid) to afford the title compound as a colourless
oil (117 mg).
[0813] MS : 389 (MNa.sup.+)
[0814] .sup.1H-NMR (CDCl.sub.3) .delta.:4.12 (2H, q), 3.48 (1H, m),
3.03 (2H, t), 2.92 (1H, dd), 2.75 (3H, m), 1.60-1.80 (7H, m),
1.10-1.35 (11 H, m), 0.82 (2H, m).
Preparation 95
tert-Butyl
(3R)-3-[({[(Z)-1-amino-2-(propylsulfonyl)ethylidene]amino}oxy)c-
arbonyl]-6-cyclohexylhexanoate
[0815] 174
[0816] A solution of
(2R)-2-(2-tert-butoxy-2-oxoethyl)-5-cyclohexylpentano- ic acid
(Preparation 1) (500 mg, 1.67 mmol), 1-(3-dimethylaminopropyl)-3-e-
thylcarbodiimide hydrochloride (354 mg, 1.85 mmol),
N-methylmorpholine (203 l, 1.85 mmol) and 1-hydroxybenzotriazole
hydrate (227 mg, 1.67 mmol) in dichloromethane (20 ml) was treated
with (1Z)-N'-hydroxy-2-(propylsulf- onyl)ethanimidamide (300 mg,
1.67 mmol) and stirred at room temperature for 18 hours. The
reaction mixture was diluted with water (10 ml) and stirred for 20
minutes. The layers were separated via a 5 micron filter cartridge.
The organic solvent was removed under reduced pressure to afford
the title compound as a yellow solid (760 mg).
[0817] MS: 483 (MNa.sup.+)
[0818] .sup.1H-NMR (CDCl.sub.3) .delta.: 5.62 (2H, brs), 3.82 (2H,
d), 3.15 (2H, m), 2.83 (1H, m), 2.65 (1H, dd), 2.42 (1H, dd), 1.90
(2H, m), 1.57-1.80 (8H, m), 1.45 (9H, s), 1.10-1.30 (7H, m), 1.08
(3H, t), 0.83 (2H, m).
Preparation 96
tert-Butyl
(3R)-6-cyclohexyl-3-{3-[(propylsulfonyl)methyl]-1,2,4-oxadiazol-
-5-yl}hexanoate
[0819] 175
[0820] A solution of tert-butyl
(3R)-3-[({[(Z)-1-amino-2-(propylsulfonyl)e-
thylidene]amino}oxy)carbonyl]-6-cyclohexylhexanoate (Preparation
95) (760 mg, 1.60 mmol) in xylene (15 ml) was heated at 130.degree.
C. for 28 hours. After cooling to room temperature the reaction
mixture was purified by column chromatography on silica gel eluting
with a gradient system of 100:0 (pentane:ethyl acetate) gradually
changing to 70:30 (pentane:ethyl acetate) to afford the title
compound as a yellow oil (400 mg).
[0821] MS: 441 (MH.sup.+)
[0822] .sup.1H-NMR (CDCl.sub.3) .delta.:4.32 (2H, s), 3.48 (1H, m),
3.13 (2H, dd), 2.80 (1H, dd), 2.65 (1H, dd), 1.92 (2H, m),
1.60-1.80 (7H, m), 1.40 (9H, s), 1.10-1.35 (11 H, m), 0.82 (2H,
m).
Preparation 97
(3R)-6-Cyclohexyl-3-{3-[(propylsulfonyl)methyl]-1,2,4-oxadiazol-5-yl}hexan-
oic acid
[0823] 176
[0824] A solution of tert-butyl
(3R)-6-cyclohexyl-3-{3-[(propylsulfonyl)me-
thyl]-1,2,4-oxadiazol-5-yl}hexanoate (Preparation 96) (371 mg, 0.84
mmol) in hydrogen chloride in 1,4-dioxan (4M) (4 ml) was stirred at
room temperature for 24 hours. The solvent was removed under
reduced pressure. The solid was dissolved in fresh hydrogen
chloride in 1,4-dioxan (4M) and stirred at room temperature for 3
hours. The solvent was removed under reduced pressure. The residue
was dissolved in ethyl acetate and washed with saturated sodium
hydrogen carbonate solution and brine, dried over anhydrous
magnesium sulphate, filtered and the solvent was removed under
reduced pressure to afford the title compound as a yellow oil (313
mg).
[0825] MS: 385 (M-H).sup.-
[0826] .sup.1H-NMR (DMSO) .delta.:4.70 (2H, s), 3.43 (1H, m), 2.75
(2H, t), 2.52 (2H, obs), 1.75 (2H, m), 1.50-1.70 (7H, m), 1.05-1.30
(9H, m), 0.97 (3H, t), 0.80 (2H, m).
Preparation 98
2-((1R)-1-{2-[(Benzyioxy)amino]-2-oxoethyl}4-cyclohexylbutyl)-N-[2-(dimeth-
ylamino)ethy]-5-methyl-1,3-oxazole-4-carboxamide
[0827] 177
[0828] A solution
2-((1R)-1-{2-[(benzyloxy)amino]-2-oxoethyl}-4-cyclohexyl-
butyl)-5-methyl-1,3-oxazole-4-carboxylic acid (Preparation 72) (200
mg, 0.47 mmol), N-methylmorpholine (77 l, 0.70 mmol),
1-hydroxybenzotriazole hydrate (63 mg, 0.47 mmol) and
1-(3-dimethylaminopropyl)-3-ethylcarbodiim- ide hydochloride (134
mg, 0.70 mmol) in dichloromethane (10 ml) was treated with
N,N-dimethylethylenediamine (56 l, 0.51 mmol) and stirred at room
temperature for 18 hours. The reaction mixture was diluted with
dichloromethane and washed with water, saturated sodium hydrogen
carbonate solution and brine, dried over anhydrous magnesium
sulphate, filtered and the solvent removed under reduced pressure.
The residue was purified by column chromatography on silica gel
eluting with a system of 94:4:0.4
(dichloromethane:methanol:ammonia) to afford the title compound as
a white solid (216 mg).
[0829] MS :499 (MH.sup.+)
[0830] Analysis: Found, C, 67.24; H, 8.60; N, 11.25%;
C.sub.28H.sub.42N.sub.4O.sub.4 requires C, 67.44; H, 8.49; N,
11.24%
[0831] .sup.1H-NMR (CDCl.sub.3) .delta.: 7.35 (5H, brs), 7.10 (1H,
br s), 4.85 (2H, s), 3.45 (2H, q), 3.35 (1H, m), 2.59 (3H, s), 2.46
(2H, t), 2.24 (6H, s), 1.60-1.75 (7H, m), 1.10-1.30 (7H, m), 0.85
(2H, m).
Preparation 99
Methyl ({[2-((1R)-1
-{2-[(benzyloxy)amino]-2-oxoethyl}4-cyclohexylbutyl)-5-
-methyl-1,3-oxazol4-yl]carbonyl}amino)acetate
[0832] 178
[0833] A solution
2-((1R)-1-{2-[(benzyloxy)amino]-2-oxoethyl}-4-cyclohexyl-
butyl)5-methyl-1,3-oxazole-4-carboxylic acid (Preparation 72) (230
mg, 0.53 mmol), N-methylmorpholine (61 l, 0.56 mmol),
1-hydroxybenzotriazole hydrate (68 mg, 0.53 mmol) and
1-(3-dimethylaminopropyl)-3-ethylcarbodiim- ide hydrochloride (107
mg, 0.56 mmol) in dichloromethane (10 ml) was treated with a
solution of glycine methyl ester hydrochloride (70 mg, 0.56 mmol)
and N-methylmorpholine (61 l, 0.56 mmol) in dichloromethane (2 ml)
and stirred at room temperature for 18 hours. The reaction mixture
was diluted with dichloromethane and washed with water, saturated
sodium hydrogen carbonate solution and brine, dried over anhydrous
magnesium sulphate, filtered and the solvent removed under reduced
pressure. The residue was purified by column chromatography on
silica gel eluting with a gradient system of 100:0:0
(dichloromethane:methanol:ammonia) gradually changing to 98:2:0.2
(dichloromethane:methanol:ammonia) to afford the title compound as
a white solid (160 mg).
[0834] MS: 522 (MNa.sup.+)
[0835] .sup.1H-NMR (CDCl.sub.3) .delta.: 7.44 (1H, br s), 7.35 (5H,
s), 5.11 (1H, s), 4.87 (2H, s), 4.12 (2H, d), 3.77 (3H, s), 3.37
(1H, m), 2.70 (1H, m), 2.56 (3H, s), 2.45 (1H, m), 1.55-1.1.80 (7H,
m), 1.05-1.40 (8H, m), 0.83 (2H, m).
Preparation 100
({[2-((1R)-1-{2-[(Benzyloxy)amino]-2-oxoethyl}4-cyclohexylbutyl)-5-methyl--
1,3-oxazol4-yl]carbonyl}amino)acetic acid
[0836] 179
[0837] A solution of methyl
({[2-((1R)-1-{2-[(benzyloxy)amino]-2-oxoethyl}-
-4-cyclohexylbutyl)-5-methyl-1,3-oxazol-4-yl]carbonyl}amino)acetate
(Preparation 99) (158 mg, 0.32 mmol) in 1,4-dioxan (5 ml) was
treated with sodium hydroxide solution (1M) (470 l, 0.47 mmol). The
mixture was stirred for 1 hour under a nitrogen atmosphere. The
reaction mixture was diluted in water and washed with ethyl
acetate. The aqueous layer was acidified with solid citric acid and
extracted with ethyl acetate. The organic layers were dried over
anhydrous magnesium sulphate, filtered and the solvent removed
under reduced pressure to afford the title compound (61 mg).
[0838] MS: 484 (M-H).sup.-
[0839] .sup.1H-NMR (CDCl.sub.3) .delta.: 7.35 (5H, br s), 4.85 (2H,
s), 4.17 (2H, d), 3.39 (1H, m), 2.60 (3H, s), 2.43 (1H, m),
1.55-1.75 (7H, m), 1.10-1.35 (9H, m), 0.85 (2H, m).
[0840] Accurate Mass:486.26 (MH.sup.+)
Preparation 101
tert-Butyl
(3R)-3-[({[(Z)-1,2-diamino-2-oxoethylidene]amino}oxy)carbonyl]--
6-cyclohexylhexanoate
[0841] 180
[0842] (R)-2-[2-(tert-Butoxy)-2-oxoethyl]-5-cyclohexylpentanoic
acid (Preparation 1) (3.47 g, 11.6 mmol) in dimethylformamide (21
ml) was treated with 1,1'-carbonyidiimidazole (1.97 g, 12.2 mmol)
and the mixture was stirred at ambient temperature for 1 hour.
N-Dimethylaminopyridine (1.43 g, 11.7 mmol) was then added in one
portion, followed by the addition of
2-amino-2-(hydroxyimino)acetamide (1.25 g, 12.1 mmol,
Helv.Chim.Acta; 47; 1964; 33-46). The resulting mixture was stirred
at room temperature for 24 hours. The mixture was partitioned
between ethyl acetate and a 10% solution of citric acid in
demineralised water. The layers were separated and the aqueous
phase was extracted with ethyl acetate. The combined organic layers
were washed with demineralised water (.times.4) and brine, dried
over anhydrous magnesium sulfate, filtered and the solvent removed
under reduced pressure. The residue was crystallised from n-hexane,
the precipitate was collected by filtration and dried in vacuo to
afford the title compound as a colourless solid (3.30 g, 74%
yield).
[0843] MS: 382 [(M-H)].sup.-
[0844] .sup.1H-NMR (CDCl.sub.3) .delta.: 7.04 (1H, br s), 5.73 (2H,
br s), 5.46 (1H, br s), 2.90 (1H, m), 2.68 (1H, dd, J=15, 12 Hz),
2.45 (1H, dd, J=18, 6 Hz), 1.63-1.74 (9H, m), 1.55-1.50 (1H, m),
1.43 (9H, s), 1.37-1.10 (5H, m), 0.93-0.82 (2H, m)
Preparation 102
tert-butyl
(3R)-6-cyclohexyl-3-[3-(1H-imidazol-2-ylmethyl)-1,2,4-oxadiazol-
-5-yl]hexanoate
[0845] 181
[0846] A solution of tert-butyl 1H-imidazole-1-carboxylate (133 mg,
0.79 mmol) in anhydrous tetrahydrofuran (2 ml), cooled to
-78.degree. C., treated with n-butyl lithium, 2.5M in hexanes (320
.mu.l, 0.79 mmol) and stirred under a nitrogen atmosphere for 1
hour. A solution of tert-butyl
(3R)-6-cyclohexyl-3-[3-(([(4-methylphenyl)sulfonyl]oxy)methyl)-1,2,4-oxad-
iazol-5- yl]hexanoate (Preparation 92) (400 mg, 0.79 mmol) in
anhydrous tetrahydrofuran (2ml) was added slowly and the mixture
was allowed to warm to room temperature and stirred at this
temperature for 18 to hours. The reaction mixture was partitioned
between ethyl acetate (30 ml) and water (30 ml). The organic layer
was washed once more with water (20 ml), dried over anhydrous
sodium sulphate, filtered and the solvent removed under reduced
pressure to afford the title compound as a pale yellow oil (307
mg).
[0847] MS: 403 (MH.sup.+)
[0848] Analysis: Found, C, 65.61; H, 8.91; N, 11.78%;
C.sub.22H.sub.34N.sub.4O.sub.3 requires C, 65.64; H, 8.51; N,
13.92%
[0849] .sup.1H-NMR (CDCl.sub.3) .delta.: 7.59 (1H, s), 7.07 (1H,
s), 7.01 (1H, s), 5.28 (2H, s), 3.40 (1H, m), 2.76 (1H, dd), 2.60
(1H, dd), 1.40-1.80 (IOH, m), 1.10-1.40 (12H, m), 0.88 (4H, m).
Preparation 103
(3R)-6-cyclohexyl-3-[3-(1H-imidazol-2-ylmethyl)-1,2,4-oxadiazol-5-yl]hexan-
oic acid
[0850] 182
[0851] tert-butyl
(3R)-6cyclohexyl-3-[3-(1H-imidazol-2-ylmethyl)-1,2,4-oxa-
diazol-5-yl]hexanoate (Preparation 102) (530 mg, 1.32 mmol) was
treated with trifluoroacetic acid (10 ml) and stirred at room
temperature for 1.5 hours. The solvent was removed under reduced
pressure and the residue azeotroped from toluene (3x30 ml) and
dichloromethane (2.times.30 ml) to afford the title compound (609
mg).
[0852] MS: 347 (MH.sup.+)
[0853] Analysis: Found, C, 48.81; H, 5.61; N, 10.40%;
C.sub.18H.sub.26N.sub.4O.sub.3. 1.5CF.sub.3CO.sub.2H. 0.13H.sub.2O
requires C, 48.52; H, 5.38; N, 10.78%
[0854] .sup.1H-NMR (CDCl.sub.3) .delta.: 8.80 (1H, s), 7.38 (1H,
s), 7.28 (1H, s), 5.41 (2H, s), 3.48 (1H, m), 2.70-2.95 (2H, m),
1.50-1.80 (7H, m), 1.10-1.40 (8H, m), 0.85 (2H, m).
Preparation 104
tert-butyl
(3R)-3-[({[(Z)-1-amino-2-(4-pyridinyi)ethylidene]amino}oxy)carb-
onyl]-6-cyclohexylhexanoate
[0855] 183
[0856] A solution of
(2R)-2-(2-tert-butoxy-2-oxoethyl)-5-cyclohexylpentano- ic acid
(Preparation 1) (500 mg, 1.67 mmol) in dichloromethane (20 ml) was
treated with 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide
hydrochloride (354 mg, 1.85 mmol), N-methylmorpholine (203 .mu.l,
1.85 mmol) and 1-hydroxybenzotriazole hydrate (227 mg, 1.67 mmol).
(1Z)-N'-hydroxy-2-(4-pyridinyl)ethanimidamide (WO 9600720) (374 mg,
1.67 mmol) followed by N-methylmorpholine (369 .mu.l, 3.34 mmol)
were added to the reaction mixture which was stirred at room
temperature for 18 hours. The reaction mixture was diluted with
water (10 ml) and stirred for 20 minutes. The layers were separated
via a 5 micron filter cartridge. The organic solvent was removed
under reduced pressure to afford the title compound as a yellow oil
(840 mg). This was used without further purification in the
following step.
[0857] MS: 454 (MNa.sup.+)
[0858] .sup.1H-NMR (CDCl.sub.3) .delta.: 8.58 (2H, d), 7.28 (2H,
d), 4.90 (1H, br s), 3.57 (2H, s), 2.85 (1H, m), 2.40 (2H, m),
1.60-1.80 (6H, m), 1.30-1.60 (12H, m), 1.10-1.30 (6H, m), 0.87 (2H,
m).
Preparation 105
tert-butyl
(3R)-6-cyclohexyl-3-[3-(4-pyridinylmethyl)-1,2,4-oxadiazol-5-yl-
]hexanoate
[0859] 184
[0860] A solution of tert-butyl
(3R)-3-[({[(Z)-1-amino-2-(4-pyridinyl)ethy-
lidene]amino}oxy)carbonyl]-6-cyclohexylhexanoate (Preparation 104)
(840 mg, 1.60 mmol) in xylene (15 ml) was heated at 130.degree. C.
for 4.5 hours. After cooling to room temperature the reaction
mixture was purified by column chromatography on silica gel eluting
with a gradient system of 90:10 (pentane:ethyl acetate) gradually
changing to 50:50 (pentane:ethyl acetate) to afford the title
compound as a yellow oil (420 mg).
[0861] MS: 436 (MNa.sup.+)
[0862] Analysis: Found, C, 69.38; H, 8.69; N, 9.72%;
C.sub.24H.sub.35N.sub.3O.sub.3. 0.1EtOAc requires C, 69.39; H,
8.45; N, 9.72%
[0863] .sup.1H-NMR (CDCl.sub.3) .delta.: 8.54 (2H, d), 7.21 (2H,
d), 4.03 (2H, s), 3.40 (1H, m), 2.77 (1H, dd), 2.60 (1H, m),
1.60-1.80 (8H, m), 1.35 (9H, s), 1.10-1.30 (7H, m), 0.82 (2H,
m).
Preparation 106
(3R)-6-cyclohexyl-3-[3-(4-pyridinylmethyl)-1,2,4-oxadiazol-5-yl]hexanoic
acid
[0864] 185
[0865] A solution of tert-butyl
(3R)-6-cyclohexyl-3-[3-(4-pyridinylmethyl)-
-1,2,4-oxadiazol-5-yl]hexanoate (Preparation 105) (400 mg, 0.97
mmol) in hydrogen chloride in 1 ,4-dioxan (4M) (5 ml) was stirred
at room temperature for 24 hours. The solvent was removed under
reduced pressure. The solid was dissolved in fresh hydrogen
chloride in 1,4-dioxan (4M) and stirred at room temperature for 3
hours. The solvent was removed under reduced pressure to afford the
title compound as a yellow oil (390 mg).
[0866] MS: 356 (M-H).sup.-
[0867] Analysis: Found, C, 59.65; H, 7.34; N, 9.62%;
C.sub.20H.sub.27N.sub.3O.sub.3. HCl. 0.35H.sub.2O. 0.25Dioxan
requires C, 59.74; H, 7.33; N, 9.95%
[0868] .sup.1H-NMR (DMSO) .delta.:8.75 (2H, d), 7.78 (2H, d), 4.40
(2H, s), 3.40 (1H, m), 2.75 (2H, m), 1.50-1.70 (7H, m), 1.00-1.25
(8H, m), 0,89 (2H, m).
Preparation 107
tert-butyl
(3R)-3-[({[(Z)-1-amino-2-(3-pyridinyl)ethylidene]amino}oxy)carb-
onyl]-6-cyclohexylhexanoate
[0869] 186
[0870] A solution of
(2R)-2-(2-tert-butoxy-2-oxoethyl)-5-cyclohexylpentano- ic acid
(Preparation 1) (500 mg, 1.67 mmol) in dichloromethane (20 ml) was
treated with 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide
hydrochloride (354 mg, 1.85 mmol), N-methylmorpholine (203 .mu.l,
1.85 mmol) and 1-hydroxybenzotriazole hydrate (227 mg, 1.67 mmol).
(1Z)-N'-hydroxy-2-(3-pyridinyl)ethanimidamide (WO 9600720) (374 mg,
1.67 mmol) followed by N-methylmorpholine (369 .mu.l , 3.34 mmol)
were added to the reaction mixture which was stirred at room
temperature for 18 hours. The reaction mixture was diluted with
water (10 ml) and stirred for 20 minutes. The layers were separated
via a 5 micron filter cartridge. The organic solvent was removed
under reduced pressure to afford the title compound as a yellow oil
(835 mg). This was used without further purification in the
following step.
[0871] MS: 432 (MH.sup.+)
[0872] .sup.1H-NMR (CDCl.sub.3) .delta.: 8.55 (2H, d), 7.70 (1H,
d), 7.28 (1H, d), 4.90 (2H, br s), 3.70 (2H, m), 2.85 (1H, m), 2.40
(2H, m), 1.60-1.80 (7H, m), 1.40 (9H, s), 1.10-1.30 (8H, m), 0.83
(2H, m).
Preparation 108
tert-butyl
(3R-6-cyclohexyl-3-[3-(3-pyridinylmethyl)-1,2,4-oxadiazol-5-yl]-
hexanoate
[0873] 187
[0874] A solution of teit-butyl
(3R)-3-[({[(Z)-1-amino-2-(3-pyridinyl)ethy-
lidene]amino}oxy)carbonyl]-6-cyclohexylhexanoate (Preparation 107)
(835 mg, 1.60 mmol) in xylene (15 ml) was heated at 130.degree. C.
for 4.5 hours. After cooling to room temperature the reaction
mixture was purified by column chromatography on silica gel eluting
with a gradient system of 90:10 (pentane:ethyl acetate) gradually
changing to 50:50 (pentane:ethyl acetate) to afford the title
compound as a yellow oil (443 mg).
[0875] MS: 436 (MNa.sup.+)
[0876] .sup.1H-NMR (CDCl.sub.3) .delta.: 8.60 (1H, s), 8.50 (1H,
d), 7.62 (1H, d), 7.21 (1H, m), 4.05 (2H, s), 3.41 (1H, m), 2.77
(1H, dd), 2.58 (1H, m), 1.60-1.75 (8H, m), 1.35 (9H, s), 1.10-1.25
(7H, m), 0.82 (2H, m).
Preparation 109
(3R)-6-cyclohexyl-3-[3-(3-pyridinylmethyl)-1,2,4-oxadiazol-5-yl]hexanoic
acid
[0877] 188
[0878] A solution of tert-butyl
(3R)-6-cyclohexyl-3-[3-(3-pyridinylmethyl)-
-1,2,4-oxadiazol-5-yl]hexanoate (Preparation 108) (408 mg, 0.99
mmol) in hydrogen chloride in 1,4-dioxan (4M) (5 ml) was stirred at
room temperature for 18 hours. T.l.c. analysis showed reaction had
gone >90%. A couple of drops of concentrated hydrochloric acid
were added and the reaction mixture was stirred at room temperature
for a further 24 hours. The solvent was removed under reduced
pressure. A fresh portion of hydrogen chloride in 1,4-dioxan (4M)
(4 ml) was added and stirred at room temperature for 4 hours. The
solvent was removed under reduced pressure and azeotroped with
ethyl acetate and diethylether. The oil was dried under reduced
pressure to afford the title compound as a yellow oil (418 mg).
[0879] MS: 356 (M-H).sup.-
[0880] Analysis: Found, C, 59.07; H, 7.43; N, 8.71%;
C.sub.20H.sub.27N.sub.3P.sub.3. HCl. 0.4H.sub.2O. 0.7Dioxan
requires C, 59.17; H, 7.49; N, 9.08%
[0881] .sup.1H-NMR (DMSO) .delta.:8.72 (1H, s), 8.63 (1H, d) 8.10
(1H, d), 7.69 (1H, t), 4.25 (2H, s), 3.38 (1H, m), 2.67 (2H, t),
1.50-1.70 (7H, m), 1.00-1.25 (8H, m), 0.79 (2H, m).
Preparation 110
(4S)-4-isopropyl-3-(4-pentenoyl)-1,3-oxazolidin-2-one
[0882] 189
[0883] A solution of (4S)-4-isopropyl-1,3-oxazolidin-2-one (26 g,
0.2 mol) in anhydrous tetrahydrofuran (600 ml) was cooled to
-78.degree. C. under a nitrogen atmosphere and treated with n-butyl
lithium, 2.5M in hexanes (80 ml, 0.2 mol) keeping the temperature
below -65.degree. C. Once the addition was complete the reaction
mixture was allowed to warm to -55.degree. C. and stirred at this
temperature for 30 minutes. The mixture was cooled back down to
-78.degree. C. and treated dropwise with a solution of 4-pentenoyl
chloride (23.7 g, 0.2 mol) in anhydrous tetrahydrofuran (100 ml).
After stirring at -78.degree. C. for 30 minutes the reaction
mixture was allowed to warm to room temperature. Saturated aqueous
ammonium chloride solution (1 L) was added and the mixture was
extracted with ethyl acetate (2.times.). The combined organic
extracts were dried over anhydrous magnesium sulphate, filtered and
the solvent removed under reduced pressure. The oil was purified by
column chromatography on silica gel eluting with a solvent gradient
of 100:0 (pentane:ethyl acetate) gradually changing to 70:30
(pentane:ethyl acetate) to afford the title compound as a yellow
oil (36.3 g).
[0884] MS: 229 (MNH.sub.4+)
[0885] .sup.1H-NMR (CDCl.sub.3) .delta.: 5.85 (1H, m), 5.10 (1H.
d), 5.02 (1H. d), 4.41 (1H. m), 4.22 (2H, m), 3.11 (1H, m), 2.98
(1H, m), 2.30-2.45 (3H, m), 0.92 (3H, d), 0.85 (3H, d).
Preparation 111
tert-butyl
(3R)-3-{[(4S)4-isopropyl-2-oxo-1,3-oxazolidin-3-yl]carbonyl)-5--
hexenoate
[0886] 190
[0887] A solution of
(4S-4-isopropyl-3-(4-pentenoyl)-1,3-oxazolidin-2-one (Preparation
110) (36 g, 0.17 mol) in anhydrous tetrahydrofuran (650 ml), under
a nitrogen atmosphere, at -78.degree. C., was treated dropwise with
sodium bis(trimethylsilyl)amide, 1M in tetrahydrofuran (188 ml,
0.19 mol) over 1.5 hours, keeping the temperature below -65.degree.
C. The mixture was stirred at -78.degree. C. for 30 minutes and
then allowed to warm to room temperature. The mixture was poured
into saturated aqueous ammonium chloride solution (500 ml) and
extracted with diethylether. The organic extract was washed with
water and brine, dried over anhydrous magnesium sulphate, filtered
and the solvent removed under reduced pressure. The brown slurry
was triturated with hexane, filtered and washed with cold hexane.
The white solid was dried under reduced pressure to afford the
title compound (35.8 g).
[0888] MS: 343 (MNH.sub.4.sup.+)
[0889] .sup.1H-NMR (CDCl.sub.3) .delta.: 5.78 (1H, m), 5.08 (1H,
d), 5.02 (1H, s), 4.40 (1H, m), 4.18-4.30 (3H, m), 2.77 (1H, dd),
2.30-2.45 (3H, m), 2.18 (1H, m), 1.41 (9H, s), 0.91 (6H, t).
Preparation 112
(2R)-2-(4,4-dimethyl-2-oxopentyl)-4-pentenoic acid
[0890] 191
[0891] A solution of tert-butyl
(3R)-3-[(4S)-4-isopropyl-2-oxo-1,3-oxazoli-
din-3-yl]carbonyl}-5-hexenoate (Preparation 111) (37.5 g, 0.11 mol)
in 1,4-dioxan (190 ml), under a nitrogen atmosphere, was treated
with a solution of lithium hydroxide monohydrate (7.3 g, 0.17 mol)
in water (75 ml). The mixture was stirred at room temperature for
18 hours. Further lithium hydroxide monhydrate (2.42 g, 0.06 mol)
was added and the mixture partitioned between ethyl acetate and
water. The aqueous layer was acidified with hydrochloric acid (2M)
(250 ml) and extracted with ethyl acetate. The organic extract was
washed with brine, dried over anhydrous magnesium sulphate,
filtered and the solvent removed under reduced pressure. The oil
was purified by column chromatography on silica gel eluting with
50:50 (pentane:ethyl acetate) to afford the title compound as a
colourless oil (14.2 g).
[0892] MS: 232 (MNH.sub.4.sup.+)
[0893] .sup.1H-NMR (CDCl.sub.3) .delta.5.77 (1H, m), 5.12 (1H, d),
5.08 (1H, s), 2.90 (1H, m), 2.60 (1H, dd), 2.25-2.55 (3H, m), 1.43
(9H, s).
Preparation 113
ethyl
(2S)-2{[(2R)-2-(4,4-dimethyl-2-oxopentyl)-4-pentenoyl]amino}3-hydrox-
ypropanoate
[0894] 192
[0895] A solution (2R)-2-(4,4-dimethyl-2-oxopentyl)-4-pentenoic
acid (Preparation 112) (15.91 g, 74.3 mmol), 1-hydroxybenzotriazole
hydrate (11.00 g, 81.4 mmol), L-serine ethylester hydrochloride
(13.84 g, 81.6 mmol) and N,N-diisopropylethylamine (27 ml, 156.0
mmol) in dichloromethane (280 ml) was treated with
1-(3-dimethylaminopropyl)-3-eth- ylcarbodiimide hydrochloride
(15.67 g, 81.7 mmol) and stirred at room temperature for 18 hours
under a nitrogen atmosphere. The reaction mixture was diluted with
dichloromethane and washed with water, aqueous citric acid (2M),
saturated sodium hydrogen carbonate solution and brine, dried over
anhydrous sodium sulphate and filtered. The solvent was removed
under reduced pressure to afford the title-compound as a yellow oil
(23.8 g).
[0896] MS: 330 (MH.sup.+)
[0897] .sup.1H-NMR (CDCl.sub.3) .delta.: 6.50 (1H, br d), 5.77 (1H,
m), 5.10 (2H, s +d), 4.59 (1H, m), 4.22 (2H, q), 4.02 (1H, br d),
3.84 (1H, br d), 2.85 (1H, br s), 2.55-2.70 (2H, m), 2.40 (2H, m),
2.18 (1H, m), 1.40 (9H, s), 1.30 (3H, t).
Preparation 114
ethyl
(4S)-2-[(1R)-1-(4,4-dimethyl-2-oxopentyl)-3-butenyl]4,5-dihydro-1,3--
oxazole4-carboxylate
[0898] 193
[0899] A solution of ethyl
(2S)-2-{[(2R)-2-(4,4-dimethyl-2-oxopentyl)-4-pe-
ntenoyl]amino-3-hydroxypropanoate (Preparation 113) (23.8 g, 72.3
mmol) in anhydrous tetrahydrofuran (300 ml), under a nitrogen
atmosphere, was treated with
(methoxycarbonylsulfamoyl)triethylammonium hydroxide triethylamine
hydrochloride salt (43.5 g, 79.4 mmol) and stirred at reflux for
1.5 hours. After allowing to cool to room temperature the mixture
was filtered, washed with ethyl acetate and the filtrate was
concentrated under reduced pressure. The oil was purified by column
chromatography on silica gel eluting with a solvent gradient of
80:20 (hexane:diethylether) gradually changing to 50:50
(hexane:diethylether) to afford the title compound as a colourless
oil (11.0 g).
[0900] MS: 312 (MH.sup.+)
[0901] .sup.1H-NMR (CDCl.sub.3) .delta.: 5.73,(1H, m), 5.05 (2H, s
+d), 4.66 (1H, m), 4.42 (1H, t), 4.38 (1H, t), 4.20 (2H, m), 3.95
(1H, m), 2.60 (1H, dd), 2.30-2.50 (3H, m), 1.41 (9H, s), 1.28 (3H,
t).
Preparation 115
ethyl
2-[(1R)-1-(4,4-dimethyl-2-oxopentyl)-3-butenyl]-1,3-oxazole4-carboxy-
late
[0902] 194
[0903] A suspension of copper (II) bromide (50.6 g, 226.6 mmol) and
hexamethylenetetramine (31.6 g, 225.5 mmol) in dichloromethane (250
ml), under a nitrogen atmosphere, was treated with
1,8-diazabicyco[5.4.0]undec- -7-ene (33 ml, 220.7 mmol). The
mixture was cooled to 0.degree. C. and treated with a solution of
ethyl (4S)-2-[(1R)-1-(4,4-dimethyl-2-oxopentyl-
)-3-butenyl]4,5-dihydro-1,3-oxazole-4-carboxylate (Preparation 114)
(17.6 g, 56.4 mmol) in dichloromethane (250 ml) via a cannula. The
reaction mixture was allowed to warm to room temperature and
stirred for 18 hours. The solvent was removed under reduced
pressure and the residue was azeotroped with ethyl acetate. The
residue was dissolved in ethyl acetate and washed with a 1:1
mixture of 0.880 ammonia solution: saturated aqueous ammonium
chloride solution (.times.2), water, hydrochloric acid (2M)
(.times.3), saturated aqueous sodium hydrogen carbonate solution
and brine. The solvent was removed under reduced pressure. The oil
was purified by column chromatography on silica gel eluting with a
solvent gradient of 95:5 (hexane:diethylether) gradually changing
to 50:50 (hexane:diethylether) to afford the title compound as a
colourless oil (11.5 g).
[0904] MS: 310 (MH.sup.+)
[0905] .sup.1H-NMR (CDCl.sub.3) .delta.: 8.10 (1H, s), 5.70 (1H,
m), 5.01 (2H, s +d), 4.37 (2H, q), 3.45 (1H, m), 2.79 (1H, dd),
2.50-2.65 (2H, m), 2.45 (1H, m), 1.30-1.45 (12H, s +t).
Preparation 116
2-[(1R)-1-(2-tert-butoxy-2-oxoethyl)-3-butenyl]-1,3-oxazole-4-carboxylic
acid
[0906] 195
[0907] Two identical reactions were put on each containing the
following:
[0908] A solution of ethyl 2-[(1R)-1
-(4,4-dimethyl-2-oxopentyl)-3-butenyl- ]-1,3-oxazole-4-carboxylate
(Preparation 115) (4.0 g, 12.9 mmol) in 1,4-dioxan (40 ml) and
water (20 ml) was cooled to 0.degree. C. and treated with lithium
hydroxide monohydrate (0.8 g, 19.0 mmol). The mixture was stirred
at 0.degree. C. for 4 hours. Further lithium hydroxide monhydrate
(0.8 g, 19.0 mmol) was added and stirred at 0.degree. C. for 10
minutes. The two reactions were combined and partitioned between
ethyl acetate and water. The aqueous layer was neutralised with
citric acid (16.3 g, 38.0 mmol) and extracted with ethyl acetate
(.times.3). The combined organic extracts were washed with water
and brine, dried over anhydrous sodium sulphate, filtered and the
solvent removed under reduced pressure to afford the title compound
as a colourless oil (8.3 g).
[0909] MS: 282 (MH.sup.+)
[0910] .sup.1H-NMR (CDCl.sub.3) .delta.: 8.10 (1H, s), 5.70 (1H,
m), 5.01 (2H, s +d), 3.47 (1H, m), 2.80 (1H, dd), 2.62 (1H, dd),
2.57 (1H, m), 2.45 (1H, m), 1.38 (9H, s).
Preparation 117
tert-butyl
(3R)-3-4-[(dimethylamino)carbonyl]-1,3-oxazol-2-yl}-5-hexenoate
[0911] 196
[0912] A solution of
2-[(1R)-1-(2-tert-butoxy-2-oxoethyl)-3-butenyl]-1,3-o-
xazole4-carboxylic acid (Preparation 116) (10.77 g, 38.3 mmol) in
dichloromethane (250 ml), at 0.degree. C., was treated with
1-hydroxybenzotriazole hydrate (5.75 g, 42.6 mmol), dimethylamine
hydrochloride (3.45 g, 42.3 mmol) and N,N-diisopropylethylamine (13
ml, 75.13 mmol) and lastly
1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (8.10
g, 42.3 mmol), allowed to warm to room temperature and stirred for
20 hours under a nitrogen atmosphere. The reaction mixture was
diluted with dichloromethane and washed with saturated sodium
hydrogen carbonate solution, aqueous citric acid (10%), water and
brine, dried over anhydrous sodium sulphate and filtered. The
solvent was removed under reduced pressure. The oil was purified by
column chromatography on silica gel eluting with a solvent gradient
of 7:3 (pentane:ethyl acetate) gradually changing to 1:1
(pentane:ethyl acetate) to afford the title compound as a
colourless oil (9.9 g).
[0913] MS: 309 (MH.sup.+)
[0914] Analysis: Found, C, 61.31; H, 7.89; N, 8.92%;
C.sub.16H.sub.24N.sub.2O.sub.4. 0.2H.sub.2O requires C, 61.60; H,
7.88; N, 8.98%
[0915] .sup.1H-NMR (CDCl.sub.3) .delta.: 8.00 (1H, s), 5.70 (1H,
m), 5.02 (2H, s +d), 3.40 (1H, m), 3.30 (3H, br s), 3.03 (3H, br
s), 2.75 (1H, m), 2.38 (1H, dd), 2.50 (1H, t), 2.43 (1H, m), 1.39
(9H, s).
Preparation 118
tert-butyl
(3R)-3-{4-[(dimethylamino)carbonyl]-1,3-oxazol-2-yl}-5-oxopenta-
noate
[0916] 197
[0917] A solution of tert-butyl
(3R)-3-{4-[(dimethylamino)carbonyl]-1,3-ox- azol-2-yl}-5-hexenoate
(Preparation 117) (1.0 g, 3.24 mmol) in acetone (20 ml) and water
(20 ml) was treated with osmium tetroxide, 2.5 % wt in tert-butanol
(500 .mu.l, 0.04 mmol) and stirred at room temperature for 5
minutes. Sodium periodate (2.13 g, 10.00 mmol) was added and the
reaction mixture stirred for 2 hours. The reaction mixture was
partitioned between ethyl acetate and aqueous sodium thiosulphate
solution (20 % wt). The organic layer was washed with water
(.times.2) and brine, dried over anhydrous sodium sulphate and
filtered. The solvent was removed under reduced pressure to afford
the title compound as a colourless oil (866 mg).
[0918] MS: 311 (MH.sup.+)
[0919] Analysis: Found, C, 56.13; H, 7.13; N, 8.66%;
C.sub.15H.sub.22N.sub.2O.sub.5. 0.6H.sub.2O requires C, 56.10; H,
7.28; N, 8.72%
[0920] .sup.1H-NMR (CDCl.sub.3) .delta.: 9.78 (1H, s), 8.01 (1H,
s), 3.87 (1H, m), 3.29 (3H, br s), 3.03 (4H, br s), 2.70-2.90 (2H,
m), 2.64 (1H, dd), 1.39 (9H, s).
Preparation 119
tert-butyl
(3R,5Z)-6-cyclobutyl-3-{4-[(dimethylamino)carbonyl]-1,3-oxazol--
2-yl}-5-hexenoate
[0921] 198
[0922] A suspension of (cyclobutylmethyl)(triphenyl)phosphonium
bromide (Preparation 122) (680 mg, 1.65 mmol) in anhydrous
tetrahydrofuran (13 ml), under a nitrogen atmosphere, at 0.degree.
C., was treated with sodium bis(trimethylsilyl)amide 1M in
tetrahydrofuran (1.65 ml, 1.65 mmol) over a period of 5 minutes.
The bright orange mixture was stirred at oaC for 1 hour. A solution
of tert-butyl (3R)-3-{4-[(dimethylamino)car-
bonyl]-1,3-oxazol-2-yl5-oxopentanoate (Preparation 118) (428 mg,
1.38 mmol) in toluene (4 ml) was added to the mixture and stirred
at 0.degree. C. for 30 minutes and allowed to warm to room
temperature for a further 30 minutes. Water (2 ml) was added and
the solvents were removed under reduced pressure. The residue was
dissolved in ethyl acetate and washed with dilute aqueous of
potassium sodium tartrate solution and brine, dried over anhydrous
sodium sulphate, filtered and the solvent removed under reduced
pressure. The oil was purified by column chromatography on silica
gel eluting with a solvent gradient of 9:1 (hexane:ethyl acetate)
gradually changing to 1:1 (hexane:ethyl acetate) to afford the
title compound as a colourless oil (282 mg).
[0923] MS: 363 (MH.sup.+)
[0924] Analysis: Found, C, 65.95; H, 8.35; N, 7.72%;
C.sub.20H.sub.30N.sub.2O.sub.4. 0.1H.sub.20 requires C, 65.95; H,
8.36; N, 7.69%
[0925] .sup.1H-NMR (CDCl.sub.3) .delta.: 8.00 (1H, s), 5.55 (1H,
t), 5.11 (1H, q), 3.00-3.40 (8H, m), 2.73 (1H, dd), 2.56 (1H, dd),
2.30-2.50 (2H, m), 2.08 (2H, m), 1.70-1.90 (4H, m), 1.38 (9H,
s).
Preparation 120
tert-butyl
(3R)-6-cyclobutyl-3-{4-[(dimethylamino)carbonyl]-1,3-oxazol-2-y-
l}hexanoate
[0926] 199
[0927] A solution of tert-butyl
(3R,5Z)-6-cyclobutyl-3-{4-[(dimethylamino)-
carbonyl]-1,3-oxazol-2-yl}-5-hexenoate (Preparation 119) (260 mg,
0.72 mmol) in ethanol (15 ml) was treated with palladium hydroxide
(250 mg) followed by ammonium formate (500 mg) and the mixture was
stirred at reflux for 3 hours. The reaction mixture was allowed to
cool to room temperature and was filtered through arbacel. The
solvent was removed under reduced pressure. The residue was
dissolved in ethyl acetate and washed with water and brine, dried
over anhydrous sodium sulphate and filtered. The solvent was
removed under reduced pressure to afford the title compound as a
colourless oil (246 mg).
[0928] MS: 366 (MH.sup.+)
[0929] .sup.1H-NMR (CDCl.sub.3) .delta.: 8.17 (1H, s), 3.31 (4H,
m), 3.05 (3H, m), 2.70 (1H, dd), 2.59 (1H, dd), 2.22 (1H, m), 2.00
(2H, m), 1.60-1.90 (4H, m), 1.55 (2H, m), 1.30-1.40 (11 H, m) 1.07
(2H, m).
Preparation 121
(3R)-6-cyclobutyl-3-4-[(dimethylamino)carbonyl]-1,3-oxazol-2-yl}hexanoic
acid
[0930] 200
[0931] A solution of tert-butyl
(3R)-6-cyclobutyl-3-{4-[(dimethylamino)car-
bonyl]-1,3-oxazol-2-yl}hexanoate (Preparation 120) (240 mg, 0.66
mmol) in dichloromethane (4 ml) was treated with trifluoroacetic
acid (1.2 ml) and stirred at room temperature for 4 hours. The
solvent was removed under reduced pressure and the residue
azeotroped from toluene and dichloromethane. The oil was purified
by column chromatography on silica gel eluting with 97:3:0.3
(dichloromethane:methanol:acetic acid) to afford the title compound
as a white solid (161 mg).
[0932] MS: 309 (MH.sup.+)
[0933] .sup.1H-NMR (CDCl.sub.3) .delta.: 8.05 (1H, s), 3.33 (4H,
m), 3.05 (3H, m), 2.87 (1H, dd), 2.67 (1H, dd), 2.20 (1H, m), 1.99
(2H, m), 1.60-1.90 (4H, m), 1.52 (2H, m), 1.36 (2H, m) 1.07 (2H,
m).
Preparation 122
(cyclobutylmethyl)(triphenyl)phosphonium bromide
[0934] 201
[0935] A solution of (bromomethyl)cyclobutane (4.5 ml, 0.04 mol) in
toluene (50 ml) was treated with triphenylphosphine (10.56 g, 0.04
mol) and the mixture stirred at reflux, under a nitrogen
atmosphere, for 2 days. The mixture was allowed to cool to room
temperature and treated with hexane (50 ml). The solvent was
decanted off and further hexane added. The solvent was decanted off
again. The solid was washed with diethyl ether (.times.2) and dried
under reduced pressure to afford the title compound as a white
solid (6.34 g).
[0936] MS: 332 (MH.sup.+)
[0937] Analysis: Found, C, 67.13; H, 5.88; N, 0.00%;
C.sub.23H.sub.24PBr requires C, 67.16; H, 5.88; N, 0.00%
[0938] .sup.1H-NMR (d.sub.6-DMSO) .delta.: 7.65-7.90 (15H, m), 3.74
(2H, m), 2.62 (1H, br m), 1.60-1.80 (6H, m).
* * * * *