U.S. patent application number 10/930901 was filed with the patent office on 2005-02-17 for compositions and methods to prevent toxicity induced by nonsteroidal antiinflammatory drugs.
This patent application is currently assigned to NitroMed, Inc.. Invention is credited to Garvey, David S., Letts, L. Gordon, Renfroe, H. Burt, Tam, Sang William.
Application Number | 20050038029 10/930901 |
Document ID | / |
Family ID | 27026571 |
Filed Date | 2005-02-17 |
United States Patent
Application |
20050038029 |
Kind Code |
A1 |
Garvey, David S. ; et
al. |
February 17, 2005 |
Compositions and methods to prevent toxicity induced by
nonsteroidal antiinflammatory drugs
Abstract
Nonsteroidal antiinflammatory drugs which have been substituted
with a nitrogen monoxide group; compositions comprising (i) a
nonsteroidal antiinflammatory drug, which can optionally be
substituted with a nitrogen monoxide group and (ii) a compound that
directly donates, transfers or releases a nitrogen monoxide group
(preferably as a charged species, particularly nitrosonium); and
methods of treatment of inflammation, pain, gastrointestinal
lesions and/or fever using the compositions are disclosed. The
compounds and compositions protect against the gastrointestinal,
renal and other toxicities that are otherwise induced by
nonsteroidal antiinflammatory drugs.
Inventors: |
Garvey, David S.; (Waltham,
MA) ; Letts, L. Gordon; (Dover, MA) ; Renfroe,
H. Burt; (Wellesley, MA) ; Tam, Sang William;
(Dover, MA) |
Correspondence
Address: |
WILMER CUTLER PICKERING HALE AND DORR LLP
THE WILLARD OFFICE BUILDING
1455 PENNSYLVANIA AVE, NW
WASHINGTON
DC
20004
US
|
Assignee: |
NitroMed, Inc.
Lexington
MA
|
Family ID: |
27026571 |
Appl. No.: |
10/930901 |
Filed: |
September 1, 2004 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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10930901 |
Sep 1, 2004 |
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10277202 |
Oct 22, 2002 |
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6790864 |
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10277202 |
Oct 22, 2002 |
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09941690 |
Aug 30, 2001 |
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6482846 |
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09941690 |
Aug 30, 2001 |
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09518541 |
Mar 3, 2000 |
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6323234 |
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09518541 |
Mar 3, 2000 |
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08931564 |
Sep 16, 1997 |
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6057347 |
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08931564 |
Sep 16, 1997 |
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08543208 |
Oct 13, 1995 |
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5703073 |
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08543208 |
Oct 13, 1995 |
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08425090 |
Apr 19, 1995 |
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6051588 |
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Current U.S.
Class: |
514/248 ;
544/236 |
Current CPC
Class: |
A61P 9/00 20180101; A61K
31/343 20130101; C07K 5/0215 20130101; A61K 31/60 20130101; A61P
43/00 20180101; A61K 31/343 20130101; A61P 25/28 20180101; A61P
19/02 20180101; C07C 381/00 20130101; A61P 1/00 20180101; A61P
29/00 20180101; C07C 327/28 20130101; C07D 417/12 20130101; C07C
203/00 20130101; A61K 2300/00 20130101; C07J 41/0055 20130101; A61P
1/04 20180101; A61K 2300/00 20130101; C07D 209/28 20130101; A61K
31/60 20130101; A61K 38/00 20130101 |
Class at
Publication: |
514/248 ;
544/236 |
International
Class: |
C07D 487/02; A61K
031/503 |
Claims
What is claimed is:
1. A non-steroidal antiinflammatory agent to which is directly or
indirectly linked at least one NO group.
2. The non-steroidal antiinflammatory agent of claim 1 which is
selected from the group consisting of compounds having the
structures: 23wherein D is selected from (i) a covalent bond; (ii)
--C(R.sub.a)--O--C(O)--Y--[C- (R.sub.b)(R.sub.c)].sub.P-T- in which
R.sub.a is lower alkyl, cycloalkyl, aryl or heteroaryl, Y is
oxygen, sulfur, or NR.sub.i in which R.sub.i is hydrogen or lower
alkyl, R.sub.b and R.sub.c are independently selected from,
hydrogen, lower alkyl, cycloalkyl, aryl, heteroaryl, arylalkyl,
alkylamino, dialkylamino or taken together are cycloalkyl or
bridged cycloalkyl, p is an integer from 1 to 6 and T is a covalent
bond, oxygen, sulfur, or nitrogen; or (iii)
--(CO)-T.sub.1-[C(R.sub.b)(R.sub.c)].sub.P-- T.sub.2- wherein
T.sub.1 and T.sub.2 are independently selected from T, and wherein
R.sub.b, R.sub.c, p and T are as defined above; Z is an aryl or
heteroaryl; and A.sub.1, A.sub.2 and A.sub.3 comprise the other
subunits of a 5- or 6-membered monocyclic aromatic ring and each is
independently selected from (1) C--R.sub.1 wherein R.sub.1 at each
occurrence is independently selected from hydrogen, lower alkyl,
lower haloalkyl, alkoxyalkyl, halogen or nitro; (2) N--R.sub.d
wherein R.sub.d at each occurrence is independently selected from a
covalent bond to an adjacent ring atom in order to render the ring
aromatic, hydrogen, lower alkyl, cycloalkyl, arylalkyl, aryl,
heteroaryl; (3) sulfur; (4) oxygen; and (5) B.sub.a=B.sub.b wherein
B.sub.a and B.sub.b are each independently selected from nitrogen
or C--R.sub.1 wherein at each occurrence R.sub.1 is as defined
above; the structures: 24wherein R.sub.b, R.sub.c, D, Z, A.sub.1,
A.sub.2 and A.sub.3 are as defined above; the structures: 25wherein
R.sub.e is hydrogen or lower alkyl; R.sub.f is selected from
26272829in which n is 0 or 1; and X is
(1)-Y-[C(R.sub.b)(R.sub.c)].sub.P-G-[C(R.sub.b)(R.sub.c).sub.p-T-NO,
wherein G is (i) a covalent bond; (ii) -T-C(O)--; (iii) --C(O)-T;
(iv) --C(Y--C(O)--R.sub.m)-- wherein R.sub.m is heteroaryl or
heterocyclic ring; and in which Y, R.sub.b, R.sub.c, p and T are as
defined above; or (2) 30in which W is a heterocyclic ring or
NR.sub.hR.sub.i wherein R.sub.h and R.sub.i are independently
selected from lower alkyl, aryl or alkenyl; and the structures:
31wherein R.sub.g is selected from 3233and X is defined as above.
Description
RELATED APPLICATIONS
[0001] This application is a continuation of U.S. application Ser.
No. 10/277,202, filed Oct. 22, 2002, which is a divisional of U.S.
application Ser. No. 09/941,690, filed Aug. 30, 2001, issued as
U.S. Pat. No. 6,482,846, which is a continuation of U.S.
application Ser. No. 09/518,541, filed Mar. 3, 2000, issued as U.S.
Pat. No. 6,323,234, which is a continuation of U.S. application
Ser. No. 08/931,564, filed Sep. 16, 1997, issued as U.S. Pat. No.
6,057,347, which is a a continuation of U.S. application Ser. No.
08/543,208, filed Oct. 13, 1995, issued as U.S. Pat. No. 5,703,073,
which is a continuation in-part of U.S. application Ser. No.
08/425,090 filed Apr. 19, 1995, issued as U.S. Pat. No. 6,051,588.
This application is also related to U.S. Pat. Nos. 5,703,073,
6,043,232, 6,043,233, 6,048,858, 6,051,588, 6,057,347, 6,083,515,
6,143,734 and 6,323,234.
FIELD OF THE INVENTION
[0002] This invention relates to the field of "aspirin-like" or
nonsteroidal antiinflammatory drug compounds and compositions that
prevent, reduce or reverse the gastrointestinal, renal, and other
toxicities associated with nonsteroidal antiinflammatory drugs.
BACKGROUND OF THE INVENTION
[0003] Arena et al., WO94/12463, discloses the chemistry and
pharmacology of nitroxybutylester ((CH2).sub.4--ONO.sub.2)
derivatives of several aryl propionic acid non-steroidal
antiinflammatory drugs including ketoprofen, flurbiprofen,
suprofen, indobufen and etodolac. Studies on nitroxybutylester
derivatives of flurbiprofen and ketoprofen are also reported in
Wallace et al., Gastroenterology, 107:173-179 (1994). See, also,
Cuzzolin et al., Pharmacol. Res., 29(1):89-97 (1994); Reuter et
al., Life Sci. (USA), 55/1(PL1-PL8) (1994); Reuter et al.,
Gastroenterology, 106(4):Suppl. A759 (1994); Wallace et al., Eur.
J. Pharmacol., 257(3):249-255 (1994); Wallace et al.,
Gastroenterology, 106(4):Suppl. A208 (1994); and Conforti et al.,
Agents-Actions, 40(3-4): 176-180 (1993). These publications
uniformly examine and rely upon the use of indirectly linked
nitrogen dioxide substitutions.
[0004] The present invention is based on the discovery by the
inventors that it is possible to link a nitrogen monoxide group,
nitric oxide (NO), to a non-steroidal antiinflammatory agent and
that the resulting compounds not only possess potent
analgesic/antiinflammatory properties but has a much reduced
potential for producing gastrointestinal lesions (ulcers).
[0005] The present invention is further based on the discovery by
the inventors that it is possible to coadminister a nonsteroidal
antiinflammatory drug (NSAID) and a compound that directly donates,
releases or transfers nitrogen monoxide (preferably as a charged
species, particularly nitrosonium) to prevent, reduce, or reverse
the gastrointestinal renal, and other toxicities induced by the
NSAID. NSAIDs are antiinflammatory, analgesic and antipyretic
compounds that act as cyclooxygenase, the enzyme responsible for
the biosyntheses of the prostaglandins and certain autocoids,
inhibitors, including inhibitors of the various isozymes of
cyclooxygenase (including but not limited to cyclooxygenase-1 and
-2) and as inhibitors of both cyclooxygenase and lipoxygenase. A
nitric oxide donor is a compound that contains a nitric oxide
moiety and which directly releases or directly chemically transfers
nitrogen monoxide (nitric oxide), preferably in its positively
charged nitrosonium form, to another molecule. Nitric oxide donors
include but are not limited to S-nitrosothiols, nitrites,
N-oxo-N-nitrosamines, and substrates of various forms of nitric
oxide synthase.
[0006] In one aspect the present invention provides a compound
comprising a non-steroidal antiinflammatory agent to which is
directly or indirectly linked at least one NO group. The
non-steroidal antiinflammatory agent can, for example, be an aryl
propionic acid or an enolic anilide. The invention also provides
compositions comprising such compounds in a pharmaceutically
acceptable carrier.
[0007] In another aspect the invention provides a composition
comprising a mixture of a therapeutically effective amount of a
nonsteroidal antiinflammatory agent and an NSAID toxicity reducing
amount of a compound that donates, transfers or releases nitric
oxide.
[0008] In another aspect the present invention provides a
composition comprising a non-steroidal antiinflammatory agent to
which is directly or indirectly linked at least one NO group and a
compound that donates, transfers or releases nitric oxide. The
non-steroidal antiinflammatory agent can, for example, be an aryl
propionic acid or an enolic anilide. The invention also provides
compositions comprising such compounds in a pharmaceutically
acceptable carrier.
[0009] In another aspect the invention provides a method for
treating inflammation, pain and/or fever in an individual in need
thereof which comprises administering to the individual a
nonsteroidal antiinflammatory agent, which may optionally be
substituted with at least one NO group, and a compound that
donates, transfers or releases nitric oxide. The NSAID or NSAID
directly or indirectly linked to at least one NO group, and nitric
oxide donor can be administered separately or as components of the
same composition.
[0010] In another aspect the invention provides a method of
treating inflammation, pain and/or fever in an individual in need
thereof which comprises administering to the individual a
composition comprising a therapeutically effective amount of an
NSAID, which may optionally be substituted with at least one NO
group, and an NSAID toxicity reducing amount of a nitric oxide
donor in a pharmaceutically acceptable carrier. Such compositions
are discussed in more detail below.
[0011] In another aspect the invention provides a method to
decrease or reverse the gastrointestinal toxicity of nonsteroidal
antiinflammatory drugs adminstered to an animal, particularly a
human, by co-administering to said animal a nitric oxide donor. The
NSAID and nitric oxide donor can be administered separately or as
components of the same composition.
[0012] In another aspect the invention provides a method to
decrease or reverse the renal toxicity of nonsteroidal
antiinflammatory drugs administered to an animal, particularly a
human, by co-administering to said animal a nitric oxide donor. The
NSAID and nitric oxide donor can be administered separately or as
components of the same composition.
[0013] In another aspect the invention provides a method to
accelerate gastrointestinal tissue repair in an animal,
particularly a human, by administering to said animal a nitric
oxide donor. The NSAID and nitric oxide donor can be administered
separately or as components of the same composition.
[0014] The compounds and compositions of the present invention are
novel and can be utilized to treat numerous inflammatory disease
states and disorders. For example, reperfusion injury to an
ischemic organ, e.g., reperfusion injury to the ischemic
myocardium, myocardial infarction, inflammatory bowel disease,
rheumatoid arthritis, osteoarthritis, hypertension, psoriasis,
organ transplant rejections, organ preservation, impotence,
radiation-induced injury, asthma, atherosclerosis, thrombosis,
platelet aggregation, metastasis, influenza, stroke, burns, trauma,
acute pancreatitis, pyelonephritis, hepatitis, autoimmune diseases,
insulin-dependent diabetes mellitus, disseminated intravascular
coagulation, fatty embolism, Alzheimer's disease, adult and
infantile respiratory diseases, carcinogenesis and hemorrhages in
neonates.
[0015] The NSAID can be nitrosylated through sites such as oxygen
(hydroxyl condensation), sulfur (sulfhydryl condensation), carbon
and nitrogen.
[0016] The term "lower alkyl"herein refers to branched or straight
chain alkyl groups comprising one to ten carbon atoms, including
methyl, propyl, isopropyl n-butyl, t-butyl, neopentyl and the
like.
[0017] The term "alkoxy" herein refers to RO-wherein R is lower
alkyl as defined above. Representative examples of alkoxy groups
include methoxy, ethoxy, t-butoxy and the like.
[0018] The term `alkoxyalkyl` herein refers to an alkoxy group as
previously defined appended to an alkyl group as previously
defined. Examples of alkoxyalkyl include, but are not limited to,
methoxymethyl, methoxyethyl, isopropoxymethyl and the like.
[0019] The term "alkenyl" herein refers to a branched or straight
chain C.sub.2-C.sub.10 hydrocarbon which also comprises one or more
carbon--carbon double bonds.
[0020] The term "amino" herein refers to --NH.sub.2.
[0021] The term "cyano" herein refers to --CN.
[0022] The term "hydroxy" herein refers to H.
[0023] The term "alkylsulfinyl" herein refers to
R.sub.50--S(O).sub.2-- wherein R.sub.50 is a branched or unbranched
lower alkyl of up to four carbons.
[0024] The term "carboxamido" herein refers to --C(O)NH.sub.2.
[0025] The term "carbamoyl" herein refers to --O--C(O)NH.sub.2.
[0026] The term "carboxyl" herein refers to CO.sub.2H.
[0027] The term "alkylamino" herein refers to R.sub.51 NH-wherein
R.sub.51 is a lower alkyl group, for example, methylamino,
ethylamino, butylamino, and the like.
[0028] The term "dialkylamino" herein refers to R.sub.52R.sub.53N--
wherein R.sub.52 and R.sub.53 are independently selected from lower
alkyl, for example dimethylamino, diethylamino, methyl propylamino,
and the like.
[0029] The term "N-alkylcarbamoyl" herein refers to
--O--C(O)N(R.sub.51)(H) wherein R.sub.51 is as previously
defined.
[0030] The term "N,N-dialkylcarbamoyl" herein refers to
--O--C(O)N(R.sub.52)(R.sub.53) wherein R.sub.52 and R.sub.53 are as
previously defined.
[0031] The term "nitroso" herein refers to the group --NO and
"nitrosylated" refers to compounds that have been substituted
therewith.
[0032] The term "aryl" herein refers to a mono- or bicyclic
carbocyclic ring system having one or two aromatic rings including,
but not limited to, phenyl, naphthyl tetrahydronaphthyl indanyl,
indenyl, and the like. Aryl groups (including bicyclic aryl groups)
can be unsubstituted or substituted with one, two or three
substituents independently selected from loweralkyl haloalkyl,
alkoxy, amino, alkylamino, dialkylamino, hydroxy, halo, and nitro.
In addition, substituted aryl groups include tetrafluorophenyl and
pentafluorophenyl.
[0033] The term "arylalkyl" herein refers to a lower alkyl radical
to which is appended an aryl group. Representative arylalkyl groups
include benzyl phenylethyl, hydroxybenzyl, fluorobenzyl,
fluorophenylethyl and the like.
[0034] The term "arylthio" herein refers to R.sub.54S-- wherein
R.sub.54 is an aryl group.
[0035] The term "cycloalkyl" herein refers to an alicyclic group
comprising from 3 to 7 carbon atoms including, but not limited to,
cyclopropyl, cyclobutyl cyclopentyl, cyclohexyl and the like.
[0036] The term "bridged cycloalkyl" herein refers to two or more
cycloalkyl radicals fused via adjacent or non-adjacent carbon
atoms, including but not limited to adamantyl and
decahydronapthyl.
[0037] The terms "halogen" or "halo" herein refer to I, Br, Cl or
F. The term "haloalkyl" herein refers to a lower alkyl radical, as
defined above, bearing at least one halogen substituent, for
example, chloromethyl, fluoroethyl or trifluoromethyl and the
like.
[0038] The term "heteroaryl" herein refers to a mono- or bi-cyclic
ring system containing one or two aromatic rings and containing at
least one nitrogen, oxygen, or sulfur atom in an aromatic ring.
Heteroaryl groups (including bicyclic heteroaryl groups) can be
unsubstituted or substituted with one, two, or three substituents
independently selected from lower alkyl, haloalkyl, alkoxy, amino,
alkylamino, dialkylamino, hydroxy, halo and nitro. Examples of
heteroaryl groups include but are not limited to pyridine,
pyrazine, pyrimidine, pyridazine, pyrazole, triazole, thiazole,
isothiazole, benzothiazole, benzoxazole, thiadiazole, oxazole,
pyrrole, imidazole, and isoxazole.
[0039] The term "heterocyclic ring" herein refers to any 3-, 4,5-,
6, or 7-membered nonaromatic ring containing at least one nitrogen
atom which is bonded to an atom which is not part of the
heterocyclic ring. In addition, the heterocyclic ring may also
contain a one additional heteroatom which may be nitrogen, oxygen,
or sulfur.
[0040] The term "heterocyclic compounds" herein refers to mono and
polycyclic compounds containing at least one heteroaryl or
heterocyclic ring.
[0041] Compounds of the invention which have one or more asymmetric
carbon atoms may exist as the optically pure enantiomers, pure
diastereomers, mixtures of enantiomers, mixtures of diastereomers,
racemic mixtures of enantiomers, diastereomeric racemates or
mixtures of diastereomeric racemates. It is to be understood that
the present invention anticipates and includes within its scope all
such isomers and mixtures thereof.
[0042] The NSAID used in the compositions of the invention can be
any of those known to the art, including those exemplified
below.
[0043] First, despite the introduction of many new drugs, aspirin
(acetylsalicylic acid) is still the most widely prescribed
antiinflammatory, analgesic and antipyretic agent and is a standard
for the comparison and evaluation of all other NSAIDs. Salicylic
acid itself is so irritating that it can only be used externally.
However, derivatives, particularly salicylate esters and salts,
have been prepared which provide ingestible forms of the
salicylates which have the desired antiinflammatory and other
properties. In addition to aspirin which is the acetate ester of
salicylic acid, are the diflurophenyl derivative (diflunisal) and
salicylsalicylic acid (salsalate). Also available are the salts of
salicylic acid, principally sodium salicylate. Sodium salicylate
and aspirin are the two most commonly used preparations for
systemic treatment. Other salicylates include salicylamide, sodium
thiosalicylate, choline salicylate and magnesium salicylate. Also
available are combinations of choline and magnesium salicylates.
Also contemplated are 5-aminosalicylic acid (mesalamine),
salicylazosulfapyridine (sulfasalazine) and methylsalicylate.
[0044] Another group of NSAID drugs included are the pyrazolon
derivatives. Included in his group are, for example,
phenylbutazone, oxyphenbutazone, antipyrine, aminopyrine, dipyrone
and apazone (azapropazone).
[0045] Another group of such NSAIDs are the para-aminophenol
derivatives. These are the so-called "coal tar" analgesics and
include phenacetin and its active metabolite acetaminophen.
[0046] Another group of compounds contemplated include
indomethacin, a methylated indole derivative, and the structurally
related compound, sulindac.
[0047] Also contemplated is a group of compounds referred to as the
fenamates which are derivatives of N-phenylanthranilic acid. The
most well known of these compounds are mefenamic, meclofenamic,
flufenamic, tolfenamic and etofenamic acids. They are used either
as the acid or as pharmaceutically acceptable salts.
[0048] Another contemplated NSAID is tolmetin which, like the other
NSAIDs discussed herein, causes gastric erosion and prolonged
bleeding time.
[0049] Another group of NSAID compounds are the propionic acid
derivatives. Principal members of this group are ibuprofen,
naproxen, flurbiprofen, fenoprofen and ketoprofen. Other members of
this group, in use or study in countries outside the U.S., include
fenbufen, pirprofen, oxaprozin, indoprofen and tiaprofenic
acid.
[0050] Also contemplated are piroxicam and amperoxicam, oxicam
derivatives which are a class of antiinflammatory enolic acids. The
other related compounds tenoxicam and tenidap are also
contemplated. Another compound that is particularly contemplated is
diclophenac, one of the series of phenylacetic acid derivatives
that have been developed as antiinflammatory agents. Other NSAIDs
which are contemplated as suitable in the compositions of the
invention include etodolac and nabumentone.
[0051] Each of the above contemplated NSAIDs is described more
fully in the literature, such as in Goodman and Gilman, The
Pharmacological Basis of Therapeutics (8th Edition), McGraw-Hill,
1993, Pgs. 638-381.
[0052] The compositions of the invention can also include NSAIDs
which have been nitrosylated through sites such as oxygen (hydroxyl
condensation), sulfur (sulfhydryl condensation), carbon and
nitrogen, including those specifically discussed below and in the
working examples that follow.
[0053] One embodiment of this aspect includes nitroso substituted
compounds of the formula: 1
[0054] wherein
[0055] D is selected from (i) a covalent bond; (ii)
--C(R.sub.a)--O--C(O)--Y-[C(R.sub.b)(R.sub.c)].sub.p-T- in which
R.sub.a is lower alkyl cycloalkyl aryl or heteroalkyl, Y is oxygen,
sulfur, or NR.sub.i in which R.sub.i is hydrogen or lower alkyl,
R.sub.b and R.sub.c are independently selected from, hydrogen,
lower alkyl cycloalkyl, aryl heteroaryl, arylalkyl alkylamino,
dialkylamino or taken together are cycloalkyl or bridged
cycloalkyl, p is an integer from 1 to 6 and T is a covalent bond,
oxygen, sulfur, or nitrogen; or (iii)-(CO)-T.sub.1-[C(R.su-
b.b)(R.sub.c)].sub.p-T.sub.2- wherein T.sub.1 and T.sub.2 are
independently selected from T, and wherein R.sub.b, R.sub.c, p and
T are as defined above;
[0056] Z is an aryl or heteroaryl; and
[0057] A.sub.1, A.sub.2 and A.sub.3 comprise the other subunits of
a 5- or 6-membered monocyclic aromatic ring and each is
independently selected from (1) C--R.sub.1 wherein R.sub.1 at each
occurrence is independently selected from hydrogen, lower alkyl
lower haloalkyl alkoxyalkyl, halogen or nitro; (2) N--R.sub.d
wherein R.sub.d at each occurrence is independently selected from a
covalent bond to an adjacent ring atom in order to render the ring
aromatic, hydrogen, lower alkyl, cycloalkyl, arylalkyl, aryl,
heteroaryl; (3) sulfur; (4) oxygen; and (5) B.sub.a=B.sub.b wherein
B.sub.a and B.sub.b are each independently selected from nitrogen
or C--R.sub.1 wherein at each occurrence R.sub.1 is as defamed
above.
[0058] Another embodiment of this aspect is nitroso substituted
compounds of the formula: 2
[0059] wherein
[0060] R.sub.b, R.sub.c, D, Z, A.sub.1, A.sub.2 and A.sub.3 are
defined as above.
[0061] Another embodiment is compounds of the formula: 3
[0062] wherein
[0063] R.sub.e is hydrogen or lower alkyl;
[0064] R.sub.f is selected from 4567
[0065] in which n is 0 or 1; and
[0066] X is (1)
--Y-[C(R.sub.b)(R.sub.c)].sub.p-G-[C(R.sub.b)(R.sub.c)].su-
b.p-T-NO, wherein G is (i) a covalent bond; (ii)-T-C(O)--; (iii)
--C(O)-T; (iv) --C(Y--C(O)--R.sub.m)-- wherein R.sub.m is
heteroaryl or heterocyclic ring; and in which Y, R.sub.b, R.sub.c,
p and T are as defined above; or (2) 8
[0067] in which W is a heterocyclic ring or NR.sub.hR.sub.i wherein
R.sub.h and R.sub.i are independently selected from lower alkyl,
aryl or alkenyl.
[0068] Another embodiment of this aspect is compounds of the
formula: 9
[0069] wherein
[0070] R.sub.g is selected from 1011
[0071] and X is defined as above.
[0072] The present invention also relates to processes for
preparing the compounds of formula (I), (II), (I) or (IV) and to
the intermediates useful in such processes.
[0073] Compounds of the present invention may be synthesized as
shown in reaction Schemes I through XI presented below, in which
R.sub.a, R.sub.b, R.sub.c, R.sub.d, R.sub.e, R.sub.f, R.sub.g,
A.sub.1, A.sub.2, A.sub.3, p, and Z are as defined above or as
depicted in the reaction schemes for formulas I, II, III or IV;
P.sup.1 is an oxygen protecting group and P.sup.2 is a sulfur
protecting group. The reactions are performed in solvents
appropriate to the reagents and materials employed are suitable for
the transformations being effected. It is understood by those
skilled in the art of organic synthesis that the functionality
present in the molecule must be consistent with the chemical
transformation proposed. This will, on occasion, necessitate
judgment by the routineer as to the order of synthetic steps,
protecting groups required, and deprotection conditions.
Substituents on the starting materials may be incompatible with
some of the reaction conditions required in some of the methods
described, bit alternative methods and substituents compatible with
the reaction conditions will be readily apparent to skilled
practitioners in the art. The use of sulfur and oxygen protecting
groups is well known in the art for protecting thiol, alcohol, and
amino groups against undesirable reactions during a synthetic
procedure and many such protecting groups are known, c.f., T. H.
Greene and P. G. M. Wuts, Protective Groups in Organic Synthesis,
John Wiley & Sons, New York (1991).
[0074] Nitroso compounds of formula (I) wherein A.sub.1, A.sub.2,
A.sub.3, R.sub.a, and Z are defined as above and an O-nitrosyated
enol is represenatative of the D group as defined above may be
prepared according to reaction Scheme I. The enolic form of the
.beta.-keto amide of the formula 1 is reacted with a suitable
nitrosylating agent such as thionyl chloride nitrite, thionyl
dinitrite [c.f., Hakimelahi et al., Helvetica Chimica Acta, 67, 967
(1984)], or nitrosium tetrafluoroborate in a suitable anhydrous
solvent such as methylene chloride, tetrahydrofuran (THF),
dimethylforamide (DMF), or acetonitrile with or without am amine
base such as pyridine or triethylamine to afford the O-nitrite IA.
12
[0075] Nitroso compounds of formula (I) wherein p, A.sub.1,
A.sub.2, A.sub.3, R.sub.a, R.sub.b, R.sub.c, and Z are defined as
above and an O-nitrosylated ester is representative of the D group
as defined above may be prepared according to Scheme II. The enolic
form of the b-keto amide of the formula 1 is converted to the ester
of the formula 2 wherein p, R.sub.b and R.sub.c are defined as
above by reaction with an appropriate protected alcohol containing
activated acylating agent wherein P.sub.1 is as defined above.
Preferred methods for the formation of enol ester are reacting the
enol with the preformed acid chloride or symmetrical anhydride of
the protected alcohol containing acid. Preferred protecting groups
for the alcohol moiety are silyl ethers such as a trimethylsilyl or
a tert-butyldimethylsilyl ether. Deprotection of the hydroxyl
moiety (fluoride ion is the preferred method for removing silyl
ether protecting groups) followed by reaction a suitable
nitrosylating agent such as thionyl chloride nitrite, thionyl
dinitrite, or nitrosium tetrafluoroborate in a suitable anhydrous
solvent such as dichloromethane, THF, DMF, or acetonitrile with or
without an amine base such as pyridine or triethylamine affords the
compound of the formula IB. 13
[0076] Nitroso compounds of formula (I) wherein p, A.sub.1,
A.sub.2, A.sub.3, R.sub.a, R.sub.b, R.sub.c, and Z are defined as
above and an 5-nitrosyated enol ester is representative of the D
group as defined above may be prepared according to reaction Scheme
III. The enolic form of the b-keto amide of the formula 1 is
converted to the ester of the formula 3 wherein p, R.sub.b and
R.sub.c are defined as above by reaction with an appropriate
protected thiol containing activated acylating agent wherein
P.sup.2 is as defined above. Preferred methods for the formation of
enol ester are reacting the enol with the preformed acid chloride
or symmetrical anhydride of the protected thiol containing acid.
Preferred protecting groups for the thiol moiety are as a thioester
such as a thioacetate or thiobenzoate, as a disulfide, as a
thiocarbamate such as N-methoxymethyl thiocarbamate, or as a
thioether such as a paramethoxybenzyl thioether, a
tetrahydropyranyl thioether or a S-triphenylmethyl thioether.
Deprotection of the thiol moiety (zinc in dilute aqueous acid,
triphenylphosphine in water and sodium borohydride are preferred
methods for reducing disulfide groups while aqueous base is
typically utilized to hydrolyze thioesters and N-methoxymethyl
thiocarbamates and mercuric trifluoroacetate, silver nitrate, or
strong acids such as trifluoroacetic or hydrochloric acid and heat
are used to remove a paramethoxybenzyl thioether, a
tetrahydropyranyl thioether or a S-triphenylmethyl thioether group)
followed by reaction with a suitable nitrosylating agent such as
thionyl chloride nitrite, thionyl dinitrite, a lower alkyl nitrite
such as tert-butyl nitrite, or nitrosium tetrafluoroborate in a
suitable anhydrous solvent such as methyene chloride, THF, DMF, or
acetonitrile with or without an amine base such as pyridine or
triethylamine affords the compound of the formula IC.
Alternatively, reacting this intermediate with a stoichiometric
quantity of sodium nitrite in aqueous acid affords the compound of
the formula IC. 14
[0077] Nitroso compounds of formula (II) wherein p, A.sub.1,
A.sub.2, A.sub.3, R.sub.b and R.sub.c, and Z are defined as above
and an O-nitrosylated ester is representative of the D group as
defined above may be prepared according to Scheme IV. The enolic
form of the .beta.-keto amide of the formula 4 is converted to the
ester of the formula 5 wherein p, R.sub.b and R.sub.c are defined
as above by reaction with an appropriate protected alcohol
containing activated acylating agent wherein P.sup.1 is as defined
above. Preferred methods for the formation of enol ester are
reacting the enol with the preformed acid chloride or symmetrical
anhydride of the protected alcohol containing acid. Preferred
protecting groups for the alcohol moiety are silyl ethers such as a
trimethylsilyl or a tert-butyldimethylsilyl ether. Deprotection of
the hydroxyl moiety (fluoride ion is the preferred method for
removing silyl ether protecting groups) followed by reaction a
suitable nitrosylating agent such as thionyl chloride nitrite,
thionyl dinitrite, or nitrosium tetrafluoroborate in a suitable
anhydrous solvent such as dichloromethane, THF, DMF, or
acetonitrile with or without an amine base such as pyridine or
triethylamine affords the compound of the formula IIA. 15
[0078] Nitroso compounds of formula (II) wherein p, A.sub.1,
A.sub.2, A.sub.3, R.sub.b, R.sub.c, and Z are defined as above and
an S-nitrosyated enol ester is represenatative of the D group as
defined above may be prepared according to reaction Scheme V. The
enolic form of the .beta.-keto amide of the formula 4 is converted
to the ester of the formula 6 wherein p, R.sub.b and R.sub.c are
defined as above by reaction with an appropriate protected thiol
containing activated acylating agent wherein P.sup.2 is as defined
above. Preferred methods for the formation of enol ester are
reacting the enol with the preformed acid chloride or symmetrical
anhydride of the protected thiol containing acid. Preferred
protecting groups for the thiol moiety are as a thioester such as a
thioacetate or thiobenzoate, as a disulfide, as a thiocarbamate
such as N-methoxymethyl thiocarbamate, or as a thioether such as a
paramethoxybenzyl thioether, a tetrahydropyranyl thioether, or a
S-triphenylmethyl thioether. Deprotection of the thiol moiety (zinc
in dilute aqueous acid, triphenylphosphine in water and sodium
borohydride are preferred methods for reducing disulfide groups
while aqueous base is typically utilized to hydrolyze thiolesters
and N-methoxymethyl thiocarbamates and mercuric trifluoroacetate,
silver nitrate, or strong acids such as trifluoroacetic or
hydrochloric acid and heat are used to remove a paramethoxybenzyl
thioether, a tetrahydropyranyl thioether or a S-triphenylmethyl
thioether group) followed by reaction a suitable nitrosylating
agent such as thionyl chloride nitrite, thionyl dinitrite, a lower
alkyl nitrite such as tert-butyl nitrite, or nitrosium
tetrafluoroborate in a suitable anhydrous solvent such as methyene
chloride, THF, DMF, or acetonitrile with or without an amine base
such as pyridine or triethylamine acid affords the compound of the
formula IIB. Alternatively, reacting this intermediate with a
stiochiometric quantity of sodium nitrite in aqueous acid affords
the compound of the formula IIB. 16
[0079] Nitroso compounds of formula (III) wherein p, R.sub.b,
R.sub.c, R.sub.e and R.sub.f are defined as above and an
O-nitrosylated ester is representative of the X group as defined
above may be prepared according to Scheme VI. An acid of the
formula 7 is converted into the ester of the formula 8 wherein p,
R.sub.b and R.sub.c are defined as above by reaction with an
appropriate monoprotected diol. Preferred methods for the
preparation of esters are initially forming the mixed anhydride via
reaction of 7 with a chloroformate such as isobutylchloroformate in
the presence of a non nucleophilic base such as triethylamine in an
anhydrous inert solvent such as dichloromethane, diethylether, or
THF. The mixed anhydride is then reacted with the monoprotected
alcohol preferably in the presence of a condensation Analyst such
as 4-dimethylamine pyridine. Alternatively, the acid 7 may be first
converted to the acid chloride by treatment with oxalyl chloride in
the presence of a catalytic amount of DMF. The acid chloride is
then reacted with the monoprotected alcohol preferably in the
presence of a condensation catalyst such as 4-dimethylamine
pyridine and a tertiary amine base such as triethyl amine to afford
the ester 8. Alternatively, the acid 7 and monoprotected diol may
be coupled to afford 8 by treatment with a dehydration agent such
as DCC. Alternatively, compound 7 may be first converted into an
alkali metal salt such as the sodium, potassium, or lithium salt,
and reacted with an alkyl halide which also contains a protected
hydroxyl group in an polar solvent such as DMF to afford 8.
Preferred protecting groups for the alcohol moiety are silyl ethers
such as a trimethylsilyl or a tert-butyldimethylsilyl ether.
Deprotection of the hydroxyl moiety (fluoride ion is the preferred
method for removing silyl ether protecting groups) followed by
reaction with a suitable nitrosylating agent such as thionyl
chloride nitrite, thionyl dinitrite, or nitrosium tetrafluoroborate
in a suitable anhydrous solvent such as dichloromethane, THF, DMF,
or acetonitrile with or without an amine base such as pyridine or
triethylamine affords the compound of the formula IIIA. 17
[0080] Nitroso compounds of formula (III) wherein p, R.sub.b,
R.sub.c, R.sub.e, and R.sub.f are defined as above and a
S-nitrosylated ester is representative of the X group as defined
above may be prepared according to Scheme VII. An acid of the
formula 7 is converted into the ester of the formula 9 by reaction
with an appropriate protected thiol containing alcohol. Preferred
methods for the preparation of esters are initially forming the
mixed anhydride via reaction of 7 with a chloroformate such as
isobutylchloroformate in the presence of a non nucleophilic base
such as triethylamine in an anhydrous inert solvent such as
diethylether or THF. The mixed anhydride is then reacted with the
thiol containing alcohol preferably in the presence of a
condensation catalyst such as 4-dimethylamine pyridine.
Alternatively, the acid 7 may be first converted to the acid
chloride by treatment with oxalyl chloride in the presence of a
catalytic amount of DMF. The acid chloride is then reacted with the
monoprotected thiol preferably in the presence of a condensation
catalyst such as 4-dimethylamine pyridine and a tertiary amine base
such as triethyl amine to afford the ester 9. Alternatively, the
acid and thiol containing alcohol may be coupled to afford 9 by
treatment with a dehydration agent such as DCC. Alternatively,
compound 7 may be first converted: into an alkali metal salt such
as the sodium, potassium, or lithium salt, and reacted with an
alkyl halide which also contains a protected thiol group in an
polar solvent such as DMF to afford 9. Preferred protecting groups
for the thiol moiety are as a thioester such as a thioacetate or
thiobenzoate, as a disulfide, as a thiocarbamate such as
N-methoxymethyl thiocarbamate, or as a thioether such as a
paramethoxybenzyl thioether, a tetrahydropyranyl thioether, or a
S-triphenylmethyl thioether. Deprotection of the thiol moiety (zinc
in dilute aqueous acid, triphenylphosphine in water and sodium
borohydride are preferred methods for reducing disulfide groups
while aqueous base is typically utilized to hydrolyze thiolesters
and N-methoxymethyl thiocarbamates and mercuric trifluoroacetate,
silver nitrate, or strong acids such as trifluoroacetic or
hydrochloric acid and heat are used to remove a paramethoxybenzyl
thioether, a tetrahydropyranyl thioether or a S-triphenylmethyl
thioether group) followed by reaction with a suitable nitrosylating
agent such as thionyl chloride nitrite, thionyl dinitrite, a lower
alkyl nitrite such as tert-butyl nitrite, or nitrosium
tetrafluoroborate in a suitable anhydrous solvent such as
methylene, chloride; THF, DMF, or acetonitrile with or without an
amine base such as pyridine or triethylamine affords the compound
of the formula IIIB. Alternatively, this intermediate may be
reacted with a stoichiometric quantity of sodium nitrite in aqueous
acid affords the compound of the formula IIIB. 18
[0081] Nitroso compounds of formula (III) wherein W, R.sub.e, and
R.sub.f are defined as above and a 6-W-substituted sydnonimine
wherein W is as defined above is representative of the X group as
defined above may be prepared according to Scheme VIII. An acid of
the formula 7 is converted into the carboximide of the formula IIIC
by reaction with a 6-W-substituted sydnonimine. Preferred methods
for the preparation of carboximides are initially forming the mixed
anhydride via reaction of 7 with a chloroformate such as
isobutylchloroformate in the presence of a non nucleophilic base
such as triethylamine in an anhydrous inert solvent such as
diethylether or THF. The mixed anhydride is then reacted with the
6-W-substituted sydnonimine to afford IIIC. Alternatively, the acid
7 may be coupled to the 6-W-substituted sydnonimine to afford IIIC
by treatment with a dehydration agent such as DCC. Alternatively,
the acid 7 may be converted into an active ester by reaction with a
suitably substituted phenol utilizing any of the conditions for
ester formation described for Scheme VI, followed by reaction with
a 6-W-substituted sydnonimine. Preferred 6-W-substituted
sydnonimines are 1,2,6,4-oxatriazolium, 6-amino-6 morpholine and
1,2,6,4-oxatriazolium, 6-(6-chloro-2-methyl-benz- ene) and
preferred active esters are para-nitrophenyl,
2,4,5-trichlorophenyl, and pentafluorophenyl. 19
[0082] Nitroso compounds of formula (IV) wherein p, R.sub.b,
R.sub.c, and R.sub.g are defined as above and an O-nitrosylated
ester is representative of the X group as defined above may be
prepared according to Scheme IX. An acid of the formula 10 is
converted into the ester of the formula 11 wherein p, R.sub.b, and
R.sub.c are defined as above, by reaction with an appropriate
monoprotected diol. Preferred methods for the preparation of esters
are initially forming the mixed anhydride via reaction of 10 with a
chloroformate such as isobutylchloroformate in the presence of a
non nucleophilic base such as triethylamine in an anhydrous inert
solvent such as dichloromethane, diethylether or THF. The mixed
anhydride is then reacted with the monoprotected alcohol preferably
in the presence of a condensation catalyst such as 4-dimethylamine
pyridine. Alternatively, the acid 10 may be first converted to the
acid chloride by treatment with oxalyl chloride in the presence of
a catalytic amount of DMF. The acid chloride is then reacted with
the monoprotected alcohol preferably in the presence of a
condensation catalyst such as 4-dimethylamine pyridine and a
tertiary amine base such as triethylamine to afford the ester 11.
Alternatively, the acid 10 and monoprotected diol may be coupled to
afford 11 by treatment with a dehydration agent such as DCC.
Alternatively, compound 10 may be first converted into an alkali
metal salt such as the sodium, potassium, or lithium salt, which is
then reacted with an alkyl halide which also contains a protected
hydroxyl group in an polar solvent such as DMF to afford 11.
Preferred protecting groups for the alcohol moiety are silyl ethers
such as a trimethylsilyl or a tert-butyldimethylsilyl ether.
Deprotection of the hydroxyl moiety (fluoride ion is the preferred
method for removing silyl ether protecting groups) followed by
reaction with a suitable nitrosylating agent such as thionyl
chloride nitrite, thionyl dinitrite, or nitrosium tetrafluoroborate
in a suitable anhydrous solvent such as methylene chloride, THF,
DMF, or acetonitrile with or without an amine base such as pyridine
or triethyl amine affords the compound of the formula IVA. 20
[0083] Nitroso compounds of formula (IV) wherein R.sub.g is defined
as above and a S-nitrosylated ester is representative of the X
group as defined above may be prepared according to Scheme X. An
acid of the formula 10 is converted into the ester of the formula
12 by reaction with an appropriate protected thiol containing
alcohol. Preferred methods for the preparation of esters are
initially forming the mixed anhydride via reaction of 10 with a
chloroformate such as isobutylchloroformate in the presence of a
non nucleophilic base such as triethylamine in an anhydrous inert
solvent such as diethylether or THF. The mixed anhydride is then
reacted with the protected thiol containing alcohol preferably in
the presence of a condensation catalyst such as 4-dimethylamine
pyridine. Alternatively, the acid 10 may be first converted to the
acid chloride by treatment with oxalyl chloride in the presence of
a catalytic amount of DMF. The acid chloride is then reacted with
the protected thiol containing alcohol preferably in the presence
of a condensation catalyst such as 4-dimethylamine pyridine and a
tertiary amine base such as triethyl amine to afford the ester 12.
Alternatively, the acid and protected thiol containing alcohol may
be coupled to afford 12 by treatment with a dehydration agent such
as DCC. Alternatively, compound 10 may be first converted into an
alkali metal salt such as the sodium, potassium, or lithium salt,
which is then reacted with an alkyl halide which also contains a
protected thiol group in an polar solvent such as DMF to afford 12
Preferred protecting groups for the thiol moiety are as a thioester
such as a thioacetate or thiobenzoate, as a disulfide, as a
thiocarbamate such as N-methoxymethyl thiocarbamate, or as a
thioether such as a paramethoxybenzyl thioether, a
tetrahydropyranyl thioether, or a S-triphenylmethyl thioether.
Deprotection of the thiol moiety (zinc in dilute aqueous acid,
triphenylphosphine in water and sodium borohydride are preferred
methods for reducing disulfide groups while aqueous base is
typically utilized to hydrolyze thiolesters and N-methoxymethyl
thiocarbamates and mercuric trifluoroacetate, silver nitrate, or
strong acids such as trifluoroacetic or hydrochloric acid and heat
are used to remove a paramethoxybenzyl thioether, a
tetrahydropyranyl thioether or a S-triphenylmethyl thioether group)
followed by reaction with a suitable nitrosylating agent such as
thionyl chloride nitrite, thionyl dinitrite, a lower alkyl nitrite
such as tert-butyl nitrite, or nitrosium tetrafluoroborate in a
suitable anhydrous solvent such as methylene chloride, THF, DMF, or
acetonitrile affords the compound of the formula IVB.
Alternatively, this intermediate may be reacted with a
stoichiometric quantity of sodium nitrite in aqueous acid affords
the compound of the formula IVB 21
[0084] Nitroso compounds of formula (IV) wherein R.sub.g is defined
as above and a 6-substituted sydnonimine is representative of the X
group as defined above may be prepared according to Scheme XI. An
acid of the formula 10 is converted into the carboximide of the
formula IVC by reaction with a 6-W-substituted sydnonimine wherein
W is as defined above. Preferred methods for the preparation of
carboximides are initially forming the mixed anhydride via reaction
of 10 with a chloroformate such as isobutylchloroformate in the
presence of a non nucleophilic base such as triethylamine in an
anhydrous inert solvent such as diethylether or THF. The mixed
anhydride is then reacted with the 6-W-substituted sydnonimine to
afford IVC. Alternatively, the acid 10 may-be coupled to the
6-W-substituted sydnonimine afford IVC by treatment with a
dehydration agent such as DCC. Alternatively, the acid 10 may be
converted into an active ester by reaction with a suitably
substituted phenol utilizing any of the conditions for ester
formation described above, followed by reaction with a
6-W-substituted sydnonimine. Preferred 6-W-substituted sydnonimines
are 1,2,6,4-oxatriazolium, 6-amino-6-morpholine and 1,2,6,4
oxatriazolium, 6-amino 6-(6-chloro-2-methyl-benzene) and preferred
active esters are para-nitrophenyl, 2,4,5-trichlorophenyl, and
pentafluorophenyl. 22
[0085] The compounds that donate, transfer or release nitric oxide
can be any of those known to the art, including those mentioned
and/or exemplified below.
[0086] Nitrogen monoxide can exist in three forms: NO.sup.-
(nitroxyl), NO. (nitric oxide) and NO.sup.+ (nitrosonium). NO. is a
highly reactive short-lived species that is potentially toxic to
cells. This is critical, because the pharmacological efficacy of NO
depends upon the form in which it is delivered. In contrast to
nitric oxide radical, nitrosonium and, nitroxyl do not react with
O.sub.2 or O.sub.2.sup.-. species, and are also resistant to
decomposition in the presence of redox metals. Consequently,
administration of NO equivalents does not result in the generation
of toxic by-products or the elimination of the active NO
moiety.
[0087] Compounds contemplated for use in the invention are nitric
oxide and compounds that release nitric oxide or otherwise directly
or indirectly deliver or transfer nitric oxide to a site of its
activity, such as on a cell membrane, in vivo. As used here, the
term `nitric oxide` encompasses uncharged nitric oxide (NO.) and
charged nitric oxide species, particularly including nitrosonium
ion (NO.sup.+) and nitroxyl ion (NO.sup.-). The reactive form of
nitric oxide can be provided by gaseous nitric oxide. The nitric
oxide releasing, delivering or transferring compounds, having the
structure F--NO wherein F is a nitric oxide releasing, delivering
or transferring moiety, include any and all such compounds which
provide nitric oxide to its intended site of action in a form
active for their intended purpose. As used here, the term "NO
adducts" encompasses any of such nitric oxide releasing, delivering
or transferring compounds, including, for example, S-nitrosothiols,
S-nitroso amino acids, S-nitroso-polypeptides, and organic
nitrites. It is contemplated that any or all of these "NO adducts"
can be mono- or poly-nitrosylated at a variety of naturally
susceptible or artificially provided binding sites for nitric
oxide.
[0088] One group of such NO adducts is the S-nitrosothiols, which
are compounds that include at least one --S--NO group. Such
compounds include S-nitroso-polypeptides (the term "polypeptide"
includes proteins and also polyamino acids that do not possess an
ascertained biological function, and derivatives thereof);
S-nitrosylated amino acids (including natural and synthetic amino
acids and their stereoisomers and racemic mixtures and derivatives
thereof); S-nitrosated sugars, S-nitrosated-modified and unmodified
oligonucleotides (preferably of at least 5, and more particularly
5-200, nucleotides); and an S-nitrosated hydrocarbon where the
hydrocarbon can be a branched or unbranched, and saturated or
unsaturated aliphatic hydrocarbon, or an aromatic hydrocarbon;
S-nitroso hydrocarbons having one or more substituent groups in
addition to the S-nitroso group; and heterocyclic compounds.
S-nitrosothiols and the methods for preparing them are described in
U.S. patent application Ser. No. 07/943,834, filed Sep. 14, 1992,
Oae et al., Org. Prep. Proc. a Int., 15(3):165-198. (1983);
Loscalzo et al., J. Pharmacol. Exp. Ther., 249(3):726-729 (1989)
and Kowaluk et al., J. Pharmacol. Exp. Ther., 256:1256-1264 (1990),
all of which are incorporated in their entirety by reference.
[0089] One particularly preferred embodiment of this aspect relates
to S-nitroso amino acids where the nitroso group is linked to a
sulfur group of a sulfur-containing amino acid or derivative
thereof. For example, such compounds include the following:
S-nitroso-N-acetylcysteine, S-nitroso-N-acetylpenicillamine,
S-nitroso-homocysteine, S-nitroso-cysteine and
S-nitroso-glutathione.
[0090] Suitable S-nitrosylated proteins include thiol-containing
proteins (where the NO group is attached to one or more sulfur
group on an amino acid or amino acid derivative thereof) from
various functional classes including enzymes, such as tissue-type
plasminogen activator(TPA) and cathepsin B; transport proteins,
such as lipoproteins, heme proteins such as hemoglobin and serum
albumin; and biologically protective proteins, such as the
immunoglobulins and the cytokines. Such nitrosylated proteins are
described in PCT Publ. Applic. No. WO 93/09806, published May 27,
1993. Examples include polynitrosylated albumin where multiple
thiol or other nucleophilic centers in the protein are
modified.
[0091] Further examples of suitable S-nitrosothiols include those
having the structures:
[0092] (i) CH3[C(R.sub.b)(R.sub.c)].sub.xSNO
[0093] wherein x equals 2 to 20 and R.sub.b and R.sub.c are as
defined above;
[0094] (ii) HS[C(R.sub.b)(R.sub.c)].sub.xSNO
[0095] wherein x equals 2 to 20; and
[0096] (iii) ONS[C(R.sub.b)(R.sub.c)].sub.xQ
[0097] wherein x equals 2 to 20 and Q is selected from the group
consisting of fluoro, alkoxy, cyano, carboxamido, cycloalkyl
arylkoxy, alkylsulfinyl arylthio, alkylamino, dialkylamino,
hydroxy, carbamoyl, N-alkylcarbamoyl, N,N-dialkylcarbamoyl, amino,
hydroxyl, carboxyl, hydrogen, nitro and aryl; and x, R.sub.b and
R.sub.c are as defined above.
[0098] Nitrosothiols can be prepared by various methods of
synthesis. In general, the thiol precursor is prepared first, then
converted to the S-nitrosothiol derivative by nitrosation of the
thiol group with NaNO.sub.2 under acidic conditions (pH is about
2.5) which yields the S-nitroso derivative. Acids which may be used
for this purpose include aqueous sulfuric, acetic and hydrochloric
acids. Alternatively, they may be nitrosated by reaction with an
organic nitrite such as tert-butyl nitrite, or an nitrosonium salt
such as nitrosonium tetraflurorborate in an inert solvent.
[0099] Another group of such NO adducts are those wherein the
compounds donate, transfer or release nitric oxide and are selected
from the group consisting of compounds that include at least one
ON--O--, ON--N-- or ON--C-- group. The compound that includes at
least one ON--O--, ON--N-- or ON--C-- group is preferably selected
from the group consisting of ON--O--, ON--N-- or ON--C-polypeptides
(the term "polypeptide" includes proteins and also polyamino acids
that do not possess an ascertained biological function, and
derivatives thereof); ON--O--, ON--N-- or ON--C-amino acids
(including natural and synthetic amino acids and their
stereoisomers and racemic mixtures); ON--O--, ON--N-- or ON-sugars;
ON--O--, ON--N-- or ON--C-modified and unmodified oligonucleotides
(preferably of at least 5, and more particularly 5-200,
nucleotides), ON--O--, ON--N-- or ON--C-hydrocarbons which can be
branched or unbranched, saturated or unsaturated aliphatic
hydrocarbons or aromatic hydrocarbons; ON--O--, ON--N-- or ON--C--
hydrocarbons having one or more subtstituent groups in addition to
the ON--O--, ON--N-- or ON--C-- group; and ON--O--, ON--N-- or
ON--C-heterocyclic compounds.
[0100] Another group of such adducts are N-oxo-N-nitrosoamines
which donate, transfer or release nitric oxide and have a
R.sub.1R.sub.2--N(O-M.sup.+)-NO group wherein R.sub.1 and R.sub.2
include polypeptides, amino ids, sugars, modified and unmodified
oligonucleotides, hydrocarbons where the hydrocarbon can be a
branched or unbranched, and saturated or unsaturated aliphatic
hydrocarbon or an aromatic hydrocarbon, hydrocarbons having one or
more substituent groups and heterocyclic compounds. M+is a metal
cation, such as, for example, a Group I metal cation.
[0101] Another group of such adducts are thionitrates which donate,
transfer or release nitric oxide and have the structure
R.sub.1--(S).sub.v--NO wherein v is an integer of at least 2.
R.sub.1 is as described above for the N-oxo-N-nitrosoamines.
Preferred are the dithiols wherein v is 2. Particularly preferred
are those compounds where R.sub.1 is a polypeptide or hydrocarbon
with a pair or pairs of thiols that are sufficiently structurally
proximate, i.e. vicinal, that the pair of thiols will be reduced to
a disulfide. Those compounds which form disulfide species release
nitroxyl ion (NO.sup.-) and uncharged nitric oxide (NO.). Those
compounds where the thiol groups are not sufficiently close to form
disulfide bridges generally only provide nitric oxide as the
NO.sup.- form but not as the uncharged NO. form.
[0102] Agents which stimulate endogenous NO synthesis such as
L-arginine, the substrate for nitric oxide synthase, are also
suitable for use in accordance with the invention.
[0103] When administered in vivo, the compositions may be
administered in combination with pharmaceutical carriers and in
dosages described herein.
[0104] The compositions of the present invention may be
administered orally, parenterally, by inhalation spray, rectally,
or topically in dosage unit formulations containing conventional
nontoxic pharmaceutically acceptable carriers, adjuvants, and
vehicles as desired. Topical administration may also involve the
use of transdermal administration such as transdermal patches or
iontophoresis devices. The term parenteral as used herein includes
subcutaneous injections, intravenous, intramuscular, intrasternal
injection, or infusion techniques.
[0105] Solid dosage forms for oral administration may include
capsules, tablets, pills, powders, granules and gels. In such solid
dosage forms, the active compounds may be admixed with at least one
inert diluent such as sucrose, lactose or starch. Such dosage forms
may also comprise, as in normal practice, additional substances
other than inert diluents, e.g., lubricating agents such as
magnesium stearate. In the case of capsules, tablets, and pills,
the dosage forms may also comprise buffering agents. Tablets and
pills can additionally be prepared with enteric coatings.
[0106] Dosage forms for topical administration of the composition
can include creams, sprays, lotions, gels, ointments and the like.
In such dosage forms the compositions of the invention can be mixed
to form white, smooth, homogeneous, opaque lotions with, for
example, benzyl alcohol 1% (wt/wt) as preservative, emulsifying
wax, glycerin, isopropyl palmitate, lactic acid, purified water,
sorbitol solution and polyethylene glycol 400. They can be mixed to
form a white, smooth, homogeneous, opaque creams with, for example,
benzyl alcohol 2% (wt/wt) as preservative, emulsifying wax,
glycerin, isopropyl palmitate, lactic acid, purified water, and
sorbitol solution. They can be mixed to form ointments with, for
example, benzyl alcohol 2% (wt/wt) as preservative, white
petrolatum, emulsifying wax, and tenox II (butylated
hydroxyanisole, propyl gallate, citric acid, propylene glycol).
Woven pads or rolls of bandaging material, e.g. gauge, can be
impregnated with the compositions in solution, lotion, cream,
ointment or other such form can also be used for topical
application. The compositions can also be applied topically using a
transdermal system, such as one of an acrylic-based polymer,
adhesive with a resinous crosslinking agent impregnated with the
composition and laminated to an impermeable backing.
[0107] Suppositories for rectal administration of the drug
composition, such as for treating pediatric fever etc., can be
prepared by mixing the drug with a suitable nonirritating excipient
such as cocoa butter and polyethylene glycols which are solid at
ordinary temperatures but liquid at the rectal temperature and will
therefore melt in the rectum and release the drug.
[0108] Injectable preparations, for example, sterile injectable
aqueous or oleaginous suspensions may be formulated according to
the known art using suitable dispersing or wetting agents and
suspending agents. The sterile injectable preparation may also be a
sterile injectable solution or suspension in a nontoxic
parenterally acceptable diluent or solvent, for example, as a
solution in 1,3-butanediol. Among the acceptable vehicles and
solvents that may be employed are water, Ringer's solution, and
isotonic sodium chloride solution. In additions sterile, fixed oils
are conventionally employed an a solvent or suspending medium.
[0109] While the compositions of the invention can be administered
as a mixture of an NSAID and a nitric oxide donor, they can also be
used in combination with one or more additional compounds which are
known to be effective against the specific disease state that one
is targeting for treatment.
[0110] The compositions of this invention can further include
conventional excipients, i.e., pharmaceutically acceptable organic
or inorganic carrier substances suitable for parenteral application
which do not deleteriously react with the active compounds.
Suitable pharmaceutically acceptable carriers include, but are not
limited to, water, salt solutions, alcohol vegetable oils,
polyethylene glycols, gelatin, lactose, amylose, magnesium
stearate, talc, silicic acid, viscous paraffin, perfume oil, fatty
acid monoglycerides and diglycerides, petroethral fatty acid
esters, hydroxymethylcellulose, polyvinylpyrrolidone, etc. The
pharmaceutical preparations can be sterilized and if desired mixed
with auxiliary agents, e.g., lubricants, preservatives,
stabilizers, wetting agents, emulsifiers, salts for influencing
osmotic pressure, buffers, colorings, flavoring and/or aromatic
substances and the like which do not deleteriously react with the
active compounds. For parenteral application, particularly suitable
vehicles consist of solutions, preferably oily or aqueous
solutions, as well as suspensions, emulsions, or implants. Aqueous
suspensions may contain substances which increase the viscosity of
the suspension and include, for example, sodium carboxymethyl
cellulose, sorbitol and/or dextran. Optionally, the suspension may
also contain stabilizers.
[0111] The composition, if desired, can also contain minor amounts
of wetting or emulsifying agents, or pH buffering agents. The
composition can be a liquid solution, suspension, emulsion, tablet,
pill, capsule, sustained release formulation, or powder. The
composition can be formulated as a suppository, with traditional
binders and carriers such as triglycerides. Oral formulations can
include standard carriers such as pharmaceutical grades of
mannitol, lactose, starch, magnesium stearate, sodium saccharine,
cellulose, magnesium carbonate, etc.
[0112] Various delivery systems are known and can be used to
administer a therapeutic compound or composition of the invention,
e.g., encapsulation in liposomes, microparticles, microcapsules and
the like.
[0113] The therapeutics of the invention can be formulated as
neutral or salt forms. Pharmaceutically acceptable salts include,
but are not limited to, those formed with free amino groups such as
those derived from hydrochloric, phosphoric, sulfuric, acetic,
oxalic, tartaric acids, etc., and those formed with free carboxyl
groups such as those derived from sodium, potassium, ammonium,
calcium, ferric hydroxides, isopropylamine, triethylamine,
2-ethylamino ethanol, histidine, procaine, etc.
[0114] The term "therapeutically effective amount," for the
purposes of the invention, refers to the amount of the nitric oxide
adduct which is effective to achieve its intended purpose. While
individual needs vary, determination of optimal ranges for
effective amounts of each nitric oxide adduct is within the skill
of the art. Generally, the dosage required to provide an effective
amount of the composition, and which can be adjusted by one of
ordinary skill in the art will vary, depending on the age, health,
physical condition, sex, weight, extent of disease of the
recipient, frequency of treatment and the nature and scope of the
disorder.
[0115] The amount of a given NSAID which will be effective in the
treatment of a particular disorder or condition will depend on the
nature of the disorder or condition, and can be determined by
standard clinical techniques. Reference is again made to Goodman
and Gilman, supra; The Physician's Desk Reference, Medical
Economics Company, Inc., Oradell, N.J., 1995; and to Drug Facts and
Comparisons, Facts and Comparisons, Inc., St. Louis, Mo., 1993. The
precise dose to be employed in the formulation will also depend on
the route of administration, and the seriousness of the disease or
disorder, and should be decided according to the judgment of the
practitioner and each patient's circumstances.
[0116] The amount of nitric oxide donor in a pharmaceutical
composition may be in amounts of 0.1-10 times the molar equivalent
of the NSAID. The usual daily doses of NSAIDs are 3-40 mg/kg body
weight and the doses of nitric oxide donors in the pharmaceutical
composition may be in amounts of 1-500 mg/kg body weight daily and
more usually about 1-50 mg/kg. Effective doses may be extrapolated
from dose-response curves derived from in vitro or animal model
test systems and are in the same ranges or less than as described
for the commercially available compounds in the Physician's Desk
Reference, supra.
[0117] The invention also provides a pharmaceutical pack or kit
comprising one or more containers filled with one or more of the
ingredients of the pharmaceutical compositions of the invention.
Associated with such container(s) can be a notice in the form
prescribed by a governmental agency regulating the manufacture, use
or sale of pharmaceuticals or biological products, which notice
reflects approval by the agency of manufacture, use or sale for
human administration.
[0118] The following non-limitative examples further describe and
enable one of ordinary skill in the art to make and use the
invention. Flash chromatography was performed on 40 micron silica
gel (Baker).
EXAMPLE 1
Cholest-5-en-3.beta.-O-nitroso alcohol
[0119] Cholesterol (0.242 g, 0.62 mmol) was dissolved in anhydrous
methylene chloride (3 mL) and pyridine (0.103 g, 3.45 mmol) was
added, followed by nitrosonium tetrafluoroborate (0.036 g, 0.31
mmol). After stirring for 1 hour at room temperature an additional
nitrosonium tetrafluoroborate (0.099 g, 0.85 mmol) was added. The
reaction mixture was stirred at room temperature for 16 hours. The
solvent was evaporated and the residue was purified by flash
chromatography on silica gel, deactivated with triethylamine,
eluted methylene chloride to give 0.165 g (64% yield) of the title
compound as a white solid. .sup.1H NMR (CDCl.sub.3), .delta.0.86
(d, 6H), 0.92 (d, 3H), 1.05-1.75 (m, 21H), 1.80-2.01 (m, 6H),
2.25-2.47 (m, 2H), 5.23 (m, 1H), 544 (m, 1H).
EXAMPLE 2
N--(N-L-.gamma.-glutamyl-S-Nitroso-L-cysteinyl)glycine
[0120] N--(N-L-.gamma.-glutamyl-L-cysteinyl)glycine (100 g, 0.325
mol) was dissolved in deoxygenated water (200 ml) and 2N HCl (162
ml) at room temperature and then the reaction mixture was cooled to
0.degree. C. With rapid stirring, a solution of sodium nitrite
(24.4 g, 0.35 mol) in water (40 ml) was added and stirring with
cooling of the reaction mixture was continued for approximately 1
hour after which time the pink precipitate which formed was
collected by vacuum filtration. The filter cake was resuspended in
chilled. 40% acetone-water (600 ml) and collected by vacumm
filtration. The filter cake was washed with acetone (2.times.200
ml) and ether (100 ml) and then dried under high vacuum at room
temperature in the dark to afford the title compound as a pink
powder. .sup.1H NMR (D.sub.2O) .delta.: 1.98 (m, 2H), 2.32 (t, 2H),
3.67 (t, 1H), 3.82 (s 2H), 3.86 (dd, 1H), 3.98 (dd, 1H), 4.53 (m,
1H).
EXAMPLE 3
S-Nitroso-triphenylmethanethiol
[0121] Triphenylmethyl mercaptan (0.050 g, 0.18 mmol) was dissolved
in anhydrous methylene chloride and cooled to 0.degree. C.
Tert-butyl nitrite (0.186 g, 1.80 mmol) was added and the resulting
mixture was stirred at 0.degree. C. for 30 min. The reaction
mixture was allowed to warm to room temperature and stirred at room
temperature for 1 hour. The solvent and excess of tert-butyl
nitrite were evaporated to give the title compound as a green solid
(0.054 g, 98%). .sup.1H NMR (CDCl.sub.3) .delta.: 7.13-7.18 (m,
4H), 7.25-7.39 (m, 11H).
EXAMPLE 4
4-O-Nitroso-1-(3-benzoyl-.alpha.-methylbenzeneacetic acid)butyl
ester
[0122] 4a. 4-Hydroxy-1-(3-benzoyl-.alpha.-methylbenzeneacetic
acid)butyl ester
[0123] 3-Benzoyl-.alpha.-methylbenzeneacetic acid (4 g, 16 mmol)
and 100 .mu.L DMF were dissolved in benzene (25 mL). Oxalyl
chloride (1.6 mL, 18 mmol) was added dropwise. Stirring was
continued for 2 hr before concentration to a syrup. Butanediol (9
mL, 100 mmol) and pyridine (1.67 mL, 21 mmol) were dissolved in
CH.sub.2Cl.sub.2 (100 mL) and dioxane (15 mL) and cooled to
0.degree. C. A solution of the acid chloride was added in
CH.sub.2Cl.sub.2 (20 mL). The reaction mixture was stirred cold for
20 min then warmed to room temperature with stirring for 2 hr. The
solution was washed 1.times.30 H.sub.2O, 1 N HCl, satd NaHCO.sub.3
and brine; dried over Na.sub.2SO.sub.4; and the volatiles were
evaporated. The residue was filtered through a pad of silica gel
eluting with 2:1 Hex:EtOAc to yield 4.8 g (91%) of hydroxy ester.
.sup.1H NMR (CDCl.sub.3) .delta. 7.41-7.81 (mult, 9H), 4.08-4.15
(mult, 2H), 3.79 (q, J=7.2 Hz, 1H), 3.59 (t, J=6.3 Hz, 2H),
1.53-1.69 (mult, 4H), 1.53 (d, J=7.2 Hz, 3H).
[0124] 4b. 4-O-Nitroso-1-(3-benzoyl-.alpha.-methylbenzeneacetic
acid)butyl ester
[0125] The product of Example 4a (1 g, 3.6 mmol) and pyridine (1.4
mL, 18 mmol) were dissolved in dichloromethane (15 mL) and cooled
to -78.degree. C. Nitrosonium tetrafluoroborate (840 mg, 7.2 mmol)
was added and the solution was kept, cold for 30 min. The reaction
was warmed to room temperature with continued stirring for 1 hr.
The mixture was diluted with dichloromethane and washed with 1N
HCl, then brine. The solution was dried over sodium sulfate and
evaporated. Chromatography on silica gel eluting with 9:1
Hexane:EtOAc gave 840 mg (76%) of the title compound. .sup.1H NMR
(CDCl.sub.3) .delta.: 7.41-7.80 (m, 9H), 4.65 (m, 1H), 4.11 (t,
J=6.0 Hz, 2H), 3.79 (q, J=7.2 Hz, 1H), 1.65-1.72 (m, 4H), 1.53 (d,
J=7.2 Hz, 3H). Anal Calcd for C.sub.20H.sub.21NO.sub.5: C, 67.59;
H. 5.96; N, 3.94. Found: C, 66.72; H, 5.95; N, 2.93
EXAMPLE 5
4-O-Nitroso-4-methyl-1-(3-benzoyl-.alpha.-methylbenzeneacetic
acid)pentyl ester
[0126] 5a. 4-Hydroxy
methyl-1-(3-benzoyl-.alpha.-methylbenzeneacetic acid)pentyl
ester
[0127] 3-Benzoyl-.alpha.-methylbenzeneacetic acid (1.99 g, 7.7
mmol) in methylene chloride (20 mL) under nitrogen and cooled over
ice was treated successively with oxalyl chloride (1.36 mL, 15.7
mmol) and dimethylformamide (5 drops). A vigorous gas evolution was
noted and the reaction mixture was stirred with slow warming and
then overnight at ambient temperature. The volatile materials were
removed in vacuo and the residue dissolved in methylene chloride
(10 mL) and added dropwise to a precooled mixture of
2-methyl-2,5-pentanediol (3.7 g, 31 mmol) and pyridine (0.69 mL,
8.6 mmol) also in methylene chloride (10 mL) under a nitrogen
atmosphere. The reaction mixture was stirred under nitrogen with
slow warming and then overnight at ambient temperature. The
solution was washed successively with 2N hydrochloric acid and 2N
sodium hydroxide, dried over anhydrous sodium sulfate, filtered and
concentrated in vacuo. The residual oil was subjected to column
chromatography using ethyl acetate/hexane (1:2). The product was
isolated as an oil in 76% yield (2.1 g). .sup.1H NMR CDCl.sub.3)
.delta.: 7.77-7.81 (nm; 3H, 7.647.43 (m, 6H), 4.18-4.03 (m, 2H),
3.80 (q, J=7.2 Hz, 1H), 1.62-1.71 (m, 2, 1.54 (d, J=7.2 Hz, 3H),
1.42-1.35 (m, 2H), 1.16 (s, 6H). Anal calcd for
C.sub.22H.sub.26O.sub.4: C, 74.55; H. 7.39.
[0128] Found: C, 74.26; H, 7.43.
[0129] 5b:
4-O-Nitroso-4-methyl-1-(3-benzoyl-.alpha.-methylbenzeneacetic
acid)pentyl ester
[0130] A solution of the product of example 5a (0.4 g, 1.13 mmol)
and pyridine (456 mL, 5.6 mmol) in methylene chloride (4 mL) was
cooled to -78 IC and nitrosonium tetrafluoroborate (262 mg, 2.26
mmol) added. The reaction mixture was stirred at -78.degree. C. for
3 hours, washed with water and dried over sodium sulfate. After
filtration and evaporation of the solvent the residual oil was
subjected to column chromatography using ethyl
acetate/hexane/triethylamine (18:80:2). The title compound was
isolated as an oil in 58% yield (0.25 g). .sup.1H NMR (CDCl.sub.3)
.delta.: 7.41-7.80 (m, 9H), 4.02-4.17 (m, 2H), 3.79 (q, J=7.2 Hz,
1H), 1.73-1.79 (m, 2H), 1.62-1.69 (m, 2H), 1.52-1.55 (m, 9H).
EXAMPLE 6
3-S-Nitroso-3-methyl-1-(3-benzoyl-.alpha.-methylbenzeneacetic
acid)butyl ester
[0131] 6a
3-Mercapto-3-methyl-1-(3-benzoyl-.alpha.-methylbenzeneacetic
acid)butyl ester
[0132] To 3-Benzoyl-.alpha.-methylbenzeneacetic acid (529 mg, 2
mmol) in benzene (5 mL) containing 5 ml of DMF was added oxalyl
chloride (200 ml 2.2 mmol) dropwise. The reaction mixture was
stirred 1.5 hr and then concentrated in vacuo to a syrup. The crude
acid chloride was dissolved in dichloromethane (10 mL) and
3-mercapto-3-methyl butanol (Sweetman et al. J. Med. Chem. 14, 868
(1971) (350 mg, 2.2 mmol) was added followed by pyridine (180 ml,
2.2 mmol). The reaction was stirred at room temperature for 1 h and
then it was diluted with dichloromethane and wash with 1N HCL
followed by saturated sodium bicarbonate, and then brine. The
organic phase was dried over sodium sulfate, concentrated in vacuo,
and the residue was chromatographed on silica gel eluting with 9:1
hexane:ethyl acetate to afford 640 mg (90%) of the product. .sup.1H
NMR (CDCl.sub.3) .delta.: 7.41-7.81 (m, 9H), 4.28 (t, j=7.1 Hz,
2H), 3.78 (q, J=7.2 Hz, 1H), 1.88 (t, J=7.0 Hz, 22H, 1.69 (s, 1H),
1.54 (d, J=7.3 Hz, 3H), 1.35 (s, 3H), 1.34 (s, 3H).
[0133] 6b.
3-S-Nitroso-3-methyl-1-(3-benzoyl-.alpha.-methylbenzeneacetic
acid)butyl ester
[0134] To a solution of the product of Example 6a (105 mg, 0.3
mmol) in dichloromethane (4 mL) was added tert-butyl nitrite (70
mg, 0.6 mmol) in a dropwise fashion. The mixture was stirred at
room temperature for 30 min. The solvent and excess reagent were
remove in vacuo to give 113 mg (quantitative) of the title
compound. .sup.1H NMR (CDCl.sub.3) .delta.: 7.44-7.81 (m, 9H), 4.29
(t, J=6.9 Hz, 2H), 3.77 (q, j=7.2 Hz, 1H), 2.51 (t, j=6.9 Hz, 2H),
1.841 (s, 3H), 1.836 (s, 3H), 1.53 (d, J=7.2 3H).
EXAMPLE 7
4-O-Nitroso-1-((S)-6-methoxy-.alpha.-methyl-2-naphthaleneacetic
acid)butyl ester
[0135] 7a. (S)-6-methoxy-.alpha.-methyl-2-naphthaleneacetic acetyl
chloride
[0136] Under a nitrogen atmosphere, oxalyl chloride (4.13 g, 30
mmol) was combined with methylene chloride (30 mL) and the
resulting mixture was cooled to 0.degree. C. Dimethylformamide (10
drops) was added and after 5 minutes of stirring, a suspension of
(S)-6-methoxy-.alpha.-methyl-2-napht- haleneacetic acid (3.00 g, 13
mmol) in methylene chloride (30 mL) was added dropwise over a 30
minute period. The reaction mixture was allowed to warm to room
temperature and stirred overnight. The solvent was evaporated in
vacuo to give the product in a quantitative yield. .sup.1H NMR
(CDCl.sub.3) .delta.: 1.5 (d, 3H), 3.91 (s, 1H), 4.21 (q, 1H),
7.09-7.14 (m, 1H), 7.15 (d, 1H), 7.42 (dd, 1H), 7.68 (s, 2H), 7.71
(s, 1H).
[0137] 7 b.
4Hydroxy-1-((S)-6-methoxy-.alpha.-methyl-2-naphthaleneacetic
acid)butyl ester
[0138] Under a nitrogen atmosphere, 1,4-butanediol (5.30 mL, 60
mmol) and pyridine (0.95 g, 12 mmol) were combined in methylene
chloride (20 mL). The resulting solution was stirred for 5 minutes
and then cooled to 0.degree. C. A solution of the product of
Example 7a (3.0 g, 12 mmol) in methylene chloride (15 ml) was added
dropwise over 30 minute period. After stirring for 20 hours at room
temperature, the reaction mixture was diluted with ethyl acetate
and washed with 1N hydrochloric acid. The organic phase was dried
over anhydrous sodium sulfate and concentrated in vacuo. The
residue was purified by flash chromatography on silica gel using
hexane/ethyl acetate (1:1 to 1:3) to afford 3.09 g (79% yield) of
the product as a colorless oil. .sup.1H NMR (CDCl.sub.3) .delta.:
1.47-1.68 (m, 4H, overlapping with a doublet at 1.57, 3H), 3.55 (t,
2H), 3.84 (q, 1H), 3.91 (s, 3H), 4.11 (t, 2H), 7.11 (m, 2H), 7.15
(d, 1H), 7.42 (dd, 1H), 7.67 (s, 1H), 7.70 (d, 2H).
[0139] 7c.
4-O-Nitroso-1-((S)-6-methoxy-.alpha.-methyl-2-naphthaleneacetic
acid)butyl ester
[0140] The product of Example 7b (0.209 g, 0.69 mmol) was dissolved
in anhydrous methylene chloride (4 mL) and pyridine (0.273 g, 3.45
mmol) was added. The resulting solution was cooled to -78.degree.
C. and nitrosonium tetrafluoroborate (0.161 g, 1.38 mmol) was added
in one portion. The reaction mixture was stirred for 1 hour at
-78.degree. C. The solvent was evaporated in vacuo and the residue
was purified by flash chromatography on silica gel, deactivated
with triethylamine, eluted with ethyl acetate/hexane (1:2) to give
0.180 g (79% yield) of the title compound as an oil. .sup.1H NMR
(CDCl.sub.3) .delta.: 1.58 (d, 3H), 1.641.69 (m, 4H), 3.85 (q, 1H),
3.92 (s, 3H), 4.11 (t, 2H), 4.60 (s, 2H), 7.10-7.13 (m, 1H), 7.15
(d, 1H), 7.39 (dd, if), 7.66 (s, 1H), 7.70 (d, 2H).
EXAMPLE 8
4-O-Nitroso-1-(1-)4-chlorobenzoyl)-5-methoxy-2-methyl-1H-indole-3-acetic
acid) butyl ester
[0141] a.
4-Hydroxy-1-(1-)4-chlorobenzoyl)-5-methoxy-2-methyl-1H-indole-3--
acetic acid)butyl ester
[0142] A stirred suspension of
1(4-chlorobenzoyl).sub.5-methoxy-2-methylin- doyl)-3-acetic acid
(3.7 g, 10.5 mmol) in methylene chloride (20 mL) under nitrogen and
cooled over ice was treated successively with oxalyl chloride (1.8
mL, 20.6 mmol) and dimethylformamide (10 drops). A vigorous gas
evolution was noted and the reaction mixture was stirred with
gradual warming to room temperature and then at ambient for a total
of 5 hours. The volatile materials were removed in vacuo and the
residue dissolved in dichloromethane (10 mL) and added dropwise to
a precooled mixture of 1,4-butanediol (4.7 g, 51.7 mmol) and
pyridine (0.92 mL, 11.4 mmol) also in methylene chloride (10 mL).
The reaction mixture was stirred with slow warming and then for 5
hours at ambient temperature under a nitrogen atmosphere. The
solution was washed with 2N hydrochloric acid, saturated sodium
bicarbonate, dried over anhydrous sodium sulfate, filtered and
concentrated in vacuo. The residual oil was subjected to column
chromatography using ethyl acetate/hexane (1:2). The product was
isolated as an oil in 75% yield (3.3 g) which solidified on
standing .sup.1H NMR (CDCl.sub.3) .delta.: 7.67 (d, J=8.4 Hz, 2H),
7.47 (d, J=8.5 Hz, 2H), 6.97 (d, J=2.5 Hz, 1H), 6.87 (d, J=9 Hz,
1H), 6.67 (dd, J=2.5 Hz, 9 Hz, 1H), 4.13 (t, J=6.4 Hz, 2H), 3.83
(s, 3H), 3.66 (s, 2H), 3.59 (t, J=6.4 Hz, 2H), 2.38 (s, 3H),
1.51-1.75 (m, 4H). Anal calcd for C.sub.23H.sub.24ClNO.sub.5: C,
64.26; H. 5.63; N, 3.26. Found: C, 64.08; H, 5.60; N, 3.18.
[0143] 8b.
4-O-Nitroso-1-(1-(4-chlorobenzoyl)-5-methoxy-2-methyl-1H-indole-
-3-acetic acid)butyl ester
[0144] A stirred solution of the product of Example 8a (1 g, 2.3
mmol), and pyridine (0.90 mL, 11.6 mmol) in methylene chloride (15
mL) at -78.degree. C. under a nitrogen atmosphere was treated with
nitrosonium tetrafluoroborate (0.54 g, 4.6 mmol). The reaction
mixture was stirred at -78.degree. C. for 3.5 hours, washed with
water, dried with anhydrous sodium sulfate and the solvent removed
in vacuo. The residual oil was subjected to column chromatography
using ethyl acetate/hexane (1:3). The product was isolated as a
yellow oil in 69% yield (0.73 g). .sup.1H NMR (CDCl.sub.3) o: 7.66
(d, J=8.5 Hz, 2 Hz), 7.47 (d, J=8.5 Hz, 2H), 6.95 (d, J=2.5 Hz,
1H), 6.85 (d, J=5 Hz, 1H), 6.66 (dd, J=2.5 Hz, 6.5 Hz, 1H), 4.66
(br s, 2H), 4.16 (t, J=6.6 Hz, 2H), 3.83 (s, 3H, 3.66 (s, 2H), 2.39
(s, 3H), 1.65-1.80 (m, 4H). Anal calcd for
C.sub.2H.sub.23ClN.sub.2O.sub.- 6 C, 60.2; H, 5.05; N, 6.1. Found:
C, 59.93; H, 4.87; N, 5.85.
EXAMPLE 9
[0145]
3-O-Nitroso-1-(1-(4-chlorobenzoyl)-5-methoxy-2-methyl-1H-indole-3-a-
cetic acid) butyl ester
[0146] 9a.
3-Hydroxy-1-(1-(4-chlorobenzoyl)-5-methoxy-2-methyl-1H-indole-3-
-acetic acid)butyl ester
[0147] A stirred suspension of
1-(4-chlorobenzoyl)-5-methoxy-2-methyl-1H-i- ndole-3-acetic acid (5
g, 13.9 mmol) in methylene chloride (25 mL) under nitrogen and
cooled over ice was treated successively with oxalyl chloride (2.44
mL, 28 mmol) and dimethylformamide (10 drops). A vigorous gas
evolution was noted and the reaction mixture was stirred with
gradual warming for a total of 5 hours. The volatile materials were
removed in vacuo and the residue dissolved in methylene chloride
(15 mL) and added dropwise to a precooled mixture
(+/-)-1,3-butanediol (8.83 g, 98 mmol) and pyridine (1.24 mL, 15.4
mmol) also in dichloromethane (10 mL). The reaction mixture was
stirred with slow warming and then over the weekend at ambient
temperature under a nitrogen atmosphere. The solution was washed
with 2N hydrochloric acid, saturated sodium bicarbonate, dried over
anhydrous sodium sulfate, filtered and concentrated in vacuo. The
residual oil was subjected to column chromatography using ethyl
acetate/hexane (1:1). The product was isolated as an oil which
solidified on standing in 75% yield (4.5 g). .sup.1H NMR indicated
that the desired product was contaminated with an isomer and so it
was recrystalised three times from diethyl ether/hexanes to give
the desired product as a solid in 15% yield (0.9 g). .sup.1H NMR
(CDCl.sub.3) .delta.: 7.66 (d, J=8.5 Hz, 2H), 7.43 (d, J=8.5 Hz,
2H), 6.95 (d, J=2.4 Hz, 1H), 6.86 (d, J=9 Hz, 1H), 6.67 (dd, J=9
Hz, 2.5 Hz), 4.30-4.39 (m, 1H), 4.15-4.4 (m, 1H), 3.83 (s, 3H),
3.75-3.85 (m, 1H), 3.67 (s, 2H), 2.38 (s, 3H), 1.95 (s, 1H),
1.65-2.8 (m, 2H), 1.16 (d, J=6.3 Hz, 3H). Anal calcd for
C.sub.23H.sub.24ClNO.sub.5: C, 64.26; H. 5.63; N, 3.26. Found: C,
64.29; H, 5.53; N, 3.18.
[0148] 9b.
3-O-Nitroso-1-(1-(4-chlorobenzoyl)-5-methoxy-2-methyl-1H-indole-
-3-acetic acid)butyl ester
[0149] A stirred solution of the product of Example 9a (0.15 g,
0.34 mmol), and pyridine (0.14 mL, 1.7 mmol) in dichloromethane (2
mL) at -78.degree. C. under a nitrogen atmosphere was treated with
nitrosonium tetrafluoroborate (0.08 g, 0.7 mmol). The reaction
mixture was stirred at -78.degree. C. for 3.5 hours, washed with
water, dried with anhydrous sodium sulfate and the solvent removed
in vacuo. The residual oil was subjected to column chromatography
using ethyl acetate/hexane (1:3). The title compound was isolated
as a yellow oil in 79% yield (0.125 g). .sup.1H NMR (CDCl.sub.3)
.delta.: 7.66 (d, J=8.5 Hz, 2H), 7.47 (d, J=8.5 Hz), 6.95 (d, J=2.3
Hz, 1H), 6.86 (d, J=9 Hz, 1H), 6.67 (dd, J=9 Hz, 2.5 Hz), 5.52
(sextet, J=6.5 Hz, 1H), 4.064.24 (m, 2H), 3.83 (s, 3H), 3.65 (s,
2H), 2.38 (s, 3H), 2.05 (q, J=4 Hz, 2H), 1.37 (d, J=6.5 Hz).
EXAMPLE 10
4-O-Nitroso-4
methyl-1-(1-(4-chlorobenzoyl)-5-methoxy-2-methyl-1H-indole-3-
-acetic acid)pentyl ester
[0150] 10a. 4-Hydroxy-4
methyl-1-(1-(4-chlorobenzoyl)-5-methoxy-2-methyl-1-
H-indole-3-acetic acid) pentyl ester
[0151] A stirred suspension of
1-(4-chlorobenzoyl)-5-methoxy-2-methyl-1H-i- ndole-3-acetic acid
(2.8 g, 7.7 mmol) in methylene chloride (25 mL) under nitrogen and
cooled over ice was treated successively with oxalyl chloride (1.36
mL, 15.7 mmol) and dimethylformamide (5 drops). A vigorous gas
evolution was noted and the reaction mixture was stirred over ice
for 30 min and then at room temperature for 3 hours. The volatile
materials were removed in vacuo and the residue dissolved in
methylene chloride (15 mL) and added dropwise to a precooled
mixture of 2-methyl-2,5-pentanediol (3.7 g, 31 mmol) and pyridine
(0.69 mL, 8.6 mmol) also in methylene chloride (10 mL). The
reaction mixture was stirred under nitrogen with slow warming and
then overnight at ambient temperature under a nitrogen atmosphere.
The solution was washed with 2N hydrochloric acid, dried over
anhydrous sodium sulfate, and filtered to give an oil which was
concentrated in vacuo. The residual oil was subjected to column
chromatography using ethyl acetate/hexane (1:2) The product was
isolated as an oil which-solidified on standing in 100% yield (3.6
g). .sup.1H NMR (CDCl.sub.3) .delta.: 7.69 (d, J=8.9 Hz, 2H), 7.47
(d, J=8.9 Hz, 2H), 6.98 (d, J=2.5 Hz, 1H), 6.87 (d, J=39 Hz, 1H),
6.67 (dd, J=9 Hz, 2.5 Hz), 4.09-4.14 (m, 2H), 3.83 (s, 3H), 3.66
(s, 3H), 2.39 (s, 3H), 1.62-1.73 (m, 2H), 1.37-1.43 (m, 2H), 1.14
(s, 6H). Anal calcd for C.sub.25H.sub.28ClNO.sub.5: C, 65.57; H.
6.16; N, 3.06.
[0152] Found: C, 65.35; H. 6.25; N, 3.10.
[0153] 10b.
4-O-Nitroso-4-methyl-1-(1-(4-chlorobenzoyl)-5-methoxy-2-methyl-
-1H-indole-3-acetic acid) pentyl ester
[0154] A solution of the product of Example 10a (0.2 g, 0.44 mmol)
and pyridine (176 mL, 2.2 mmol) in methylene chloride (2 mL) was
cooled over dry ice and nitrosonium tetrafluoroborate (101 mg, 0.87
mmol) added. The reaction mixture was stirred at -78.degree. C. for
3 hours, allowed to stand at the same temperature overnight, washed
with water and dried over sodium sulfate. After filtration and
evaporation of the solvent the residual oil was subjected to column
chromatography (twice) using ethyl acetate/hexanes/triethylamine
(25:73:2). The product was isolated as an oil in 42% yield (0.09
g). .sup.1H NMR (CDCl.sub.3) .delta.: 7.66 (d, J=7.5 Hz, 2H), 7.47
(d, J=7.5 Hz, 2H), 6.96 (d, J=2.5 Hz, 1H), 6.86 (d, J=9 Hz, 1H),
6.66 (dd, J=7.5 Hz, 2.5 Hz), 4.11 (t, J=6 Hz, 2H), 3.83 (s, 3H),
3.66 (s, 2H), 2.39 (s, 3H), 1.75-1.81 (m, 2H), 1.64-1.72 (m, 2H),
1.51 (s, 6H).
EXAMPLE 11
3-S-Nitroso-3-methyl-1-(.alpha.-methyl-4-(2-methylpropyl)benzeneacetic
acid)butyl ester
[0155] A solution of .alpha.-methyl-4-(2-methylpropyl)benzeneacetic
acid (1.52 g, 7.4 mmol) in methylene chloride (15 mL) cooled over
ice and under nitrogen, was treated successively with oxalyl
chloride (1.29 mL, 1.88 g, 14.8 mmol) and dimethylformamide (5
drops). The resultant solution was stirred over ice for 30 min and
then at ambient temperature for 2 hours. The excess volatile
materials were removed in vacuo and the residue, dissolved in
methylene chloride (5 mL), added to a precooled solution of
pyridine (0.54 mL, 6.7 mmol) and 3-mer 3-methylbutanol (0.8 g, 6.7
mmol) in methylene chloride (15 mL). The reaction mixture was
stirred over ice for 30 min and then at ambient temperature for 3
hours. The solution was then diluted with additional methylene
chloride and washed with 2N hydrochloric acid, saturated sodium
bicarbonate and brine and the organic phase dried with sodium
sulfate, filtered and the solvent removed in vacuo. The residual
oil was subjected to column chromatography using ethyl
acetate/hexane (1:3). The product was isolated as an oil in 68%
yield (1.4 g). .sup.1H NMR (CDCl.sub.3) .delta.: 7.18 (d, 3=7.5 Hz,
2H), 7.09 (d, J=7.5 Hz, 2H), 4.25 (t, J=6.5 Hz, 2H), 3.67 (q, J=7
Hz, 1H), 2.44 (d, J=7.8 Hz, 2H), 1.77-1.9 (m, 3H) 1.48 (d, J=7 Hz,
3H), 1.32 (s, 6H), 0.89 (d, J=6.6 Hz, 6H).
[0156] 11b.
3-S-Nitroso-3-methyl-1-(.alpha.-methyl-4-(2-methylpropyl)benze-
neacetic acid)butyl ester
[0157] A solution of the product of Example 11a (0.4 g, 1.2 mmol)
in methylene chloride (8 mL) under nitrogen was treated with tert
butyl mite (0.62 mL, 0.53 g, 5 mmol). After stirring for 1 hour at
ambient temperature the volatile materials were removed in vacuo.
The residual green oil was subjected to column chromatography using
ethyl acetate/hexanes (1:19). The product was isolated as green oil
in 65% yield (0.25 g). .sup.1H NMR (CDCl.sub.3) .delta.: 7.0 (d,
J=7.5 Hz, 2), 7.10 (d, J=7.5 Hz, 2H1, 4.27 (t, J=6.9 Hz, 2H), 3.66
(q, J=7.2 Hz, 1H), 2.49 (t, 3=6.6 Hz, 2H), 2.44 (d, J=7.2 Hz, 2H),
1.8-1.9 (m, 1H), 1.81 (s, 3H), 1.80 (s, 3H), 1.48 (d, J=7.2 Hz,
3H), 0.89 (d, J=6.6 Hz, 6H).
EXAMPLE 12
[0158]
4-O-Nitroso-1-(.alpha.-methyl-4-(2-methylpropyl)benzeneacetic
acid)butyl ester
[0159] 12a.
4Hydroxy-1-(.alpha.-methyl-4-(2-methylpropyl)benzeneacetic
acid)butyl ester
[0160] .alpha.-Methyl-4-(2-methylpropyl)benzeneacetic acid (4 g, 19
mmol) and 10 uL DMF were dissolved in benzene (30 mL). Oxalyl
chloride was added dropwise. Stirring was continued for 2 hr before
concentration to a syrup. Butanediol (9 mL, 100 mmol) and pyridine
(1.67 mL, 21 mmol) were dissolved in dichloromethane (100 mL) and
dioxane (15 mL) and cooled to 0.degree. C. A solution of the acid
chloride was added in dichloromethane (20 mL). The reaction mixture
was stirred cold for 20 min then warmed to room temperature with
stirring for 2 hr. The solution was washed H.sub.2O, 1 N HCl, satd
Sodium bicarbonate and finally brine; dried over sodium sulfate;
and evaporated. The residue was filtered through silica gel eluting
with 2:1 hexane:EtOAc to yield 4.8 g (91%) of the product. .sup.1H
NMR (CDCl.sub.3) .delta.: 7.19 (d, J=6.2 Hz, 2H), 7.08 (d, J=8.2
Hz, 2H), 4.074.12 (m, 2H), 3.68 (q, J=7.1 Hz 1H), 3.58 (t, J=6.3
Hz, 1H), 2.44 (d, J=7.2 Hz, 2H), 1.84 (sept, J=6.8 Hz, 1H),
1.50-1.69 (m, 4H), 1.48 (d, J=7.2 Hz, 3H), 0.88 (d, J=6.6 Hz, 6H).
Anal Calcd for C.sub.17H.sub.26O.sub.3: C, 73.34; H, 9.41. Found:
C, 73.17; H. 9.67
[0161] 12b.
4-O-Nitroso-1-(.alpha.-methyl-4-(2-methylpropyl)benzeneacetic
acid)butyl ester
[0162] The product of Example 12a (1 g, 3.6 mmol) and pyridine (1.4
mL, 18 mmol) were dissolved in dichloromethane (15 mL) and cooled
to -78.degree. C. Nitrosonium tetrafluoroborate (840 mg, 7.2 mmol)
was added and the solution was kept cold for 30 min. The reaction
was warmed to room temperature with continued stirring for 1 hr.
The mixture was diluted with dichloromethane and washed
successively with 1N HCl, H.sub.2O, and brine. The solution was
dried over sodium sulfate and evaporated. Chromatography on silica
gel eluting with 9:1 hexane:EtOAc gave 840 mg (76%) of the title
compound. .sup.1H NMR (CDCl.sub.3) .delta.: 7.18 (d, J=8.1 Hz, 21),
7.08 (d, J=8.1 Hz, 2H), 4.62 (m, 2H), 4.074.12 (m, 2H), 3.68 (q,
J=7.1 Hz, 1H), 2.44 (d, J=7.2 Hz, 2H), 1.84 (sept, J=6.7 Hz, 1H),
1.64-1.68 (m, 4H), 1.48 (d, J=7.2 Hz, 3H), 0.88 (d, J=6.6 Hz,
6H).
EXAMPLE 13
4-O-Nitroso-1-(2-Fluoro-.alpha.-methyl-biphenylacetic acid)butyl
ester
[0163] 13a. 2-Fluoro-.alpha.-methyl-biphenylacetic acid
chloride
[0164] Under a nitrogen atmosphere, oxalyl chloride (3.8 g, 30
mmol) was combined with methylene chloride (30 mL). The resulting
mixture was cooled to 0 IC and dimethylformamide (10 drops) was
added. After 5 minutes of stirring a solution of
2-fluoro-.alpha.-methyl-biphenylacetic acid (3.0 g, 12 mmol) in
methylene chloride (30 mL) was added dropwise over a 30 minute
period. The reaction mixture was allowed to warm to room
temperature and stirred overnight. The solvent was evaporated to
give the product in a quantitative yield as a yellow solid. .sup.1H
NMR (CDCl.sub.3) .delta.: 1.58 (d, 3H), 4.20 (q, 1H), 7.11 (t, 2H),
7.33-7.47 (m, 4H), 7.54 (d, 2H).
[0165] 13b. 4-Hydroxy-1-(2-Fluoro-.alpha.-methyl-biphenylacetic
acid)butyl ester
[0166] Under a nitrogen atmosphere, 1,4-butanediol (5.30 mL, 60
mmol) and pyridine (0.95 g, 12 mmol) were combined in methylene
chloride (20 mL). The resulting solution was stirred for 5 minutes
and then cooled to 0.degree. C. A solution of the product of
Example 13a (3.0 g, 12 mmol) in methylene chloride (15 ml) was
added dropwise over 30 minute period. After stirring for 20 hours
at room temperature, the reaction mixture was diluted with ethyl
acetate and washed with 1N hydrochloric acid. The organic phase was
dried over anhydrous sodium sulfate and concentrated in vacuo. The
residue was purified by flash chromatography on silica-gel eluting
with methylene chloride/hexane (2:1) to give 1.66 g (44%) of the
product as a colorless oil. .sup.1H NMR (CDCl.sub.3) .delta.: 1.56
(d, 3H), 1.61-1.77 (m, 4H), 3.63 (t, 2H), 3.75 (q, 1H), 4.14 (t,
2H), 7.14 (t, 2H), 1.27-7.45 (m, 4H), 7.53 (d, 2H).
[0167] 13c. 4-O-Nitroso-1-(2-1-Fluoro-.alpha.-methyl-biphenylacetic
acid)butyl ester
[0168] The product of Example 13b (0.190 g, 0.60 mmol) was
dissolved in anhydrous methylene chloride (4 mL) and pyridine
(0.237 g, 3.00 mmol) was added. The resulting solution was cooled
to -78.degree. C. and nitrosonium tetrafluoroborate (0.084 g, 0.72
mmol) was added. The reaction mixture was stirred for 1 hour at
-78.degree. C. and an additional nitrosonium tetrafluoroborate
(0.047 g, 0.40 mmol) was added. After 30 minutes of stirring at
-78.degree. C., the solvent was evaporated in vacuo and the residue
was purified by flash chromatography on silica gel, deactivated
with triethylamine, eluted with methylene chloride/hexane (3:1) to
give 0.117 g (57% yield) of the title compound. .sup.1H NMR
(CDCl.sub.3) .delta.: 1.54 (d, 3H), 1.68-1.83 (m, 4H), 3.75 (q,
1H), 4.14 (t, 2H), 4.67 (s, 2H), 7.14 (t, 2H), 7.34-7.48 (m, 4H),
7.54 (d, 2H).
EXAMPLE 14
4-O-Nitroso-1-(2-Fluoro-.alpha.-methyl-biphenylacetic
acid)thiobutyl ester
[0169] 14a. 1-tert-Butyldimethylsilyloxy-4-chloro-butanol
[0170] 4Chloro-1-butanol (5.43 g, 50 mmol) was dissolved in
dimethylformamide (50 mL) and tert-butyldimethylsilylchloride (7.54
g, 50 mmol) was added, followed by imidazole (3.4 g, 50 mmol).
After 24 hours of stirring at room temperature, the reaction
mixture was diluted with hexane, washed with water and brine and
dried over anhydrous sodium sulfate. The solvent was evaporated to
give colorless liquid which was purified by chromatography on
silica gel eluting with hexane/ethyl acetate (30:1) to give the
product (7.26 g, 56%). .sup.1H NMR (CDCl.sub.3) .delta.: 0.05 (s,
6H), 0.89 (s, 9H), 1.64-1.68 (m, 2H), 1.82-1.86 (m, 2H), 3.57 (t,
2H), 3.64 (t, 2H).
[0171] 14b. 4-tert-Butyldimethylsilyloxy-1-acetyl-thiobutyl
ester
[0172] Under a nitrogen atmosphere, potassium thioacetate (0.53 g,
4.7 mmol) was dissolved in dimethylformamide (12 mL) and cooled to
0.degree. C. A solution of the product of Example 14a (1.01 g, 3.91
mmol) in dimethylformamide (14 mL) was added. After 0.24 hours of
stirring at room temperature, the solvent was evaporated and the
residue was partitioned between hexane and water (1:3). The organic
layer was concentrated in vacuo to give the product (0.820 &
71%) as a yellow liquid. .sup.1H NMR (CDCl.sub.3) .delta.: 0.04 (s,
6H), 0.88 (s, 9H), 1.57-1.64 (m, 4H), 2.32 (s, 3H), 2.89 (t, 2H),
3.61 (t, 2H).
[0173] 14c. 4-tert-Butyldimethylsilyloxy-1-butane thiol
[0174] The product of Example 14b (5.7 g, 19.2 mmol) was dissolved
in methanol (30 mL) and degassed with nitrogen gas for 30 minutes.
Potassium carbonate (2.92 g, 21.1 mmol) was added in one portion.
After 1 hour of stirring at room temperature, the solvent was
evaporated and the residue was partioned between hexane and water.
The organic layer was dried over anhydrous sodium sulfate and
concentrated in vacuo to give the product (3.2 g, 66%). .sup.1H NMR
(CDCl.sub.3) .delta.: 0.05 (s, 6H), 0.89 (s, 9H), 1 34 (t, 1H),
1.61-1.68 (m, 4H), 2.51-2.57 (q, 2H), 3.62 (t, 2H).
[0175] 14d.
4-tert-Butyldimethylsilyloxy-1-(2-Fluoro-.alpha.-methyl-biphen-
ylacetic acid) thiobutyl ester
[0176] The product of Example 14c (1.37 g, 5.4 mmol) was combined
with pyridine (0:142 g, 1.8 mmol) in methylene chloride (5 mL) and
the resulting solution was cooled to 0.degree. C. A solution of the
product of Example 13a (0.500 g, 1.8 mmol) in methylene chloride (4
mL) was added dropwise. After 22 hours of stirring at room
temperature, the reaction mixture was diluted with ethyl acetate
and washed with 1N hydrochloric acid. The organic layer was dried
over anhydrous sodium sulfate and concentrated in vacuo to give the
product (0.526 g, 59%). .sup.1H NMR (CDCl.sub.3) .delta.: 0.04 (s,
6H), 0.89 (s, 9H), 1.56 (d, 3H), 1.57-1.62 (m, 4H), 1.88-2.29 (M,
2H), 3.61 (t, 2H), 7.15 (t, 2H), 7.37-7.44 (m, 4H), 7.54 (d,
2H).
[0177] 14e. 4-Hydroxy-1-(2-Fluoro-.alpha.-methyl-biphenylacetic
acid)thiobutyl ester
[0178] The product of Example 14d (0.320 g, 0.64 mmol) was
dissolved in the mixture of glacial acetic acid (0.5 mL), water (1
mL), and tetrahydrofuran (5 mL). The resulting solution was stirred
for 24 hours at room temperature. The solvent was evaporated and
the residue was partioned between methylene chloride and water. The
organic layer was washed with saturated sodium bicarbonate solution
and brine, and dried over anhydrous sodium sulfate. The solvent was
evaporated to give the product (0.235 g, 100%). .sup.1H NMR
(CDCl.sub.3) o: 1.57 (d, 3H), 1.58-1.69 (m, 4H), 2.87-2.93 (m, 2H),
3.63 (t, 2H), 3.84-3.92 (q, 1H), 7.14 (t, 2H), 7.37-7.44 (m, 4H),
7.54 (d, 2H).
[0179] 14f. 4-O-Nitroso-1-(2-Fluoro-.alpha.-methyl-biphenylacetic
acid)thiobutyl ester
[0180] The product of Example 14e (0.235 g, 0.61 mmol) was
dissolved in anhydrous methylene chloride (3 mL) and pyridine
(0.097 g, 1.23 mmol) was added. The resulting solution was cooled
to -78.degree. C. and nitrosonium tetrafluoroborate (0.144 g, 1.23
mmol) was added in one portion. The reaction mixture was stirred
for 1 hour at -78.degree. C., the solvent was evaporated, and the
residue was purified by chromatography on silica gel eluted with
hexane/ethyl acetate (10:1) to give the title compound (0.110 g,
44%). .sup.1H NMR (CDCl.sub.3) .delta.: 157 (d, 3H), 1.58-1.80 (m,
4H), 3.85-3.93 (q, 1H), 4.69 (t, 2H), 7.14 (t, 2H), 7.37-7.44 (m,
4H), 7 55 (d, 2H).
EXAMPLE 15
4-O-Nitroso-2-methyl-N-2-pyridinyl-2-H-1,2-benzothiazine-2-carboxamide-1,1-
-dioxide
[0181]
4-Hydroxy-2-methyl-N-2-pyridinyl-2-H-1,2-benzothiazine-2-carboxamid-
e-1,1-dioxide (10.0 g, 30 mmol) was dissolved in anhydrous
methylene chloride and cooled to 0 C. Nitrosonium tetrafluoroborate
(4.407 g, 38 mmol) was added in one portion, followed by pyridine
(2.98 g, 38 mmol). The reaction mixture was stirred at room
temperature for 7 days and then additional nitrosonium
tetrafluoroborate (0.571 g, 1.72 mmol) was added. After stirring
for 14 days at room temperature, the reaction mixture was poured
into saturated sodium bicarbonate solution and extracted with
methylene chloride. The solvent was evaporated, the residue was
treated with ethyl acetate and filtered. The precipitate was
dissolved in the mixture of methylene chloride/ethylacetate (1:1),
and the solution was treated with decolorising charcoal, filtered
and concentrated in vacuo to give the title compound as a solid
(1.56 g, 14%). .sup.1H NMR (CDCl.sub.3, 300 MHz), .delta.: 2.96 (s,
3H), 6.84 (t, 1H), 7.17 (t, 1H), 7.60-7.86 (m, 5H), 8.22 (d,
1H).
EXAMPLE 16
[0182]
4-O-Nitroso-hydroxymethylene-(1-(3-benzoyl-.alpha.-methylbenzeneace-
tic acid))benzyl ester
[0183] 16a. 3-benzoyl-.alpha.-methylbenzeneacetic acid chloride
[0184] 3-Benzoyl-.alpha.-methylbenzeneacetic acid (3.2 g, 12.6
mmol) was treated in the same manner as set forth in Example 13a.
Evaporation of the solvent, afforded the product as a yellow oil in
a quantitative yield. .sup.1H NMR (CDCl.sub.3), .delta.: 1.64 (d,
3H), 4.21 (q, 1H), 7.45-7.51 (m, 4H), 7.62 (d, 1H), 7 72-7.82 (m,
4H).
[0185] 16b.
4-Hydroxymethylene-(1-(3-benzoyl-.alpha.-methylbenzeneacetic
acid))benzyl ester
[0186] Under a nitrogen atmosphere, 1,4-benzenedimethanol (0.507 g,
3.67 mmol) and pyridine (0.145 g, 1.83 mmol) were combined in
methylene chloride (5 mL). The resulting solution was stirred for 5
minutes and then cooled to 0.degree. C. A solution of the product
of Example 16a (0.500 g, 1.83 mmol) in methylene chloride (5 mL)
was added dropwise over 15 minutes. The reaction mixture was
allowed to warm to room temperature and was then stirred over 2
days period. The solvent was evaporated and the residue was
dissolved in ethyl acetate, washed with 1N hydrochloric acid and
saturated sodium bicarbonate solution. The organic phase was dried
over anhydrous sodium sulfate and concentrated in vacuo. The
residue was purified by flash chromatography on silica-gel eluting
with hexane/ethyl acetate (5:1 to 2:1) to give 0.092 g (42%) of the
product H NMR (CDCl.sub.3) o: 1.60 (d, 3H), 2.19 (s, 1H), 3.90 (q,
1H), 4.71 (s, 2H), 5.17 (s, 2H), 7.32 (dd, 4H), 7 45-7u.82 (m, 7H),
7 84 (d, 2H).
[0187] 16c.
4-O-Nitroso-hydroxymethylene-(1-(3-benzoyl-methylbenzeneacetic
acid)) benzyl ester
[0188] The product of Example 16b (0.090 g, 0.24 mmol) was treated
in the same manner as set forth in Example 7c. Purification of the
crude product was accomplished using flash chromatography on silica
gel eluted with hexane/ethyl acetate (1:2) to give 0.069 g (71%) of
the title compound as a yellow oil. .sup.1H NMR (CDCl.sub.3, 300
MHz), .delta.: 1.55 (3H), 3.85 (q, 1H), 5.11 (s, 2H), 5.67 (s, 2H),
7.27-7.80 (m, 9H).
EXAMPLE 17
[0189]
3-O-Nitroso-hydroxymethylene-(1-(3-benzoyl-.alpha.-methylbenzeneace-
tic acid)) benzyl ester
[0190] 17a.
3-Hydroxymethylene-(1-(3-benzoyl-.alpha.-methylbenzeneacetic
acid))benzyl ester
[0191] Under a nitrogen atmosphere, 1,3-benzenedimethanol (0.500 g,
3.62 mmol) and pyridine (0.193 g, 2.44 mmol) were combined in
methylene chloride (7 mL). The resulting solution was stirred for 5
minutes and then cooled to 0 C. A solution of the the product of
Example 16a (0.665 g, 2.44 mmol) in methylene chloride (5 mL) was
added dropwise over 15 minutes. The reaction mixture was stirred 2
h 30 min at 0.degree. C., concentrated in vacuo, diluted with ethyl
acetate, washed with 1N hydrochloric acid and saturated sodium
bicarbonate solution. The organic phase was dried over anhydrous
sodium sulfate and concentrated in vacuo. The residue was purified
by flash chromatography on silica gel eluting with hexane/ethyl
acetate (2:1) to give 0.530 g (58%) of the product .sup.1H NMR
(CDCl.sub.3) .delta.: 1.55 (.delta.: 3H), 3.85 (q, 1H), 4.64 (s,
2H), 5.12 (d, 2H), 7.13-7.18 (m, 1H), 7.22 (s, 1H), 7.26-7.30 (m,
2H), 7.40-7.67 (m, 6H), 7 73-7.78 (m, 3H).
[0192] 17b.
3-O-Nitroso-hydroxymethylene-(1-(3-benzoyl-.alpha.-methylbenze-
neacetic acid)) benzyl ester
[0193] The product of Example 17a (0.74 g, 0.198 mmol) was treated
in the same manner as set forth in Example 7c. Purification of the
crude product was accomplished using flash chromatography on silica
gel eluted with hexane/ethyl acetate (2:1) to give 0.046 g (71%) of
the title compound. .sup.1H NMR (CDCl.sub.3) .delta.: 1.55 (d, 3H),
3.85 (q, 1H), 5.12 (s, 2H, 5.65 (s, 2H), 7.18-7.31 (m; 4H),
7.40-7.75 (m, 6H), 7.767.79 (m, 3H).
EXAMPLE 18
[0194]
3-O-Nitroso-hydroxymethylene-1-(1-(3-benzoyl-.alpha.-methylbenzenea-
cetic acid))-hydroxymethyladamantyl ester
[0195] 18a. 1,3-Dicarboxymethyl adamantane
[0196] 1,3-adamantanedicarboxylic acid (1.5 g, 5.95 mmol) was
dissolved in methanol (30 mL) and concentrated sulfuric acid (0.5
mL, 8.90 mmol) was added. The reaction mixture was stirred at room
temperature for 20 hours. After concentration in vacuo, the residue
was dissolved in methylene chloride, washed with water/brine (1:1),
and dried over anhydrous sodium sulfate. The solvent was evaporated
to give the product as a white solid in a quantitative yield.
.sup.1H NMR (CDCl.sub.3) .delta.: 1.65-1.71 (m, 2H), 1.76-1.82 (m,
8H), 1.98-2.03 (m, 2H), 2.07-2.18 (m, 2H), 3.66 (s, 6H).
[0197] 18b. 1,3-Dihydroxymethyl adamantane
[0198] Under a nitrogen atmosphere, the product of Example 18a
(1.33 g, 5.95 mmol) was dissolved in tetrahydrofuran (20 mL) and
lithium aluminum hydride (0.316 g, 8.33 mmol) was added in one
portion. The reaction mixture was allowed to reflux for 30 minutes,
and was then quenched with water (0.316 mL, 8.33 mmol), 15% sodium
hydroxide solution (0.316 mL), and water (0.95 mL). After 15 hours
of stirring at room temperature, the reaction mixture was filtered
through PTFE and filtrate was partitioned between ethyl acetate and
brine. The organic phase was dried over anhydrous sodium sulfate,
filtered through PTFE and concentrated in vacuo to give the product
(0.370 g, 28%) as a white solid: .sup.1H NMR (CDCl.sub.3) .delta.:
1.24-1.29 (m, 2H), 1.42-1.52 (m, 8H), 1.61-1.68 (m, 2H), 2.07-2.16
(m, 2H), 3.25 (s, 4H).
[0199] 18c.
3-Hydroxymethylene-1-(1-(3-benzoyl-.alpha.-methylbenzeneacetic
acid))-hydroxymethyladamantyl ester
[0200] The product of Example 18b (0.199 g, 0.54 mmol) was
dissolved in tetrahydrofuran (10 mL) and pyridine (0.047 g, 0.59
mmol) was added. A solution of the product of Example 16a (0.161 g,
0.59 mmol) in chloroform (3 mL) was added dropwise. The reaction
mixture was stirred at room temperature for 40 hours. The solvent
was evaporated, the residue was dissolved in methylene chloride,
washed with 1N hydrochloric acid, saturated sodium bicarbonate
solution and brine, and dried over anhydrous sodium sulfate. The
solvent was evaporated and the residue was purified by flash
chromatography on silica gel eluted with hexane/ethyl acetate (2:1)
to give the product (0.102 g, 28%) as a colorless oil. .sup.1H NMR
(CDCl.sub.3) .delta.: 1.13-1.17 (m, 2H), 1.18-1.55 (m, 10H),
1.98-2.02 (m, 2H), 3.18 (s, 2H), 3.66 (d, 1H), 3.77 (d, 1H), 3.83
(q, 1H), 7.43-7.68 (m, 6H), 7.76-7.81 (m, 3H).
[0201] 18d.
3-O-Nitroso-hydroxymethylene-1-(1-(3-benzoyl-.alpha.-methylben-
zeneacetic acid))-hydroxymethyladamantyl ester
[0202] The product of Example 18c (0.056 g, 0.083 mmol) was
dissolved in anhydrous methylene chloride (2 mL) and pyridine (2
drops) was added. The resulting solution was cooled to -78.degree.
C. and nitrosonium tetrafluoroborate was added in one portion. The
reaction mixture was stirred for 3 hours at -78.degree. C., washed
with water, brine, dried over anhydrous sodium sulfate and
concentrated in vacuo. The residue was purified by chromatography
on silica gel eluted with hexane/ethyl acetate (15:1) to give the
title compound as a colorless oil. .sup.1H NMR (CDCl.sub.3)
.delta.: 1.15-1.19 (m, 2H), 1.29-1.61 (m, 10H), 1.98-2.03 (m, 2H),
3.65 (d, 1H), 3.77 (d, 1H), 3.82 (q, 1H), 4.33 (s, 2H), 7.43-7.68
(m, 6H), 7.76-7.81 (m, 3H).
EXAMPLE 19
Comparative In Vivo Analgesic, Antiinflammatory and Gastric Lesion
Activities
[0203] The phenylbenzoquinone-induced writhing test in mice was
used to measure analgesic activity. The ability of the compounds to
inhibit phenylbenzoquinone-induced writhing in mice was measured
using the method of Siegmund et. al. Proc. Soc. Exp. Biol. Med. 95:
729-131, 1957. Male CD-1 mite (Charles River Laboratories,
Wilmington, Mass.) weighing 20-25 g were fasted overnight. Vehicle
or compounds were administered by oral gavage. 1 hour prior to i.p.
injection of 2 mg/kg of phenylbenzoquinone. In the case of a nitric
oxide adduct being given in combination with a NSAID, the nitric
oxide adduct was administered immediately before the NSAID. Five
minutes after the i.p. injection of phenylbenzoquinone, the number
of writhes in a 5 minute period was counted.
[0204] The rat paw edema test was used to measure antiinflammatory
activity. The rat paw edema test was performed according to the
method of Winter et al., Proc. Soc. Exp. Biol. Med. 111: 544-547,
1962. Male Sprague-Dawley rats (250-275 g) were fasted overnight
and dosed by oral gavage with vehicle or suspensions of compound
one hour prior to the subplantar injection of 50 ml of 1%
suspension of carrageenin. Three hours later, the paw volume was
measured and compared with the initial volume measured immediately
after carrageenin injection.
[0205] The rat gastric lesion test (Kitagawa et al., J. Pharmacol.
Exp Ther. 253:1133-1137, 1990; Al-Ghamdi et al., J Int. Med. Res.,
19: 2242, 1991) was used to evaluate the potential of compounds to
produce gastric lesion. Male Sprague Dawley rats (Charles River
Laboratories, Wilmington, Mass.) weighing 230-250 g were used for
the experiments. The rats were housed with laboratory chow and
water ad libitum prior to the study. The rats were fasted for 24-30
hours with free access to water and then dosed by oral gavage with
vehicle or with drugs given at a volume of 0.5 mL/100 g. For the
unmodified NSAIDs being given in combination with a nitric oxide
adduct (NO-adduct), the NO-adduct was administered by oral gavage
immediately prior to the administration of NSAID by oral gavage.
Food was withheld for 18 hours after the inital dosing. For acute
studies, rats were euthanized by CO.sub.2 eighteen hours after
dosing and the stomachs were dissected. For the multiple dosing
studies, the results of which are in Table 3, food was given
eighteen hours after the first dose and the rats were maintained on
food and water ad libitum while receiving a single daily dose for
the remainder of the experiment. For the multiple dosing studies,
the results of which are in Table 4, the rats were either fasted
24-30 hours before the first dosing and for 4 hours after the first
dosing, (4 day study with ketoprofen, Example 4, and Example 6);
allowed access to food and water ad libitum before as well as
during the experiment, (7 day study with ketoprofen and Example 4);
or fasted 24-30 hours prior to the first dosing and for 18 hours
after the first dosing, (7 day study with ibuprofen, Example 11,
and Example 12). The stomachs were dissected along the greater
curvature, washed with a directed stream of 0.9% saline and pinned
open on a sylgard based petridish for examination of the
hemorrhagic lesion. Gastric lesion score was expressed in mm and
calculated by summing the length of each lesion.
[0206] Table 1 shows the relative activities of compounds in the
analgesic, antiinflammatory and gastric lesion tests, and are
expressed, for each novel NSAID compound, as described according to
the general formulas (I), (II), (III) and (IV), or NSAID
coadministered with an NO-adduct, as the ratio of activitiy
relative to the parent NSAID.
1 TABLE 1 Relative Activity Compound Analgesia Antiinflammation
Gastric Lesion Ketoprofen 1 1 1 Example 4 1.6 0.58 0.03 Example 6 1
ND ND Example 5 1.1 ND ND Example 16 1.1 ND ND Flurbiprofen 1 1 1
Example 13 0.31 1.83 0.5 Indomethacin 1 1 1 Example 8 1 1 0.08
Ibuprofen ND 1 1 Example 12 ND 1 <0.03 Example 11 ND 1 <0.05
Piroxicam 1 ND 1 Piroxicam + Example 2 2.3 ND 0.08 ND--not
determined
[0207] Table 2 shows the results of single dose treatment studies
in which various NO-adducts were administered in combination with
various NSAIDs. The combinations are able to protect against the
NSAID induced gastric toxicity.
2TABLE 2 Molar Dose Gastric Ratio Lesion NSAID (mg/kg) NO-Adduct
NSAID:NO-Adduct Protection Piroxicam 16 Example 2 1:1 +++ Piroxicam
8 Example 2 1:1 +++ Piroxicam 8 Isoamyl nitrite 1:3 +++ Piroxicam 8
Isosorbide 1:3 +++ dinitrate Piroxicam 8 Example 1 1:2 ++
Flurbiprofen 16 Example 2 1:1 ++ Tenidap 16 Example 2 1:1 ++
70-100% Protection = +++; 40-69% Protection = ++; 20-39% Protection
= +
[0208] Table 3 shows the results of multiple dose treatment studies
in which various NO-adducts were administered in combination with
various NSAIDs. The combinations are able to protect against the
NSAID induced gastric toxicity.
3TABLE 3 Molar Dose Treatment Ratio Gastric Lesion (Days) NSAID
(mg/kg) NO-Donor NSAID:NO-Adduct Protection 3 Piroxicam 16 Example
2 1:1 +++ 14 Piroxicam 16 Example 2 1:1 ++ 7 Ibuprofen 40 Example 2
1:1 + 14 Ibuprofen 30 Example 2 1:1 ++ 70-100% Protection = +++;
40-69% Protection = ++; 20-39% Protection = +
[0209] Table 4 shows the results of multiple dose treatment studies
in which various novel NSAID compounds directly or indirectly
linked to various NO-adducts were administered. The modified NSAIDs
containing NO-adducts produced significantly less gastric
toxicity.
4 TABLE 4 Treatment Relative Gastric Compound (mg/kg) (Days) Lesion
Activity Ketoprofen 10 4 +++++ Example 4 14 4 + Example 6 15 4 ++
Ketoprofen 10 7 +++++ Example 4 14 7 + Ibuprofen 30 7 +++++ Example
11 50 7 + Example 12 45 7 + Vehicle 7 + 100% of the gastric toxcity
induced by the parent NSAID = +++++ 21-40% of the gastric toxcity
induced by the parent NSAID = ++ 1-20% of the gastric toxcity
induced by the parent NSAID = +
* * * * *