U.S. patent application number 12/472958 was filed with the patent office on 2009-09-17 for biaryl substituted nitrogen containing heterocycle inhibitors of lta4h for treating inflammation.
This patent application is currently assigned to DECODE GENETICS EHF. Invention is credited to Mark E. Gurney, Mahnaz Keyvan, Michael David Krohn, Vincent Sandanayaka, Jasbir Singh.
Application Number | 20090233969 12/472958 |
Document ID | / |
Family ID | 37603154 |
Filed Date | 2009-09-17 |
United States Patent
Application |
20090233969 |
Kind Code |
A1 |
Sandanayaka; Vincent ; et
al. |
September 17, 2009 |
BIARYL SUBSTITUTED NITROGEN CONTAINING HETEROCYCLE INHIBITORS OF
LTA4H FOR TREATING INFLAMMATION
Abstract
The present invention relates to a chemical genus of biaryl
substituted nitrogen-attached heterocycles that are inhibitors of
LTA4H (leukotriene A4 hydrolase). The compounds have the general
formula: ##STR00001## They are useful for the treatment and
prevention and prophylaxis of inflammatory diseases and
disorders.
Inventors: |
Sandanayaka; Vincent;
(Northboro, MA) ; Singh; Jasbir; (Naperville,
IL) ; Keyvan; Mahnaz; (Plainfield, IL) ;
Krohn; Michael David; (Romeoville, IL) ; Gurney; Mark
E.; (Grand Rapids, MI) |
Correspondence
Address: |
HESLIN ROTHENBERG FARLEY & MESITI PC
5 COLUMBIA CIRCLE
ALBANY
NY
12203
US
|
Assignee: |
DECODE GENETICS EHF
Reykjavik
IS
|
Family ID: |
37603154 |
Appl. No.: |
12/472958 |
Filed: |
May 27, 2009 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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11462296 |
Aug 3, 2006 |
|
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12472958 |
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60752274 |
Dec 21, 2005 |
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Current U.S.
Class: |
514/327 ;
435/184; 514/424; 546/216; 546/221; 548/541 |
Current CPC
Class: |
C07D 207/12 20130101;
C07D 211/46 20130101; A61P 29/00 20180101 |
Class at
Publication: |
514/327 ;
548/541; 514/424; 546/216; 546/221; 435/184 |
International
Class: |
A61K 31/445 20060101
A61K031/445; C07D 207/12 20060101 C07D207/12; A61K 31/40 20060101
A61K031/40; C07D 211/46 20060101 C07D211/46; A61P 29/00 20060101
A61P029/00; C12N 9/99 20060101 C12N009/99 |
Claims
1. A compound of formula: ##STR00028## wherein Ar is selected from
the group consisting of aryl, aryl substituted with from one to
three substituents independently selected from the group consisting
of halogen, loweralkyl, loweracyl, loweralkoxy, fluoroloweralkyl,
fluoroloweralkoxy, formyl, cyano, benzyl, benzyloxy, phenyl,
substituted phenyl, heteroaryl, heterocyclylalkyl and nitro; and
heteroaryl, heteroaryl substituted with from one to three
substituents independently selected from the group consisting of
halogen, loweralkyl, loweracyl, loweralkoxy, fluoroloweralkyl,
fluoroloweralkoxy, formyl, cyano, sulphonamide, amido, phenyl,
heteroaryl, heterocyclylalkyl and nitro; X is selected from the
group consisting of direct bond, SO, S(O.sub.2), NR.sup.10,
CH.sub.2CH.sub.2, CH.sub.2NH, NHCH.sub.2, CH.dbd.CH,
CH.sub.2C.dbd.O, OCR.sup.1aR.sup.1b and CR.sup.1aR.sup.1bO;
R.sup.10 is selected separately in each occurrence from the group
consisting of H and lower alkyl; R.sup.1a and R.sup.1b are selected
from the group consisting of H and lower alkyl, or R.sup.1a and
R.sup.1b taken together may form a 3-6 membered ring, which may
optionally contain a heteroatom chosen from O, S, SO, SO.sub.2, and
NR.sup.10; m is zero, 1 or 2; n is an integer chosen from 1, 2 and
3; p is an integer from 0-3; Y is selected from H, H; O; and H,
R.sup.3; R.sup.3 is lower alkyl; taken together ZW is H or Z is
(CH.sub.2).sub.1-10; in which one or two (CH.sub.2) may optionally
be replaced by a C.sub.3-C.sub.6 carbocycle, a C.sub.3-C.sub.6
heterocycle, --O--, --NR.sup.10--, --SO--, --S(O).sub.2--,
--C(.dbd.O)-- or --C.dbd.O(NH)--, provided that said --O--,
--NR.sup.10--, --SO--, --S(O).sub.2--, --C(.dbd.O)-- or
--C.dbd.O(NH)-- are not at the point of attachment to nitrogen and
are separated by at least one --(CH.sub.2).sub.2--; W is selected
from acyl, hydroxyl, carboxyl, amino, carboxamido, sulphonamide,
aminoacyl, --COOalkyl, --CHO, --C(O)fluororalkyl,
--C(O)CH.sub.2C(O)Oalkyl, --C(O)CH.sub.2C(O)Ofluoroalkyl, --SH,
--C(O)NH(OH), --C(O)N(OH)R, --N(OH)C(O)OH, --N(OH)C(O)R.sup.4,
heterocyclyl, substituted aryl, and substituted heterocyclyl; and
R.sup.4 is selected from the group consisting of H and lower
alkyl.
2. A compound according to claim 1, wherein Ar is phenyl or
substituted phenyl and m is zero of formula: ##STR00029## wherein
R.sup.1 or R.sup.2 are independently chosen from hydrogen, halogen,
loweralkyl, loweracyl, loweralkoxy, fluoroloweralkyl,
fluoroloweralkoxy, formyl, cyano, benzyl, benzyloxy, sulphonamide,
amido, phenyl, substituted phenyl, heteroaryl, heterocyclylalkyl
and nitro; or R.sup.1 and R.sup.2 taken together may form a 5-6
membered ring, which may optionally contain an oxygen.
3. A compound according to claim 2, wherein n is 1 or 2; p is 1 or
2; and ZW is H or Z is (CH.sub.2).sub.1-10 and W is selected from
acyl, hydroxyl, carboxyl, amino, carboxamido, aminoacyl, and
--COOalkyl.
4. (canceled)
5. A pharmaceutical composition comprising a pharmaceutically
acceptable carrier and a therapeutically effective amount of at
least one compound according to claim 1.
6. A method for inhibiting leukotriene A4 hydrolase comprising
contacting the LTA4H enzyme with a therapeutically effective amount
of a compound according to claim 1.
7. A method for treating a disorder associated with leukotriene A4
hydrolase comprising administering to a mammal a therapeutically
effective amount of a compound or a salt, hydrate or ester thereof
according to claim 1.
8. A method according to claim 7 wherein said disorder is
associated with inflammation.
9. A method according to claim 8 wherein said disorder is selected
from allergic inflammation, acute inflammation and chronic
inflammation.
10. A method according to claim 7, wherein said disorder is chosen
from asthma, chronic obstructive pulmonary disease (COPD),
rheumatoid arthritis, multiple sclerosis, inflammatory bowel
diseases (IBD), ulcerative colitis, and psoriasis.
11. A method according to claim 7, wherein said disorder is chosen
from hypercholesterolemia, atherosclerosis, thrombosis, stroke,
acute coronary syndrome, stable angina, peripheral vascular
disease, critical leg ischemia, intermittent claudication,
abdominal aortic aneurysm and myocardial infarction.
12. A compound of formula: ##STR00030## wherein Ar is selected from
the group consisting of aryl other than phenyl, aryl other than
phenyl substituted with from one to three substituents
independently selected from the group consisting of halogen,
loweralkyl, loweracyl, loweralkoxy, fluoroloweralkyl,
fluoroloweralkoxy, formyl, cyano, benzyl, benzyloxy, phenyl,
substituted phenyl, heteroaryl, heterocyclylalkyl and nitro; and
heteroaryl, heteroaryl substituted with from one to three
substituents independently selected from the group consisting of
halogen, loweralkyl, loweracyl, loweralkoxy, fluoroloweralkyl,
fluoroloweralkoxy, formyl, cyano, sulphonamide, amido, phenyl,
heteroaryl, heterocyclylalkyl and nitro; X is selected from the
group consisting of direct bond, O, SO, S(O.sub.2), NR.sup.10,
CH.sub.2, CF.sub.2, CH.sub.2CH.sub.2, CH.sub.2NH, NHCH.sub.2,
CH.dbd.CH, C.dbd.O, CH.sub.2C.dbd.O, CR.sup.1aR.sup.1b,
OCR.sup.1aR.sup.1b and CR.sup.1aR.sup.1bO; R.sup.10 is selected
separately in each occurrence from the group consisting of H and
lower alkyl; R.sup.1a and R.sup.1b are selected from the group
consisting of H and lower alkyl, or R.sup.1a and R.sup.1b taken
together may form a 3-6 membered ring, which may optionally contain
a heteroatom chosen from O, S, SO, SO.sub.2, and NR.sup.10; m is
zero, 1 or 2; n is an integer chosen from 1, 2 and 3; p is an
integer from 0-3; Y is selected from H, H; O; and H, R.sup.3;
R.sup.3 is lower alkyl; taken together ZW is H or Z is
(CH.sub.2).sub.1-10; in which one or two (CH.sub.2) may optionally
be replaced by a C.sub.3-C.sub.6 carbocycle, a C.sub.3-C.sub.6
heterocycle, --O--, --NR.sup.10--, --SO--, --S(O).sub.2--,
--C(.dbd.O)-- or --C.dbd.O(NH)--, provided that said --O--,
--NR.sup.10--, --SO--, --S(O).sub.2--, --C(.dbd.O)-- or
--C.dbd.O(NH)-- are not at the point of attachment to nitrogen and
are separated by at least one --(CH.sub.2).sub.2--; W is selected
from acyl, hydroxyl, carboxyl, amino, carboxamido, sulphonamide,
aminoacyl, --COOalkyl, --CHO, --C(O)fluororalkyl,
--C(O)CH.sub.2C(O)Oalkyl, --C(O)CH.sub.2C(O)Ofluoroalkyl, --SH,
--C(O)NH(OH), --C(O)N(OH)R, --N(OH)C(O)OH, --N(OH)C(O)R.sup.4,
heterocyclyl, substituted aryl, and substituted heterocyclyl; and
R.sup.4 is selected from the group consisting of H and lower
alkyl.
13. A pharmaceutical composition comprising a pharmaceutically
acceptable carrier and a therapeutically effective amount of at
least one compound according to claim 12.
14. A method for inhibiting leukotriene A4 hydrolase comprising
contacting the LTA4H enzyme with a therapeutically effective amount
of a compound according to claim 12.
15. A method for treating a disorder associated with leukotriene A4
hydrolase comprising administering to a mammal a therapeutically
effective amount of a compound or a salt, hydrate or ester thereof
according to claim 12.
16. A method according to claim 15 wherein said disorder is
associated with inflammation.
17. A method according to claim 16 wherein said disorder is
selected from allergic inflammation, acute inflammation and chronic
inflammation.
18. A method according to claim 15, wherein said disorder is chosen
from asthma, chronic obstructive pulmonary disease (COPD),
rheumatoid arthritis, multiple sclerosis, inflammatory bowel
diseases (IBD), ulcerative colitis, and psoriasis.
19. A method according to claim 15, wherein said disorder is chosen
from hypercholesterolemia, atherosclerosis, thrombosis, stroke,
acute coronary syndrome, stable angina, peripheral vascular
disease, critical leg ischemia, intermittent claudication,
abdominal aortic aneurysm and myocardial infarction.
Description
CROSS-REFERENCE TO RELATED APPLICATION
[0001] This application claims priority from U.S. Provisional
Application Ser. No. 60/752,274, filed Dec. 21, 2005, the entire
contents of which are incorporated herein by reference.
FIELD OF THE INVENTION
[0002] The present invention relates to a chemical genus of
biphenyl nitrogen containing heterocyclic derivative inhibitors of
LTA4H (leukotriene A4 hydrolase) useful for the treatment and
prevention and prophylaxis of inflammatory diseases and
disorders.
BACKGROUND OF THE INVENTION
[0003] The end products of the leukotriene pathway are potent
inflammatory lipid mediators derived from arachidonic acid. They
can potentially contribute to development of atherosclerosis and
destabilization of atherosclerotic plaques through lipid oxidation
and/or proinflammatory effects. As described elsewhere, a gene on
chromosome 13q12 has been identified as playing a major role in
myocardial infarction (MI), [Helgadottir et al., Nature Genetics
doi:10.1038/ng1311, 8 Feb. 2004]. This gene (ALOX5AP), herein after
referred to as an MI disease gene, comprises nucleic acid that
encodes 5-lipoxygenase activating protein (FLAP), herein after
referred to as FLAP. DNA variants in the FLAP gene increase risk
for myocardial infarction by 1.8 fold and for stroke by 1.7 fold.
The leukotriene pathway, through FLAP, leads to the production of
leukotriene B4 by the enzyme leukotriene A4 hydrolase (LTA4H).
Leukotriene B4 is one of the most potent chemokine mediators of
arterial inflammation. Particular DNA variants in the gene encoding
LTA4H also elevate risk for MI and stroke, as described elsewhere
[Hakonarsson et al., J. Am. Med. Assoc. 293, 2245-2256 (2005)].
Individuals with a prior history of MI produce more leukotriene B4
when their isolated neutrophils are stimulated with ionomycin.
Increased LTB4 production is particularly marked in male patients
with a prior history of MI who carry risk variants in the FLAP gene
[Helgadottir et al.]. The treatment (prophylactic and/or
therapeutic) of certain diseases and conditions (e.g., MI, acute
coronary syndrome (ACS), stroke, atherosclerosis) associated with
FLAP or with LTA4H can be accomplished by inhibiting LTA4H.
Inhibiting LTA4H is advantageous for methods of treatment for MI or
susceptibility to MI; for ACS (e.g. unstable angina,
non-ST-elevation myocardial infarction (NSTEMI) or ST-elevation
myocardial infarction (STEMI)); for decreasing risk of a second MI;
for stroke (including transient ischemic attack) or susceptibility
to stroke; for atherosclerosis, such as for patients requiring
treatment (e.g. angioplasty, stents, coronary artery bypass graft)
to restore blood flow in coronary arteries, such as patients
requiring treatment for peripheral vascular disease including
peripheral occlusive arterial disease, critical limb ischemia (e.g.
gangrene, ulceration), and intermittent claudication to restore
blood flow in the lower limbs; for atherosclerotic reno-vascular
disease; for abdominal aortic aneurysm; and/or for decreasing
leukotriene synthesis (e.g. for treatment of MI).
[0004] US Patent Application Publication No. 20050043378 and
20050043379, relate to benzoxazol-2-yl, benzothiazol-2-yl and
1H-benzimidazol-2-yl compounds and derivatives thereof useful as
leukotriene A4 hydrolase (LTA4H) inhibitors in treating
inflammation and disorders associated with inflammation. These
disclosures are incorporated herein by reference as they relate to
utility.
SUMMARY OF THE INVENTION
[0005] The present invention relates to compounds exhibiting LTA4H
enzyme inhibition, having general formula:
##STR00002##
wherein Ar is selected from the group consisting of aryl, aryl
substituted with from one to three substituents independently
selected from the group consisting of halogen, loweralkyl,
loweracyl, loweralkoxy, fluoroloweralkyl, fluoroloweralkoxy,
formyl, cyano, benzyl, benzyloxy, phenyl, substituted phenyl,
heteroaryl, heterocyclylalkyl and nitro; heteroaryl, and heteroaryl
substituted with from one to three substituents independently
selected from the group consisting of halogen, loweralkyl,
loweracyl, loweralkoxy, fluoroloweralkyl, fluoroloweralkoxy,
formyl, cyano, sulphonamide, amido, phenyl, heteroaryl,
heterocyclylalkyl and nitro; X is selected from the group
consisting of direct bond, O, SO, S(O.sub.2), NR.sup.10, CH.sub.2,
CF.sub.2, CH.sub.2CH.sub.2, CH.sub.2NH, NHCH.sub.2, CH.dbd.CH,
C.dbd.O, CH.sub.2C.dbd.O; CR.sup.1aR.sup.1b, OCR.sup.1aR.sup.1b and
CR.sup.1aR.sup.1bO; R.sup.10 is selected separately in each
occurrence from the group consisting of H and lower alkyl; R.sup.1a
and R.sup.1b are selected from the group consisting of H and lower
alkyl, or R.sup.1a and R.sup.1b taken together may form a 3-6
membered ring, which may optionally contain a heteroatom chosen
from O, S, SO, SO.sub.2, and NR.sup.10; m is zero, 1 or 2; n is an
integer chosen from 1, 2, or 3; p is an integer from 0-3;
Y is H, H; O; or H, R.sup.3;
[0006] R.sup.3 is lower alkyl; taken together ZW is H or Z is
(CH.sub.2).sub.1-10; in which one or two (CH.sub.2) may optionally
be replaced by a C.sub.3-C.sub.6 carbocycle, a C.sub.3-C.sub.6
heterocycle, --O--, --NR.sup.10--, --SO--, --S(O).sub.2--,
--C(.dbd.O)-- or --C.dbd.O(NH)--, provided that said --O--,
--NR.sup.10--, --SO--, --S(O).sub.2--, --C(.dbd.O)-- or
--C.dbd.O(NH)-- are not at the point of attachment to nitrogen and
are separated by at least one --(CH.sub.2)--; W is selected from
acyl, hydroxyl, carboxyl, amino, carboxamido, sulphonamide,
aminoacyl, --COOalkyl, --CHO, --C(O)fluororalkyl,
--C(O)CH.sub.2C(O)Oalkyl, --C(O)CH.sub.2C(O)Ofluoroalkyl, --SH,
--C(O)NH(OH), --C(O)N(OH)R, --N(OH)C(O)OH, --N(OH)C(O)R.sup.4,
heterocyclyl, substituted aryl, and substituted heterocyclyl; and
R.sup.4 is selected from the group consisting of H and lower
alkyl.
[0007] In a second aspect the invention relates to a pharmaceutical
composition comprising a pharmaceutically acceptable carrier and a
therapeutically effective amount of a compound as described
herein.
[0008] In a third aspect, the invention relates to methods for the
treatment and prevention or prophylaxis of a disease, condition or
disorder associated with leukotriene A4 hydrolase. The methods
comprise administering to a mammal a therapeutically effective
amount of a compound described above. The disease or condition may
be related to allergic, acute or chronic inflammation. The disease
may be for example contact and atopic dermatitis, arthritis,
allergic rhinitis, asthma or an autoimmune diseases such as Crohn's
disease, psoriasis, ulcerative colitis, hypercholesterolemia,
inflammatory bowel disease, multiple sclerosis, ankylosing
spondylitis, and the like. Similarly, the compounds defined above
can be used in preventing recurring inflammatory attacks. The
compounds are also useful for treating and preventing
atherosclerosis, thrombosis, stroke, acute coronary syndrome,
stable angina, peripheral vascular disease, critical leg ischemia,
intermittent claudication, abdominal aortic aneurysm and myocardial
infarction.
DETAILED DESCRIPTION OF THE INVENTION
[0009] Throughout this specification the substituents are defined
when introduced and retain their definitions.
[0010] In one aspect the invention relates to biphenyl heterocycle
derivatives useful as LTA4H enzyme inhibitors, having the general
formula:
##STR00003##
[0011] In some embodiments, Ar is phenyl or substituted phenyl and
m is zero. These embodiments are illustrated by the formula:
##STR00004##
wherein R.sup.1 and R.sup.2 are independently chosen from hydrogen,
halogen, loweralkyl, loweracyl, loweralkoxy, fluoroloweralkyl,
fluoroloweralkoxy, formyl, cyano, benzyl, benzyloxy, sulphonamide,
amido, phenyl, substituted phenyl, heteroaryl, heterocyclylalkyl
and nitro; or R.sup.1 and R.sup.2 taken together may form a 5-6
membered ring, which may optionally contain an oxygen.
[0012] In some embodiments, n is 1 or 2, and p is 1 or 2;
ZW is H or
Z is (CH.sub.2).sub.1-10;
Y is H, H; O; or H and R.sup.3; and
[0013] W is selected from acyl, hydroxyl, carboxyl, amino,
carboxamido, aminoacyl, and --COOalkyl.
[0014] Examples of the substituent Y can be represented by the
structures below wherein the wavy lines indicate ring bonds:
##STR00005##
[0015] In some embodiments, n is 1 or 2, and p is 1 or 2;
ZW is H or
[0016] Z is (CH.sub.2).sub.1-10, in which one or two (CH.sub.2) may
optionally be replaced by a C.sub.3-C.sub.6 carbocycle or a
C.sub.3-C.sub.6 heterocycle;
Y is H, H; O; or H, R.sup.3; and
[0017] W is selected from acyl, hydroxyl, carboxyl, amino,
carboxamido, aminoacyl, and --COOalkyl.
[0018] Examples where one or two (CH.sub.2) linkers of Z are
optionally replaced by a C.sub.3-C.sub.6 carbocycle or a
C.sub.3-C.sub.6 heterocycle include but are not limited to the
structures below wherein the wavy lines indicate ring bonds:
##STR00006##
[0019] In some embodiments, p is one and Y is H, H. These
embodiments are illustrated by the formula:
##STR00007##
wherein R.sup.1 and R.sup.2 are chosen from hydrogen,
trifluoromethyl, methyl, methoxy, halogen, phenyl, cyano and nitro,
or R.sup.1 and R.sup.2 taken together may form a 5-6 membered ring,
which ring may optionally contain an oxygen; X is chosen from --O--
and --CH.sub.2--;
ZW is H or
Z is (CH.sub.2).sub.1-3 and
W is COOH.
[0020] In another aspect the present invention provides a
pharmaceutical composition comprising a pharmaceutically acceptable
carrier and a therapeutically effective amount of at least one
compound as described above.
[0021] Methods of the invention parallel the compositions and
formulations. The methods comprise administering to a patient in
need of treatment a therapeutically effective amount of a compound
according to the invention.
[0022] The present invention provides a method for inhibiting
leukotriene A4 hydrolase comprising contacting the LTA4H enzyme
with a therapeutically effective amount of a compound according to
the general formula
##STR00008##
[0023] Furthermore, the present invention provides a method for
treating a disorder associated with leukotriene A4 hydrolase
comprising administering to a mammal a therapeutically effective
amount of a compound or a salt, hydrate or ester thereof according
to the general formula given above. It may be found upon
examination that additional species and genera not presently
excluded are not patentable to the inventors in this application.
In either case, the exclusion of species and genera in applicants'
claims are to be considered artifacts of patent prosecution and not
reflective of the inventors' concept or description of their
invention.
[0024] The invention, in a composition aspect, is all compounds of
the general formula above, except those that are in the public's
possession. The invention, in a method aspect, is a method
employing compounds of the general formula above, except those
methods that are in the public's possession.
[0025] In some embodiments the disorder is associated with
inflammation. In some embodiments the disorder is selected from
allergic inflammation, acute inflammation and chronic
inflammation.
[0026] Compounds of the genus represented by the general formula
above are inhibitors of LTA4H enzyme. As such they have utility in
treating and preventing inflammatory diseases and disorders, as
described above, particularly for such conditions as asthma,
chronic obstructed pulmonary disease (COPD), atherosclerosis,
rheumatoid arthritis, multiple sclerosis, hypercholoesterolemia,
inflammatory bowel diseases (IBD; including Crohn's disease and
ulcerative colitis), or psoriasis, which are each characterized by
excessive or prolonged inflammation at some stage of the
disease.
[0027] Recent research indicates that the compounds are also useful
for treating and preventing atherosclerosis, thrombosis, stroke,
acute coronary syndrome, stable angina, peripheral vascular
disease, critical leg ischemia, intermittent claudication,
abdominal aortic aneurysm and myocardial infarction
atherosclerosis, thrombosis, stroke, acute coronary syndrome and
myocardial infarct.
[0028] The compounds may be presented as salts. The term
"pharmaceutically acceptable salt" refers to salts whose counter
ion derives from pharmaceutically acceptable non-toxic acids and
bases. Suitable pharmaceutically acceptable base addition salts for
the compounds of the present invention include, but are not limited
to, metallic salts made from aluminum, calcium, lithium, magnesium,
potassium, sodium and zinc or organic salts made from lysine,
N,N-dialkyl amino acid derivatives (e.g. N,N-dimethylglycine,
piperidine-1-acetic acid and morpholine-4-acetic acid),
N,N'-dibenzylethylenediamine, chloroprocaine, choline,
diethanolamine, ethylenediamine, meglumine (N-methylglucamine) and
procaine. When the compounds contain a basic residue, suitable
pharmaceutically acceptable base addition salts for the compounds
of the present invention include inorganic acids and organic acids.
Examples include acetate, benzenesulfonate (besylate), benzoate,
bicarbonate, bisulfate, carbonate, camphorsulfonate, citrate,
ethanesulfonate, fumarate, gluconate, glutamate, bromide, chloride,
isethionate, lactate, maleate, malate, mandelate, methanesulfonate,
mucate, nitrate, pamoate, pantothenate, phosphate, succinate,
sulfate, tartrate, p-toluenesulfonate, and the like.
[0029] For convenience and clarity certain terms employed in the
specification, examples and claims are described herein.
[0030] Alkyl is intended to include linear, branched, or cyclic
hydrocarbon structures and combinations thereof. Lower alkyl refers
to alkyl groups of from 1 to 6 carbon atoms. Examples of lower
alkyl groups include methyl, ethyl, propyl, isopropyl, butyl, s-
and t-butyl and the like. Preferred alkyl groups are those of
C.sub.20 or below. Cycloalkyl is a subset of alkyl and includes
cyclic hydrocarbon groups of from 3 to 8 carbon atoms. Examples of
cycloalkyl groups include c-propyl, c-butyl, c-pentyl, norbornyl
and the like.
[0031] C.sub.1 to C.sub.20 hydrocarbon includes alkyl, cycloalkyl,
alkenyl, alkynyl, aryl, arylalkyl and combinations thereof.
Examples include phenethyl, cyclohexylmethyl, camphoryl, adamantyl
and naphthylethyl.
[0032] Alkoxy or alkoxyl refers to groups of from 1 to 8 carbon
atoms of a straight, branched, cyclic configuration and
combinations thereof attached to the parent structure through
oxygen. Examples include methoxy, ethoxy, propoxy, isopropoxy,
cyclopropyloxy, cyclohexyloxy and the like. For the purposes of
this application, alkoxy also includes methylenedioxy and
ethylenedioxy. Lower-alkoxy refers to groups containing one to four
carbons.
[0033] Alkoxyalkyl refers to ether groups of from 3 to 8 atoms of a
straight, branched, cyclic configuration and combinations thereof
attached to the parent structure through an alkyl. Examples include
methoxymethyl, methoxyethyl, ethoxypropyl, and the like.
[0034] Alkoxyaryl refers to alkoxy substituents attached to an
aryl, wherein the aryl is attached to the parent structure.
Arylalkoxy refers to aryl substituents attached to an oxygen,
wherein the oxygen is attached to the parent structure. Substituted
arylalkoxy refers to a substituted aryl substituent attached to an
oxygen, wherein the oxygen is attached to the parent structure.
[0035] Acyl refers to groups of from 1 to 8 carbon atoms of a
straight, branched, cyclic configuration, saturated, unsaturated
and aromatic and combinations thereof, attached to the parent
structure through a carbonyl functionality. One or more carbons in
the acyl residue may be replaced by nitrogen, oxygen or sulfur as
long as the point of attachment to the parent remains at the
carbonyl. Examples include acetyl, benzoyl, propionyl, isobutyryl,
t-butoxycarbonyl, benzyloxycarbonyl and the like. Lower-acyl refers
to groups containing one to four carbons.
[0036] Aryl and heteroaryl mean a 5- or 6-membered aromatic or
heteroaromatic ring containing 0-3 heteroatoms selected from O, N,
or S; a bicyclic 9- or 10-membered aromatic or heteroaromatic ring
system containing 0-3 heteroatoms selected from O, N, or S; or a
tricyclic 13- or 14-membered aromatic or heteroaromatic ring system
containing 0-3 heteroatoms selected from O, N, or S. The aromatic
6- to 14-membered carbocyclic rings include, e.g., benzene and
naphthalene, and according to the invention benzoxalane and
residues in which one or more rings are aromatic, but not all need
be. The 5- to 10-membered aromatic heterocyclic rings include,
e.g., imidazole, pyridine, indole, thiophene, benzopyranone,
thiazole, furan, benzimidazole, quinoline, isoquinoline,
quinoxaline, pyrimidine, pyrazine, tetrazole and pyrazole.
[0037] Arylalkyl refers to a substituent in which an aryl residue
is attached to the parent structure through alkyl. Examples are
benzyl, phenethyl and the like. Heteroarylalkyl refers to a
substituent in which a heteroaryl residue is attached to the parent
structure through alkyl. Examples include, e.g., pyridinylmethyl,
pyrimidinylethyl and the like. Heterocyclylalkyl refers to a
substituent in which a heterocyclyl residue is attached to the
parent structure through alkyl. Examples include morpholinoethyl
and pyrrolidinylmethyl.
[0038] Heterocycle means a cycloalkyl or aryl residue in which from
one to three carbons is replaced by a heteroatom selected from the
group consisting of N, O and S. The nitrogen and sulfur heteroatoms
may optionally be oxidized, and the nitrogen heteroatom may
optionally be quaternized. Examples of heterocycles include
pyrrolidine, pyrazole, pyrrole, indole, quinoline, isoquinoline,
tetrahydroisoquinoline, benzofuran, benzodioxan, benzodioxole
(commonly referred to as methylenedioxyphenyl, when occurring as a
substituent), benzo-[1,4]-dioxane (commonly referred to as
ethylenedioxyphenyl, when occurring as a substitutent), tetrazole,
morpholine, thiazole, pyridine, pyridazine, pyrimidine, thiophene,
furan, oxazole, oxazoline, isoxazole, dioxane, tetrahydrofuran and
the like. It is to be noted that heteroaryl is a subset of
heterocycle in which the heterocycle is aromatic. Examples of
heterocyclyl residues additionally include piperazinyl,
2-oxopiperazinyl, 2-oxopiperidinyl, 2-oxo-pyrrolidinyl,
2-oxoazepinyl, azepinyl, 4-piperidinyl, pyrazolidinyl, imidazolyl,
imidazolinyl, imidazolidinyl, pyrazinyl, oxazolidinyl,
isoxazolidinyl, thiazolidinyl, isothiazolyl, quinuclidinyl,
isothiazolidinyl, benzimidazolyl, thiadiazolyl, benzopyranyl,
benzothiazolyl, tetrahydrofuryl, tetrahydropyranyl, thienyl,
benzothienyl, thiamorpholinyl, thiamorpholinylsulfoxide,
thiamorpholinylsulfone, oxadiazolyl, triazolyl and
tetrahydroquinolinyl.
[0039] An oxygen heterocycle is a heterocycle containing at least
one oxygen in the ring; it may contain additional oxygens, as well
as other heteroatoms. A sulphur heterocycle is a heterocycle
containing at least one sulphur in the ring; it may contain
additional sulphurs, as well as other heteroatoms. A nitrogen
heterocycle is a heterocycle containing at least one nitrogen in
the ring; it may contain additional nitrogens, as well as other
heteroatoms. Oxygen heteroaryl is a subset of oxygen heterocycle;
examples include furan and oxazole. Sulphur heteroaryl is a subset
of sulphur heterocycle; examples include thiophene and thiazine.
Nitrogen heteroaryl is a subset of nitrogen heterocycle; examples
include pyrrole, pyridine and pyrazine. A saturated nitrogenous
heterocycle is a subset of nitrogen heterocycle. Saturated
nitrogenous heterocycle contain at least one nitrogen and may
contain additional nitrogens, as well as other heteroatoms.
Examples include pyrrolidine, pyrazolidine, piperidine, morpholine,
and thiomorpholine.
[0040] Substituted alkyl, aryl, cycloalkyl, heterocyclyl etc. refer
to alkyl, aryl, cycloalkyl, or heterocyclyl wherein up to three H
atoms in each residue are replaced with halogen, haloalkyl,
hydroxy, loweralkoxy, carboxy, carboalkoxy (also referred to as
alkoxycarbonyl), carboxamido (also referred to as
alkylaminocarbonyl), cyano, carbonyl, nitro, amino, alkylamino,
dialkylamino, mercapto, alkylthio, sulfoxide, sulfone, acylamino,
amidino, phenyl, benzyl, heteroaryl, phenoxy, benzyloxy, or
heteroaryloxy.
[0041] The terms "halogen" and "halo" refer to fluorine, chlorine,
bromine or iodine.
[0042] The term "hydroxamate" refers to hydroxamic acid and its
salts and esters:
##STR00009##
[0043] The term "prodrug" refers to a compound that is made more
active in vivo. Activation in vivo may come about by chemical
action or through the intermediacy of enzymes. Microflora in the GI
tract may also contribute to activation in vivo.
[0044] It will be recognized that the compounds of this invention
can exist in radiolabeled form, i.e., the compounds may contain one
or more atoms containing an atomic mass or mass number different
from the atomic mass or mass number usually found in nature.
Radioisotopes of hydrogen, carbon, phosphorous, fluorine, and
chlorine include .sup.2H, .sup.3H, .sup.13C, .sup.14C, .sup.15N,
.sup.35S, .sup.18F, and .sup.36Cl, respectively. Compounds that
contain those radioisotopes and/or other radioisotopes of other
atoms are within the scope of this invention. Tritiated, i.e.
.sup.3H, and carbon-14, i.e., .sup.14C, radioisotopes are
particularly preferred for their ease in preparation and
detectability. Radiolabeled compounds of formula .PSI. of this
invention and prodrugs thereof can generally be prepared by methods
well known to those skilled in the art. Conveniently, such
radiolabeled compounds can be prepared by carrying out the
procedures disclosed in the Examples and Schemes by substituting a
readily available radiolabeled reagent for a non-radiolabeled
reagent.
[0045] As used herein, and as would be understood by the person of
skill in the art, the recitation of "a compound" is intended to
include salts, solvates, co-crystals and inclusion complexes of
that compound.
[0046] The term "solvate" refers to a compound of formula I in the
solid state, wherein molecules of a suitable solvent are
incorporated in the crystal lattice. A suitable solvent for
therapeutic administration is physiologically tolerable at the
dosage administered. Examples of suitable solvents for therapeutic
administration are ethanol and water. When water is the solvent,
the solvate is referred to as a hydrate. In general, solvates are
formed by dissolving the compound in the appropriate solvent and
isolating the solvate by cooling or using an antisolvent. The
solvate is typically dried or azeotroped under ambient conditions.
Co-crystals are combinations of two or more distinct molecules
arranged to create a unique crystal form whose physical properties
are different from those of its pure constituents. Pharmaceutical
co-crystals have recently become of considerable interest for
improving the solubility, formulation and bioavailability of such
drugs as itraconazole [see Remenar et al. J. Am. Chem. Soc. 125,
8456-8457 (2003)] and fluoxetine. Inclusion complexes are described
in Remington: The Science and Practice of Pharmacy 19.sup.th Ed.
(1995) volume 1, page 176-177. The most commonly employed inclusion
complexes are those with cyclodextrins, and all cyclodextrin
complexes, natural and synthetic, with or without added additives
and polymer(s), as described in U.S. Pat. Nos. 5,324,718 and
5,472,954, are specifically encompassed within the claims. The
disclosures of Remington and the '718 and '954 patents are
incorporated herein by reference.
[0047] The compounds described herein may contain asymmetric
centers and may thus give rise to enantiomers, diastereomers, and
other stereoisomeric forms. Each chiral center may be defined, in
terms of absolute stereochemistry, as (R)- or (S)-. The present
invention is meant to include all such possible isomers, as well
as, their racemic and optically pure forms. Optically active (R)-
and (S)-isomers may be prepared using chiral synthons or chiral
reagents, or resolved using conventional techniques. The prefix
"rac" refers to a racemate. When the compounds described herein
contain olefinic double bonds or other centers of geometric
asymmetry, and unless specified otherwise, it is intended that the
compounds include both E and Z geometric isomers. The
representation of the configuration of any carbon-carbon double
bond appearing herein is selected for convenience only, and unless
explicitly stated, is not intended to designate a particular
configuration. Thus a carbon-carbon double bond depicted
arbitrarily as E may be Z. E, or a mixture of the two in any
proportion. Likewise, all tautomeric forms are also intended to be
included.
[0048] The graphic representations of racemic, ambiscalemic and
scalemic or enantiomerically pure compounds used herein are taken
from Maehr J. Chem. Ed. 62, 114-120 (1985): solid and broken wedges
are used to denote the absolute configuration of a chiral element;
wavy lines and single thin lines indicate disavowal of any
stereochemical implication which the bond it represents could
generate; solid and broken bold lines are geometric descriptors
indicating the relative configuration shown but denoting racemic
character; and wedge outlines and dotted or broken lines denote
enantiomerically pure compounds of indeterminate absolute
configuration.
[0049] Terminology related to "protecting", "deprotecting" and
"protected" functionalities occurs throughout this application.
Such terminology is well understood by persons of skill in the art
and is used in the context of processes that involve sequential
treatment with a series of reagents. In that context, a protecting
group refers to a group, which is used to mask a functionality
during a process step in which it would otherwise react, but in
which reaction is undesirable. The protecting group prevents
reaction at that step, but may be subsequently removed to expose
the original functionality. The removal or "deprotection" occurs
after the completion of the reaction or reactions in which the
functionality would interfere. Thus, when a sequence of reagents is
specified, as it is in the processes of the invention, the person
of ordinary skill can readily envision those groups that would be
suitable as "protecting groups". Suitable groups for that purpose
are discussed in standard textbooks in the field of chemistry, such
as Protective Groups in Organic Synthesis by T. W. Greene [John
Wiley & Sons, New York, 1991], which is incorporated herein by
reference.
[0050] A comprehensive list of abbreviations utilized by organic
chemists appears in the first issue of each volume of the Journal
of Organic Chemistry. The list, which is typically presented in a
table entitled "Standard List of Abbreviations", is incorporated
herein by reference.
[0051] In general, the compounds of the present invention may be
prepared by the methods illustrated in the general reaction schemes
as, for example, described below, or by modifications thereof,
using readily available starting materials, reagents and
conventional synthesis procedures. In these reactions, it is also
possible to make use of variants that are in themselves known, but
are not mentioned here. The starting materials, for example in the
case of suitably substituted benzimidazole ring compounds, are
either commercially available, synthesized as described in the
examples or may be obtained by the methods well known to persons of
skill in the art.
[0052] LTA4H inhibitors have been shown to be effective
anti-inflammatory agents in pre-clinical studies. For example, oral
administration of LTA4H inhibitor SC57461 to rodents resulted in
the inhibition of ionophore-induced LTB4 production in mouse blood
ex vivo and in rat peritoneum in vivo (Kachur et al., 2002, J.
Pharm. Exp. Ther. 300(2), 583-587). Furthermore, eight weeks of
treatment with the same inhibitor compound significantly improved
colitis symptoms in a primate model (Penning, 2001, Curr. Pharm.
Des. 7(3): 163-179). The spontaneous colitis that develops in these
animals is very similar to human IBD. Therefore persons of skill in
the art accept that positive results in LTA4H models are predictive
of therapeutic utility in this and other human inflammatory
diseases.
[0053] The inflammatory response is characterized by pain,
increased temperature, redness, swelling, or reduced function, or
by a combination of two or more of these symptoms. The terms
inflammation, inflammatory diseases or inflammation-mediated
diseases or conditions include, but are not limited to, acute
inflammation, allergic inflammation, and chronic inflammation.
[0054] Autoimmune diseases are associated with chronic
inflammation. There are about 75 different autoimmune disorders
known that may be classified into two types, organ-specific
(directed mainly at one organ) and non-organ-specific (affecting
multiple organs).
[0055] Examples of organ-specific autoimmune disorders are
insulin-dependent diabetes (Type I) which affects the pancreas,
Hashimoto's thyroiditis and Graves' disease which affect the
thyroid gland, pernicious anemia which affects the stomach,
Cushing's disease and Addison's disease which affect the adrenal
glands, chronic active hepatitis which affects the liver;
polycystic ovary syndrome (PCOS), celiac disease, psoriasis,
inflammatory bowel disease (IBD) and ankylosing spondylitis.
[0056] Examples of non-organ-specific autoimmune disorders are
rheumatoid arthritis, multiple sclerosis, systemic lupus and
myasthenia gravis.
[0057] Furthermore, the compounds, compositions and methods of the
present invention are useful in treating cancer. Leukotriene
synthesis has been shown to be associated with different types of
cancer including esophageal cancer, brain cancer, pancreatic
cancer, colon cancer.
[0058] The terms "methods of treating or preventing" mean
amelioration, prevention or relief from the symptoms and/or effects
associated with lipid disorders. The term "preventing" as used
herein refers to administering a medicament beforehand to forestall
or obtund an acute episode. The person of ordinary skill in the
medical art (to which the present method claims are directed)
recognizes that the term "prevent" is not an absolute term. In the
medical art it is understood to refer to the prophylactic
administration of a drug to substantially diminish the likelihood
or seriousness of a condition, and this is the sense intended in
applicants' claims. As used herein, reference to "treatment" of a
patient is intended to include prophylaxis. Throughout this
application, various references are referred to. The disclosures of
these publications in their entireties are hereby incorporated by
reference as if written herein.
[0059] The term "mammal" is used in its dictionary sense. Humans
are included in the group of mammals, and humans would be the
preferred subjects of the methods of While it may be possible for
the compounds of formula .PSI. to be administered as the raw
chemical, it is preferable to present them as a pharmaceutical
composition. According to a further aspect, the present invention
provides a pharmaceutical composition comprising a compound of
formula I, or a pharmaceutically acceptable salt or solvate
thereof, together with one or more pharmaceutically carriers
thereof and optionally one or more other therapeutic ingredients.
The carrier(s) must be "acceptable" in the sense of being
compatible with the other ingredients of the formulation and not
deleterious to the recipient thereof.
[0060] The formulations include those suitable for oral, parenteral
(including subcutaneous, intradermal, intramuscular, intravenous
and intraarticular), rectal and topical (including dermal, buccal,
sublingual and intraocular) administration. The most suitable route
may depend upon the condition and disorder of the recipient. The
formulations may conveniently be presented in unit dosage form and
may be prepared by any of the methods well known in the art of
pharmacy. All methods include the step of bringing into association
a compound of formula .PSI. or a pharmaceutically acceptable salt
or solvate thereof ("active ingredient") with the carrier, which
constitutes one or more accessory ingredients. In general, the
formulations are prepared by uniformly and intimately bringing into
association the active ingredient with liquid carriers or finely
divided solid carriers or both and then, if necessary, shaping the
product into the desired formulation.
[0061] Formulations of the present invention suitable for oral
administration may be presented as discrete units such as capsules,
cachets or tablets each containing a predetermined amount of the
active ingredient; as a powder (including micronized and
nanoparticulate powders) or granules; as a solution or a suspension
in an aqueous liquid or a non-aqueous liquid; or as an oil-in-water
liquid emulsion or a water-in-oil liquid emulsion. The active
ingredient may also be presented as a bolus, electuary or
paste.
[0062] A tablet may be made by compression or molding, optionally
with one or more accessory ingredients. Compressed tablets may be
prepared by compressing in a suitable machine the active ingredient
in a free-flowing form such as a powder or granules, optionally
mixed with a binder, lubricant, inert diluent, lubricating, surface
active or dispersing agent. Molded tablets may be made by molding
in a suitable machine a mixture of the powdered compound moistened
with an inert liquid diluent. The tablets may optionally be coated
or scored and may be formulated so as to provide sustained, delayed
or controlled release of the active ingredient therein.
[0063] The pharmaceutical compositions may include a
"pharmaceutically acceptable inert carrier", and this expression is
intended to include one or more inert excipients, which include
starches, polyols, granulating agents, microcrystalline cellulose,
diluents, lubricants, binders, disintegrating agents, and the like.
If desired, tablet dosages of the disclosed compositions may be
coated by standard aqueous or nonaqueous techniques,
"Pharmaceutically acceptable carrier" also encompasses controlled
release means.
[0064] Compositions of the present invention may also optionally
include other therapeutic ingredients, anti-caking agents,
preservatives, sweetening agents, colorants, flavors, desiccants,
plasticizers, dyes, and the like. Any such optional ingredient
must, of course, be compatible with the compound of the invention
to insure the stability of the formulation. The dose range for
adult humans is generally from 0.1 .mu.g to 10 g/day orally.
Tablets or other forms of presentation provided in discrete units
may conveniently contain an amount of compound of the invention
which is effective at such dosage or as a multiple of the same, for
instance, units containing 0.1 mg to 500 mg, usually around 5 mg to
200 mg. The precise amount of compound administered to a patient
will be the responsibility of the attendant physician. However, the
dose employed will depend on a number of factors, including the age
and sex of the patient, the precise disorder being treated, and its
severity. The frequency of administration will depend on the
pharmacodynamics of the individual compound and the formulation of
the dosage form, which may be optimized by methods well known in
the art (e.g. controlled or extended release tablets, enteric
coating etc.).
[0065] Combination therapy can be achieved by administering two or
more agents, each of which is formulated and administered
separately, or by administering two or more agents in a single
formulation. Other combinations are also encompassed by combination
therapy. For example, two agents can be formulated together and
administered in conjunction with a separate formulation containing
a third agent. While the two or more agents in the combination
therapy can be administered simultaneously, they need not be. For
example, administration of a first agent (or combination of agents)
can precede administration of a second agent (or combination of
agents) by minutes, hours, days, or weeks. Thus, the two or more
agents can be administered within minutes of each other or within
any number of hours of each other or within any number or days or
weeks of each other. In some cases even longer intervals are
possible.
[0066] While in many cases it is desirable that the two or more
agents used in a combination therapy be present in within the
patient's body at the same time, this need not be so. Combination
therapy can also include two or more administrations of one or more
of the agents used in the combination. For example, if agent X and
agent Y are used in a combination, one could administer them
sequentially in any combination one or more times, e.g., in the
order X-Y-X, X-X-Y, Y-X-Y, Y-Y-X, X-X-Y-Y, etc.
[0067] As LTA4H inhibitors, the compounds of formula .PSI. have
utility in treating and preventing inter alia inflammation. The
compounds and compositions can be used advantageously in
combination with other agents useful in treating and preventing
inflammatory conditions and for treating and preventing
atherosclerosis, thrombosis, stroke, acute coronary syndrome,
stable angina, peripheral vascular disease, critical leg ischemia,
intermittent claudication, abdominal aortic aneurysm and myocardial
infarction.
[0068] In general, the compounds of the present invention may be
prepared by the methods illustrated in the general reaction schemes
as, for example, described below, or by modifications thereof,
using readily available starting materials, reagents and
conventional synthesis procedures. The following specific
non-limiting examples are illustrative of the invention.
##STR00010##
##STR00011##
Scheme 1: (i) diisopropylazodicarboxylate, PPh.sub.3, THF; (ii)
Br(CH.sub.2).sub.nCO.sub.2R.sub.2, Et.sub.3N; (iii) 2N NaOH or 4M
HCl
Example 1
##STR00012##
[0070] (S) 3-(4-Benzyl-phenoxy)-pyrrolidine hydrochloride salt: To
a solution of (R)-3-hydroxy-pyrrolidine carboxylic acid-t-butyl
ester (187 mg, 1.0 mmol) in anhydrous THF (1 mL) was added
4-hydroxydiphenyl methane (240 mg, 1.3 mmol) in THF (1 mL) and
triphenyl phosphine (315 mg, 1.2 mmol) in THF (1 mL). The resulting
mixture was cooled to 0.degree. C. using ice-water bath and purged
with nitrogen. Diisopropyl azodicarboxylate (242 mg, 1.2 mmol) was
dissolved in 2 mL of THF and added to above solution dropwise over
a period of 15 min under nitrogen. Reaction then was heated at
70.degree. C. for 16 h. THF was removed in vacuo and crude mixture
was purified by silica gel flash chromatography (10% EtOAc/Hexane)
to obtain the product. To a solution of the product in dioxane (1
mL) was added 4M HCl in dioxane (3 mL) at rt and the resulting
mixture was stirred for 30 min at that temperature. The solvent was
removed in vacuo to obtain thick oil. The oil was triturated with
ether to obtain a white solid (143 mg, 49%): MS; m/z 254.5 (M+H);
.sup.1H NMR (400 MHz, DMSO-d.sub.6); .delta. 2.09-2.18 (m, 2H),
3.23-3.31 (m, 4H), 3.40-3.44 (m, 1H), 3.87 (s, 2H), 5.07 (m, 1H),
6.88 (dd, 2H, J1=6.4 Hz, J2=2 Hz), 7.16-7.22 (m, 5H), 7.26-7.29 (m,
2H), 9.30 (s, 2H); HPLC (UV); 98%.
[0071] Elemental analysis: Calc C, 70.46; H, 6.96; N, 4.83. Found
C, 70.64; H, 7.06; N, 4.90
Example 2
##STR00013##
[0073] (R) 3-(4-Benzyl-phenoxy)-pyrrolidine hydrochloride salt: To
a solution of (S)(+)-1BOC-3-hydroxy pyrrolidine (936 mg, 5.0 mmol)
in anhydrous THF (5 mL) was added 4-hydroxydiphenyl methane (1197
mg, 6.5 mmol) in THF (5 mL) and triphenyl phosphine (1574 mg, 6
mmol) in THF (5 mL). The resulting mixture was cooled to 0.degree.
C. using ice-water bath and purged with nitrogen. Diisopropyl
azodicarboxylate (1213 mg, 6 mmol) was dissolved in 5 mL of THF and
added to above solution drop wise over period of 30 min under
nitrogen. Reaction then was heated at rt for 3 hrs and 70.degree.
C. for 16 h. THF was removed in vacuo and crude mixture was
purified by silica gel flash chromatography (10% EtOAc/Hexane) to
obtain the product. To a solution of the product in dioxane (4 mL)
was added 4M HCl in dioxane (10 mL) at rt and the resulting mixture
was stirred for 3 h at that temperature. The solvent was removed in
vacuo and residue was triturated with ether to obtain a white
crystalline solid (788 mg, 63%): MS; m/z 254.4 (M+H); .sup.1H NMR
(400 MHz, DMSO-d.sub.6); .delta. 2.09-2.17 (m, 2H), 3.19-3.33 (m,
4H), 3.40-3.44 (m, 1H), 3.87 (s, 2H), 5.07 (m, 1H), 6.88 (dd, 2H,
J1=6.4 Hz, J2=2 Hz), 7.15-7.22 (m, 5H), 7.26-7.29 (m, 2H), 9.44 (s,
2H); HPLC (UV); 99.1%.
Example 3
##STR00014##
[0075] 4-(4-Phenoxy-phenoxy)-piperidine hydrochloride salt: To a
solution of t-butyl-4-hydroxy-1-piperidine carboxylate (1.0 g, 5.0
mmol) in anhydrous THF (5 mL) was added 4-phenoxyphenol (1.21 g,
6.5 mmol) in THF (5 mL) and triphenyl phosphine (1.57 g, 6 mmol) in
THF (5 mL). The resulting mixture was cooled to 0.degree. C. using
ice-water bath and purged with nitrogen. Diisopropyl
azodicarboxylate (1.21 g, 6 mmol) was dissolved in 5 mL of THF and
added to above solution drop wise over period of 20 min under
nitrogen. Reaction then was heated at rt for 3 h and 70.degree. C.
for 48 h. THF was removed in vacuo and crude mixture was purified
by silica gel flash chromatography (10% EtOAc/Hexane) to obtain the
product. To a solution of the product in dioxane (4 mL) was added
4M HCl in dioxane (10 mL) at rt and the resulting mixture was
stirred for 3 h at that temperature. The solvent was removed in
vacuo and residue was triturated with ether to obtain a white
crystalline solid (918 mg, 68%): MS; m/z 270.5 (M+H); .sup.1H NMR
(400 MHz, DMSO-d.sub.6); .delta. 1.81-1.87 (m, 2H), 2.07-2.12 (m,
2H), 3.02-3.08 (m, 2H), 3.19-3.25 (m, 2H), 3.56 (s, 1H), 4.57-4.61
(m, 1H), 6.92-7.10 (m, 7H), 7.33-7.37 (m, 2H), 8.91 (s, 2H); HPLC
(UV); 96%. Elemental analysis: Calc C, 66.77; H, 6.59; N, 4.58.
Found C, 65.82; H, 6.70; N, 4.77.
Example 4
##STR00015##
[0077] Step 1
[0078] 4-[(R)-3-(4-Benzyl-phenoxy)-pyrrolidin-1-yl]-butyric acid
methyl ester: To a solution of (R)-3-(4-Benzyl-phenoxy)-pyrrolidine
(126 mg, 0.5 mmol) in anhydrous CH.sub.2Cl.sub.2 (0.5 mL) was added
methyl-4-bromobutyrate (99 mg, 0.55 mmol) in CH2Cl2 (0.5 mL) and
triethylamine (101 mg, 1.0 mmol. The resulting mixture was purged
with nitrogen and stirred at rt for 16 h. CH.sub.2Cl.sub.2 was
removed in vacuo and crude mixture was partioned between EtOAc and
water. EtOAc layer was removed, washed with brine, dried over
anhydrous MgSO4 and concentrated. The crude mixture was purified by
silica gel flash chromatography (40% EtOAc/Hexane) to obtain the
product as yellow solid (71 mg, 40%): MS; m/z 354.5 (M+H); .sup.1H
NMR (400 MHz, DMSO-d.sub.6); .delta. 1.65-1.72 (m, 2H), 2.18-2.39
(m, 4H), 2.54-2.63 (m, 5H), 2.78-2.82 (m, 1H), 3.57 (s, 3H), 3.85
(s, 2H), 5.78 (m, 1H), 6.78 (d, 2H, J=8.8 Hz), 7.11 (d, 2H, J=8.4
Hz), 7.16-7.29 (m, 5H); HPLC (ELSD); 99%.
[0079] Step 2
[0080] 4-[(R)-3-(4-Benzyl-phenoxy)-pyrrolidin-1-yl]-butyric acid
hydrochloride salt: To a solution of the product (65 mg, 0.184
mmol) from step 1 in 4:1 mixture of MeOH/water (1 mL) was added 2M
solution of NaOH (110 uL, 0.221 mmol). Reaction mixture was heated
at 50.degree. C. for 16 h. Solvent was removed in vacuo, residue
was dissolved in water and pH was adjusted to 2 with 1M HCl
solution. Then product was extracted with EtOAc, washed with water,
brine, dried over anhydrous MgSO4 and concentrated to give the
title compound (43 mg, 70%): MS; m/z 340.5 (M+H); .sup.1H NMR (400
MHz, DMSO-d.sub.6); .delta. 1.85-1.91 (m, 2H), 2.08 (br, 1H), 2.33
(m, 3H), 3.14-3.18 (m, 6H), 3.88 (s, 2H), 5.07 (s, 1H), 6.87-6.89
(m, 2H), 7.15-7.22 (m, 5H), 7.26-7.29 (m, 2H); HPLC (ELSD); 99%.
Elemental analysis: Calc C, 67.10; H, 6.97; N, 3.73. Found C,
66.99; H, 7.03; N, 3.79.
Example 5
##STR00016##
[0082] Step 1
[0083] 3-[4-(4-Phenoxy-phenoxy)-piperidin-1-yl]-propionic acid
methyl ester: To a solution of 4-(4-Phenoxy-phenoxy)-piperidine
(152 mg, 0.5 mmol) in anhydrous CH.sub.2Cl.sub.2 (0.5 mL) was added
methyl-3-bromopropionate (91 mg, 0.55 mmol) in CH.sub.2Cl.sub.2
(0.5 mL) and triethylamine (101 mg, 1.0 mmol). The resulting
mixture was purged with nitrogen and stirred at rt for 48 h. CH2Cl2
was removed in vacuo and crude mixture was partioned between EtOAc
and water. EtOAc layer was removed, washed with saturated NaHCO3,
dried over anhydrous MgSO4 and concentrated. The crude mixture was
purified by silica gel flash chromatography (50% EtOAc/Hexane) to
obtain the product (146 mg, 82%): MS; m/z 356.5 (M+H); .sup.1H NMR
(400 MHz, DMSO-d.sub.6); .delta. 1.56-1.60 (m, 2H), 1.89-1.92 (m,
2H), 2.19-2.49 (m, 2H), 2.46-2.49 (m, 2H), 2.58 (t, 2H, J=6.8 Hz),
2.60-2.69 (m, 2H), 3.59 (s, 3H), 4.28-4.30 (m, 1H), 6.91-6.96 (m,
6H), 7.04-7.08 (m, 1H), 7.32-7.36 (m, 2H); HPLC (UV); 94.7%.
[0084] Step 2
[0085] 3-[4-(4-Phenoxy-phenoxy)-piperidin-1-yl]-propionic acid: To
a solution of the product (143 mg, 0.4 mmol) from step 1 in 4:1
mixture of MeOH/water (2 mL) was added 2M solution of NaOH (241 uL,
0.48 mmol). Reaction mixture was heated at 50.degree. C. for 16 h.
Solvent was removed in vacuo, residue was washed with water,
dissolved in MeOH and filtered. Filtrate was concentrated to give
the product as white solid (155 mg, 100%): MS; m/z 342.5 (M+H);
.sup.1H NMR (400 MHz, DMSO-d.sub.6); .delta. 1.99 (br, 2H),
2.17-2.21 (m, 2H), 2.84-2.88 (m, 2H), 3.25-3.32 (m, 7H), 6.92-7.10
(m, 7H), 7.33-7.37 (m, 2H); HPLC (UV); 93%.
Example 6
##STR00017##
[0087] Step 1
[0088] 4-[4-(4-Phenoxy-phenoxy)-piperidin-1-yl]-butyric acid methyl
ester: To a solution of 4-(4-Phenoxy-phenoxy)-piperidine (152 mg,
0.5 mmol) in anhydrous CH2Cl2 (0.5 mL) was added
methyl-3-bromopropionate (99 mg, 0.55 mmol) in CH2Cl2 (0.5 mL) and
triethylamine (101 mg, 1.0 mmol). The resulting mixture was purged
with nitrogen and stirred at rt for 48 h. CH2Cl2 was removed in
vacuo and crude mixture was partioned between EtOAc and water.
EtOAc layer was removed, washed with brine, dried over anhydrous
MgSO4 and concentrated. The crude mixture was purified by silica
gel flash chromatography (40-70% EtOAc/Hexane) to obtain the
product as yellow oil (125 mg, 68%): .sup.1H NMR (400 MHz,
DMSO-d.sub.6); .delta. 1.54-1.69 (m, 4H), 1.89-1.92 (m, 2H),
2.14-2.18 (m, 2H), 2.26-2.33 (m, 4H), 2.64-2.67 (m, 2H), 3.58 (s,
3H), 4.27-4.31 (m, 1H), 6.91-6.96 (m, 6H), 7.04-7.08 (m, 1H),
7.32-7.36 (m, 2H).
[0089] Step 2
[0090] 4-[4-(4-Phenoxy-phenoxy)-piperidin-1-yl]-butyric acid
hydrochloride salt: To a solution of the product (120 mg, 0.32
mmol) from step 1 in 4:1 mixture of MeOH/water (2 mL) was added 2M
solution of NaOH (195 uL, 0.39 mmol). Reaction mixture was heated
at 50.degree. C. for 16 h. Solvent was removed in vacuo, residue
was dissolved in water and pH was adjusted to 2 with 1M HCl
solution. Then product was extracted with EtOAc, washed with water,
brine, dried over anhydrous MgSO4 and concentrated to give the
title compound as white solid (65 mg, 58%): MS; m/z 356.6 (M+H);
.sup.1H NMR (400 MHz, DMSO-d.sub.6); .delta. 1.87-1.95 (m, 5H),
2.15 (br, 2H), 2.34 (t, 2H, J=7.2 Hz), 3.06-3.1 (m, 4H), 3.26-3.36
(m, 2H), 6.92-7.01 (m, 7H), 7.33-7.36 (m, 2H); HPLC (ELSD);
99%.
Example 7
##STR00018##
[0092] 4-[4-(4-Trifluoromethyl-phenoxy)-phenoxy]-piperidine
hydrochloride salt: To a solution of t-butyl-4-hydroxy-1-piperidine
carboxylate (1.0 g, 5.0 mmol) in anhydrous THF (5 mL) was added
4-[(4-trifluoromethyl)phenoxy]phenol (1.65 g, 6.5 mmol) in THF (5
mL) and triphenyl phosphine (1.57 g, 6 mmol) in THF (5 mL). The
resulting mixture was cooled to 0.degree. C. using ice-water bath
and purged with nitrogen. Diisopropyl azodicarboxylate (1.21 g, 6
mmol) was dissolved in 5 mL of THF and added to above solution drop
wise over a period of 30 min under nitrogen. Reaction then was
stirred at 0.degree. C. for 0.5 h heated at 70.degree. C. for 16 h.
THE was removed in vacuo and crude mixture was purified by silica
gel flash chromatography (10% EtOAc/Hexane) to obtain the product.
To a solution of the product in dioxane (4 mL) was added 4M HCl in
dioxane (10 mL) at rt and the resulting mixture was stirred for 2 h
at that temperature. The solvent was removed in vacuo and residue
was triturated with ether to obtain a white solid (1.21 g, 65%):
MS; m/z 338.4 (M+H), .sup.1H NMR (400 MHz, DMSO-d.sub.6); .delta.
1.83-1.90 (m, 2H), 2.09-2.14 (m, 2H), 3.03-3.09 (m, 2H), 3.20-3.26
(m, 2H), 3.32 (s, 1H), 4.62-4.66 (m, 1H), 7.06-7.10 (m, 7H), 7.70
(d, 2H, J=8.8 Hz), 9.08 (s, 2H); HPLC (UV); 97%.
Example 8
##STR00019##
[0094] Step 1
[0095] 4-(4-Methoxy-phenoxy)-biphenyl: A mixture of 4-bromobiphenyl
(1.16 g, 5 mmol), 4-methoxyphenol (413 mg, 3.33 mmol), Cs2CO3 (2.17
g, 6.66 mmol) in anhydrous dioxane (10 mL) was purged with nitrogen
for 10 min. Then copper (I) iodide (18 mg, 0.096 mmol) and
N,N-dimethylglycine HCl (41 mg, 0.29 mmol) were added. Reaction was
stirred at 90.degree. C. over night. Reaction mixture was cooled
and poured over 1:1 mixture of EtOAc/water. Organic layer was
separated, washed with water, brine, dried over anhydrous MgSO4 and
concentrated to give the crude product. It was used in next step
without purification (1.26 g, 91%).
[0096] Step 2
[0097] 4-(Biphenyl-4-yloxy)-phenol: 4-(4-Methoxy-phenoxy)-biphenyl
(1.0 g, 3.6 mmol) in anhydrous CH2Cl2 (10 mL) was cooled to
-78.degree. C. and 1M boron tribromide (10.8 mL, 10.8 mmol) in
CH2Cl2 was added dropwise over period of 20 min under nitrogen. The
resulting mixture was stirred at -78.degree. C. for 2 h and at rt
for 1 h. Then it was cooled to 0.degree. C. and poured slowly over
1:1 mixture of CH2Cl2/water. CH2Cl2 layer was separated, washed
with water, brine, dried over anhydrous MgSO4 and concentrated. The
crude product was purified using silica gel flash chromatography
(20% EtOAc/Hexane) to obtain the product as yellow solid (265 mg,
28%): MS; m/z 260.9 (M-H); .sup.1H NMR (400 MHz, DMSO-d.sub.6);
.delta. 6.79-6.82 (m, 1H), 6.91-6.98 (m, 2H), 7.30-7.49 (m, 4H),
7.60-7.68 (m, 6H), 8.36 (s, 1H).
[0098] Step 3
[0099] 4-[4-(Biphenyl-4-yloxy)-phenoxy]-piperidine hydrochloride
salt: To a solution of t-butyl-4-hydroxy-1-piperidine carboxylate
(201 mg, 1 mmol) in anhydrous THF (1 mL) was added
4-(biphenyl-4-yloxy)-phenol (262 mg, 1 mmol) in THF (1 mL) and
triphenyl phosphine (314 mg, 1.2 mmol) in THF (1 mL). The resulting
mixture was cooled to 0.degree. C. using ice-water bath and purged
with nitrogen. Diisopropyl azodicarboxylate (242 mg, 1.2 mmol) was
dissolved in 2 mL of THF and added to above solution drop wise over
a period of 15 min under nitrogen. Reaction then was stirred at
0.degree. C. for 2 h heated at 70.degree. C. for 16 h. THE was
removed in vacuo and crude mixture was purified by silica gel flash
chromatography (10% EtOAc/Hexane) to obtain the product. To a
solution of the product in dioxane (1 mL) was added 4M HCl in
dioxane (5 mL) at rt and the resulting mixture was stirred for 2 h
at that temperature. Precipitated white solid was removed by
filtration, washed with ether and dried in vacuum (185, 43%): MS;
m/z 346.4 (M+H); .sup.1H NMR (400 MHz, DMSO-d.sub.6); .delta.
1.82-1.88 (m, 2H), 2.08-2.2.13 (m, 2H), 3.03-3.09 (m, 2H),
3.21-3.32 (m, 2H), 4.59-4.62 (m, 1H), 7.00-7.06 (m, 6H), 7.32-7.36
(m, 1H), 7.42-7.47 (m, 2H), 7.61-7.66 (m, 4H), 8.87 (s, 1H); HPLC
(UV); 99%.
Example 9
##STR00020##
[0101] 4-[4-(4-Trifluoromethoxy-phenoxy)-phenoxy]-piperidine
hydrochloride: To a solution of
t-butyl-4-hydroxy-1-piperidinecarboxylate (0.40 g, 1.99 mmol) in
anhydrous THF (8 mL) was added
4-(4-Trifluoromethoxy-phenoxy)-phenol (0.58 g, 2.65 mmol) and
triphenylphosphine (0.63 g, 2.40 mmol). The resulting mixture was
cooled to 0.degree. C. Diisopropylazodicarboxylate (0.47 mL, 2.43
mmol) was added portionwise over a ten minute period. The reaction
mixture was warmed to ambient temperature for an hour and then
heated to 60.degree. C. for 48 h. The solvent was removed in vacuo.
The crude residue was purified by silica gel flash chromatography
using hexane/EtOAc (gradient system) to give a yellow oil. To the
oil was added 4 M HCl in dioxane (7 mL). The resulting mixture was
stirred at ambient temperature 2 h. The solvent was removed under
reduced pressure to obtain crude product. The residue was
triturated with ether to afford the title compound (98 mg, 14%);
.sup.1H NMR (400 MHz, DMSO-d.sub.6); .delta. 1.85 (m, 2H), 2.10 (m,
2H), 3.06 (m, 2H), 3.23 (m, 2H), 4.61 (m, 1H), 7.02 (d, J=8.8 Hz,
2H), 7.06 (s, 3H), 7.35 (d, J=8.8 Hz, 2H), 8.92 (br s, 2H); MS
(m/z) 354.4 (M+1); LC (97.5%).
Example 10
##STR00021##
[0103] Step 1
[0104] 4-[4-(4-Chloro-phenoxy)-phenoxy]-piperidine: To a solution
of t-butyl-4-hydroxy-1-piperidinecarboxylate (0.40 g, 1.99 mmol) in
anhydrous THF (8 mL) was added 4-(4-chloro-phenoxy)-phenol (0.58 g,
2.61 mmol) and triphenylphosphine (0.63 g, 2.40 mmol). The
resulting mixture was cooled to 0.degree. C.
Diisopropylazodicarboxylate (0.47 mL, 2.43 mmol) was added
portionwise over a ten minute period. The reaction mixture was
warmed to ambient temperature for an hour and then heated to
47.degree. C. for 64 h. The solvent was removed in vacuo. The crude
residue was purified by silica gel flash chromatography using
hexane/EtOAc (gradient system) to give the title compound (0.93 g,
100%) as a yellow oil.
[0105] Step 2
[0106] 4-[4-(4-Chloro-phenoxy)-phenoxy]-piperidine hydrochloride:
To the product from step 1 (0.88 g, 2.17 mmol) was added 4 M HCl in
dioxane (10 mL). The resulting mixture was stirred at ambient
temperature for 2 hours. The solvent was removed under reduced
pressure. The residue was triturated with ether to afford the title
compound (0.34 g, 47%) as a white solid; .sup.1H NMR (400 MHz,
DMSO-d.sub.6); .delta. 1.83 (m, 2H), 2.09 (m, 2H), 3.06 (m, 2H),
3.23 (m, 2H), 4.60 (m, 1H), 6.94 (d, J=9.2 Hz, 2H), 7.01-7.06 (m,
4H), 7.40 (d, J=9.2 Hz, 2H), 8.81 (br s, 2H); MS (m/z) 304.4 (M+1);
LC (100%).
Example 11
##STR00022##
[0108] (R)-3-(4-Phenoxy-phenoxy)-pyrrolidine hydrochloride:
N-t-butylcarbonyl-(R)-(-)-3-pyrrolidinol (1 g, 5.34 mmol) was taken
into anhydrous tetrahydrofuran (2 mL) in a nitrogen flushed 20 mL
vial. 4-phenoxyphenol (1.29 g, 6.94 mmol) was added to the mixture
followed by triphenylphosphine (1.86 g, 6.41 mmol). The reaction
was cooled to 0.degree. C. and a solution of diisopropyl
azodicarboxylate (1.3 g, 6.41 mmol) in anhydrous tetrahydrofuran (2
mL) was slowly added to the reaction over 5 min. The mixture was
then allowed to warm to room temperature overnight. The reaction
was concentrated to dryness under vacuum and the residue was
purified by flash silica chromatography (20:1 silica packing ratio,
eluted with 1% methanol in dichloromethane). The enriched product
was then taken into 4N HCl in dioxane, and stirred at room
temperature overnight. The mixture was then concentrated to dryness
and triturated with diethyl ether to provide the title product
(1.24 g, 80%); LCMS; m/z 256 (M+1 of free amine). .sup.1H NMR (400
MHz, DMSO); .delta. 2.13-2.17 (.quadrature..quadrature.m, 2H),
3.25-3.47 (m, 4H), 5.10 (s, 1H), 6.93-6.95 (m, 2H), 7.02 (s, 4H),
7.09-7.11 (m, 1H), 7.34-7.38 (m, 2H), 9.54 (s, 2H).
Example 12
##STR00023##
[0110] Step 1
[0111] 4-[(R)-3-(4-Phenoxy-phenoxy)-pyrrolidin-1-yl]-butyric acid
methyl ester (R)-3-(4-Phenoxy-phenoxy)-pyrrolidine (1.24 g, 4.25
mmol) was taken into anhydrous DMF (10 mL) in a nitrogen flushed
100 mL, 1-neck round bottom flask. Methyl-4-bromobutyrate (846 mg,
4.68 mmol) was added to the mixture followed by potassium carbonate
(1.18 g, 8.5 mmol). The reaction was heated at 60.degree. C. for 24
h. The mixture was then cooled to room temperature and partitioned
between ethyl acetate and water. The water layer was washed with
ethyl acetate (3.times.). The combined ethyl acetate layers were
then washed with brine, dried over anhydrous sodium sulfate,
filtered, and concentrated to dryness. The resulting residue was
then purified by silica gel flash chromatography (20:1 silica
ratio, eluted with 1:1 ethyl acetate and hexane) to obtain the
title product (894.4 mg, 59%) LCMS; m/z 356 (M+1); .sup.1H NMR (400
MHz, DMSO) .delta. 1.65-1.78 (m, 3H), 2.21-2.26 (m, 1H), 2.32-2.35
(m, 2H), 2.37-2.42 (m, 3H), 2.58-2.61 (m, 1H), 2.63-2.69 (m, 1H),
2.80-2.84 (m, 1H), 3.58 (s, 3H), 4.80 (s, 1H), 6.89-6.98 (m, 6H),
7.07-7.09 (m, 1H), 7.32-7.36 (m, 2H).
[0112] Step 2
[0113] 4-[(R)-3-(4-Phenoxy-phenoxy)-pyrrolidin-1-yl]-butyric acid
hydrogen chloride
[0114] The product from step 1 (200 mg, 0.56 mmol), was taken into
1.029N NaOH (aq) (1.0933 mL, 1.125 mmol), and methanol (3 mL). The
reaction was heated to 60.degree. C. and run for 24 h. The mixture
was concentrated to dryness and taken into 1N HCl in dioxane (1.125
mL, 0.1.125 mmol), and was left to react for 24 h at room
temperature. The mixture was concentrated to dryness and taken into
dichloromethane. The solution was filtered to remove salts, and
then concentrated to dryness. The residue was then taken into an
excess of 1N HCl in dioxane and left to react for 24 h. at room
temperature. The mixture was then concentrated to dryness to yield
the title product (123.5 mg, 0.327 mmol, 58% yield); LC/MS; m/z 343
(m+2 of free amine); .sup.1H NMR (400 MHz, DMSO) .delta. 1.89-1.93
(m, 2H), 2.14 (s, 1H), 2.33-2.49 (m, 3H), 3.17-3.21 (m, 3H), 3.57
(s, 2H), 5.10 (s, 1H), 6.92-6.96 (m, 2H), 7.02 (s, 4H), 7.07-7.11
(m, 1H), 7.34-7.38 (m, 2H).
Example 13
##STR00024##
[0116] 4-[(R)-3-(4-Phenoxy-phenoxy)-pyrrolidin-1-yl]-butyramide:
4-[(R)-3-(4-Phenoxy-phenoxy)-pyrrolidin-1-yl]-butyric acid methyl
ester (200 mg, 0.563 mmol) was taken into 7N Ammonia in methanol
solution (5 mL) in a pressure tube. The tube was sealed and heated
to 100.degree. C. with stirring and left to react for 48 h. The
reaction was concentrated to dryness, and purified by flash silica
chromatography (20:1 silica ratio, eluted with a gradient of 10%
methanol in dichloromethane to 15% methanol in dichloromethane) to
provide the title product (141 mg, 74%); LCMS; m/z 341 (M+1);
.sup.1H NMR (400 MHz, DMSO) .delta. 1.62-1.68 (m, 2H), 1.75-1.78
(m, 1H), 2.06-2.10 (m, 2H), 2.22-2.25 (m, 1H), 2.35-2.43 (m, 3H),
2.59-2.68 (m, 2H), 2.80-2.84 (m, 1H), 4.80-4.83 (m, 1H), 6.69 (s,
1H), 6.89-6.93 (m, 4H), 6.94-6.98 (m, 2H), 7.05-7.09 (m, 1H), 7.24
(s, 1H), 7.32-7.36 (m, 2H).
Example 14
##STR00025##
[0118] Step 1
[0119] (R)-3-(4-Iodo-phenoxy)-pyrrolidine-1-carboxylic acid
tert-butyl ester: N-tert-butyl-butoxy
carbonyl-(R)-(-)-3-pyrrolidinol (3 g, 16 mmol), 4-iodophenol (4.58
g, 21 mmol), and triphenyl phosphine (5.04 g, 19 mmol), were taken
into anhydrous tetrahydrofuran (10 mL) in a nitrogen flushed 200 mL
round bottom flask. The reaction was cooled to 0.degree. C. and
diisopropyl diazodicarboxylate (1.044 mL, 19 mmol) taken into
anhydrous tetrahydrofuran (10 mL) was added to the reaction
dropwise over 10 minutes. The reaction was run at 0.degree. C. for
30 minutes and then allowed to warm to room temperature over 12 h.
The reaction was then heated to 70.degree. C. and left to react for
12 h. The mixture was concentrated to dryness, and the compound was
purified by flash column chromatography (40:1 silica ratio, eluted
with a gradient of dichlormethane to 10% methanol in
dichloromethane) to obtain the title product (5.18 g, 83%); LCMS;
m/z 390 (M+1) 95% pure. .sup.1H NMR (400 MHz, DMSO);
.delta..quadrature. 1.40 (s, 9H), 2.01-2.03 (m, 2H), 3.03-3.42 (m,
3H), 3.43-3.54 (m, 1H), 4.98 (s, 1H), 6.80 (d, 2H, J=8.8), 7.60 (d,
2H, J=8.8).
[0120] Step 2
[0121]
(R)-3-[4-(4-Chloro-phenoxy)-phenoxy]-pyrrolidine-1-carboxylic acid
tert-butyl ester: 4-chlorophenol (495 mg, 3.85 mmol), the product
from step 1 (1 g. 2.57 mmol), cesium carbonate (1.67 g, 5.14 mmol),
copper iodide (51 mg, 0.27 mmol), and N,N-dimethylglycine HCl (35
mg, 0.249 mmol), were taken into anhydrous dioxane (12 mL) in a
nitrogen flushed 20 mL vial. The reaction was heated to 90.degree.
C. for 24 h. The mixture was concentrated to dryness, and the
resulting oil was partitioned between water and ethyl acetate. The
aqueous layer was washed with ethyl acetate 3.times.. The combined
organic layers were washed with brine, dried over sodium sulfate
and concentrated to dryness. The compound was purified by flash
column chromatography (20:1 silica ratio, eluted with 20% ethyl
acetate in hexane) to obtain the title product (1 g, .about.90%);
LCMS; m/z 390 (M) 88% pure; .sup.1H NMR (400 MHz, DMSO);
.delta..quadrature. 1.41 (s, 9H), 2.00-2.20 (m, 2H), 3.32-3.60 (m,
4H), 4.98 (s, 1H), 6.76 (d, 2H, J=8.8), 6.94-7.03 (m, 4H), 7.19 (d,
2H, J=8.8), 9.70 (s, 1H).
[0122] Step 3
[0123] (R)-3-[4-(4-Chloro-phenoxy)-phenoxy]-pyrrolidine:
(R)-3-[4-(4-Chloro-phenoxy)-phenoxy]-pyrrolidine-1-carboxylic acid
tert-butyl ester (1 g, 2.56 mmol) was taken into 4N HCl in dioxane
(10 mL), and the reaction was run at room temperature for 24 h. The
mixture was concentrated to dryness, and the resulting oil was
triturated with diethyl ether to obtain the title product (344 mg,
41%); LCMS; m/z 290 (M+1, free amine) 81% pure; .sup.1H NMR (400
MHz, DMSO); .delta. 2.13-2.16 (m, 2H), 3.24-3.33 (m, 4H), 5.11 (s,
1H), 6.95 (d, J=8.8, 2H), 7.04 (s, 4H), 7.40 (d, J=8.8, 2H), 9.66
(s, 2H).
[0124] Step 4
[0125]
4-{(R)-3-[4-(4-Chloro-phenoxy)-phenoxy]-pyrrolidin-1-yl}-butyric
acid methyl ester: (R)-3-[4-(4-Chloro-phenoxy)-phenoxy]-pyrrolidine
(344 mg, 1.05 mmol) was taken into anhydrous dimethylformamide (5
mL), and methyl-4-bromobutyrate (209 mg, 1.16 mmol), potassium
carbonate (290 mg, 2.1 mmol) was added. The reaction was sealed and
heated to 60.degree. C. and left to react for 24 h. The mixture was
then partitioned between ethyl acetate and water. The aqueous layer
was washed with ethyl acetate 3.times.. The combined organic layers
were washed with brine, dried over sodium sulfate and concentrated
to dryness. The residue was purified by flash column chromatography
(50:1 packing ration, eluted with 5% methanol in dichloromethane)
to obtain the title product (86.8 mg, 21%); LCMS; m/z 390 (M) 71%
pure; .sup.1H NMR (400 MHz, DMSO); .delta..quadrature. 1.66-1.80
(m, 3H), 2.22-2.26 (m, 1H), 2.31-2.35 (m, 2H), 2.37-2.41 (m, 3H),
2.58-2.67 (m, 2H), 2.81-2.84 (m, 1H), 3.58 (s, 3H), 4.80-4.83 (m,
1H), 6.91-6.95 (m, 4H), 6.98-7.00 (m, 2H), 7.37-7.39 (m, 2H).
[0126] Step 5
[0127]
4-{(R)-3-[4-(4-Chloro-phenoxy)-phenoxy]-pyrrolidin-1-yl}-butyric
acid HCl
4-{(R)-3-[4-(4-Chloro-phenoxy)-phenoxy]-pyrrolidin-1-yl}-butyric
acid methyl ester (86.8 mg, 0.222 mmol) was taken into 1.029N NaOH
solution in water (0.433 mL, 0.445 mmol), and methanol (2 mL). The
reaction was sealed and heated to 60.degree. C. and left to react
for 24 h. The reaction was concentrated to dryness and taken into
4N HCl in dioxane (0.111 ml, 0.445 mmol) and dichloromethane (1
mL). The reaction was stirred for 15 minutes, and then concentrated
to dryness. The residue was taken into dichloromethane and filtered
through a syringe filter. The filtrate was then treated with excess
4N HCl in dioxane, stirred for 15 minutes and then concentrated to
dryness. The crude product was purified by reverse phase semi-prep
HPLC to obtain the title product (43 mg, 47%); LCMS; m/z 376 (M,
free amine); .sup.1H NMR (400 MHz, DMSO); .delta. 1.63-1.70 (m,
2H), 1.74-1.82 (m, 1H), 2.21-2.29 (m, 3H), 2.42-2.45 (m, 3H),
2.62-2.73 (m, 2H), 2.85-2.89 (m, 1H), 4.85 (s, 1H), 6.91-6.96 (m,
4H), 6.98-7.01 (m, 2H), 7.36-7.40 (m, 2H), 8.21 (s, 1H).
Example 15
##STR00026##
[0129] 4-[(R)-3-(4-Benzyl-phenoxy)-pyrrolidin-1-yl]-butyramide:
4-[(R)-3-(4-Benzyl-phenoxy)-pyrrolidin-1-yl]-butyric acid methyl
ester (200 mg, 0.566 mmol) was taken into 7N ammonia in methanol
solution (5 mL) in a pressure tube. The reaction was sealed and
heated at 100.degree. C. for 48 h. The mixture was then cooled to
room temperature and concentrated to dryness. The resulting residue
was then purified by silica gel flash chromatography (20:1 silica
ratio, eluted with a gradient of 10% methanol in dichloromethane to
15% methanol in dichloromethane) to obtain the title product (145
mg, 75%); LCMS; m/z 340 (M+1); .sup.1H NMR (400 MHz, DMSO); .delta.
1.58-1.74 (.quadrature..quadrature.m, 3H), 2.04-2.08 (m, 2H),
2.19-2.24 (m, 1H), 2.33-2.41 (m, 3H), 2.54-2.58 (m, 1H), 2.61-2.65
(m, 1H), 2.78-2.82 (m, 1H), 3.85 (s, 2H), 4.77-4.80 (m, 1H), 6.68
(s, 1H), 6.78 (d, J=8.8, 2H), 7.11 (d, J=8.4, 2H), 7.17-7.22 (m,
3H), 7.25-7.29 (m, 1H).
Assays to Determine Potency of LTA4 Hydrolase Inhibitors
[0130] (1) In Vitro Assay Testing Inhibitory Activity Against
Purified Recombinant Human LTA.sub.4 Hydroase:
[0131] A human LTA4 hydrolase full-length cDNA clone
(NM.sub.--000895) was purchased from OriGene Technologies
(Rockville, Md.). The gene was amplified by polymerase chain
reaction and transferred via pDONR201 into the bacterial expression
vector pDEST17 by recombination (both plasmids from Invitrogen,
Carlsbad, Calif.). The resulting construct was transformed into
Escherichia coli BL21-AI (Invitrogen), and expression was induced
by chemical induction with arabinose. The recombinant enzyme was
purified by chromatography on a FPLC system (Amersham Biosciences,
Uppsala, Sweden) using immobilized metal affinity chromatography
(Ni-NTA Superflow, Qiagen, Hilden, Germany) and anion exchange
chromatography (MonoQ HR 10/10, Amersham Biosciences).
[0132] The compounds of the invention were incubated in a series of
dilutions with 200 nM of recombinant enzyme in assay buffer (100 mM
Tris-HCl, 100 mM NaCl, 5 mg/ml fatty-acid free BSA, 10% DMSO, pH
8.0) for 10 min at room temperature to allow binding between LTA4
hydrolase and the inhibitors. LTA4 was prepared by alkaline
hydrolysis of LTA4 methyl ester (Biomol, Plymouth Meeting, Pa., or
Cayman Chemicals, Ann Arbor, Mich.). A solution of 10 .mu.g of the
ester was dried under a nitrogen stream and redissolved in 60 .mu.l
of a solution of 80% acetone and 20% 0.25 M NaOH. After incubation
for 40 min at room temperature the resulting approximately 500
.mu.M tock of LTA4 was kept at -80.degree. C. for no more than a
few days prior to use.
[0133] Immediately before the assay, LTA4 was diluted to a
concentration of 10 .mu.M in assay buffer (without DMSO) and added
to the reaction mixture to a final concentration of 2 .mu.M to
initiate the enzyme reaction. After incubation for 120 sec at room
temperature, the reaction was stopped by adding 2 volumes of
chilled quenching buffer, containing acetonitril with 1% acetic
acid and 225 nM LTB.sub.4-d.sub.4 (Biomol). The samples were then
kept at 4.degree. C. over night to complete protein precipitation
and centrifuged for 15 min at 1800 g. LTB.sub.4 formed was measured
by LC-MS/MS using LTB.sub.4-d.sub.4 as an internal standard and an
external LTB.sub.4 standard (Biomol) as reference. Briefly, the
analyte was separated from LTB.sub.4 isomers formed by spontaneous
hydrolysis of LTA4 using isocratic elution (modified protocol from
Mueller et al. (1996), J. Biol. Chem. 271, 24345-24348) on a HPLC
system (Waters, Milford, Mass.) and analyzed on a tandem quadrupole
mass spectrometer (Waters). MRM transitions followed on 2 channels
were 335.2>195.3 (LTB.sub.4) and 339.2>197.3
(LTB.sub.4-d.sub.4). Based on the amounts of LTB.sub.4 found at
each inhibitor concentration, a dose-response curve was fitted to
the data and an IC.sub.50 value was calculated.
[0134] (2) Ex Vivo Assay Testing Inhibitory Activity in Human Whole
Blood after Stimulation with Calcium Ionophor:
[0135] Human blood was collected in heparin-containing Vacutainer
tubes. For each sample, 200 .mu.l of blood were dispensed into a
pre-warmed plate and 188 .mu.l of RPMI-1640 medium (Invitrogen)
containing 20 .mu.g/ml Indomethacin (Sigma, St. Louis, Mo.) were
added. Then 4 .mu.l of a series of compound dilutions (in DMSO)
were added, followed by a 15 min incubation at 37.degree. C. with
gentle shaking. After that, blood samples were stimulated by adding
Ionomycin (Calbiochem) to a final concentration of 20 .mu.M. After
another incubation at 37.degree. C. for 30 min, samples were
centrifuged for 5 min at 1800 g and 4.degree. C. Supernatants were
taken and LTB.sub.4 concentrations were determined using a
commercially available enzyme-linked immunoassay (R&D Systems,
Minneapolis, Minn.) according to the manufacturer's instructions.
Results obtained for different concentrations of hydrolase
inhibitor were then used to fit a dose-response curve and calculate
an IC.sub.50 value.
TABLE-US-00001 ##STR00027## IC50 (.mu.M) IC50 (.mu.M) Ex. R1 X * n
R2 (hLTA4H) (hWB) 1 H CH2 S 1 H A A 2 H CH2 R 1 H A A 3 H O N/A 2 H
A A 4 H CH2 R 1 (CH2)3CO2H A ND 5 H O N/A 2 (CH2)2CO2H A ND 6 H O
N/A 2 (CH2)3CO2H A ND 7 CF3 O N/A 2 H A ND 8 Ph O N/A 2 H A A 9
OCF3 O N/A 2 H B ND 10 Cl O N/A 2 H A ND 11 H O S 1 H A ND 12 H O S
1 (CH2)3CO2H A ND 13 H O S 1 (CH2)3CONH2 A ND 14 Cl O S 1
(CH2)3CO2H A ND 15 H CH2 S 1 (CH2)3CONH2 A ND A = <5 uM; B =
5-20 uM; ND = Not Determined
* * * * *