Nitrosated and nitrosylated nonsteroidal antiinflammatory compounds, compositions and methods of use

The present invention describes novel nitrosated and/or nitrosylated nonsteroidal antiinflammatory compounds, and novel compositions comprising at least one nitrosated and/or nitrosylated nonsteroidal antiinflammatory compound, and, optionally, at least one compound that donates, transfers or releases nitric oxide, elevates endogenous levels of endothelium-derived relaxing factor, stimulates endogenous synthesis of nitric oxide or is a substrate for nitric oxide synthase. The present invention also provides methods for treating, preventing and/or reducing inflammation, pain, and fever; decreasing or reversing the gastrointestinal, renal and other toxicities resulting from the use of nonsteroidal antiinflammatory drugs; treating and/or preventing gastrointestinal disorders; treating inflammatory disease states and disorders; and treating and/or preventing ophthalmic diseases or disorders.

FIELD OF INVENTION

The present invention describes novel nitrosated and/or nitrosylated nonsteroidal antiinflammatory drugs, and novel compositions comprising at least one nitrosated and/or nitrosylated nonsteroidal antiinflammatory drug, and, optionally, at least one compound that donates, transfers or releases nitric oxide, elevates endogenous levels of endothelium-derived relaxing factor, stimulates endogenous synthesis of nitric oxide or is a substrate for nitric oxide synthase. The present invention also provides methods for treating, preventing and/or reducing inflammation, pain, and fever; decreasing or reversing the gastrointestinal, renal and other toxicities resulting from the use of nonsteroidal antiinflanmatory compounds; treating and/or preventing gastrointestinal disorders; treating inflammatory disease states and disorders; and treating and/or preventing ophthalmic diseases or disorders.

BACKGROUND OF THE INVENTION

The chemistry and pharmacology of nitroxybutylester ((CH 2 ) 4 ONO 2 ) derivatives of several aryl propionic acid nonsteroidal antiinflammatory compounds, including ketoprofen, flurbiprofen, suprofen, indobufen and etodolac, was described in PCT Application No. WO 94/12463. 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. U.S. Pat. No. 5,703,073 describes nonsteroidal antiinflammatory compounds containing a nitrogen monoxide group indirectly linked to the nonsteroidal antiinflammatory compound and their protection against gastrointestinal, renal and other toxicities normally induced by nonsteroidal antiinflammatory compounds. The compounds described in U.S. Pat. No. 5,703,073 all contain a heteroatom flanked by a carbonyl group in the form of an ester, amide or thioester in the main chain of the linker.

The use of nonsteroidal antiinflammatory compounds for the treatment and/or prevention of ophthalmic diseases or disorders such as glaucoma, inflammations of the eye and elevation of intraocular pressure has been described. For example, U.S. Pat. No. 5,474,985 describes the use of nonsteroidal antiinflammatory compounds to treat or prevent non-inflammatory induced, elevated intraocular pressure associated with the administration of corticosteroids; U.S. Pat. Nos. 5,674,888 and 5,599,535 describe the use of nonsteroidal antiinflammatory compounds to treat loss of trabecular meshwork resulting from aging, exposure to toxic substances, environmental stresses, such as oxidative or phagocytic injury, or glucocorticoid exposure; U.S. Pat. No. 5,814,655 describes topical ophthalmic compositions comprising nonsteroidal antiinflammatory compounds; Wiederholt et al., Invest. Opthalmol. Vis. Sci., 2515-2520 (1994) describes the use of nitric oxide donors to relax trabecular meshwork and ciliary muscle; Behar-Cohen et al., Invest. Opthalmol. Vis. Sci ., describes the use of nitric oxide donors to decrease intraocular pressure.

There is a need in the art for nonsteroidal antiinflammatory compounds that do not have the adverse side effects associated with prior art compounds. There is also a need for new and improved treatments of inflammatory disease states and disorders; and ophthalmic diseases and disorders. The present invention is directed to these, as well as other important ends.

SUMMARY OF THE INVENTION

The present invention is based on the discovery that it is possible to link a nitrogen monoxide group (NO), and/or a nitrogen dioxide group (NO 2 ) (i.e., nitrosylated and/or nitrosated group, respectively) to a nonsteroidal antiinflammatory compound and that the resulting compounds have good bioavailibility, possess potent analgesic and antiinflammatory properties and have an unexpectedly reduced potential for producing gastrointestinal lesions (ulcers). The novel compounds also have unexpected properties in the treatment and/or prevention of ophthalmic diseases and disorders.

The present invention is also based on the discovery that it is possible to administer at least one nitrosated and/or nitrosylated nonsteroidal antiinflammatory compound (NSAID) and at least one nitric oxide donor to prevent, reduce, or reverse gastrointestinal, renal, and other toxicities induced by the NSAID. NSAIDs are antiinflammatory, analgesic and antipyretic compounds that act at cyclooxygenase, the enzyme responsible for the biosyntheses of the prostaglandins and certain autocoid 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 releases or chemically transfers nitric oxide to another molecule. Nitric oxide donors include, for example, S-nitrosothiols, nitrites, N-oxo-N-nitrosamines, and substrates of the various isozymes of nitric oxide synthase.

One aspect of the present invention provides novel nitrosated and/or nitrosylated nonsteroidal antiinflammatory compounds. The nonsteroidal antiinflammatory compound can be nitrosated and/or nitrosylated through one or more sites such as oxygen (hydroxyl condensation), sulfur (sulfhydryl condensation), carbon and/or nitrogen. The nonsteroidal antiinflammatory compound can be, for example, an aryl propionic acid, an aryl acetic acid or an enolic anilide. The present invention also provides compositions comprising such compounds in a pharmaceutically acceptable carrier.

Another aspect of the invention provides compositions comprising a therapeutically effective amount of at least one nitrosated and/or nitrosylated nonsteroidal antiinflammatory compound and at least one compound that donates, transfers or releases nitrogen monoxide as a charged species, i.e., nitrosonium (NO ) or nitroxyl (NO ), or as the neutral species, nitric oxide (NO ), and/or stimulates endogenous production of nitric oxide or endothelium-derived relaxing factor (EDRF) in vivo and/or is a substrate for nitric oxide synthase. The nitrosated and/or nitrosylated nonsteroidal antiinflammatory compounds can be, for example, aryl propionic acids, aryl acetic acids, or enolic anilides. The invention also provides for such compositions in a pharmaceutically acceptable carrier.

Yet another aspect of the present invention provides kits comprising at least one nitrosated and/or nitrosylated nonsteroidal antiinflammatory compound, and, optionally, at least one compound that donates, transfers or releases nitrogen monoxide as a charged species, i.e., nitrosonium (NO ) or nitroxyl (NO ), or as the neutral species, nitric oxide (NO ), and/or stimulates endogenous production of nitric oxide or EDRF in vivo and/or is a substrate for nitric oxide synthase. The nitrosated and/or nitrosylated NSAID and the nitric oxide donor can be separate components in the kit or can be in the form of a composition.

The present invention also provides methods for treating and/or preventing inflammation, pain and fever; decreasing and/or reversing gastrointestinal, renal and other toxicities resulting from the use of nonsteroidal antiinflammatory compounds; and treating and/or preventing gastrointestinal disorders in a patient in need thereof which comprises administering to the patient a therapeutically effective amount of at least one nitrosated and/or nitrosylated nonsteroidal antiinflammatory compound, and, optionally, at least one compound that donates, transfers or releases nitrogen monoxide as a charged species, i.e., nitrosonium (NO ) or nitroxyl (NO ), or as the neutral species, nitric oxide (NO ), and/or stimulates endogenous production of nitric oxide or endothelium-derived relaxing factor (EDRF) in vivo and/or is a substrate for nitric oxide synthase. The nitrosated and/or nitrosylated NSAID and nitric oxide donor can be administered separately or as components of the same composition.

The present invention also provides methods to treat inflammatory disease states and disorders by administering to a patient in need thereof a therapeutically effective amount of at least one nitrosated and/or nitrosylated nonsteroidal antiinflammatory compound, and, optionally, at least one nitric oxide donor. The nitrosated and/or nitrosylated NSAID and nitric oxide donor can be administered separately or as components of the same composition. Such inflammatory disease states and disorders include, 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 rejection, organ preservation, a female or male sexual dysfunctions, radiation-induced injury, asthma, atherosclerosis, thrombosis, platelet aggregation, restenosis, metastasis, influenza, incontinence, stroke, burn, trauma, acute pancreatitis, pyelonephritis, hepatitis, an autoimmune disease, an immunological disorder, senile dementia, insulin-dependent diabetes mellitus, disseminated intravascular coagulation, fatty embolism, Alzheimer's disease, adult or infantile respiratory disease, carcinogenesis or a hemorrhage in a neonate.

The present invention also provides methods to treat and/or prevent ophthalmic diseases and disorders by administering to a patient in need thereof a therapeutically effective amount of at least one nitrosated and/or nitrosylated nonsteroidal antiinflammatory compound, and, optionally, at least one nitric oxide donor. The ophthalmic diseases and disorders include glaucoma, inflammation of the eye and elevation of intraocular pressure. The nitrosated and/or nitrosylated NSAID and nitric oxide donor can be administered separately or as components of the same composition.

These and other aspects of the present invention are explained in detail below.

DETAILED DESCRIPTION OF THE INVENTION

As used throughout the disclosure, the following terms, unless otherwise indicated, shall be understood to have the following meanings.

Gastrointestinal disorder refers to any disease or disorder of the upper gastrointestinal tract of a patient including, for example, peptic ulcers, stress ulcers, gastric hyperacidity, dyspepsia, gastroparesis, Zollinger-Ellison syndrome, gastroesophageal reflux disease, short-bowel (anastomosis) syndrome, hypersecretory states associated with systemic mastocytosis or basophilic leukemia and hyperhistaminemia, and bleeding peptic ulcers that result, for example, from neurosurgery, head injury, severe body trauma or burns.

Upper gastrointestinal tract refers to the esophagus, the stomach, the duodenum and the jejunum.

Ulcers refers to lesions of the upper gastrointestinal tract lining that are characterized by loss of tissue. Such ulcers include gastric ulcers, duodenal ulcers and gastritis.

NSAID refers to a nonsteroidal anti-inflammatory compound or a nonsteroidal anti-inflammatory drug. NSAIDs inhibit cyclooxygenase, the enzyme responsible for the biosyntheses of the prostaglandins and certain autocoid 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.

Patient refers to animals, preferably mammals, more preferably humans.

Transdermal refers to the delivery of a compound by passage through the skin and into the blood stream.

Transmucosal refers to delivery of a compound by passage of the compound through the mucosal tissue and into the blood stream.

Penetration enhancement or permeation enhancement refers to an increase in the permeability of the skin or mucosal tissue to a selected pharmacologically active compound such that the rate at which the compound permeates through the skin or mucosal tissue is increased.

Carriers or vehicles refers to carrier materials suitable for compound administration and include any such material known in the art such as, for example, any liquid, gel, solvent, liquid diluent, solubilizer, or the like, which is non-toxic and which does not interact with any components of the composition in a deleterious manner.

Nitric oxide adduct or NO adduct refers to compounds and functional groups which, under physiological conditions, can donate, release and/or directly or indirectly transfer any of the three redox forms of nitrogen monoxide (NO , NO , NO ), such that the biological activity of the nitrogen monoxide species is expressed at the intended site of action.

Nitric oxide releasing or nitric oxide donating refers to methods of donating, releasing and/or directly or indirectly transferring any of the three redox forms of nitrogen monoxide (NO , NO , NO ), such that the biological activity of the nitrogen monoxide species is expressed at the intended site of action.

Nitric oxide donor or NO donor refers to compounds that donate, release and/or directly or indirectly transfer a nitrogen monoxide species, and/or stimulate the endogenous production of nitric oxide or endothelium-derived relaxing factor (EDRF) in vivo and/or elevate endogenous levels of nitric oxide or EDRF in vivo. NO donor also includes compounds that are substrates for nitric oxide synthase.

Alkyl refers to a lower alkyl group, a haloalkyl group, an alkenyl group, an alkynyl group, a bridged cycloalkyl group, a cycloalkyl group or a heterocyclic ring, as defined herein.

Lower alkyl refers to branched or straight chain acyclic alkyl group comprising one to about ten carbon atoms (preferably one to about eight carbon atoms, more preferably one to about six carbon atoms). Exemplary lower alkyl groups include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, t-butyl, pentyl, neopentyl, iso-amyl, hexyl, octyl, and the like.

Haloalkyl refers to a lower alkyl group, an alkenyl group, an alkynyl group, a bridged cycloalkyl group, a cydoalkyl group or a heterocyclic ring, as defined herein, to which is appended one or more halogens, as defined herein. Exemplary haloalkyl groups include trifluoromethyl, chloromethyl, 2-bromobutyl, 1-bromo-2-chloro-pentyl, and the like.

Alkenyl refers to a branched or straight chain C 2 -C 10 hydrocarbon (preferably a C 2 -C 8 hydrocarbon, more preferably a C 2 -C 6 hydrocarbon) which can comprise one or more carbon carbon double bonds. Exemplary alkenyl groups include propylenyl, buten-1-yl, isobutenyl, penten-1-yl, 2,2-methylbuten-1-yl, 3-methylbuten-1-yl, hexan-1-yl, hepten-1-yl, octen-1-yl, and the like.

Heterocyclic compounds refer to mono- and polycyclic compounds comprising at least one aryl or heterocyclic ring.

Alkylaryl refers to an alkyl group, as defined herein, to which is appended an aryl group, as defined herein. Exemplary alkylaryl groups include benzyl, phenylethyl, hydroxybenzyl, fluorobenzyl, fluorophenylethyl, and the like.

Arylalkyl refers to an aryl radical, as defined herein, attached to an alkyl radical, as defined herein.

Cycloalkylalkyl refers to a cycloalkyl radical, as defined herein, attached to an alkyl radical, as defined herein.

Heterocyclicalkyl refers to a heterocyclic ring radical, as defined herein, attached to an alkyl radical, as defined herein.

Arylheterocyclic ring refers to a bi- or tricyclic ring comprised of an aryl ring, as defined herein, appended via two adjacent carbon atoms of the aryl ring to a heterocyclic ring, as defined herein. Exemplary arylheterocyclic rings include dihydroindole, 1,2,3,4-tetra-hydroquinoline, and the like.

Alkoxy refers to R 50 O , wherein R 50 is an alkyl group, as defined herein. Exemplary alkoxy groups include methoxy, ethoxy, t-butoxy, cyclopentyloxy, and the like.

Arylalkoxy or alkoxyaryl refers to an alkoxy group, as defined herein, to which is appended an aryl group, as defined herein. Exemplary arylalkoxy groups include benzyloxy, phenylethoxy, chlorophenylethoxy, and the like.

Alkoxyalkyl refers to an alkoxy group, as defined herein, appended to an alkyl group, as defined herein. Exemplary alkoxyalkyl groups include methoxymethyl, methoxyethyl, isopropoxymethyl, and the like.

Alkoxyhaloalkyl refers to an alkoxy group, as defined herein, appended to a haloalkyl group, as defined herein. Exemplary alkoxyhaloalkyl groups include 4-methoxy-2-chlorobutyl and the like.

Cycloalkoxy refers to R 54 O , wherein R 54 is a cycloalkyl group or a bridged cycloalkyl group, as defined herein. Exemplary cycloalkoxy groups include cyclopropyloxy, cyclopentyloxy, cyclohexyloxy, and the like.

Haloalkoxy refers to a haloalkyl group, as defined herein, to which is appended an alkoxy group, as defined herein. Exemplary haloalkyl groups include 1,1,1-trichloroethoxy, 2-bromobutoxy, and the like.

Hydroxy refers to OH.

Oxo refers to O.

Oxy refers to O R 77 wherein R 77 is an organic or inorganic cation.

Hydroxyalkyl refers to a hydroxy group, as defined herein, appended to an alkyl group, as defined herein.

Amino refers to NH 2 .

Nitrate refers to O NO 2 .

Nitrite refers to O NO.

Thionitrate refers to S NO 2 .

Thionitrite and nitrosothiol refer to S NO.

Nitro refers to the group NO 2 and nitrosated refers to compounds that have been substituted therewith.

Nitroso refers to the group NO and nitrosylated refers to compounds that have been substituted therewith.

Nitrile and cyano refer to CN.

Alkylamino refers to R 50 NH , wherein R 50 is an alkyl group, as defined herein. Exemplary alkylamino groups include methylamino, ethylamino, butylamino, cyclohexylamino, and the like.

Arylamino refers to R 55 NH , wherein R 55 is an aryl group, as defined herein.

Dialkylamino refers to R 52 R 53 N , wherein R 52 and R 53 are each independently an alkyl group, as defined herein. Exemplary dialkylamino groups include dimethylamino, diethylamino, methyl propargylamino, and the like.

Diarylamino refers to R 55 R 56 N , wherein R 55 and R 60 are each independently an aryl group, as defined herein.

Alkylarylamino refers to R 52 R 55 N , wherein R 52 is an alkyl group, as defined herein, and R 55 is an aryl group, as defined herein.

Aminoalkyl refers to an amino group, an alkylamino group, a dialkylamino group, an arylamino group, a diarylamino group, an alkylarylamino group or a heterocyclic ring, as defined herein, to which is appended an alkyl group, as defined herein.

Aminoaryl refers to an amino group, an alkylamino group, a dialkylamino group, an arylamino group, a diarylamino group, an alkylarylamino group or a heterocyclic ring, as defined herein, to which is appended an aryl group, as defined herein.

Sulfonyl refers to S(O) 2 OR 58 , wherein R 58 is an alkyl group, an aryl group, an alkylaryl group or an aryl heterocyclic ring, as defined herein.

Sulfonic acid refers to S(O) 2 OR 76 , wherein R 76 is a hydrogen, an organic cation or an inorganic cation.

Alkylsulfonic acid refers to a sulfonic acid group, as defined herein, appended to an alkyl group, as defined herein.

Arylsulfonic acid refers to an sulfonic acid group, as defined herein, appended to an aryl group, as defined herein

Sulfonic ester refers to S(O) 2 OR 58 , wherein R 58 is an alkyl group, an aryl group, an alkylaryl group or an aryl heterocyclic ring, as defined herein.

Sulfonamido refers to S(O) 2 N(R 51 )(R 57 ), wherein R 51 and R 57 are each independently a hydrogen atom, an alkyl group, an aryl group, an alkylaryl group, or an arylheterocyclic ring, as defined herein, and R 51 and R 57 when taken together are a heterocyclic ring, a cycloalkyl group or a bridged cycloalkyl group, as defined herein.

Alkylsulfonamido refers to a sulfonamido group, as defined herein, appended to an alkyl group, as defined herein.

Arylsulfonamido refers to a sulfonamido group, as defined herein, appended to an aryl group, as defined herein.

Alkylthio refers to R 50 S , wherein R 50 is an alkyl group, as defined herein.

Arylthio refers to R 55 S , wherein R 55 is an aryl group, as defined herein.

Alkylsulfinyl refers to R 50 S(O) , wherein R 50 is an alkyl group, as defined herein.

Alkylsulfonyl refers to R 50 S(O) 2 , wherein R 50 is an alkyl group, as defined herein.

Arylsulfinyl refers to R 55 S(O) , wherein R 55 is an aryl group, as defined herein.

Arylsulfonyl refers to R 55 S(O) 2 , wherein R 55 is an aryl group, as defined herein.

Amidyl refers to R 51 C(O)N(R 57 ) wherein R 51 and R 57 are each independently a hydrogen atom, an alkyl group, an aryl group, an alkylaryl group, or an arylheterocyclic ring, as defined herein.

Ester refers to R 51 C(O)O wherein R 51 is a hydrogen atom, an alkyl group, an aryl group, an alkylaryl group, or an arylheterocyclic ring, as defined herein.

Carbamoyl refers to O C(O)N(R 51 )(R 57 ), wherein R 51 and R 57 are each independently a hydrogen atom, an alkyl group, an aryl group, an alkylaryl group or an arylheterocyclic ring, as defined herein, or R 51 and R 57 taken together are a heterocyclic ring, a cycloalkyl group or a bridged cycloalkyl group, as defined herein.

Carboxyl refers to C(O)OR 76 wherein R 76 is a hydrogen, an organic cation or an inorganic cation.

Carboxylic ester refers to C(O)OR 58 , wherein R 58 is an alkyl group, an aryl group, an alkylaryl group or an aryl heterocyclic ring, as defined herein.

Alkylcarboxylic acid and alkylcarboxyl refer to an alkyl group, as defined herein, appended to a carboxyl group, as defined herein.

Alkylcarboxylic ester refers to an alkyl group, as defined herein, appended to a carboxylic ester group, as defined herein.

Arylcarboxylic acid refers to an aryl group, as defined herein, appended to a carboxyl group, as defined herein.

Arylcarboxylic ester refers to an aryl group, as defined herein, appended to a carboxylic ester group, as defined herein.

Carboxamido refers to C(O)N(R 51 )(R 57 ), wherein R 51 and R 57 are each independently a hydrogen atom, an alkyl group, an aryl group, an alkylaryl group or an arylheterocyclic ring, as defined herein, and R 51 and R 57 when taken together are a heterocyclic ring, a cycloalkyl group or a bridged cycloalkyl group, as defined herein.

Alkylcarboxamido refers to an alkyl group, as defined herein, appended to a carboxamido group, as defined herein.

Arylcarboxamido refers to an aryl group, as defined herein, appended to a carboxamido group, as defined herein.

Urea refers to N(R 58 ) C(O)N(R 51 )(R 57 ) wherein R 51 , R 57 , and R 58 are each independently a hydrogen atom, an alkyl group, an aryl group, an alkylaryl group, or an arylheterocyclic ring, as defined herein, or R 51 and R 57 taken together are a heterocyclic ring, a cycloalkyl group or a bridged cycloalkyl group, as defined herein.

Phosphoryl refers to P(R 70 )(R 71 )(R 72 ), wherein R 70 is a lone pair of electrons, thial or oxo, and R 71 and R 72 are each independently a covalent bond, a hydrogen, a lower alkyl, an alkoxy, an alkylamino, a hydroxy, an oxy or an aryl, as defined herein.

Silyl refers to Si(R 73 )(R 74 )(R 75 ), wherein R 73 , R 74 and R 75 are each independently a covalent bond, a lower alkyl, an alkoxy, an aryl or an arylalkoxy, as defined herein.

The NSAIDs that are nitrosated and/or nitrosylated in accordance with the invention and/or are included in the compositions of the invention can be any of those known in the art, including those exemplified below.

Despite the introduction of many new drugs, aspirin (acetylsalicylic acid) is still the most widely prescribed antiinflammatory, analgesic and antipyretic compound 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 for use in the present invention are 5-aminosalicylic acid (mesalamine), salicylazosulfapyridine (sulfasalazine) and methylsalicylate.

Another group of NSAIDs are the para-aminophenol derivatives, which are the so-called coal tar analgesics, including, for example, phenacetin and its active metabolite acetaminophen.

Another group of compounds for use in the present invention include indomethacin, a methylated indole derivative, and the structurally related compound sulindac.

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 is mefenamic, meclofenamic, flufenamic, tolfenamic and etofenamnic acids. They are used either as the acid or as pharmaceutically acceptable salts.

Another contemplated NSAID is tolmetin which, like the other NSAIDs discussed herein, causes gastric erosion and prolonged bleeding time.

Another group of NSAIDs are the propionic acid derivatives. Principal members of this group are, for example, ibuprofen, naproxen, flurbiprofen, fenoprofen and ketoprofen. Other members of this group, in use or study in countries outside the U.S., include, for example, fenbufen, pirprofen, oxaprozin, indoprofen and tiaprofenic acid.

Also contemplated for use in the present invention are piroxicam and ampiroxicam, oxicam derivatives which are a class of antiinflammatory enolic acids. The other related compounds, tenoxicam and tenidap, can also be used. Another compound that is particularly preferred in the present invention is diclofenac, one of the series of phenylacetic acid derivatives that have been developed as antiinflammatory compounds. Other NSAIDs which are contemplated as suitable in the present invention include etodolac and nabumentone.

Each of the above NSAIDs is described more fully in the literature, such as in Goodman and Gilman, The Pharmacological Basis of Therapeutics (9th Edition), McGraw-Hill, 1995, Pgs. 617-657; the Merck Index on CD-ROM, Twelfth Edition, Version 12:1, 1996.

In one embodiment, the present invention describes nitrosated and/or nitrosylated NSAIDs of Formula (I):

wherein

R g is a hydrogen atom or a lower alkyl group;

R h is:

n is an integer of 0 or 1;

(i) -T-B l W B t -T-NO s ;

wherein

s is an integer of 1 or 2;

T at each occurrence is independently a covalent bond, a carbonyl, an oxygen, S(O) o or N(R a )(R i ) ;

o is an integer from 0 to 2;

R a is a lone pair of electrons, a hydrogen or an alkyl group;

R i is a hydrogen, an alkyl group, an aryl group, an alkylcarboxylic acid group, an aryl carboxylic acid group, an alkylcarboxylic ester group, an arylcarboxylic ester group, an alkylcarboxamido, an arylcarboxamido, an alkylaryl, an alkylsulfinyl, an alkylsulfonyl, an arylsulfinyl, an arylsulfonyl, a sulfonamido, a carboxamido, a carboxylic ester, an amino alkyl, an amino aryl, CH 2 C (T-Q)(R e )(R f ), or (N 2 O 2 ) M , wherein M is an organic or inorganic cation,

q is an integer from 1 to 5;

B at each occurrence is independently an alkyl group, an aryl group, (C(R e )(R f )) p , a heterocyclic ring, an aryl heterocyclic ring, or (CH 2 CH 2 O) q ;

p is an integer from 1 to 10;

R e and R f are each independently a hydrogen, an alkyl, a cycloalkoxy, a halogen, a hydroxy, an hydroxyalkyl, an alkoxyalkyl, an arylheterocyclic ring, an alkylaryl, a cycloalkylalkyl, a heterocyclicalkyl, an alkoxy, a haloalkoxy, an amino, an alkylamino, a dialkylamino, an arylamino, a diarylamino, an alkylarylamino, an alkoxyhaloalkyl, a haloalkoxy, a sulfonic acid, an alkylsulfonic acid, an arylsulfonic acid, an arylalkoxy, an alkylthio, an arylthio, a cyano, an aminoalkyl, an aminoaryl, an alkoxy, an aryl, an arylalkyl, an alkylaryl, a carboxamido, a alkyl carboxamido, an aryl carboxamido, an amidyl, a carboxyl, a carbamoyl, an alkylcarboxylic acid, an arylcarboxylic acid, an ester, a carboxylic ester, an alkylcarboxylic ester, an arylcarboxylic ester, a haloalkoxy, a sulfonamido, an alkylsulfonamido, an arylsulfonamido, a urea, a nitro, -T-NO s , or (C(R e )(R f )) k -T-NOR s , or R e and R f taken together with the carbon atoms to which they are attached are a heterocyclic ring, a cycloalkyl group or a bridged cycloalkyl group;

R b and R c are each independently a haloalkyl, an alkenyl group, an alkynyl group, a bridged cycloalkyl group, a heterocyclic ring, a cycloalkoxy, a halogen, a hydroxy, an hydroxyalkyl, an alkoxyalkyl, an arylheterocyclic ring, an alkylaryl, a cycloalkylalkyl, a heterocyclicalkyl, an alkoxy, a haloalkoxy, an amino, an alkylamino, a dialkylamino, an arylamino, a diarylamino, an alkylarylamino, an alkoxyhaloalkyl, a haloalkoxy, a sulfonic acid, an alkylsulfonic acid, an arylsulfonic acid, an arylalkoxy, an alkylthio, an arylthio, a cyano, an aminoalkyl, an aminoaryl, an alkoxy, an arylalkyl, an alkylaryl, a carboxamido, an alkyl carboxamido, an aryl carboxamido, an amidyl, a carboxyl, a carbamoyl, an alkylcarboxylic acid, an arylcarboxylic acid, an ester, a carboxylic ester, an alkylcarboxylic ester, an arylcarboxylic ester, a haloalkoxy, a sulfonamido, an alkylsulfonamido, an arylsulfonamido, a urea, a nitro, -T-NO s , or (C(R e )(R f )) k -T-NO s , or R b and R c taken together with the carbon atoms to which they are attached are a carbonyl, a methanthial, a heterocyclic ring, a cloalkyl group or a bridged cycloalkyl group;

J is a carbonyl, a phosphoryl or a silyl;

k, l, t and z are each independently an integer from 1 to 3;

y is an integer from 1 to 3;

x and r are each independently an integer from 0 to 3;

with the proviso that when R i is CH 2 C(T-NO s )(R e )(R f ) or (N 2 O 2 ) M , or R b , R c , R e or R f are T-NO s or (C(R e )(R f )) k -T-NO s , then the -T-NO s subgroup designated in X can be a hydrogen, an alkyl, an alkoxy, an alkoxyalkyl, an aminoalkyl, a hydroxy, a heterocyclic ring or an aryl group.

In cases where multiple designations of variables which reside in sequence are chosen as a covalent bond or the integer chosen is 0, the intent is to denote a single covalent bond connecting one radical to another. For example, B 0 would denote a covalent bond, while B 2 denotes (B B) and (C(R e )(R f )) 2 denotes C(R e )(R f ) C(R e )(R f ) .

Another embodiment of the present invention describes nitrosated and/or nitrosylated NSAIDs of Formula (II):

wherein

R k is:

and X is as defined herein.

Another embodiment of the present invention describes nitrosated and/or nitrosylated NSAIDs of Formula (III)

wherein

X is as defined herein;

R i at each occurrence is independently R i , wherein R i is as defined herein;

Z is an aryl group; and

A 1 , A 2 and A 3 comprise the other subunits of a 5- or 6-membered monocyclic aromatic ring and each of A 1 , A 2 and A 3 is independently:

(1) C R o , wherein R o at each occurrence is independently a hydrogen, an alkyl, an alkoxyalkyl, a halogen or a nitro group;

(2) N R p , wherein R p at each occurrence is independently a covalent bond to an adjacent ring atom in order to render the ring aromatic, a hydrogen, an alkyl, an arylalkyl, an aryl or a heteroaryl group;

(4) an oxygen atom; or

(5) B a B b , wherein B a and B b are each independently a nitrogen atom or C R o wherein R o is as defined herein.

Another embodiment of the present invention describes nitrosated and/or nitrosylated NSAIDs of Formula (IV):

wherein

R m is an alkyl group or an aryl group; and X, Z, A 1 , A 2 and A 3 are as defined herein.

Compounds of the present invention which have one or more asymmetric carbon atoms can 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.

Another aspect of the present invention provides processes for making the novel compounds of the invention and to the intermediates useful in such processes. The compounds of the present invention for Formulas (I), (II), (III) and (IV) can be synthesized by one skilled in the art following the methods and examples described herein. The reactions are performed in solvents appropriate to the reagents and materials used are suitable for the transformations being effected. It is understood by one skilled in the art of organic synthesis that the functionality present in the compound 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, but alternative methods and substituents compatible with the reaction conditions will be readily apparent to one skilled in the art. The use of sulfur and oxygen protecting groups is well known in the art for protecting thiol and alcohol groups against undesirable reactions during a synthetic procedure and many such protecting groups are known, such as those described by T. H. Greene and P. G. M. Wuts, Protective Groups in Organic Synthesis , John Wiley & Sons, New York (1991), the disclosure of which is incorporated by reference herein in its entirety.

The chemical reactions described above are generally disclosed in terms of their broadest application to the preparation of the compounds of this invention. Occasionally, the reactions may not be applicable as described to each compound included within the disclosed scope. The compounds for which this occurs will be readily recognized by one skilled in the art. In all such cases, either the reactions can be successfully performed by conventional modifications known to one skilled in the art, e.g., by appropriate protection of interfering groups, by changing to alternative conventional reagents, by routine modification of reaction conditions, and the like, or other reactions disclosed herein or otherwise conventional, will be applicable to the preparation of the corresponding compounds of this invention. In all preparative methods, all starting materials are known or readily preparable from known starting materials.

Nitroso compounds of Formula (I), wherein R g and R h are as defined herein, and an O-nitrosylated NSAID ester in which 2 4- 2-(nitrosooxy)ethyl piperazinyl ethan-1-ol is representative of the X group as defined herein may be prepared as described below. An appropriate acid (i.e., Formula (I) where X is substituted with hydroxyl) is converted into the ester by reaction with an appropriate monoprotected diol. Preferred methods for the preparation of esters are initially forming the mixed anhydride via reaction of the acid 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 may first be 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 produce the ester. Alternatively, the acid and monoprotected diol may be coupled to produce the ester by treatment with a dehydration agent such as DCC. Alternatively, the acid may first be converted into an alkali metal salt such as the sodium, potassium or lithium salt, and reacted with an alkyl halide that also contains a protected hydroxyl group in a polar solvent such as DMF to produce the ester. 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 dichlormethane, THF, DMF or acetonitrile produces the compound of Formula (I).

Nitroso compounds of Formula (I), where R g and R h are as defined herein, and a S-nitrosylated NSAID ester in which 2- methyl 2-methyl-2-(nitrosothiol) propyl amino ethan-1-ol is representative of the X group as defined herein may be synthesized as described below. An appropriate acid (i.e., Formula (I) where X is substituted with hydroxyl) is converted into the ester 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 the acid with a chloroformate such as isobutylchloroformate in the presence of a non-nucelophilic 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-dimethylamnine pyridine. Alternatively, the acid may first be 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 produce an ester. Alternatively, the appropriate acid and protected thiol-containing alcohol may be coupled to produce the ester by treatment with a dehydration agent such as DCC. Alternatively, the acid may first be 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 a polar solvent such as DMF to produce the ester. Preferred protecting groups for the thiol moiety are as a thioester such as thioacetate or thiobenzoate, as a disulfide, as a thiocarbamate such as N-methoxymethyl thiocarbamate, or as a thioether such as 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 used 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 methylene chloride, THF, DMF or acetonitrile produces the compound of Formula (I). Alternatively, a stoichiometric quantity of sodium nitrite in aqueous acid produces the compound of Formula (I).

Nitroso compounds of Formula (II), where R k is defined herein and a S-nitrosylated NSAID ester in which 2- methyl 2-methyl-2-(nitrosothiol)propyl amino ethan-1-ol is representative of the X group as defined herein may be synthesized as described below. An appropriate acid (i.e., Formula (II) where X is substituted with hydroxyl) is converted into the ester 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 the acid with a chloroformate such as isobutylchloroformate in the presence of a non-nucelophilic 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 may first be 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 produce an ester. Alternatively, the appropriate acid and protected thiol-containing alcohol may be coupled to produce the ester by treatment with a dehydration agent such as DCC. Alternatively, the acid may first be 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 a polar solvent such as DMF to produce the ester. Preferred protecting groups for the thiol moiety are as a thioester such as thioacetate or thiobenzoate, as a disulfide, as a thiocarbamate such as N-methoxymethyl thiocarbamate, or as a thioether such as 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 used 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 methylene chloride, THF, DMF or acetonitrile produces the compound of Formula (II). Alternatively, a stoichiometric quantity of sodium nitrite in aqueous acid produces the compound of Formula (II).

Nitroso compounds of Formula (II) where R k is as defined herein and an O-nitrosylated NSAID ester in which 2 4- 2-(nitrosooxy)ethyl piperazinyl ethan-1-ol is representative of the X group as defined herein may be prepared as described below. An appropriate acid (i.e., Formula (II) where X is substituted with hydroxyl) is converted into the ester by reaction with an appropriate monoprotected diol. Preferred methods for the preparation of esters are initially forming the mixed anhydride via reaction of the acid 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 may first be converted to the acid chloride 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 produce the ester. Alternatively, the acid and monoprotected diol may be coupled to produce the ester by treatment with a dehydration agent such as DCC. Alternatively, the acid may first be converted into an alkali metal salt such as the sodium, potassium or lithium salt, and reacted with an alkyl halide that also contains a protected hydroxyl group in a polar solvent such as DMF to produce the ester. Preferred protecting groups for the alcohol moiety are silyl ethers such as 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 produces the compound of Formula (II).

Nitroso compounds of Formula (III) wherein A 1 , A 2 , A 3 , R i and Z are as defined herein and an S-nitrosylated enol ester in which 2- methyl 2-methyl-2-nitrosothiol)propyl amino acetyl is representative of the X group as defined herein may be prepared as described below. The enolic form of the -keto amide of Formula (III) where X is substituted with hydrogen is converted to the ester by reaction with an appropriate protected thiol containing activated acylating agent. Preferred methods for the formation of an 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 used 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 methylene chloride, THF, DMF or acetonitrile with or without an amine base such as pyridine or triethylamine acid produces the compound of Formula (III). Alternatively, a stoichi metric quantity of sodium nitrite in aqueous acid produces the compound of Formula (III).

Nitroso compounds of Formula (III) wherein A 1 , A 2 , A 3 , R i and Z are as defined herein and an O-nitrosylated enol ester in which 2- methyl 2-methyl-2-nitrosooxy)ethyl amino acetyl is representative of the X group as defined herein may be prepared as described below. The enolic form of the -keto amide of Formula (III) where X is substituted by hydrogen is converted to the ester by reaction with an appropriate protected alcohol containing activated acylating agent. 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 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 triethylamnine produces the compound of Formula (III).

Nitroso compounds of Formula (IV) wherein A 1 , A 2 , A 3 , R m and Z are as defined herein and an S-nitrosylated enol ester in which 2- methyl 2-methyl-2-nitrosothiol)propyl amino acetyl is representative of the Y group as defined herein may be prepared as described below. The enolic form of the -keto amide of Formula (IV) where X is substituted with hydrogen is converted to the ester by reaction with an appropriate protected thiol-containing alcohol activated acylating agent. Preferred methods for the formation of an 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 used 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 methylene chloride, THF, DMF or acetonitrile with or without an amine base such as pyridine or triethylamine acid produces the compound of Formula (IV). Alternatively, a stoichiometric quantity of sodium nitrite in aqueous acid produces the compound of Formula (IV).

Nitroso compounds of Formula (IV) wherein A 1 , A 2 , A 3 , R m and Z are as defined herein and an O-nitrosylated enol ester in which 2- methyl 2-methyl-2-nitrosooxy)ethyl amino acetyl is representative of the X group as defined herein may be prepared as described below. The enolic form of the -keto amide of Formula (IV) where X is substituted by hydrogen is converted to the ester by reaction with an appropriate protected alcohol containing activated acylating agent. 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 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 produces the compound of Formula (IV).

The compounds of the present invention include NSAIDs, including those described herein, which have been nitrosated and/or nitrosylated through one or more sites such as oxygen (hydroxyl condensation), sulfur (sulfflydryl condensation), carbon and/or nitrogen. The nitrosated and/or nitrosylated NSAIDs of the present invention donate, transfer or release a biologically active form of nitrogen monoxide (i.e., nitric oxide).

Nitrogen monoxide can exist in three forms: NO (nitroxyl), NO (uncharged nitric oxide) and NO (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 the nitric oxide radical (NO ), nitrosonium (NO ) does not react with O 2 or O 2 species, and functionalities capable of transferring and/or releasing NO and NO are also resistant to decomposition in the presence of many redox metals. Consequently, administration of charged NO equivalents (positive and/or negative) is a more effective means of delivering a biologically active NO to the desired site of action.

Compounds contemplated for use in the present invention (e.g., nitrosated and/or nitrosylated NSAIDs) are, optionally, used in combination with nitric oxide and compounds that release nitric oxide or otherwise directly or indirectly deliver or transfer a biologically active form of nitrogen monoxide to a site of its intended activity, such as on a cell membrane in vivo.

The term nitric oxide encompasses uncharged nitric oxide (NO) and charged nitrogen monoxide species, preferably charged nitrogen monoxide species, such as nitrosonium ion (NO ) and nitroxyl ion (NO ). The reactive form of nitric oxide can be provided by gaseous nitric oxide. The nitrogen monoxide releasing, delivering or transferring compounds have the structure F-NO, wherein F is a nitrogen monoxide releasing, delivering or transferring moiety, and include any and all such compounds which provide nitrogen monoxide to its intended site of action in a form active for its intended purpose. The term NO adducts encompasses any nitrogen monoxide releasing, delivering or transferring compounds, including, for example, S-nitrosothiols, nitrites, nitrates, S-nitrothiols, sydnonimines, 2-hydroxy-2-nitrosohydrazines (NONOates), (E)-alkyl-2- (E)-hydroxyimino -5-nitro-3-hexene amines or amnides, nitrosoamines, furoxans as well as substrates for the endogenous enzymes which synthesize nitric oxide. The NO adducts can be mono-nitrosylated, poly-nitrosylated, mono-nitrosated and/or poly-nitrosated at a variety of naturally susceptible or artificially provided binding sites for biologically active forms of nitrogen monoxide.

One group of NO adducts is the S-nitrosothiols, which are compounds that include at least one S NO group. These compounds include S-nitroso-polypeptides (the term polypeptide includes proteins and 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-nitrosylated sugars; S-nitrosylated, modified and unmodified, oligonucleotides (preferably of at least 5, and more preferably 5-200 nucleotides); straight or branched, saturated or unsaturated, aliphatic or aromatic, substituted or unsubstituted S-nitrosylated hydrocarbons; and S-nitroso heterocyclic compounds. S-nitrosothiols and methods for preparing them are described in U.S. Pat. Nos. 5,380,758 and 5,703,073; WO 97/27749; WO 98/19672; and Oae et al, Org. Prep. Proc. Int., 15(3):165-198 (1983), the disclosures of each of which are incorporated by reference herein in their entirety.

Another embodiment of the present invention is S-nitroso amino acids where the nitroso group is linked to a sulfur group of a sulfur-containing amino acid or derivative thereof. Such compounds include, for example, S-nitroso-N-acetylcysteine, S-nitroso-captopril, S-nitroso-N-acetylpenicillamine, S-nitroso-homocysteine, S-nitroso-cysteine and S-nitroso-glutathione.

Suitable S-nitrosylated proteins include thiol-containing proteins (where the NO group is attached to one or more sulfur groups 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 immunoglobulins, antibodies and cytokines. Such nitrosylated proteins are described in WO 93/09806, the disclosure of which is incorporated by reference herein in its entirety. Examples include polynitrosylated albumin where one or more thiol or other nucleophilic centers in the protein are modified.

Other examples of suitable S-nitrosothiols include:

(ii) ONS(C(R e )(R f )) m R e ; and

wherein m is an integer from 2 to 20; R e and R f are each independently a hydrogen, an alkyl, a cycloalkoxy, a halogen, a hydroxy, an hydroxyalkyl, an alkoxyalkyl, an arylheterocyclic ring, an alkylaryl, a cycloalkylalkyl, a heterocyclicalkyl, an alkoxy, a haloalkoxy, an amino, an alkylamino, a dialkylamino, an arylamino, a diarylamino, an alkylarylamino, an alkoxyhaloalkyl, a haloalkoxy, a sulfonic acid, an alkylsulfonic acid, an arylsulfonic acid, an arylalkoxy, an alkylthio, an arylthio, a cyano, an aminoalkyl, an aminoaryl, an alkoxy, an aryl, an arylalkyl, an alkylaryl, a carboxamido, a alkyl carboxamido, an aryl carboxamido, an amidyl, a carboxyl, a carbamoyl, an alkylcarboxylic acid, an arylcarboxylic acid, an ester, a carboxylic ester, an alkylcarboxylic ester, an arylcarboxylic ester, a haloalkoxy, a sulfonamido, an alkylsulfonamido, an arylsulfonamido, a urea, a nitro, or -T-Q; or R e and R f taken together are a carbonyl, a methanthial, a heterocyclic ring, a cycloalkyl group or a bridged cycloalkyl group; Q is NO or NO 2 ; and T is independently a covalent bond, a carbonyl, an oxygen, S(O) o or N(R a )R I , wherein o is an integer from 0 to 2, R a is a lone pair of electrons, a hydrogen or an alkyl group; R i is a hydrogen, an alkyl, an aryl, an alkylcarboxylic acid, an aryl carboxylic acid, an alkylcarboxylic ester, an arylcarboxylic ester, an alkylcarboxamido, an arylcarboxamido, an alkylaryl, an alkylsulfinyl, an alkylsulfonyl, an arylsulfinyl, an arylsulfonyl, a sulfonamido, a carboxamido, a carboxylic ester, an amino alkyl, an amino aryl, CH 2 C(T-Q)(R e )(R f ), or (N 2 O 2 ) M , wherein M is an organic or inorganic cation; with the proviso that when R i is CH 2 C(T-Q)(R e )(R f ) or (N 2 O 2 ) M ; then -T-Q can be a hydrogen, an alkyl group, an alkoxyalkyl group, an aminoalkyl group, a hydroxy group or an aryl group.

In cases where R e and R f are a heterocyclic ring or taken together R e and R f are a heterocyclic ring, then R i can be a substituent on any disubstituted nitrogen contained within the radical wherein R i is as defined herein.

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 2 under acidic conditions (pH is about 2.5) which yields the S-nitroso derivative. Acids which can be used for this purpose include aqueous sulfuric, acetic and hydrochloric acids. The thiol precursor can also be nitrosylated by reaction with an organic nitrite such as tert-butyl nitrite, or a nitrosonium salt such as nitrosonium tetraflurorborate in an inert solvent.

Another group of NO adducts for use in the present invention include nitrates that donate, transfer or release nitric oxide, such as compounds comprising at least one O 2 N O , O 2 N N , O 2 N S or O 2 N C group. Preferred among these compounds are O 2 N O , O 2 N N , O 2 N S or O 2 N C polypeptides (the term polypeptide includes proteins and also polyamino acids that do not possess an ascertained biological function, and derivatives thereof); O 2 N O , O 2 N N , O 2 N S or O 2 N C amino acids (including natural and synthetic amino acids and their stereoisomers and racemic mixtures); O 2 N O , O 2 N N , O 2 N S or O 2 N C-sugars; O 2 N O , O 2 N N , O 2 N S or O 2 N C modified and unmodified oligonucleotides (comprising at least 5 nucleotides, preferably 5-200 nucleotides); O 2 N O , O 2 N N , O 2 N S or O 2 N C straight or branched, saturated or unsaturated, aliphatic or aromatic, substituted or unsubstituted hydrocarbons; and O 2 N O , O 2 N N , O 2 N S or O 2 N C heterocyclic compounds. Preferred examples of compounds comprising at least one O 2 N O , O 2 N N , O 2 N S or O 2 N C group include isosorbide dinitrate, isosorbide mononitrate, clonitrate, erythrityltetranitrate, mannitol hexanitrate, nitroglycerin, pentaerythritoltetranitrate, pentrinitrol and proparylnitrate.

Another group of NO adducts are N-oxo-N-nitrosoamines that donate, transfer or release nitric oxide and are represented by the formula: R 1 R 2 N(O-M ) NO, where R 1 and R 2 are each independently a polypeptide, an amino acid, a sugar, a modified or unmodified oligonucleotide, a straight or branched, saturated or unsaturated, aliphatic or aromatic, substituted or unsubstituted hydrocarbon, or a heterocyclic group, and where M is an organic or inorganic cation, such as, for example, an alkyl substituted ammonium cation or a Group I metal cation.

Another group of NO adducts are thionitrates that donate, transfer or release nitric oxide and are represented by the formula: R 1 (S) NO 2 , where R 1 is a polypeptide, an amino acid, a sugar, a modified or unmodified oligonucleotide, a straight or branched, saturated or unsaturated, aliphatic or aromatic, substituted or unsubstituted hydrocarbon, or a heterocyclic group. Preferred are those compounds where R 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. Compounds which form disulfide species release nitroxyl ion (NO ) and uncharged nitric oxide (NO ). Compounds where the thiol groups are not sufficiently close to form disulfide bridges generally provide nitric oxide as the NO form and not as the uncharged NO form.

The present invention is also directed to compounds that stimulate endogenous NO or elevate levels of endogenous endothelium-derived relaxing factor (EDRF) in vivo or are substrates for nitric oxide synthase. Such compounds include, for example, L-arginine, L-homoarginine, and N-hydroxy-L-arginine, including their nitrosated and nitrosylated analogs (e.g., nitrosated L-arginine, nitrosylated L-arginine, nitrosated N-hydroxy-L-arginine, nitrosylated N-hydroxy-L-arginine, nitrosated L-homoarginine and nitrosylated L-homoarginine), precursors of L-arginine and/or physiologically acceptable salts thereof, including, for example, citrulline, ornithine or glutamine, inhibitors of the enzyme arginase (e.g., N-hydroxy-L-arginine and 2(S)-amino-6-boronohexanoic acid) and the substrates for nitric oxide synthase, cytokines, adenosin, bradykinin, calreticulin, bisacodyl, and phenolphthalein. EDRF is a vascular relaxing factor secreted by the endothelium, and has been identified as nitric oxide (NO) or a closely related derivative thereof (Palmer et al, Nature, 327:524-526 (1987); Ignarro et al, Proc. Natl. Acad. Sci. USA, 84:9265-9269 (1987)).

The present invention is also based on the discovery that the administration of a therapeutically effective amount of the compounds and compositions described herein is effective for treating inflammation, pain and fever. For example, the patient can be administered a therapeutically effective amount of at least one nitrosated and/or nitrosylated NSAID of the present invention. In another embodiment, the patient can be administered a therapeutically effective amount of at least one nitrosated and/or nitrosylated NSAID, and, at least one compound that donates, transfers or releases nitric oxide, or elevates levels of endogenous EDRF or nitric oxide, or is a substrate for nitric oxide synthase. The compounds can be administered separately or in the form of a composition.

Another aspect of the invention provides methods to decrease or reverse gastrointestinal, renal and other toxicity (such as, for example, kidney toxicity) resulting from the use of nonsteroidal antiinflammatory drugs by administering to a patient in need thereof a therapeutically effective amount of the compounds and/or compositions described herein. For example, the patient can be administered a therapeutically effective amount of at least one nitrosated and/or nitrosylated NSAID, and, optionally, at least one compound that donates, transfers or releases nitric oxide, or elevates levels of endogenous EDRF or nitric oxide, or is a substrate for nitric oxide synthase. The nitrosated and/or nitrosylated NSAID and nitric oxide donor can be administered separately or as components of the same composition.

Another aspect of the invention provides methods for decreasing and/or preventing gastrointestinal disorders by administering to the patient in need thereof a therapeutically effective amount of the compounds and/or compositions described herein. For example, the patient can be administered a therapeutically effective amount of at least one nitrosated and/or nitrosylated NSAID, and, optionally, at least one compound that donates, transfers or releases nitric oxide, or elevates levels of endogenous EDRF or nitric oxide, or is a substrate for nitric oxide synthase. The nitrosated and/or nitrosylated NSAID and nitric oxide donor can be administered separately or as components of the same composition. Such gastrointestinal disorders include, for example, peptic ulcers, stress ulcers, gastric hyperacidity, dyspepsia, gastroparesis, Zollinger-Ellison syndrome, gastroesophageal reflux disease, short-bowel (anastomosis) syndrome, hypersecretory states associated with systemic mastocytosis or basophilic leukemia and hyperhistaminemia, and bleeding peptic ulcers that result, for example, from neurosurgery, head injury, severe body trauma or burns.

Another aspect of the invention provides methods for treating inflammatory disease states and disorders by administering to the patient in need thereof a therapeutically effective amount of at least one nitrosated and/or nitrosylated nonsteroidal antiinflammatory compound, and, optionally, at least one nitric oxide donor. Such inflammatory disease states and disorders include, 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 rejection, organ preservation, a female or male sexual dysfunction, radiation-induced injury, asthma, atherosclerosis, thrombosis, platelet aggregation, restenosis, metastasis, influenza, incontinence, stroke, bur, trauma, acute pancreatitis, pyelonephritis, hepatitis, an autoimmune diseases, an immunological disorder, senile dementia, insulin-dependent diabetes mellitus, disseminated intravascular coagulation, fatty embolism, Alzheimer's disease, adult or infantile respiratory disease, carcinogenesis or a hemorrhage in a neonate. The compounds and compositions of the present invention can also be administered in combination with other medications used for the treatment of these disorders.

Another aspect of the invention provides methods for treating and/or preventing ophthalmic diseases and disorders in a patient by administering to the patient a therapeutically effect amount of at least one nitrosated and/or nitrosylated nonsteroidal antiinflammatory compound, and optionally at least one nitric oxide donor. For example, the patient can be administered a therapeutically effective amount of at least one nitrosated and/or nitrosylated NSAID, and, optionally, at least one compound that donates, transfers or releases nitric oxide, or elevates levels of endogenous EDRF or nitric oxide, or is a substrate for nitric oxide synthase. The nitrosated and/or nitrosylated NSAID and nitric oxide donor can be administered separately or as components of the same composition. Such ophthalmic diseases and disorders include, for example, glaucoma, inflammation of the eye and elevation of intraocular pressure.

When administered in vivo, the compounds and compositions of the present invention can be administered in combination with pharmaceutically acceptable carriers and in dosages described herein. When the compounds and compositions of the present invention are administered as a mixture of at least one nitrosated and/or nitrosylated NSAID and at least one 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 targeted for treatment. The nitric oxide donors and/or other additional compounds can be administered simultaneously with, subsequently to, or prior to administration of the nitrosated and/or nitrosylated NSAID.

The compounds and compositions of the present invention can be administered by any available and effective delivery system including, but not limited to, orally, bucally, parenterally, by inhalation spray, by topical application, by injection, transdermally, or rectally (e.g., by the use of suppositories) in dosage unit formulations containing conventional nontoxic pharmaceutically acceptable carriers, adjuvants, and vehicles, as desired. Parenteral includes subcutaneous injections, intravenous, intramuscular, intrasternal injection, or infusion techniques.

Transdermal compound administration, which is known to one skilled in the art, involves the delivery of pharmaceutical compounds via percutaneous passage of the compound into the systemic circulation of the patient. Topical administration can also involve the use of transdermal administration such as transdermal patches or iontophoresis devices. Other components can be incorporated into the transdermal patches as well. For example, compositions and/or transdermal patches can be formulated with one or more preservatives or bacteriostatic agents including, but not limited to, methyl hydroxybenzoate, propyl hydroxybenzoate, chlorocresol, benzalkonium chloride, and the like. Dosage forms for topical administration of the compounds and compositions can include creams, sprays, lotions, gels, ointments, eye drops, nose drops, ear drops, and the like. In such dosage forms, the compositions of the invention can be mixed to form white, smooth, homogeneous, opaque cream or lotion with, for example, benzyl alcohol 1% or 2% (wt/wt) as a preservative, emulsifying wax, glycerin, isopropyl palmitate, lactic acid, purified water and sorbitol solution. In addition, the compositions can contain polyethylene glycol 400. 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., gauze, 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.

Solid dosage forms for oral administration can include capsules, tablets, effervescent tablets, chewable tablets, pills, powders, sachets, granules and gels. In such solid dosage forms, the active compounds can be admixed with at least one inert diluent such as sucrose, lactose or starch. Such dosage forms can 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, effervescent tablets, and pills, the dosage forms can also comprise buffering agents. Soft gelatin capsules can be prepared to contain a mixture of the active compounds or compositions of the present invention and vegetable oil. Hard gelatin capsules can contain granules of the active compound in combination with a solid, pulverulent carrier such as lactose, saccharose, sorbitol, mannitol, potato starch, corn starch, amylopectin, cellulose derivatives of gelatin. Tablets and pills can be prepared with enteric coatings.

Liquid dosage forms for oral administration can include pharmaceutically acceptable emulsions, solutions, suspensions, syrups, and elixirs containing inert diluents commonly used in the art, such as water. Such compositions can also comprise adjuvants, such as wetting agents, emulsifying and suspending agents, and sweetening, flavoring, and perfuming agents.

Suppositories for vaginal or rectal administration of the compounds and compositions of the invention, such as for treating pediatric fever and the like, can be prepared by mixing the compounds or compositions with a suitable nonirritating excipient such as cocoa butter and polyethylene glycols which are solid at room temperature but liquid at rectal temperature, such that they will melt in the rectum and release the drug.

Injectable preparations, for example, sterile injectable aqueous or oleaginous suspensions can be formulated according to the known art using suitable dispersing agents, wetting agents and/or suspending agents. The sterile injectable preparation can 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 can be used are water, Ringer's solution, and isotonic sodium chloride solution. Sterile fixed oils are also conventionally used as a solvent or suspending medium.

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, for example, water, salt solutions, alcohol, vegetable oils, polyethylene glycols, gelatin, lactose, amylose, magnesium stearate, talc, surfactants, silicic acid, viscous paraffin, perfume oil, fatty acid monoglycerides and diglycerides, pentoethral fatty acid esters, hydroxymethyl-cellulose, polyvinylpyrrolidone, and the like. 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.

The composition, if desired, can also contain minor amounts of wetting agents, emulsifying agents and/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, and the like.

Various delivery systems are known and can be used to administer the compounds or compositions of the present invention, including, for example, encapsulation in liposomes, microbubbles, emulsions, microparticles, microcapsules and the like.

The bioavailabilty of the compositions can be enhanced by micronization of the formulations using conventional techniques such as grinding, milling, spray drying and the like in the presence of suitable excipients or agents such as phospholipids or surfactants.

The compounds and compositions of the present invention can be formulated as neutral or pharmaceutically acceptable salt forms. Pharmaceutically acceptable salts include, for example, those formed with free amino groups such as those derived from hydrochloric, hydrobromic, phosphoric, sulfuric, acetic, citric, benzoic, fumaric, glutamic, lactic, malic, maleic, nitric, succinic, tartaric p-toluene-sulfonic, methanesulfonic, acids, gluconic acid, and the like, 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, and the like.

Therapeutically effective amount refers to the amount of the nitrosated and/or nitrosylated NSAID and nitric oxide donor that is effective to achieve its intended purpose. While individual patient needs may vary, determination of optimal ranges for effective amounts of each of the compounds and compositions 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 the dysfunction of the recipient, frequency of treatment and the nature and scope of the dysfunction or disease.

The amount of a given nitrosated and/or nitrosylated 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, including reference to Goodman and Gilman, supra; The Physician's Desk Reference, Medical Economics Company, Inc., Oradell, N.J., 1995; and Drug Facts and Comparisons, Inc., St. Louis, Mo., 1993. The precise dose to be used in the formulation will also depend on the route of administration, and the seriousness of the disease or disorder, and should be decided by the physician and the patient's circumstances.

The amount of nitric oxide donor in a pharmaceutical composition can be in amounts of about 0.1 to about 10 times the molar equivalent of the NSAID. The usual daily doses of NSAIDs are about 3 to about 40 mg/kg of body weight and the doses of nitric oxide donors in the pharmaceutical composition can be in amounts of about 1 to about 500 mg/kg of body weight daily, preferably about 1 to about 50 mg/kg of body weight daily. 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.

The present invention also provides pharmaceutical kits comprising one or more containers filled with one or more of the ingredients of the pharmaceutical compositions of the present invention, including, at least, one or more of the nitrosated and/or nitrosylated NSAIDs described herein and one or more of the NO donors described herein. Associated with such kits 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.

EXAMPLES

The following non-limiting examples further describe and enable one of ordinary skill in the art to make and use the present invention. In each of the examples, flash chromatography was performed on 40 micron silica gel (Baker).

To a stirred suspension of 4-piperidone (10.0 g, 65.0 mmol) and bromobenzyl acetate (14.9 g, 65.2 mmol) in acetone (100 ml) was added K 2 CO 3 (9.0 g) and Et 3 N (9.1 ml, 65.2 mmol). The reaction mixture was stirred at room temperature for two days, and then the solvent was evaporated. The residue was partitioned between EtOAc and water. The aqueous layer was extracted with EtOAc. The organic extracts were combined and dried over Na 2 SO 4 . The solvent was evaporated to afford the title compound (13.3 g, 53.8 mmol, 83%) as a thick oil. 1 H NMR (300 MHz, CDCl 3 ) 7.33-7.37 (m, 5H), 5.18 (s, 2H), 3.42 (s, 2H), 2.91 (t, J 6.1 Hz, 4H), 2.50 (t, J 6.1 Hz, 4H).

To an ice-cooled solution of LiAlH 4 (24.0 ml in 1M tetrahydrofuran, 24.0 mmol) was added dropwise a solution of the product of Example 1c (5.32 g, 19.2 mmol) in 20 ml tetrahydrofuran. The solution was stirred cold for half an hour after the addition was complete. Water was added dropwise to quench the reaction. 5% methanol/dichloromethane solution was added and the mixture was filtered through Celite. The filtrate was concentrated to give an oil, which was dissolved in ether (20 ml). HCl in ether was added to precipitate the salt, which was filtered and washed thoroughly with ether to remove the last trace of benzyl alcohol. The amine was liberated by adding 10% ammonium hydroxide solution followed by extraction with EtOAc. The organic extracts were combined and dried over Na 2 SO 4 . The solvent was evaporated to afford the title compound (2.1 g, 12.0 mmol, 62%) as a clear oil. 1 H NMR (300 MHz, CDCl 3 ) 3.60 (t, J 5.4 Hz, 2H), 2.45-2.67 (m, 4H), 2.57 (t, J 5.4 Hz, 2H), 1.64-1.76 (m, 4H), 1.46 (s, 3H).

The product of Example 1e (287 mg, 0.63 mmol) was dissolved in ether and HCl in ether was added dropwise. The white solid thus formed was collected and washed thoroughly with ether and vacuum dried to give the HCl salt (270 mg, 0.55 mmol) as a white solid. The salt (200 mg, 0.41 mmol) was dissolved in dichloromethane (4 ml). The solution was cooled to 78 C. t-Butyl nitrite (54 L, 0.41 mmol) was added. The cold bath was then removed. Ten minutes later, the solvent was evaporated to give a green solid, which was converted to the free amine by treatment with saturated aqueous K 2 CO 3 and then extracted with EtOAc. The EtOAc extracts were combined and dried over Na 2 SO 4 . The solvent was evaporated and the crude product was chromatographed on silica gel eluting with 1:1 EtOAc/hexanes to give the title compound (135 mg, 0.28 mmol, 69%) as a thick oil. 1 H NMR (300 MHz, CDCl 3 ) 7.33-7.36 (m, 2H), 7.21-7.24 (m, 1H), 7.08-7.14 (m, 1H), 6.88-7.01 (m, 3H), 6.53-6.56 (m, 1H), 4.29 (t, J 5.8 Hz, 2H), 3.82 (s, 2H), 2.66-2.76 (m, 2H), 2.68 (t, J 5.8 Hz, 2H), 2.33-2.40 (m, 2H), 2.08-2.17 (m, 2H), 1.94 (s, 3H).

A mixture of the product of Example 2 a (10 g, 34.91 mmol), ethanol amine (4.26 g, 69.82 mmol) and MgSO 4 (10 g) in dry CHCl 3 (100 ml) was heated under reflux for 8 hours. The solid was filtered and the solvent was evaporated under reduced pressure to yield a viscous yellow liquid. The crude product was dissolved in methanol (125 ml) and NaBH 4 (3.3 g, 87.25 mmol) was added portionwise over 10 mm. The resulting solution was stirred at room temperature for 1 hour. Methanol was evaporated and the crude material was partitioned between a mixture of water (200 ml) and ethyl acetate (100 ml). The organic layer was separated and the aqueous layer was extracted with ethyl acetate (100 ml). The combined organic layers were dried over Na 2 SO 4 and evaporated under reduced pressure to give a colorless viscous liquid. This product was then further purified by dissolving in ether (50 ml) followed by the dropwise addition of HCl in ether to form a white salt. The salt was washed with ether (2 50 ml) and then the solid was dissolved in water (100 ml). The aqueous layer was washed with ether (100 ml) and the ether layer was discarded. The aqueous layer was basified with 15% ammonium hydroxide (10 ml) to form a white suspension which was extracted with ethyl acetate (2 50 ml). The organic layer was dried over Na 2 SO 4 and concentrated under reduced pressure to give the title compound (12.2 g, 93%) as a viscous liquid. 1 H NMR (300 MHz, CDCl 3 ) 1.20-1.95 (in, 20 H), 2.66 (s, 4 H), 2.78 (t, J 5.2 Hz, 4 H), 3.61 (t, J 5.2 Hz, 4 H); 13C NM (75 MHz, CDCl 3 ) 22.7, 25.8, 34.3, 51.5, 54.7, 60.6, 68.2.

Ammonia (20 mL) was condensed into a 3-neck flask fitted with a dry ice condenser. The product of Example 4c (340 mg, 1 mmol) was added in ETOH (4 mL) followed by metallic sodium (76 mg, 3.3 mmol) until the blue color persisted. A small amount of NH 4 Cl was added to discharge the blue color and the ammonia was allowed to evaporate under a stream of nitrogen. The residue was partitioned between Et 2 O and 1 N HCl. The aqueous layer was extracted with Et 2 O (1 20 mL). The combined organic layers were washed with brine (1 10 mL) and dried over Na 2 SO 4 . Evaporation of the solvent gave the title compound (210 mg, 80%) as an oil. 1 H NMR (300 MHz, CDCl 3 ) 3.95 (t, J 6.7 Hz, 2 H), 3.80-3.90 (mult, 2 H), 3.20-3.35 (mult, 2 H), 1.95 (t, J 6.7 Hz, 2 H), 1.60-1.80 (mult, 4 H), 1.46 (s, 9 H).

A mixture of isobutylene epoxide (25.0 g, 346 mmol), water (50 ml), and KSCN (67.2 g, 692 mmol) was stirred at room temperature for 20 hours. The organic layer was separated and dried over Na 2 SO 4 . The solid was filtered off to give the title compound (26.4 g, 87%) as a clear oil. 1 H NMR (300 MHz, CDCl 3 ) 2.41(s, 2H), 1.62 (s, 6H).

The product of Example 8b (5.9 g, 27.1 nmmol) in CH 2 Cl 2 (100 ml) was treated with 1 N HCl Et 2 O (70 ml). The solvent was removed to give a white solid. The solid was dissolved in EtOH (30 ml) and water (20 ml) and added dropwise to a stirred solution of t-BuONO (6.2 g, 54.1 mmol) in EtOH (10 ml). The reaction was kept at room temperature for one hour after which the volatiles were evaporated. The residue was partitioned between satd NaHCO 3 and EtOAc. The aqueous layer was extracted with EtOAc. The organic extracts were combined and dried over Na 2 SO 4 . The volatiles were evaporated. The residue was chromatographed on silica gel eluting with MeOH:CH 2 Cl 2 1:19 to give the title compound (3.15 g, 47%) as a green oil. 1 H NMR (300 MHz, CDCl 3 ) 3.67 (t, J 5.3, 2 H), 3.00 (s, 2 H), 2.62-2.67 (m, 4 H), 2.48-2,54 (m, 6 H), 1.88 (s, 6 H); 13 C NMR (75 MHz, CDCl 3 ) 68.1, 59.1, 58.8, 57.6, 55.4, 53.0, 27.0; mass spectrum (m/e): 248 (MH ).

Dihydroxy acetone dimer (7.5 g, 41.46 mmol) was added to a stirred solution of TBDMSCI (25.0 g, 166 mmol) in dry pyridine (100 mL). The resulting solution was stirred at room temperature for 12 hours. Ethyl acetate (100 mL) was added and the solution was washed with 10% HCl (3 50 mL) and water (200 mL). The organic phase was dried over Na 2 SO 4 and evaporated to give the title compound (25.0 g, 94%) as a viscous oil. 1 H NMR (300 MHz, CDCl 3 ) 4.45 (s, 4H), 0.94 (s, 18H), 0.11 (s, 12H).

The product of Example 9b (5.1 g, 13.1 mmol) and benzylmercaptan (1.53 mL, 13.1 mmol) in piperidine (50 mL) was heated at 100 C. for 4 hours and then cooled to room temperature. Water (50 mL) was added and the aqueous layer was extracted with EtOAc (3 50 mL). The combined organic layers were dried over Na 2 SO 4 . The solvent was evaporated and the residue was purified by chromatography on silica gel eluting with 5:95 EtOAc:hexane to afford the title compound (4.6 g, 68%) as a viscous liquid. The viscous liquid (10.0 g, 19.5 mmol) was dissolved in CH 3 CN (10 mL) and 48% HF (10 mL) was added. The solution was stirred at room temperature for 2 hours. Satd NaHCO 3 (100 mL) was added. The solution was extracted with EtOAc (3 100 mL). The combined organics were dried over Na 2 SO 4 . The solvent was evaporated and the residue was chromatographed on silica gel eluting with 1:2 hexane:EtOAc to give the title compound (4.7 g, 95%). 1 H NMR (300 MHz, CDCl 3 ) 7.22-7.30 (mult, 5H), 3.75 (s, 2H), (ABq, J 9.9 Hz, 2H), 3.60 (d, J 4.8 Hz, 2H), 2.04-2.08 (mult, 1H), (ABq, J 17.8 Hz, 2H).

A mixture of the product of Example 2a (20 g, 69.8 mmol) and propanol amine (10.5 g, 140 mmol) in CHCl 3 (150 mL) were heated at 65 C. for 8 hours. The solvent was evaporated to obtain a viscous yellow liquid which was dissolved in MeOH (200 mL). NaBH 4 (5.3 g, 140 mmol) was added portionwise over 10 minutes and the resulting solution was stirred at room temperature for 1 hour. The solvent was evaporated and the residue was partitioned between water (200 mL) and EtOAc (100 mL). The organic layer was separated and the aqueous layer was extracted with EtOAc (2 50 mL). The combined organic layers were dried over Na 2 SO 4 and concentrated to afford the title compound (27.5 g, 97%) as a colorless viscous oil. 1 H NMR (300 MHz, CDCl 3 ) 3.84 (t, J 5.3 Hz, 4H), 2.91 (t, J 5.5 Hz, 4H), 2.66 (s, 4H), 1.20-1.80 (mult, 24H); 13 C NMR (75 MHz, CDCl 3 ) 64.8, 57.1, 54.4, 50.6, 34.4, 30.4, 25.8, 22.2.

A 1M solution of sodium hexamethyldisilazane (NaHMDS, 350 mL, 0.35 mol) in THF was added slowly to a suspension of (methoxymethyl)triphenyl phosphonium chloride (120 g, 0.35 mol,) in THF (100 mL) at 78 C. under N 2 . The resulting brown solution was stirred at 78 C. for 20 minutes and then 1-carbethoxy-4-piperidone (50 g, 0.292 mol) in THF (50 mL) was added dropwise. The mixture was stirred at 78 C. for 5 minutes and then for 2 hours at room temperature. Water (200 mL) was added and the layers were separated. The aqueous layer was extracted with EtOAc and the combined organic extracts were dried over Na 2 SO 4 . The solvent was evaporated to give an orange solid which was suspended in Et 2 O (200 mL). The solid was removed by filtration and the filtrate was concentrated to give a yellow oil which was triturated with hexane (200 mL). The white solid which precipitated was removed by filtration. The filtrate was concentrated in vacuo and this was procedure repeated twice more to give the title compound (52 g, 89%) as a pale yellow oil. TLC R f 0.72 (EtOAc:hexane, 1:2); 1 H NMR (CDCl 3 ) 5.84 (s, 1H), 4.11 (q, J 7 Hz, 2H), 3.54 (s, 3 H), 3.37-3.42 (mult, 4 H), 2.24 (t, J 5.6 Hz, 2H), 1.99 (t, J 5.6 Hz, 2H), 1.24 (t, J 7 Hz, 3H); 13 C NMR (CDCl 3 ) 155.6, 140.8, 113.4, 61.3, 59.5, 45.8, 44.6, 29.6, 25.2, 14.8; mass spectrum (API-TIS) m/z 200 (M H). Anal Calcd for C 10 H 17 N 1 O 3 : C, 60.28; H, 8.60; 7.03. Found: C, 60.29; H, 8.63; N, 6.81.

To a stirred solution of the product of Example 12b in CCl 4 (120 mL) was added dropwise S 2 Cl 2 (13.43 mL, 0.168 mol) over a period of 5 minutes at 50 C. After a short lag period (10-15 minutes), evolution of HCl gas was observed. After the gas evolution had ceased, the mixture was stirred at 55 C. for 1 hour and then cooled to room temperature. The solvent was evaporated to give a yellow oil which was purified by flash chromatography on silica gel eluting with 1:2 EtOAc:hexane to give a pale yellow oil which was dried in vacuo to give the title compound (76% based on 12a) as a sticky oil which solidified on standing at room temperature. 1 H NMR (CDCl 3 ) 9.04, 1H), 4.11 (q, J 7 Hz, 2H), 3.65-3.85 (mult, 2H), 3.14-3.20 (mult, 2H), 2.01-2.07 (mult, 2H), 1.71-1.80 (mult, 2H), 1.25 (t, J 7 Hz, 3H); 13 C NMR (CDCl 3 ) 193.2, 155.3, 61.8, 59.6, 40.8, 29.5, 14.7; mass spectrum (API-TIS) m/z 450 (M NH 4 ).

A mixture of product of Example 12c (7.0 g, 16.18 mmol) and propanol amine (2.91 g, 38.8 mmol) in dry CHCl 3 (50 mL) was heated at 65 C. for 8 hours. The solvent was evaporated to obtain a viscous yellow liquid which was dissolved in MeOH (30 mL). NaBH 4 (1.5 g, 38.83 mmol) was added portionwise over 10 minutes and the resulting solution was stirred at room temperature for 1 hour. Formaldehyde 38% (30 mL) was added and the resulting cloudy solution was stirred 2 hours at room temperature. The solvent was evaporated and the residue was partitioned with a mnixture of water (100 mL) and EtOAc (50 mL). The organic extracts were separated and the aqueous layer was extracted with EtOAc (2 50 mL). The combined organic layers were dried over Na 2 SO 4 and evaporated to give a colorless viscous oil (9.2 g). The colorless oil (8.2 g) in THF (50 mL) was added to a stirred solution of lithium aluminum hydride (1M, 42 mL, 42 mmol) at room temperature under N 2 . The resulting clear solution was stirred at room temperature for 3 hours and the excess lithium aluminum hydride was destroyed by portionwise addition of solid Na 2 SO 4 10H 2 O ( 10 g). The precipitate was removed by filtration and washed with 10% MeOH in CH 2 Cl 2 (2 50 mL). The combined filtrate was dried over Na 2 SO 4 and concentrated to give a viscous liquid (5.1 g). The viscous liquid (5 g) was dissolved in MeOH (30 mL) and cooled to 0 C. and conc HCl (3 mL) was added. t-BuONO (3.2 mL, 26.8 mmol,) was then added via syringe and the resulting green solution was stirred for 20 minutes at room temperature. The solution was poured onto crushed ice ( 10 g) and made basic with 10% Na 2 CO 3 (10 mL). The green aqueous solution was extracted with EtOAc (3 50 mL). The combined organics were dried over Na 2 SO 4 and concentrated. The residue was purified by chromatography on silica gel eluting with 1:4 MeOH:CH 2 Cl 2 to afford the title compound (1.2 g). 1 H NMR (CDCl 3 ): 3.70 (t, J 5.5 Hz, 2H), 3.23 (s, 2H), 2.74-2.80 (mult, 2H), 2.69 (t, J 6.1 Hz, 2H), 2.50-2.67 (mult, 2H), 2.34 (s, 3H), 2.31 (s, 3H), 2.25-2.45 (mult, 4H), 1.67 (p, J 5.9 Hz, 2H); 13 C NMR (CDCl 3 ) 69.9, 62.9, 59.8, 51.4, 46.0, 44.3, 34.1, 29.0; mass spectrum (API-TIS) m/z 26 (M H).

A solution of DIBAL in hexane (83 mL, 83 mmol) was added to a stirred solution of the product of Example 13b (11.7 g, 38.74 mmol) in THF (40 mL) at 78 C. under N 2 . The cold bath was removed and the mixture was stirred 1.5 hours. Solid Na 2 SO 4 10H 2 O (3 g) was added portionwise with stirring until a thick precipitate was formed. 10% MeOH in CH 2 Cl 2 (100 mL) was added and the mixture was filtered. The solid was washed with additional 10% MeOH in CH 2 Cl 2 (100 mL) and the solvent was evaporated. The residue was chromatographed on silica gel eluting with 1:9 MeOH:CH 2 Cl 2 to give the title compound (5.2 g, 50.6%) as a solid. 1 H NMR (300 MHz, CDCl 3 ) 7.20-7.35 (mult, 5H), 3.86 (t, J 6.4 Hz, 2H), 3.66 (s, 2H), 2.50-2.57 (mult, 4H), 2.29 (s, 3H), 1.88 (t, J 6.5 Hz, 2H), 1.65-1.84 (mult, 4H).

The product of Example 13c (7.8 g, 29.38 mmol) was dissolved in THF (50 mL) and cooled to 78 C. and liquid NH 3 (100 mL) was added. Small pieces of Na (2 g) were added until the blue color persisted for 10 mninutes. Solid NH 4 Cl ( 5 g) was added to discharge the color and the cold bath was removed and NH 3 was evaporated (12 hours). Ether (100 mL) was added to the pale yellow solid and HCl in Et 2 O (10 mL) was added until the solution became acidic. The mixture was left in a freezer for 30 minutes. The solid which formed was removed by filtration and washed with Et 2 O (50 mL). The residue was triturated with MeOH (100 mL) and the undissolved solid was removed by filtration. The solvent was concentrated to 25 mL and conc HCl (2 mL) was added. 90% t-BuONO (3.1 mL, 23.7 mmol) was added via a syringe. The resulting olive green solution was stirred at room temperature for 20 minutes and then poured onto crushed ice (5 g). 10% Na 2 CO 3 (10 mL) was added and the mixture was extracted with EtOAc (3 50 mL). The combined organics were dried over Na 2 SO 4 and concentrated to give the title compound (3.6 g, 60%) as green oil. 1 H NMR (300 MHz, CDCl 3 ) 3.88, J 6.9 Hz, 2H), 2.25-2.95 (mult, 13H), 2.30 (s, 3H); 13 C NMR (75 MHz, CDCl 3 ) 62.5, 57.8, 51.5, 46.1, 36.4.

BOC anhydride (14.83 g, 67.96 mmol) was added to a stirred solution of the product of Example 18a (13.6 g, 67.96 mmol) in THF (100 mL) and the mixture was stirred at room temperature for 2 hours. Water (200 mL) and EtOAc (100 mL) were added. The organic layer was isolated, dried over Na 2 SO 4 , and concentrated to give the title compound as a viscous oil (20 g, 98%). 1 H NMR (300 MHz, CDCl 3 ) 2.35-3.75 (br mult, 8H), 1.45 (s, 9H); mass spectrum (API-TIS) m/z 301 (M H).

A mixture of the product of Example 18b (20 g, 66.6 mmol) and solid K 2 CO 3 (5 g) in MeOH (50 mL) and water (10 mL) were heated at 60 C. for 18 hours. The solvent was evaporated to give a viscous oil which was extracted with EtOAc (5 50 mL). The combined organics were washed with water (50 mL). The organic phase was dried over Na 2 SO 4 and the solvent was evaporated to afford the title compound (10 g, 66%) as an oil. 1 H NMR (300 MHz, CDCl 3 ) 3.74 (mult, 2H), 3.30-3.50 (mult, 3H), 2.90-3.10 (mult, 3H), 1.46 (s, 9H); mass spectrum (API-TIS) m/z 205 (M H).

A mixture of the product of Example 2a (5.84 g, 20.4 mmol) and the product of Example 18c (10 g, 49.01 mmol) in dry CHCl 3 (50 mL) were heated at 65 C. for 16 hours. The solvent was evaporated to obtain a viscous yellow liquid which was dissolved in MeOH (50 mL). NaBH 4 (1.8 g, 47.3 mmol) was added portionwise over 10 minutes and the resulting solution was stirred at room temperature for 1 hour. Formaldehyde 38% (20 mL) was added and the resulting cloudy solution was stirred for 2 hours at room temperature. The solvent was evaporated and the residue was partitioned between water (100 mL) and EtOAc (50 mL). The organic layer was separated and the aqueous layer was extracted with EtOAc (2 50 mL). The combined organic layers were dried over Na 2 SO 4 and concentrated to give (16 g) a colorless viscous oil. This colorless oil in THF (50 mL) was added in a dropwise fashion to a stirred solution of lithium aluminum hydride (1M, 60 mL, 60 mmol) at room temperature under N 2 . The resulting clear solution was stirred at room temperature for 4 hours. The excess lithium aluminum hydride was destroyed by portionwise addition of Na 2 SO 4 10H 2 O (10 g). The precipitate was removed by filtration and the solid was washed with 10% MeOH in CH 2 Cl 2 (2 50 mL). The combined filtrate was dried over Na 2 SO 4 and concentrated to give a viscous liquid (10 g). This viscous liquid (10 g) was dissolved in MeOH (30 mL) and cooled to 0 C. Concentrated HCl (5 mL) was added. 90% t-BuONO (5.4 mL, 38.4 mmol) was added via a syringe and the resulting green solution was stirred for 20 minutes at room temperature. The solution was poured onto crushed ice (10 g) and the resulting mixture was made basic with 10% Na 2 CO 3 (10 mL). The green aqueous solution was extracted with EtOAc (3 50 mL). The organic layer was dried over Na 2 SO 4 and concentrated. The residue was purified by chromatography on silica gel eluting with 1:9 MeOH:EtOAc to give the title compound (4.3 g). 1 H NMR (CDCl 3 ) 3.57 (t, J 5.3 Hz, 2H), 3.19 (s, 2H), 3.05 (br s, 1H), 2.38 (s, 3H), 2.27 (s, 3H), 2.11-2.80 (mult, 8H), 1.40-1.85 (mult, 6H); 13 C NMR (75 MHz, CDCl 3 ) 69.3, 64.4, 58.9, 58.5, 58.4, 55.3, 45.3, 42.2, 34.2, 25.5, 22.2.

N-Methyl ethylenediamine (15 g, 202.3 mmol) was added dropwise to a stirred solution of ethyl trifluoroacetate (28.7 g, 204.34 mmol) in dry Et 2 O (50 mL) at 0 C. The resulting solution was stirred at room temperature for 2 hours. Hexane (75 mL) was added and the solution was left at 20 C. for 16 hours to produce a white precipitate which was removed by filtration, washed with Et 2 O (100 mL) and dried in vacuo for 3 hours to afford the title compound (29.4 g, 85%). 1 H NMR (300 MHz, CDCl 3 ) 3.42 (t, J 5.8 Hz, 2H), 2.80 (t, J 5.9 Hz, 2H), 2.43 (s, 3H); mass spectrum (API-TIS) 171 (M H).

BOC anhydride (37.2 g, 170.4 mmol) was added to a stirred solution of the product of Example 20a (29.0 g, 170.45 mmol) in THF (100 mL) and the mixture was stirred at room temperature for 2 hours. Water (200 mL) and EtOAc (100 mL) were added. The organic layer was isolated and dried over Na 2 SO 4 . The solvent was evaporated to give the title compound as a viscous oil (45 g, 98%). 1 H NMR (300 MHz, CDCl 3 ) 3.48 (br s, 4H), 2.90 (s, 3H), 1.45 (s, 9H); mass spectrum (API-TIS) m/z 271 (M H).

A mixture of the product of Example 2a (5.34 g, 18.5 mmol) and the product of Example 20c (7.80 g, 44.76 mmol) in dry CHCl 3 (75 mL) were heated at 65 C. for 16 hours. The solvent was evaporated to obtain a viscous yellow liquid which was dissolved in MeOH (50 mL). NaBH 4 (3.4 g, 89.5 mmol) was added portionwise over 10 minutes and the resulting solution was stirred at room temperature for 1 hour. The solvent was evaporated and the residue was dissolved in water (100 mL). The mixture was extracted with EtOAc (3 75 mL). The combined organic extracts were dried over Na 2 SO 4 and concentrated to give a colorless viscous oil. The colorless oil (10.2 g) was dissolved in CH 3 CN (100 mL) and tert-butyl bromoacetate (20 g, 102.5 mmol) and solid K 2 CO 3 (10 g) were subsequently added. The resulting suspension was stirred at room temperature for 12 hours. The solid was removed by filtration and washed with CH 3 CN (50 mL). The filtrate was concentrated and the residue was chromatographed on silica gel eluting with 1:9 EtOAc:hexane to give the title compound (6.3 g, 41% based on cyclohexanecarboxaldehyde disulfide) and an unidentified lower Rf product (2.2 g). 1 H NMR (300 MHz, CDCl 3 ) 3.38 (br s, 4H), 3.26 (br s, 4H), 2.87 (s, 10H), 2.84 (s,4H), 1.47 (s, 18H), 1.45 (s, 18H), 1.17-1.80 (mult, 20H); 13 C NMR (75 MHz, CDCl 3 ) 171.3, 155.5, 80.8, 79.1, 65.6, 57.1, 56.1, 54.4, 47.7, 34.7, 32.7, 28.4 (3C), 28.2 (3C), 25.6, 22.3 (2C); mass spectrum (API-TIS) m/z 831 (M H).

To a stirred solution of lithium aluminum hydride (1M, 23 mL, 23 mmol) was added the product of Example 20d (6.30 g, 7.58 mmol) in THF (50 mL) dropwise at room temperature under N 2 . The resulting clear solution was stirred at room temperature for 1 hour and then heated at 70 C. for 12 hours and cooled to room temperature again. The excess lithium aluminum hydride was carefully destroyed by portionwise addition of solid Na 2 SO 4 10H 2 O (10 g). The precipitate was removed by filtration and washed with 10% MeOH in CH 2 Cl 2 (2 50 mL). The combined filtrate was dried over Na 2 SO 4 and concentrated to give a viscous liquid (3.2 g). The viscous liquid (3 g) was dissolved in MeOH (25 mL) and cooled to 0 C. Concentrated HCl (5 mL) was added. A solution of 90% t-BuONO (2.2 mL) was added via a syringe and the resulting green solution was stirred for 20 minutes at room temperature. The solution was then poured onto crushed ice (10 g) and the resulting mixture was made basic with 10% Na 2 CO 3 (10 mL). The green aqueous solution was extracted with EtOAc (3 50 mL), dried over Na 2 SO 4 , and concentrated. The residue was purified by chromatography on silica gel eluting with 1:9 MeOH:CH 2 Cl 2 to give the title compound (0.7 g) (substantial decomposition occurred during chromatography). 1 H NMR (CDCl 3 ): 3.55, J 5.3 Hz, 2H), 3.35 (s, 2H), 2.70-2.82 (mult, 4H), 2.50-2.60 (mult, 2H), 2.22 (s, 6H), 2.10-2.40 (mult, 4H), 1.30-1.72 (mult, 6H); mass spectrum (API-TIS) m/z 290 (M H).

The product of Example 21a was dissolved in Et 2 O and HCl in Et 2 O was added dropwise. The white solid which formed was collected and washed thoroughly with Et 2 O and vacuum dried to give the HCl salt (400 mg, 0.94 mmol) as a white solid. The white solid (400 mg, 0.94 mmol) was dissolved in CH 2 Cl 2 (4 ml) and cooled to 0 C. t-Butyl nitrite (187 L, 1.42 mmol) was added. After 30 minutes, the solvent was evaporated to give a green solid which was partitioned between satd K 2 CO 3 and EtOAc. The EtOAc extracts were combined and dried over Na 2 SO 4 . The solvent was evaporated and the residue was chromatographed on silica gel eluting with 1:1 EtOAc:hexane to give the title compound as green solid. mp 131 C.; 1 H NMR (300 MHz, CDCl 3 ) 7.38-7.41 (mult, 1H), 7.09-7.15 (mult, 2H), 4.13-4.30 (mult, 2H), 3.91 (s, 3H), 3.85 (q, J 7.2 Hz, 1H), 2.54-2.63 (mult, 4H), 2.19-2.27 (mult, 4H), 1.96-2.04 (mult, 2H), 1.87 (s, 3H), 1.58 (d, J 7.1 Hz, 3H).

To a suspension of NaH (3.13 g, 0.13 mol) in DMSO was added trimethylsulfoxonium iodide (28.7 g, 0.13 mol) in several portions. The mixture was stirred for 30 minutes. tert-Butyl-4-oxopiperidinecarboxyxlate (20.0 g, 0.10 mmol) was added at once and the mixture was heated at 60 C. for an hour. The reaction mixture was cooled to room temperature and poured into water. The mixture was extracted with EtOAc (2 ). The combined organic layers were dried over Na 2 SO 4 and then concentrated to give the title compound as a white solid (20.2 g, 9.46 mmol, 94%). 1 H NMR (300 MHz, CDCl 3 ) 3.67-3.75 (mult, 2H), 3.37-3.46 (mult, 2H), 2.68 (s, 2H), 1.74-1.83 (mult, 2H), 1.46 (s, 9H), 1.39-1.45 (mult, 2H).

To a solution of the product of Example 22c (3.92 g, 12.89 mmol) in THF (38 ml) was added lithium aluminum hydride (1M, 19.3 mL, 19.3 mmol) in THF. The mixture was refluxed overnight. The reaction was cooled to room temperature. Methanol was added to quench the reaction until no more bubbles were observed, followed by the addition of H 2 O until no more bubbles were formed. The mixture was filtered through celite and washed with 5:95 MeOH:CH 2 Cl 2 . The filtrate was concentrated to give the title compound (2.5 g, 11.46 mmol, 88%) which was used without further purification.

To a solution of the product of Example 23b (6.2 g, 31.1 mmol) in MeOH (90 ml) was added thiourea (2.85 g, 37.4 mmol). The reaction mixture was heated at 45 C. for 3 hours. The solvent was evaporated and the residue was triturated with Et 2 O and filtered. The filtrate was concentrated and again triturated with hexane and filtered. Evaporation of the solvent gave the title compound (5.06 g, 23.5 mmol, 76%) as a clear oil. 1 H NMR (300 MHz, CDCl 3 ) 3.69 (s, 3H), 2.71-2.80 (mult, 4H), 2.34-2.56 (mult, 4H), 2.34 (s, 2H), 2.10-2.20 (mult, 2H), 1.52-1.60 (mult, 2H).

To a mixture of the product of Example 23d (1.21 g, 6.40 mmol) in CH 2 Cl 2 was added HCl in Et 2 O. The solvent was evaporated to give a solid which was dissolved in EtOH (10 ml) and H 2 O (2 ml). This homogeneous solution was added slowly to a stirred solution of t-BuONO (0.94 ml, 8.0 mmol) in EtOH (10 ml) over 10 minutes. The reaction mixture was stirred for 1 hour. The solvent was evaporated and the residue was dissolved in CH 2 Cl 2 and washed with satd Na 2 CO 3 . The organic layer was dried over Na 2 SO 4 and concentrated to give the title compound as a green oil (1.25 g, 5.73 mmol, 90%). 1 H NMR (300 MHz, CDCl 3 ) 3.82 (t, J 5.2 Hz, 2H), 2.88-2.93 (mult, 2H), 2.65 (t, J 5.7 Hz, 2H), 2.47-2.53 (mult, 2H), 2.30-2.42 (mult, 2H), 2.21-2.28 (mult, 2H), 2.00 (s, 2H), 1.70-1.76 (mult, 2H).

To a solution of the product of Example 25b (1.0 g, 4.0 mmol) in CH 2 Cl 2 (50 ml) was added ethanolamine (0.27 g, 4.42 mmol) followed by hydroxysuccinamide (509 mg, 4.4 mmol). DCC (824 mg, 4.0 mmol) in CH 2 Cl 2 (4 mL) was then added and the reaction was stirred at room temperature for 0.5 hours. The reaction mixture was then poured into water (50 ml) and extracted with EtOAc (6 ). The solvent was evaporated to give the title compound which was used for the next reaction without further purification.

A mixture of 3-methyl-3-(phenylmethylthio)butanoic acid (1 g, 4.6 mmol) and hexachloroacetone were dissolved in CH 2 Cl 2 (20 mL) and cooled to 78 C. Triphenyl phosphine was added and the mixture was stirred for 30 minutes. Hydroxyethyl piperazine (550 L, 4.5 mmol) was added in CH 2 Cl 2 (10 mL) dropwise. Triethylamine (630 L, 4.5 mmol) was added in CH 2 Cl 2 (10 mL) dropwise. The cold bath was removed and the solution was stirred for 24 hours. The solvent was evaporated. The crude mixture was poured into 1N HCl (100 mL) and washed with Et 2 O (2 50 mL). The aqueous layer was made basic with 10% K 2 CO 3 in a brine solution. The product was extracted with EtOAc (3 100 mL), dried over Na 2 SO 4 , and concentrated. This gave the title compound which was used immediately in the next reaction. 1 H NMR (300 MHz, CDCl 3 ) 7.18-7.37 (mult, 5H), 3.8 (s, 2H), 3.6 (t, J 5 Hz, 4H), 3.4 (t, J 5 Hz, 2H), 2.45-2.65 (mult, 8H), 1.5 (s, 6H).

Ammonia (100 mL) was condensed into a 3 neck flask at 78 C. The product of Example 29a was added to the flask in a minimum amount of Et 2 O. The solution was stirred for 20 mninutes. Sodium was added in pea sized chunks until the solution remained a blue color for greater than 10 minutes. The solution was stirred for an additional 30 minutes. The ice bath was removed and the ammonia was allowed to evaporate at room temperature. This gave the title compound (600 mg, 55% over 2 steps). 1 H NMR (300 MHz, CDCl 3 ) 3.63 (t, J 5 Hz, 4H), 3.52 (t, J 5 Hz, 2H), 2.63 (s, 2H), 2.38-2.57 (mult, 6H), 1.51 (s, 6H).

To a stirred solution of ethyl N-(t-butoxycarbonyl)isonipecotate (1.06 g, 4.12 mmol) in THF (8 mL) at 78 C. was added LDA (1.5M, 2.75 mL, 4.12 mmol) dropwise, and the mixture was stirred for 30 minutes before addition of a solution of dicyclopropylthioketone (415 mg, 3.30 mmol) in THF (1 mL). After the addition, the reaction mixture was warmed to room temperature over 2 hours, quenched with satd aq NH 4 Cl, and extracted with EtOAc. The combined organic extracts were dried over Na 2 SO 4 , filtered, and concentrated to afford the title compound as a viscous oil (1.53 g, 96%), which was used in the next step without further purification.

A solution of the product of Example 31a (7.43 g, 28.6 mmol), di-t-butyl dicarbonate (15.6 g, 71.5 mmol), DMAP (12 mg), and Et 3 N (20 mL, 143 mmol) in CH 2 Cl 2 (100 mL) were stirred at room temperature for 15 hours. After being diluted with CH 2 Cl 2 (200 mL), the mixture was washed with 1N HCl, dried over Na 2 SO 41 filtered, and concentrated. This gave the title compound (12.5 g) as an oil which was used in the next step without further purification.

The title compound was synthesized from the product of Example 2a in using a sequence analogous to the preparation of the product of Example 31c. 1 H NMR (300 MHz, CDCl 3 ) 2.88 (t, J 6.6 Hz, 2H), 2.77 (t, J 7.7 Hz, 2H), 2.54 (s, 2H), 2.38 (s, 3H), 1.9-1.4 (mult, 12H).

The product of Example 42b (4.50 g, 3.83 mmol) was dissolved in HOAc (22 mL) and powdered zinc (9 g) was added. The resulting suspension was stirred at room temperature for 12 hours. The solid was removed by filtration and washed with HOAc (25 mL). The filtrate was made basic with concentrated NH 4 OH in crushed ice (100 g) and extracted with EtOAc (3 50 mL). The combined organic layers were dried over Na 2 SO 4 and filtered. The solvent was evaporated to give a white foam (4 g), which was

To a stirred solution of the 1-(8-aza-1,4-dioxaspiro 4.5 dec-8-yl)-2-methylpropane-2-thiol (Synthesis, 1999, 7, 1106) (1.15 g, 4.98 mmol) in THF (12 mL) was added 6 N HCl (12 mL). The mixture was heated at 70 C. overnight, then poured into saturated Na 2 CO 3 . The mixture was extracted with EtOAc. The combined organic extracts were dried over Na 2 SO 4 and evaporated. The residue was dissolved in EtOAc and acidified by adding HCl/EtOAc until no more solid formed. The solvent was decanted and the solid was then partitioned between EtOAc and satd Na 2 CO 3 . The aqueous layer was extracted with EtOAc and the combined organic extracts were dried over Na 2 SO 4 and evaporated to give the title compound (0.90 g, 4.81 mmol, 97%) as a green oil. 1 H NMR (300 MHz, CDCl 3 ) 2.96 (t, J 6.0, 4H), 2.52 (s, 2H), 2.41 (t, J 6.0, 4H), 1.34 (s, 6H).

A suspension of (2S)-2-amino-3-methyl-3-sulfanylbutanoic acid (5.0 g, 703 mnuol) in CH 2 Cl 2 (150 mL) was cooled to 0 C. Trifluoroacetic acid (54 mL, 703 mmol) was added dropwise over a period of 5 min. 2,4,6-trimethoxybenzyl alcohol (6.64 g, 34 mmol) in CH 2 Cl 2 (137 mL) was added dropwise at 0 C. with stirring. The stirring was continued for 1 hour at 0 C. and 2 hours at room temperature, the solvent removed in vacuo and the residue was dried under high vacuum for 3 hours. The crude red solid was recrystallized from 1:1:1 CH 2 Cl 2 :MeOH:EtOAc to give the title compound 10.5 g (95%) as a white solid which was used for the next step without further purification. 1 H NMR (300 MHz, CDCl 3 ) 6.10 (s, 2H), 3.84 (s, 6H), 3.76 (s, 3H), 3.40 4.10 (mult, 3H), 1.69 (s, 3H), 1.23 (s, 3H); mass spectrum (API-TIS) m/z 330 (M H).

To a stirred solution of the product of Example 44a (10.5 g, 32 mmol) in THF (80 mL) was added dropwise lithium aluminum hydride (1 M in THF, 64 mL, 64 mmol) at 0 C. under nitrogen. The resulting solution was stirred at 0 C. for 1 hour and then at room temperature for 2 hours. The excess reducing agent was destroyed carefully by portionwise addition of Na 2 SO 4 10H 2 O at 0 C. The granular white precipitate was removed by filtration and washed with 30% MeOH in CH 2 Cl 2 . The combined filtrate was dried over Na 2 SO 4 , filtered and evaporated to give the title compound 7.6 g (76%) as a yellow oil which was used for the next step without further purification. 1 H NMR (300 MHz, CDCl 3 ) 6.10 (s, 2H), 3.80 (s, 6 H), 3.77 (s, 3H), 3.74 (s, 2H), 3.60-3.40 (mult, 2H), 3.36-3.43 (mult, 1H), 2.93-2.97 (m, 1H), 1.45 (s, 3H), 1.30 (s, 3H); mass spectrum (API-TIS) m/z 316 (M H).

m-Chloroperoxybenzoic acid (57-86%, 24.24 g, 80-121 mmol) was added to an ice-cooled solution of 4-chloro-2-methylthiopyrimidine (6.41 g, 39.9 mmol) in CH 2 Cl 2 (120 mL). The reaction was stirred in the ice-bath for 10 min and at room temperature for 3 hours. The reaction was quenched with 6% Na 2 S 2 O 3 (50 mL). To the resulting mixture was carefully added saturated NaHCO 3 (100 mL). The organic layer was separated and the aqueous layer was extracted with CH 2 Cl 2 (3 50 mL). The combined organic extracts were washed with saturated NaHCO 3 (50 mL), water, brine, dried over Na 2 SO 4 , concentrated under reduced pressure. The crude product was dissolved in CH 2 Cl 2 (20 mL) and triturated with hexane (about 120 mL) to precipitate the title compound. The white solid was collected on a sintered glass funnel and washed with hexane (50 mL). The filtrate was concentrated and the residue was treated as above to yield a second crop. The solid was dried under vacuum to give the title compound as a white powder, 5.8 g (86%). 1 H NMR (300 MHz, CDCl 3 ) 8.87 (d, J 5.4 Hz, 1H), 7.56 (d, J 5.4 Hz, 1H), 3.39 (s, 3H); 13 C NMR (75 MHz, CDCl 3 ) 166.1, 163.3, 159.4, 124.7, 97.15, 39.1; mass spectrum (API-TIS) m/z 193 (M H). Anal Calcd for C 5 H 5 ClN 2 O 2 S: C, 31.18; H, 2.62; N, 14.54. Found: C, 31.21; H, 2.63; N, 14.55.

A suspension of (2S)-amino-3-methyl-3-sulfanylbutanoic (11.5 g, 77.2 mmol) in CH 2 Cl 2 (60 mL) was treated with trifluoroacetic acid (13.7 mL) and stirred to dissolve at room temperature. The mixture was then cooled to 10 C. under nitrogen. A solution of 4-methoxybenzyl chloride (10.5 mL, 77.25mmol) in CH 2 Cl 2 (90 mL) was added dropwise through an additional funnel over a period of 1.5 hours. Stirring was continued for 1.5 hours at room temperature. Methanol (10 mL) was added to dissolve the precipitate. The crude reaction was concentrated in vacuo. The residue was dissolved in CH 2 Cl 2 (40 mL) and extracted with water (7 50 mL). The combined aqueous extracts were frozen and lyophilized. The residue was dissolved in methanol/water (1:3, 200 mL) and brought to pH 6-7 with sodium bicarbonate. The white solid was isolated by filtration, rinsed with MeOH/water (1:3), and dried to give the title compound (6.92 g, 33%). 1 H NMR (300 MHz, CD 3 OD) 7.28 (d, J 7.0 Hz, 2H), 6.85 (d, J 8.4 Hz, 2H), 3.79-3.76 (m, 5H), 3.51 (s, 1H), 1.63 (s, 3H), 1.33 (s, 3H).

To a stirred suspension of methyl (2S)-2-amino-3-hydroxypropanoate hydrogen chloride (1.73 g, 11.1 mmol) in CHCl 3 (50 mL) at 78 C. under nitrogen was added a solution of triethylamine (3.9 mL, 27.8 mmol) and the product of Example 47b in THF (30 mL). The resulting solution was stirred at 78 C. for 4 hours and then warmed to room temperature overnight. The solvents were removed in vacuo. The residue was partitioned between Et 2 O and H 2 O. The organic phase was washed with brine, dried over MgSO 4 , filtered and concentrated to give the title compound (3.17 g), which was used for next step without further purification. Mass spectrum (API-TIS) m/z 371 (M H).

The product of Example 48a (5.3 g, 21.6 mmol) was dissolved in ethanol (25 mL) and a solution of sodium hydroxide (3.1 g, 77.9 mmol) in water (30 mL) was added. The reaction mixture was stirred at room temperature for 5 hours. The reaction mixture was concentrated under reduced pressure and concentrated HCl was added to pH 5.6. Ethanol was added and the volatiles were evaporated. The residue was suspended in EtOAc and filtered. The filter cake was washed with CH 2 Cl 2 and the filtrate was concentrated in vacuo to give 4.5 g (96%) of the title compound. 1 H NMR (300 MHz, CD 3 OD) 3.13 (d, 2H), 2.55-2.68 (m, 4H), 2.17-2.26 9m, 1H), 1.77-1.92 (m, 5H), 1.32 (s, 6H).

The product of Example 48b (1.31 g, 6.04 mmol) was dissolved in anhydrous methanol (20 mL) and 2N HCl (12 mL, 24 mmol) was added. The resulting mixture was cooled to 0 C. and a solution of sodium nitrite (1.66 g, 24.1 mmol) in water (5 mL) was added. The reaction mixture was stirred at 0 C. for 40 min. Ethanol (30 mL) was added and the volatiles were evaporated. The residue was dissolved in ethanol and sodium chloride was removed by filtration. The filtrate was concentrated in vacuo and the residue was purified by flash chromatography on silica gel, eluting with 100:1 to 40:1 CH 2 Cl 2 :MeOH to give 0.73 g (49%) of the title compound as a green oil. 1 H NMR (300 MHz, CDCl 3 ) 3.04 (s, 2H), 2.87-2.93 (m, 2H), 2.46 (t, 2H), 2.30-2.34 (m, 1H), 1.89 (s, 6H), 1.68-1.87 (m, 4H),

Comparative In Vivo Analgesic, Antiinflammatory and Gastric Lesion Activities

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: 729-731, 1957. Male CD-1 mice (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. Five minutes after the i.p. injection of phenylbenzoquinone, the number of writhes in a 5 minute period was counted.

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 l of 1% suspension of carrageenan. Three hours later, the paw volume was measured and compared with the initial volume measured immediately after carrageenan injection.

The rat gastric lesion test, described by Kitagawa et al, J. Phirmacol. Exp. Ther., 253:1133-1137 (1990), and Al-Ghamdi et al, J. Int. Med. Res., 19:2242 (1991), was used to evaluate the activity 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 hours with free access to water and then dosed by oral gavage with vehicle or with test compounds given at a volume of 0.5 mL/100 g. Food was withheld for 18 hours after the initial dosing. Rats were euthanized by CO 2 eighteen hours after dosing. The stomachs were dissected along the greater curvature, washed with a directed stream of 0.9% saline and pinned open on a sylgard based petri dish for examination of the hemorrhagic lesion. Gastric lesion score was expressed in mm and calculated by summning the length of each lesion.

Table 1 shows the relative activities of compounds in the analgesic, antiinflammatory and gastric lesion tests, and are expressed as the ratio of activity relative to the parent NSAID. The results show that the nitrosylated NSAIDs have either comparable or enhanced analgesic and antiinflammatory activities compared to their parent NSAID molecule. Table 1 also shows that the nitrosylated NSAIDs of the present invention have significantly and unexpectedly decreased gastric lesion activities.

Although the invention has been set forth in detail, one skilled in the art will appreciate that numerous changes and modifications can be made to the invention, and that such changes and modifications may be made without departing from the spirit and scope of the present invention.