Amide therapeutics for the treatment of inflammatory bowel disease

Disclosed are methods for treating or preventing inflammatory bowel disease (IBD) using amide and related compounds. Pharmaceutical compositions containing amide compounds which are useful for the treatment or prophylaxis of IBD are also disclosed.

BACKGROUND OF THE INVENTION
 1. Field of the Invention
 This invention relates to the treatment of inflammatory bowel disease
 (IBD). More specifically, this invention is directed to methods for
 treating or preventing IBD using amide compounds. This invention is also
 directed to pharmaceutical compositions containing amide compounds which
 are useful for the treatment or prophylaxis of IBD.
 2. State of the Art
 The term inflammatory bowel disease ("IBD") describes a group of chronic
 inflammatory disorders of unknown causes involving the gastrointestinal
 tract ("GI tract"). The prevalence of IBD in the US is estimated to be
 about 200 per 100,000 population or approximately 500,000 people. Patients
 with IBD can be divided into two major groups, those with ulcerative
 colitis ("UC") and those with Crohn's disease ("CD").
 In patients with UC, there is an inflammatory reaction primarily involving
 the colonic mucosa. The inflammation is typically uniform and continuous
 with no intervening areas of normal mucosa. Surface mucosal cells as well
 as crypt epithelium and submucosa are involved in an inflammatory reaction
 with neutrophil infiltration. Ultimately, this situation typically
 progresses to epithelial damage with loss of epithelial cells resulting in
 multiple ulcerations, fibrosis, dysplasia and longitudinal retraction of
 the colon.
 CD differs from UC in that the inflammation extends through all layers of
 the intestinal wall and involves mesentery as well as lymph nodes. CD may
 affect any part of the alimentary canal from mouth to anus. The disease is
 often discontinuous, i.e., severely diseased segments of bowel are
 separated from apparently disease-free areas. In CD, the bowel wall also
 thickens which can lead to obstructions. In addition, fistulas and
 fissures are not uncommon.
 Clinically, IBD is characterized by diverse manifestations often resulting
 in a chronic, unpredictable course. Bloody diarrhea and abdominal pain are
 often accompanied by fever and weight loss. Anemia is not uncommon, as is
 severe fatigue. Joint manifestations ranging from arthralgia to acute
 arthritis as well as abnormalities in liver function are commonly
 associated with IBD. Patients with IBD also have an increased risk of
 colon carcinomas compared to the general population. During acute
 "attacks" of IBD, work and other normal activity are usually impossible,
 and often a patient is hospitalized.
 Although the cause of IBD remains unknown, several factors such as genetic,
 infectious and immunologic susceptibility have been implicated. IBD is
 much more common in caucasians, especially those of Jewish descent. The
 chronic inflammatory nature of the condition has prompted an intense
 search for a possible infectious cause. Although agents have been found
 which stimulate acute inflammation, none has been found to cause the
 chronic inflammation associated with IBD. The hypothesis that IBD is an
 autoimmune disease is supported by the previously mentioned
 extraintestinal manifestation of IBD as joint arthritis, and the known
 positive response to IBD by treatment with therapeutic agents such as
 adrenal glucocorticoids, cyclosporine and azathioprine, which are known to
 suppress immune response. In addition, the GI tract, more than any other
 organ of the body, is continuously exposed to potential antigenic
 substances such as proteins from food, bacterial byproducts (LPS), etc.
 Once the diagnosis has been made, typically by endoscopy, the goals of
 therapy are to induce and maintain a remission. The least toxic agents
 which patients are typically treated with are the aminosalicylates.
 Sulfasalazine (Azulfidine), typically administered four times a day,
 consists of an active molecule of aminosalicylate (5-ASA) which is linked
 by an azo bond to a sulfapyridine. Anaerobic bacteria in the colon split
 the azo bond to release active 5-ASA. However, at least 20% of patients
 cannot tolerate sulfapyridine because it is associated with significant
 side-effects such as reversible sperm abnormalities, dyspepsia or allergic
 reactions to the sulpha component. These side effects are reduced in
 patients taking olsalazine. However, neither sulfasalazine nor olsalazine
 are effective for the treatment of small bowel inflammation. Other
 formulations of 5-ASA have been developed which are released in the small
 intestine (e.g. mesalamine and asacol). Normally it takes 6-8 weeks for
 5-ASA therapy to show full efficacy.
 Patients who do not respond to 5-ASA therapy, or who have a more severe
 disease, are prescribed corticosteroids. However, this is a short term
 therapy and cannot be used as a maintenance therapy. Clinical remission is
 achieved with corticosteroids within 2-4 weeks, however the side effects
 are significant and include a Cushing goldface, facial hair, severe mood
 swings and sleeplessness. The response to sulfasalazine and
 5-aminosalicylate preparations is poor in Crohn's disease, fair to mild in
 early ulcerative colitis and poor in severe ulcerative colitis. If these
 agents fail, powerful immunosuppressive agents such as cyclosporine,
 prednisone, 6-mercaptopurine or azathioprine (converted in the liver to
 6mercaptopurine) are typically tried. For Crohn's disease patients, the
 use of corticosteroids and other immunosuppressives must be carefully
 monitored because of the high risk of intra-abdominal sepsis originating
 in the fistulas and abscesses common in this disease. Approximately 25% of
 IBD patients will require surgery (colectomy) during the course of the
 disease.
 Oxygen-derived free radicals such as HO., the superoxide anion and other
 reactive oxygen species such as HOCl, have emerged as a common pathway of
 tissue injury in a wide variety of diseases whose underlying cause is an
 inappropriately vigorous and sustained immune response (failure to control
 or down regulate response to the initial, appropriate stimulus). Examples
 of other diseases, in addition to IBD and arthritis, where this mechanism
 appear to be the operative cause are ARDS, septic shock, asthma, diabetes,
 multiple sclerosis, uveitis, etc. Typically, both a cytokine-mediated
 immune response and a nonspecific inflammatory cascade are involved in the
 primary inappropriate response with both responses mediated through active
 oxygen species (oxidative stress). The inappropriate secondary response,
 also mediated through oxidative stress) may involve tissue damaging
 oxidation by neutrophils and tissue macrophages.
 Various approaches have been taken to suppress this inappropriate
 inflammatory response. Small molecule inhibitors of the various
 leukotriene, PAF and cyclooxygenase pathways have shown only limited
 efficacy, perhaps because blocking only one of many pathways does not
 provide a sufficiently large decrease in overall oxidative stress. Another
 approach has been the use of antibodies or cloned receptor molecules which
 target specific proteins in the inflammatory cascade such as IL-1, IL-6 or
 TNF-.alpha.. However, this approach is practical only for acute
 conditions, like septic shock or ARDS, where IV administration and
 antibody formation against the therapeutic protein is less of a concern.
 For a chronic condition like IBD, an orally active small molecule that is
 fully active when dosed once-a-day would be the preferred method of
 treatment.
 Another approach to mitigating the oxidative stress resulting from an
 inflammatory response is to employ nitrone-related therapeutics (NRTs).
 The prototype NRT is .alpha.-phenyl-t-butyl nitrone (PBN) shown below.
 ##STR1##
 NRTs represent a new category of therapeutics with the inherent capacity to
 overcome the shortcomings of other previously studied compounds. Among
 other properties, NRTs such as PBN are believed to trap free radicals (R.)
 by adding the radical to form a more unreactive nitroxyl free radical.
 Nitrones were first used as analytical tools capable of reacting with
 highly reactive radicals to yield free radical adducts that are much less
 reactive. In many cases, the free radical/nitrone-adduct complex is stable
 enough to allow in vivo isolation and quantitation using electron spin
 resonance (ESR). The concept of using nitrones as therapeutics in, for
 example, neurodegenerative diseases resulted from the observations that
 nitrones, such as PBN, trap reactive oxygen species and/or secondary free
 radicals following ischemia. The therapeutic effects of nitrones may
 result because the nitrones convert highly reactive radicals into much
 less reactive products. Certain NRTs have been shown to protect
 experimental animals from ischemia/reperfusion injury (stroke). NRTs,
 administered chronically, reverse the age-associated increase in
 oxidatively damaged protein and the age-associated decrease in the
 activity of the oxidative-sensitive enzyme, glutamine synthetase, in the
 brain.
 Accompanying the NRT-mediated changes in oxidized protein and glutamine
 synthetase activity is a significant improvement in the performance of
 animals in behavioral tests measuring short-term spatial memory. For
 example, it has been shown that prototype NRTs mitigate the effects of
 this inflammatory cascade in a number of in vivo models. Of particular
 interest is the consistent and well documented protection shown by PBN
 against the lethality induced by LPS in various rodent models of septic
 shock. Remarkably, PBN has also been shown to increase the life span of
 senescence-accelerated mice by one third, perhaps by mitigating free
 radical damage. PBN has also been shown to block inducible nitric oxide
 synthetase ("iNOS"), the enzyme responsible for producing large amounts of
 the highly damaging NO. Thus, PBN can both trap HO. and suppress formation
 of NO., potentially neutralizing the effects of the two agents considered
 to be the most damaging to tissue.
 When evaluating the prospects of using an antioxidant to successfully treat
 IBD, it is perhaps also useful to consider that the anti-oxidant defense
 of the human colon is relatively deficient compared to human liver
 (mucosal levels of SOD, catalase and GSH representing 8%, 4% and 40%,
 respectively of liver levels), thus leaving the colon particularly
 sensitive to oxidative stress. A considerable number of chemical
 modifications have been made to increase NRTs suitability as therapeutic
 agents. The effects of intrinsic chemical reactivity and radical trapping
 ability have been examined by substituting the phenyl ring with electron
 donating or electron withdrawing substituents. More water soluble
 analogues have also been made which, for example, have a carboxylate or
 sodium sulfonate group on the phenyl ring. In addition, lipophilic
 analogues have been made with functional group substitutions on either the
 phenyl ring or the nitronyl nitrogen. The alkyl nitrogen substituent has
 also been varied through the standard straight chain and branched C.sub.3
 -C.sub.5 substituents. Nitrone isosteres and related compounds have also
 targeted and examined for efficacy. This approach has led to various
 classes of compounds, such as substituted ureas, amides, thioamides, azoxy
 derivatives, sulphones, and hydroxamic acids. Among these, some benzamide
 compounds substantially similar in structure to some nitrones, such as
 PBN, have been shown to have activity in the treatment of Parkinson's
 disease, HIV dementia, and related conditions.
 As a final aspect of background, in evaluating the effectiveness of
 compounds in the treatment of IBD, an in vivo model based upon
 trinitrobenzene sulfonic acid ("TNBS") is used.
 References relating to the above-mentioned subjects include:
 Glickman, R M (1994) Inflammatory Bowel Disease in Harrison's Principles of
 Internal Medicine (McGraw Hill, New York, N.Y.) Chapter 255: 1403-1416.
 Calkins, B M, Mendeloff, Al (1986) Epidemiology of Inflammatory Bowel
 Disease, Epidemiology Review 8: 60-90.
 Levin, B. (1992) Inflammatory Bowel Disease and Colon Cancer, Cancer
 (Supplement), 70: 1313-1316.
 Crotty, B. (1994) Ulcerative Colitis and Xenobiotic Metabolism, Lancet,
 343: 35-38.
 Hanauer, S B, Baert, F. (1994) Medical Therapy of Inflammatory Bowel
 Disease, Med Clin North Am, 78: 1413-1426.
 MacDermott, R P (1994) Alterations in the Mucosal System in Ulcerative
 Colitis and Crohn's Disease, Med Clin North Am, 78: 1207-1231.
 Hanauer, B. (1993) Medical Therapy of Ulcerative Colitis, Lancet, 342:
 412-417.
 Winrow, V R, Winyard, P G, Morris, C J, Blake, D R (1993) Free radicals in
 Inflammation: Second Messengers and Mediators of Tissue Destruction, Br
 Med Bull 49: 506-522.
 Floyd, R A and Carney, J., Nitrone Radical Traps (NRTs) Protect in
 Experimental Neurodegenerative Diseases, in Neuroprotective Approaches to
 the Treatment of Parkinson's Disease and Other Neurodegenerative Disorders
 (Olanow, C W, Jenner, P and Youssim E, Eds.) Academic Press, New York,
 N.Y., in press.
 Cao, X. and Phillis, J W (1994) a-Phenyl-N-tert-butyl-nitrone Reduces
 Cortical Infarct and Edema in Rats Subjected to Focal Ischemia. Brain Res.
 644: 267-272.
 Zhao, Q., Pahlmark, K., Smith, M.-J., and Siesjo, B. (1994) Delayed
 Treatment with the Spin Trap a-phenyl-n-tert-butyl nitrone (PBN) Reduces
 Infarct Size Following Transient Middle Cerebral Artery Occlusion in Rats.
 Acta Physiol. Scad. 152: 349-350.
 Oliver, C N, Starke-Reed, P E, Stadtman, E R, Carney, J M and Floyd, R A
 (1990) Oxidative Damage to Brain Proteins, Los of Glutamine Synthetase
 Activity and Production of Free Radicals During Ischemia Induced Injury to
 Gerbil Brain. Proc. Natl. Acad. Sci. USA 87: 5144-5147.
 Carney, J M, Starke-Reed, P E Oliver, C N, Landrum, R W, Cheng, M S, Wu, J
 F and Floyd, R A (1991) Reversal or age-related increase in brain protein
 oxidation in enzyme activity, and loss in temporal and spatial memory by
 chronic administration of the spin-trapping compound
 N-tert-butyl-.alpha.-phenylnitrone. Proc. Natl. Acad. Sci., 88: 3633-3636.
 Novelli, G P (1992) Oxygen Radicals in Experimental Shock: Effects of
 Spin-Trapping Nitrones in Ameliorating Shock Pathophysiology, Critical
 Care Medicine, 20: 499-507.
 Hamburger, S A, McCay, P B (1989) Endotoxin-Induced Mortality in Rats is
 Reduced by Nitrones, Circulatory Shock, 29: 329-334.
 Progrebniak, H W, Merino, M J, Hahn, S M, Mitchell, J B, Pass, H I (1992)
 Spin Trap Salvage from Endotoxemia: The Role of Cytokine Down-Regulation,
 Surgery, 112: 130-139.
 McKechnie, K., Furman, B L, Paratt J R (1986), Modification by Oxygen Free
 Radical Scavengers of the Metabolic and Cardiovascular Effects of
 Endotoxin Infusion in Conscious Rats, Circulatory Shock 19: 429-439.
 Edamatsu, R, Mori, A., Packer, L (1995) The Spin Trap
 N-tert-.alpha.-phenyl-butylnitrone Prolongs the Life Span of the
 Senescence Accelerated Mouse, Biochem Biophys Res Comm 211: 847-849.
 Miyajima, T., Kotake, Y. (1995) Spin Trapping Agent, Phenyl N-Tert_Butyl
 Nitrone, Inhibits Induction of Nitric Oxide Synthase in Endotoxin-Induced
 Shock in Mice, Biochem Biophys Res Commun, 215: 114-121.
 Boettner, G R (1987) ESR Parameters of Spin Adducts, Free Radical Biology,
 3: 259-303.
 Harris, M L, Schiller, H J, Reilly, P M, Donowitz, M, Grisham, M B, Bulkley
 (1992), Free Radicals and Other Reactive Oxygen Metabolites in
 Inflammatory Bowel Disease: Cause, Consequence or Epiphenomenom,
 Pharmacol. Ther., 53: 375-408.
 Grisham M B, MacDermott, R P, Deitch E A (1990), Oxidant Defence Mechanisms
 in the Human Colon, Inflammation, 14: 669-680.
 Elson, C O, Startor, R B, Tennyson, G S, Ridell, R H (1995), Experimental
 Models of Inflammatory Bowel Disease, Gastroenterology, 109: 1344-1367.
 Yamada, T, Marshall, S, Specian, R D, Grisham, M B (1992) A Comparative
 Analysis of Two Models of Colitis in Rats, Gastroenterology, 102:
 1524-1534.
 Wallace, J A, MacNaughton, W K, Morris, G P, Beck P L (1989) Inhibition of
 Leulotriene Synthesis Markedly Accelerates Healing in a Rat Model of
 Inflammatory Bowel Disease, Gastroenterology, 95: 29-35.
 Higa, A. McKnight, G W, Wallace, J L (1993) Attenuation of Epithelial
 Injury in Acute Experimental Colitis by Immunomodulators, Eur. J.
 Pharmacol. 239: 171-178.
 Castro, G A, Roy, S A, Stockstill, R D (1974) Trichinella Spiralis:
 Peroxidase Activity in Isolated Cells from the Rat Intestine, Exp.
 Parasitol., 36: 307-315.
 SUMMARY OF THE INVENTION
 It has now been found that certain amide compounds are effective for the
 treatment and prophylaxis of IBD. It has also been discovered that certain
 related compounds are effective for the treatment and prophylaxis of IBD.
 Accordingly, in one of its composition aspects, this invention provides a
 pharmaceutical composition for the treatment or prophylaxis of
 inflammatory bowel disease comprising a pharmaceutically acceptable
 carrier and an effective inflammatory bowel disease-treating amount of a
 compound of formula I:
 ##STR2##
 wherein
 A is selected from the group consisting of alkylene and cycloalkylene;
 R.sup.1 is naphthyl or a group having the formula:
 ##STR3##
 wherein each R.sup.a is independently selected from the group consisting
 of aminocarbonyl, hydroxy and trifluoromethyl;
 R.sup.b is hydrogen or acyl;
 R.sup.2 is selected from the group consisting of hydrogen, alkyl, aralkyl
 and cycloalkyl;
 R.sup.3 is selected from the group consisting of alkyl, aralkyl and
 cycloalkyl;
 m is an integer from 0 to 3; and n is 0 or 1;
 and pharmaceutically acceptable salts thereof;
 with the proviso that:
 when R.sup.2 is hydrogen, R.sup.3 is tert-butyl and n is 0, then R.sup.1 is
 not a 4-hydroxyphenyl group or a 4-tert-butylaminocarbonyl group.
 When present, A is preferably alkylene having from about 2 to about 4
 carbon atoms or cycloalkylene having from about 3 to about 5 carbon atoms.
 More preferably, A is ethylene, i.e., --CH.sub.2 CH.sub.2 --, or
 trans-cyclopropylene, i.e.,
 ##STR4##
 Preferably, R.sup.1 is phenyl, 4-trifluoromethylphenyl, 2-hydroxyphenyl,
 2-naphthyl, indol-5-yl, 1-acetylindol-5-yl, and the like.
 R.sup.2 is preferably hydrogen or methyl, more preferably R.sup.2 is
 hydrogen.
 Preferably, R.sup.3 is alkyl or cycloalkyl having 3 to 6 carbon atoms. More
 preferably, R.sup.3 is alkyl having 3 to 6 carbon atoms. Still more
 preferably, R.sup.3 is n-butyl, tert-butyl or cyclohexyl.
 Preferably, m is 0 or 1.
 In another of its composition aspects, this invention provides a
 pharmaceutical composition for the treatment or prophylaxis of
 inflammatory bowel disease comprising a pharmaceutically acceptable
 carrier and an effective inflammatory bowel disease-treating amount of a
 compound selected from the group consisting of:
 N-n-butyl 3-phenylpropionamide,
 N-n-butyl trans-2-phenyl-1-cyclopropanecarboxamide,
 N-tert-butyl 4-trifluoromethylbenzamide,
 N-tert-butyl 2-hydroxybenzamide,
 N-tert-butyl naphthylene-2-carboxamide,
 N-tert-butyl indole-5-carboxamide,
 N-tert-butyl 1-acetylindole-5-carboxamide,
 and pharmaceutically acceptable salts thereof.
 It has also been discovered that certain structurally-related compounds are
 effective for the treatment and prophylaxis of IBD. Accordingly, in yet
 another of its composition aspects, this invention provides a
 pharmaceutical composition for the treatment or prophylaxis of
 inflammatory bowel disease comprising a pharmaceutically acceptable
 carrier and an effective inflammatory bowel disease-treating amount of a
 compound selected from the group consisting of:
 4-(acetamido)benzoic acid,
 N-tert-butyl 4-[2-(4-(3-methoxyphenyl)piperazin-1-yl)acetamido)]benzamide,
 7-nitroindole,
 and pharmaceutically acceptable salts thereof.
 Another aspect of this invention is directed to methods for treating a
 patient suffering from or susceptible to an inflammatory bowel condition.
 Accordingly, this invention provides a method for treating a patient
 suffering from or susceptible to an inflammatory bowel condition
 comprising administering to said patient a pharmaceutical composition
 comprising a pharmaceutically acceptable carrier and an effective
 inflammatory bowel condition-treating amount of a compound of formula I:
 ##STR5##
 wherein
 A is selected from the group consisting of alkylene and cycloalkylene;
 R.sup.1 is naphthyl or a group having the formula:
 ##STR6##
 wherein each R.sup.a is independently selected from the group consisting
 of aminocarbonyl, hydroxy and trifluoromethyl;
 R.sup.b is hydrogen or acyl;
 R.sup.2 is selected from the group consisting of hydrogen, alkyl, aralkyl
 and cycloalkyl;
 R.sup.3 is selected from the group consisting of alkyl, aralkyl and
 cycloalkyl;
 m is an integer from 0 to 3; and n is 0 or 1;
 and pharmaceutically acceptable salts thereof;
 with the proviso that:
 when R.sup.2 is hydrogen, R.sup.3 is tert-butyl and n is 0, then R.sup.1 is
 not a 4-hydroxyphenyl group or a 4-tert-butylaminocarbonyl group.
 Preferably, A, R.sup.1, R.sup.2, R.sup.3, n and m are as described above.
 In another of its method aspects, this invention provides a method for
 treating or preventing inflammatory bowel disease comprising:
 (a) identifying a patient suffering from or susceptible to an inflammatory
 bowel condition; and
 (b) administering to said patient a pharmaceutical composition comprising a
 pharmaceutically acceptable carrier and an effective inflammatory bowel
 condition-treating amount of a compound of formula I above.
 In still another of its method aspects, this invention provides a method
 for treating a patient suffering from or susceptible to an inflammatory
 bowel condition comprising administering to said patient a pharmaceutical
 composition comprising a pharmaceutically acceptable carrier and an
 effective inflammatory bowel condition-treating amount of a compound
 selected from the group consisting of:
 N-n-butyl 3-phenylpropionamide,
 N-n-butyl trans-2-phenyl-1-cyclopropanecarboxamide,
 N-tert-butyl 4-trifluoromethylbenzamide,
 N-tert-butyl 2-hydroxybenzamide,
 N-tert-butyl naphthylene-2-carboxamide,
 N-tert-butyl indole-5-carboxamide,
 N-tert-butyl 1-acetylindole-5-carboxamide,
 and pharmaceutically acceptable salts thereof.
 In yet another of its method aspects, this invention provides a method for
 treating a patient suffering from or susceptible to an inflammatory bowel
 condition comprising administering to said patient a pharmaceutical
 composition comprising a pharmaceutically acceptable carrier and an
 effective inflammatory bowel condition-treating amount of a compound
 selected from the group consisting of:
 4-(acetamido)benzoic acid,
 N-tert-butyl 4-[2-(4-(3-methoxyphenyl)piperazin-1-yl)acetamido)]benzamide,
 7-nitroindole,
 and pharmaceutically acceptable salts thereof.
 In still another of its method aspects, this invention provides a method
 for treating or preventing inflammatory bowel disease comprising:
 (a) identifying a patient suffering from or susceptible to an inflammatory
 bowel condition; and
 (b) administering to said patient a pharmaceutical composition comprising a
 pharmaceutically acceptable carrier and an effective inflammatory bowel
 condition-treating amount of a compound selected from the group consisting
 of:
 4-(acetamido)benzoic acid,
 N-tert-butyl 4-[2-(4-(3-methoxyphenyl)piperazin-1-yl)acetamido)]benzamide,
 7-nitroindole,
 and pharmaceutically acceptable salts thereof.
 In the methods of this invention, the pharmaceutical compositions may be
 administered orally, parenterally, or rectally. The methods of this
 invention are be effective where the inflammatory bowel condition is
 ulcerative colitis or Crohn's disease.
 In one embodiment of the above methods, the pharmaceutical composition is
 preferably administered as an oral dose in an amount of from 0.1 to about
 150 mg/kg of patient weight.
 In another embodiment of the above methods, the pharmaceutical composition
 is preferably administered intravenously in an amount of from about 0.01
 mg/kg/hour to about 100 mg/kg/hour of patient weight for at least about 1
 hour.
 In still another embodiment of the above methods, the pharmaceutical
 composition is preferably administered rectally in an amount of from 1 to
 about 150 mg/kg of patient weight.
 In one of its composition aspects, this invention is also directed to novel
 amide compounds. Accordingly, this invention is directed to each of the
 following compounds:
 N-n-butyl 3-phenylpropionamide,
 N-n-butyl trans-2-phenyl-1-cyclopropanecarboxamide,
 N-tert-butyl 4-trifluoromethylbenzamide,
 N-tert-butyl 2-hydroxybenzamide,
 N-tert-butyl naphthylene-2-carboxamide,
 N-tert-butyl indole-5-carboxamide,
 N-tert-butyl 1-acetylindole-5-carboxamide,
 N-tert-butyl 4-[2-(4-(3-methoxyphenyl)piperazin-1-yl)acetamido)]benzamide,
 and pharmaceutically acceptable salts thereof.
 DETAILED DESCRIPTION OF THE INVENTION
 The treatment methods and pharmaceutical compositions of this invention
 employ one or more amides or related compounds as the active agent. For
 the purposes of this invention, the amide compounds are named using
 conventional amide nomenclature, i.e., the substituents on the amide
 nitrogen atom are given the N-prefix. For example, N-tert-butyl
 4-trifluoromethylbenzamide has the formula:
 ##STR7##
 In some cases, the amides of this invention may contain one or more chiral
 centers. Typically, such compounds will be prepared as a racemic mixture.
 If desired, however, such compounds can be prepared or isolated as pure
 stereoisomers, i.e., as individual enantiomers or diastereomers, or as
 stereoisomer-enriched mixtures. All such stereoisomers (and enriched
 mixtures) of the amide of formula I are included within the scope of this
 invention. Pure stereoisomers (or enriched mixtures) may be prepared
 using, for example, optically active starting materials or stereoselective
 reagents well known in the art. Alternatively, racemic mixtures of such
 compounds can be separated using, for example, chiral column
 chromatography, chiral resolving agents and the like.
 Additionally, all geometric isomers of the amide compounds of formula I are
 included within the scope of this invention including, for example, all
 cis and trans isomers of the amides of formula I wherein A is
 cycloalkylene.
 Definitions
 When describing the compounds, pharmaceutical compositions and methods of
 this invention, the following terms have the following meanings unless
 otherwise specified.
 "Acyl" refers to the group "--C(O)R" where R is alkyl or aryl.
 "Acylamino" refers to the group "--NRC(O)R" where each R is independently
 hydrogen, alkyl, aralkyl or cycloalkyl.
 "Acetamido" refers to the group "--NHC(O)CH.sub.3 ".
 "Alkoxy" refers to the group "--OR" where R is alkyl.
 "Alkyl" refers to monovalent alkyl groups preferably having from 1 to about
 12 carbon atoms, more preferably 1 to 8 carbon atoms and still more
 preferably 1 to 6 carbon atoms. This term is exemplified by groups such as
 methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl,
 n-hexyl, n-octyl, tert-octyl and the like. The term "lower alkyl" refers
 to alkyl groups having 1 to 6 carbon atoms.
 "Alkylene" refers to divalent alkylene groups preferably having from 1 to
 12 carbon atoms and more preferably 1 to 6 carbon atoms which can be
 straight chain or branched. This term is exemplified by groups such as
 methylene (--CH.sub.2 --), ethylene (--CH.sub.2 CH.sub.2 --), the
 propylene isomers (e.g., --CH.sub.2 CH.sub.2 CH.sub.2 -- and
 --CH(CH.sub.3)CH.sub.2 --) and the like.
 "Aminocarbonyl" refers to the group "--C(O)NRR" where each R is
 independently hydrogen, alkyl, aralkyl or cycloalkyl.
 "Aralkyl" refers to "aryl-alkylene-" groups preferably having from 1 to 10
 carbon atoms in the alkylene moiety and from 6 to 14 carbon atoms in the
 aryl moiety. Such aralkyl groups are exemplified by benzyl, phenethyl, and
 the like.
 "Aryl" refers to an unsaturated aromatic carbocyclic group of from 6 to 14
 carbon atoms having a single ring (e.g., phenyl) or multiple condensed
 rings (e.g., naphthyl or anthryl). Preferred aryls include phenyl,
 naphthyl and the like. Unless otherwise indicated, such aryl groups can
 optionally be substituted with from 1 to 5 substituents, preferably 1 to 3
 substituents, selected from the group consisting of alkyl, alkoxy,
 alkoxycarbonyl, carboxyl, cyano, halo, hydroxy, nitro, thioalkoxy and the
 like.
 "Carboxyl" refers to the group "--C(O)OH" and salts thereof.
 "Cyano" refers to the group "--CN".
 "Cycloalkyl" refers to cyclic alkyl groups of from 3 to 10 carbon atoms
 having a single cyclic ring or multiple condensed rings which can be
 optionally substituted with from 1 to 3 alkyl groups. Such cycloalkyl
 groups include, by way of example, single ring structures such as
 cyclopropyl, cyclobutyl, cyclopentyl, cyclooctyl, 1-methylcyclopropyl,
 2-methylcyclopentyl, 2-methylcyclooctyl, and the like, or multiple ring
 structures such as adamantanyl, and the like.
 "Cycloalkylene" refers to divalent cyclic alkyl groups of from 3 to 10
 carbon atoms having a single cyclic ring which can be optionally
 substituted with from 1 to 3 alkyl groups. Such cycloalkyl groups include,
 by way of example, cyclopropylene, cyclobutylene, cyclopentylene and the
 like.
 "Halo" or "halogen" refers to fluoro, chloro, bromo and iodo.
 "Hydroxy" refers to the group "--OH".
 "Nitro" refers to the group "--NO.sub.2 ".
 "Sulfonate" or "sulfo" refers to the group "--SO.sub.3 H" and salts
 thereof.
 "Thioalkoxy" or "alkylthioether" refers to "alkyl-S--" groups. Preferred
 thioalkoxy groups include, by way of example, thiomethoxy, thioethoxy,
 n-thiopropoxy, isothiopropoxy, n-thiobutoxy and the like.
 "Trifluoromethyl" refers to the group "--CF.sub.3 ".
 "Pharmaceutically acceptable salt" refers to salts which are acceptable for
 administration to mammals including, by way of illustration, alkali and
 alkaline earth metal salts and addition salts of free acids and amines.
 Such pharmaceutically acceptable salts may be derived from a variety of
 organic and inorganic counter-ions well known in the art and include, by
 way of example only, sodium, potassium, calcium, magnesium, ammonium,
 tetraalkylammonium, and the like; and when the molecule contains a basic
 functionality, salts of organic or inorganic acids, such as hydrochloride,
 hydrobromide, tartrate, mesylate, acetate, maleate, oxalate and the like.
 The term "pharmaceutically acceptable cation" refers to a pharmaceutically
 acceptable cationic counterion of an acidic functional group. Such cations
 are exemplified by sodium, potassium, calcium, magnesium, ammonium,
 tetraalkylammonium cations, and the like.
 General Synthetic Procedures
 The compounds of this invention can be prepared from readily available
 starting materials using the following general methods and procedures. It
 will be appreciated that where typical or preferred process conditions
 (i.e., reaction temperatures, times, mole ratios of reactants, solvents,
 pressures, etc.) are given, other process conditions can also be used
 unless otherwise stated. Optimum reaction conditions may vary with the
 particular reactants or solvent used, but such conditions can be
 determined by one skilled in the art by routine optimization procedures.
 Additionally, as will be apparent to those skilled in the art, conventional
 protecting groups may be necessary to prevent certain functional groups
 from undergoing undesired reactions. The choice of a suitable protecting
 group for a particular functional group as well as suitable conditions for
 protecting and deprotecting various functional groups are well known in
 the art. For example, numerous protecting groups, and their introduction
 and removal, are described in T. W. Greene and G. M. Wuts, Protecting
 Groups in Organic Synthesis, Second Edition, Wiley, New York, 1991, and
 references cited therein.
 In a preferred method of synthesis, the amide compounds of this invention
 are prepared by coupling a carboxylic acid halide of formula II:
 ##STR8##
 wherein A, R.sup.1 and n are as defined above, and X is halo, preferably
 chloro or bromo, with an amine of formula III:
 R.sup.2 --NH--R.sup.3 III
 wherein R.sup.2 and R.sup.3 are as defined above, under conventional
 reaction conditions.
 The coupling reaction is typically conducted by contacting the carboxylic
 acid halide II with an excess, preferably about 1.1 to about 3 equivalents
 per carbonyl halide group, of amine III. This reaction is typically
 conducted at a temperature of from about -10.degree. C. to about
 30.degree. C. for about 1 to about 24 hours. Typically, the reaction is
 conducted in an inert diluent such as dichloromethane, chloroform,
 benzene, toluene, acetonitrile, tetrahydrofuran and the like.
 Preferably, the coupling reaction is conducted in the presence of a
 suitable base to scavenge the acid generated during the reaction. Suitable
 bases include, by way of example, triethylamine, diisopropylethylamine,
 N-methylmorpholine and the like. Alternatively, an excess of amine III may
 be used to scavenge the acid generated during the reaction.
 Upon completion of the coupling reaction, the amide is recovered by
 conventional methods including precipitation, chromatography, filtration,
 distillation and the like. If a protecting group has been employed, the
 protecting group is then removed using conventional procedures. For
 example, when 2-acetoxybenzoyl chloride is coupled with an amine, the
 acetyl group of the resulting amide may be removed to re-generate a
 hydroxyl group by treating the amide with 5% hydrochloric acid in an inert
 diluent.
 The carboxylic acid halides employed in the coupling reaction are either
 commercially available or can be prepared from commercially available
 starting materials and reagents using conventional procedures. For
 example, carboxylic acid halides can be readily prepared from the
 corresponding carboxylic acid by contacting the carboxylic acid with an
 inorganic acid halide, such as thionyl chloride, phosphorous trichloride,
 phosphorous tribromide or phosphorous pentachloride, or alternatively,
 with oxalyl chloride under conventional conditions. Generally, this
 reaction is conducted using about 1 to 5 molar equivalents of the
 inorganic acid halide or oxalyl chloride, either neat or in an inert
 solvent, such as carbon tetrachloride, at temperature in the range of
 about 0.degree. C. to about 80.degree. C. for about 1 to about 48 hours. A
 catalyst, such as N,N-dimethylformamide, may also be used in this
 reaction.
 Preferred carboxylic acid halides for use in this invention
 3-phenylpropionyl chloride, trans-2-phenyl-1-cyclopropanecarbonyl
 chloride, 4-trifluoromethylbenzoyl chloride, 2-acetoxybenzoyl chloride,
 terephthaloyl chloride, 2-naphthoyl chloride and the like.
 The amines of formula III are also either known compounds or compounds that
 can be prepared from known starting material and reagents by conventional
 procedures. Examples of suitable amines for use in this reaction include,
 but are not limited to, methylamine, ethylamine, n-propylamine,
 isopropylamine, n-butylamine, isobutylamine, sec-butylamine,
 tert-butylamine, n-pentylamine, cyclopentylamine, n-hexylamine,
 cyclohexylamine, n-octylamine, tert-octylamine, dimethylamine,
 diethylamine, di-n-propylamine, diisopropylamine, di-n-butylamine,
 diisobutylamine, di-sec-butylamine, di-n-hexylamine, methylethylamine,
 methyl-n-propylamine, methylisopropylamine, methyl-n-butylamine,
 methyl-tert-butylamine, methyl-tert-octylamine, methylcyclopentylamine,
 methylcyclohexylamine, ethyl-n-propylamine, ethylisopropylamine,
 ethyl-n-butylamine, ethylcyclohexylamine, benzylamine, pyrrolidine,
 piperidine and the like. Preferred amine include n-butylamine and
 tert-butylamine.
 Alternatively, the amides of formula I can be prepared by coupling a
 carboxylic acid (i.e., a compound of formula II where X is OH) with an
 amine of formula III using conventional coupling reagent, such as
 dicyclohexylcarbodiimide and the like, or N,N'-carbonyldiimidazole. This
 reaction can be conducted with or without the use of well known additives
 such as N-hydroxysuccinimide, 1-hydroxybenzotriazole, etc. which are known
 to facilitate the coupling of carboxylic acids and amines.
 The 4-acetamidobenzoic acid employed in the pharmaceutical compositions and
 methods of this invention is commercially available from, for example,
 Aldrich Chemical Company.
 Pharmaceutical Compositions
 When used as pharmaceuticals, the compounds employed in this invention are
 typically administered in the form of a pharmaceutical composition. Such
 compositions can be prepared using procedures well known in the
 pharmaceutical art and comprise at least one active compound.
 Generally, the compounds of this invention are administered in a
 pharmaceutically effective amount. The amount of the compound actually
 administered will typically be determined by a physician, in the light of
 the relevant circumstances, including the condition to be treated, the
 chosen route of administration, the actual compound administered, the age,
 weight, and response of the individual patient, the severity of the
 patient's symptoms, and the like.
 The compound(s) is typically formulated into a pharmaceutical composition
 suitable for oral, parenteral (e.g. intravenous or intramuscular
 injection), or rectal (e.g. suppository) administration.
 The compositions for oral administration can take the form of liquid
 solutions or suspensions, powders, tablets, capsules or the like. In such
 compositions, the amide compound is usually a minor component (0.1 to
 about 50% by weight) with the remainder being various vehicles or carriers
 and processing aids helpful for forming the desired dosing form. A liquid
 form may include a suitable aqueous or nonaqueous vehicle with buffers,
 suspending dispensing agents, colorants, flavors and the like.
 A solid form may include, for example, any of the following ingredients, or
 compounds of a similar nature: a binder such as microcrystalline
 cellulose, gum tragacanth or gelatin; an excipient such as starch or
 lactose; a disintegrating agent such as alginic acid, Primogel, or corn
 starch; a lubricant such as magnesium stearate; a glidant such as
 colloidal silicon dioxide; a sweetening agent such as sucrose or
 saccharin; or a flavoring agent such as peppermint, sugar, methyl
 salicylate, or orange flavoring.
 Injectable compositions are commonly based upon injectable sterile saline
 or phosphate-buffered saline or other injectable carriers known in the
 art. Again, the active amide compound is typically a minor component,
 often being from about 0.05 to 10% by weight, with the remainder being the
 injectable carrier and the like.
 Rectal administration is usually by suppository. Suppositories are
 generally made with a base component of cocoa butter, glycerinated
 gelatin, hydrogenated vegetable oils, mixtures of polyethylene glycols of
 various molecular weights, or fatty acid esters of polyethylene glycol.
 The active amide compound is usually a minor component, often from about
 0.05 to 20% by weight, with the remainder being the base component.
 The components for orally administrable, injectable compositions and
 suppositories are merely representative. Other materials as well as
 processing techniques and the like are set forth in Part 8 of Remington's
 Pharmaceutical Sciences, 18th edition, 1990, Mack Publishing Company,
 Easton, Pa., 18042, which is incorporated herein by reference.
 One can also administer the compounds of the invention in sustained release
 forms or from sustained release drug delivery systems. A description of
 representative sustained release materials can be found in the
 incorporated materials in Remington's Pharmaceutical Sciences.
 Conditions Treated and Treatment Regimens
 The conditions treated with the pharmaceutical compositions of this
 invention generally include IBD and the various symptoms which fall within
 a definition of IBD. The formulations are administered to achieve a
 therapeutic effect. For those compounds that exhibit a long residency in
 the body, a once-a-day regimen is possible. Alternatively, multiple doses,
 such as up to three doses per day, typically, may offer more effective
 therapy. Thus, a single dose or a multidose regimen may be used.
 In any event, the pharmaceutical composition is administered in such a
 manner so that compound is delivered into the patient's bloodstream. One
 excellent mode for accomplishing this is intravenous administration.
 Intravenous dose levels for treating IBD range from about 0.01 mg/kg/hour
 of active amide compound to about 100 mg/kg/hour, all for from about 1 to
 about 120 hours and especially 1 to 96 hours. A preloading bolus of from
 about 50 to about 5000 mg may also be administered to achieve adequate
 steady state levels. Other forms of parenteral administration, such as
 intramuscular injection can be used, as well. In this case, similar dose
 levels are employed.
 With oral dosing, one to three oral doses per day, each from about 0.1 to
 about 150 mg/kg of active compound are employed, with preferred doses
 being from about 0.15 to about 100 mg/kg.
 With rectal dosing, one to three rectal doses per day, each from about 1 to
 about 150 mg/kg of active compound are employed, with preferred doses
 being from about 1 to about 100 mg/kg.
 In any treatment regimen, the health care professional should assess the
 patient's condition and determine whether or not the patient would benefit
 from treatment. Some degree of routine dose optimization may be required
 to determine an optimal doing level and pattern.
 A positive dose-response relationship has been observed. As such and
 bearing in mind the severity of the side effects and the advantages of
 providing maximum possible amelioration of symptoms, it may be desired in
 some settings to administer large amounts of active compound, such as
 those described above.

The following formulation examples illustrate representative pharmaceutical
 compositions of this invention. The present invention, however, is not
 limited to the following pharmaceutical compositions.
 FORMULATION 1
 Tablets
 A compound of formula I is admixed as a dry powder with a dry gelatin
 binder in an approximate 1:2 weight ratio. A minor amount of magnesium
 stearate is added as a lubricant. The mixture is formed into 240-270 mg
 tablets (80-90 mg of active compound per tablet) in a tablet press.
 FORMULATION 2
 Capsules
 A compound of formula I is admixed as a dry powder with a starch diluent in
 an approximate 1:1 weight ratio. The mixture is filled into 250 mg
 capsules (125 mg of active compound per capsule).
 FORMULATION 3
 Liquid
 A compound of formula I (125 mg), sucrose (1.75 g) and xanthan gum (4 mg)
 are blended, passed through a No. 10 mesh U.S. sieve, and then mixed with
 a previously made solution of microcrystalline cellulose and sodium
 carboxymethyl cellulose (11:89, 50 mg) in water. Sodium benzoate (10 mg),
 flavor, and color are diluted with water and added with stirring.
 Sufficient water is then added to produce a total volume of 5 mL.
 FORMULATION 4
 Injection
 The compound of formula I is dissolved in a buffered sterile saline
 injectable aqueous medium to a concentration of approximately 5 mg/mL.
 The following synthetic and biological examples are offered to illustrate
 this invention and are not to be construed in any way as limiting the
 scope of this invention.
 EXAMPLES
 In the examples below, all temperatures are in degrees Celsius (unless
 otherwise indicated). Examples 1-8 describe the synthesis of various
 amides; and the Bioassay Examples describe the testing of such compounds.
 In the examples below, the following abbreviations have the following
 meanings. Abbreviations not defined below have their generally accepted
 meaning.
 bd=broad doublet
 bs=broad singlet
 d=doublet
 dd=doublet of doublets
 dec=decomposed
 dH.sub.2 O=distilled water
 EtOAc=ethyl acetate
 EtOH=ethanol
 g=grams
 h=hours
 Hz=hertz
 L=liter
 m=multiplet
 min=minutes
 M=molar
 MeOH=methanol
 mg=milligram
 MHz=megahertz
 mL=milliliter
 mmol=millimole
 m.p.=melting point
 N=normal
 q=quartet
 quint.=quintet
 s=singlet
 t=triplet
 THF=tetrahydrofuran
 tlc=thin layer chromatography
 .mu.g=microgram
 .mu.L=microliter
 UV=ultraviolet
 Example 1
 Synthesis of N-tert-Butyl 4-Trifluoromethylbenzamide
 To N-tert-butylamine (3.50 g, 0.0048 mol) in 75 mL of benzene was added
 4-trifluoromethylbenzoyl chloride (5.00 g, 0.024 mol). The temperature was
 maintained below 15.degree. C. during the addition and then the reaction
 mixture was allowed to warm to ambient temperature and stirred for 3
 hours. The solvent was then stripped and the residue was dissolved in 75
 mL of dichloromethane. This solution was washed with 5% hydrochloride acid
 (2.times.75 mL), water (2.times.75 mL), dried over MgSO.sub.4 and then the
 solvent was removed in vacuo to provide 5.05 g (85-9% yield) of the title
 compound, m.p. 146-148.degree. C.
 Example 2
 Synthesis of N-n-Butyl 3Phenylpropionamide
 Following the procedure of Example 1 above and using 3-phenylpropionyl
 chloride and n-butylamine (and ethyl acetate in place of benzene), the
 title compound was prepared as a white solid, m.p. 26.7-33.1.degree. C.
 Example 3
 Synthesis of N-n-Butyl trans-2-Phenyl-1-cyclopropanecarboxamide
 Following the procedure of Example 1 above and using
 trans-2-phenyl-1-cyclopropanecarbonyl chloride and n-butylamine (and ethyl
 acetate in place of benzene), the title compound was prepared as a waxy
 white solid.
 Example 4
 Synthesis of N-tert-Butyl Naphthalene-2-carboxamide
 Following the procedure of Example 1 above and using 2-naphthoyl chloride
 and tert-butylamine (and ethyl acetate in place of benzene), the title
 compound was prepared as a white solid, m.p. 154.8-159.2.degree. C.
 Example 5
 Synthesis of N-tert-Butyl 2-Hydroxybenzamide
 Step A
 Synthesis of N-tert-Butyl 2-Acetoxybenzamide
 Following the procedure of Example 1 above and using 2-acetoxybenzoyl
 chloride and tert-butylamine, the title compound was prepared.
 Step B
 Synthesis of N-tert-Butyl 2-Hydroxybenzamide
 The product from Step A (2.35 g, 0.010 mol) was dissolved in THF wand 10 mL
 of water was added to form a clear suspension. Hydrochloric acid (5%) was
 added until the solution reached a pH.about.1. The reaction mixture was
 then refluxed for 24 hours. TLC indicated that about half of the starting
 material remained, so an additonal 10 mL of 5% hydrochloric acid was added
 and the reaction mixture was refluxed for 24 hours. The THF was then
 removed by rotoevaporation and the remaining aqueous mixture was extracted
 with hexanes (2.times.20 mL). The combined hexane layers were washed with
 water (2.times.40 mL), dried over MgSO.sub.4 and the hexane removed in
 vacuo to provide 1.56 g (80.8% yield) of the title compound as a white
 crystalline solid, m.p. 78-79.5.degree. C.
 Example 6
 Synthesis of N-tert-Butyl Indole-5carboxamide
 To a cooled to -30.degree. C. solution of indole-5-carboxylic acid and
 triethylamine in DMF was added methyl sulfonyl chloride in portions to
 avoid temperature elevation. Following the addition, solution was warmed
 to room temperature and was stirred for 1 hour and 15 minutes. Reaction
 mixture was then cooled to -20.degree. C. and N-tert-butyl-amine was added
 in two portions so the temperature remained at -10.degree. C. After 13
 hours of stirring, the solution was diluted with water and saturated with
 sodium chloride. The solution was extracted with ethyl acetate and the
 organic phase was dried over magnesium sulfate, filtered and concentrated
 under vacuum to afford the title compound as a thick brown oil (NMR showed
 34% DMF) (28% yield), melting point=176.4.degree. C.
 Spectroscopic data was as follows:
 .sup.1 H NMR (CDCl.sub.3, 270 MHZ): .delta.=10.48 ppm (1H, s), 8.11 ppm
 (1H, m) 7.65 ppm (1H, dd, J=8.66; 1.73), 7.42 ppm (1H, dt, J=8.66; 0.74),
 7.38 ppm (1H, dd, J=2.94), 6.98 ppm (1H, s), 6.53 ppm (1H, ddd, J=2.94;
 2.10; 0.74) 1.48 ppm (9H, s).
 Example 7
 Synthesis of N-tert-Butyl 1-Acetylindole-5-carboxamide
 N-tert-Butyl indole-5-carboxamide was dissolved in dry THF and solution was
 cooled to -78.degree. C. Butyllithium was added dropwise with continuous
 stirring. The reaction mixture was stirred at -78.degree. C. for 40
 minutes followed by dropwise addition of acetyl chloride. After addition
 was complete, the reaction mixture was stirred at -78.degree. C. for 30
 minutes and then stirred for 1 hour at room temperature. The reaction
 mixture was diluted with water and organic phase separated. The aqueous
 phase was saturate with sodium chloride and was extracted with ethyl
 acetate. The extracts were combined, dried over magnesium sulfate,
 filtered and concentrated under vacuum. The crude product was purified by
 column chromatrography to afford the title compound as a white solid
 (14.1% yield), melting point=186.8.degree. C.
 Spectroscopic data was as follows:
 .sup.1 H NMR (CDCl.sub.3, 270 MHZ): .delta.=8.38 ppm (1H, d, J=8.66), 8.09
 ppm (1H, d, J=1.49), 7.82 ppm (2H, dd, J=8.66; 1.49), 7.81 ppm (2H, d,
 J=3.71), 7.14 ppm (1H, s), 6.74 ppm (1H, d, J=3.71), 2.69 ppm (3H, s),
 1.48 ppm (9H, s).
 Example 8
 Synthesis of N-tert-butyl
 4-[2-(4-(3-methoxyphenyl)piperazin-1-yl)acetamido)]benzamide
 To a cooled to -20.degree. C. solution of tert-butyl 4-aminobenzamide and
 triethylamine (1.30 eq.) in ethyl acetate was added chloroacetyl chloride
 portion-wise with continuous stirring. The reaction mixture was stirred
 for a total of 4 hours at room temperature. The solution was then diluted
 with water and extracted with ethyl acetate and the organic phases were
 combined, washed with water and brine, and dried over magnesium sulfate.
 The organic phases were concentrated under vacuum to give N-tert-butyl
 4-(2-chloroacetamido)benzamide as a grayish solid (74% yield).
 N-tert-Butyl 4-(2-chloroacetamido)benzamide, 1-(3-methoxyphenyl)piperazine
 dichloride (1.1 eq.) and sodium bicarbonate (4.0 eq) were all dissolved in
 DMF and stirred at 85.degree. C. for 10 hours. The reaction mixture was
 diluted with methylene chloride/water and the organic phase was washed
 with water/brine and was dried over magnesium sulfate. Evaporation of the
 solvent gave gray-brownish solid which was purified by dry flash column
 chromatography (ethyl acetate; methylene chloride=1:1) to give the title
 compound as a white solid (53.4% yield), melting point=183.2.degree. C.
 Spectroscopic data was as follows:
 .sup.1 H NMR (CDCl.sub.3, 270 MHZ): .delta.=9.28 ppm (1H, s), 7.71 ppm (2H,
 d, J=8.91), 7.62 ppm (2H, d, J=8.91), 7.19 ppm (1H, m), 6.56 ppm (1H, ddd,
 J=8.23; 2.23; 0.74), 6.49 ppm (2H, m), 6.44 (1H, d, J=2.47), 5.99 ppm (1H,
 s), 3.79 ppm (3H, s), 3.27 ppm (4H, t, J=4.89), 3.20 ppm (2H, s), 2.77 ppm
 (4H, t, J=4.89), 1.47 ppm (9H, s).
 Bioassy Example 1
 Evaluation of Compounds in TNBS Model for IBD
 In this experiment, the ability of compounds of this invention to reduce
 colonic inflammation is demonstrated using the trinitrobenzene sulphonic
 acid ("TNBS") model for IBD. The TNBS model is one of the standard IBD
 models used in IBD discovery research and it has been extensively
 evaluated in rodents. See, for example, C. O. Elson et al. (1995),
 Experimental Models of Inflammatory Bowel Disease, Gastroenterology, 109:
 1344-1367 and references cited therein. In this model, a single enema of
 TNBS induces a prolonged colonic inflammatory response (up to several
 weeks) that is transmural and is accompanied by oxidative damage as
 evidenced by an increase in myeloperoxidase ("MPO") activity.
 Additionally, the inflammation is characterized by discrete areas of acute
 necrosis, inflammation and muscle thickening. Agents with
 anti-inflammatory effects in patients with IBD show efficacy in this
 model. Although the mechanism by which TNBS induces an inflammatory
 response is unknown, it is thought to have an immunological basis.
 Induction of Colitis
 Male Sprague-Dawley rats (200-250 g) were housed in standard cages (2 per
 cage) and fed rat chow and tap water ad libitum. After an overnight fast,
 rats were brought into the laboratory and randomized into treatment
 groups. Colitis was induced by intrarectal administration of 0.5 ml of
 TNBS solution (50 mg/kg in 50% ethanol) using a 1 mL syringe attached to a
 5 cm polyethylene catheter. Control animals received saline (0.9%) or a 1%
 methyl cellulose suspension at identical time points.
 Tissue Analysis
 Three days after TNBS administration, the rats were sacrificed and the
 colons excised and opened longitudinally. In 5 cm segments of colon, gross
 morphology was determined using the following scale:

Grade Finding
 0 No damage
 1 One area of Inflammation (red), no ulcers
 2 Ulcers, no area of inflammation
 3 Ulcers, one area of inflammation
 4 More than 2 ulcers, inflammation at one site
 5 More than 2 ulcers, inflammation &gt;1 cm
 The weights of each 5 cm colonic segment were also recorded to assess
 inflammatory induced edema.
 Dosing Regimen
 Each of the compounds from Examples 1-6 were tested in the TNBS model at 10
 mg/kg p.o. (oral) dosing. Each of the test compounds was administered by
 oral gavage as a 1% carboxy methyl cellulose ("CMC") suspension 1 hour
 prior to the administration of TNBS. Control rats were given CMC only.
 Results
 Each of the test compounds reduced TNBS-induced damage compared to the
 controls. The reduction in TNBS-induced damage ranged from about 25% to
 about 71% (average scores). Surprisingly, N-tert-butyl 4-hydroxybenzamide,
 a positional isomer of N-ten-butyl 2-hydroxybenzamide, was found to
 exacerbated the TNBS-induced damage compared to the controls.
 Bioassay Example 2
 Mouse Dextran Sulfate IBD Model
 Another model used for screening candidate IBD-treating compounds is the
 Dextran Sulfate ("DSS") model. Similar to the TNBS model, DSS induced
 colitis is widely used as a screening tool for IBD therapeutics. When
 administered orally, DSS induces IBD-like symptoms in Swiss-Webster mice.
 This model can be used to determine the effectiveness of compounds of this
 invention when such compounds are administered orally (p.o.).
 Individually housed 30-40 g male Swiss-Webster mice (B & K Universal,
 Fremont, Calif.) receive 3% DSS (Sigma Chemicals, St. Louis, Mo.) in their
 drinking water for 7 days. All animals receive food and water ad libitum.
 Two groups of mice are dosed orally with either the test compound in a
 dosing vehicle (1% methyl cellulose, dose range of 10 mg/kg to 30 mg/kg)
 or dosing vehicle alone (control).
 Clinical signs of colitis are assessed by a disease activity index ("DAI")
 consisting of changes in stool characteristics, fecal occult bleeding and
 body weight loss. The DAI is very similar to the Crohn's Disease Activity
 Index used in clinical trials to evaluate new agents to prevent/treat IBD.
 The DAI data are analyzed using Proc Anova in SAS with a Bonferoni
 post-hoc analysis, and Model 108 in WinNonlin.TM. (Professional Version
 1.5, Scientific Consulting, Apex, N.C.) for the ED.sub.50 and E.sub.max
 values. The wet weight and myeloperoxidase ("MPO") data (collected only on
 Day 7) are analyzed by Proc TTest in SAS. MPO is a marker for neutrophil
 infiltration. The following criteria are employed in this assay:
 DAI Scoring (Daily)
 Stool Characteristic: 0=normal, 2=loose and 4=diarrhea
 Fecal Occult Blood: 0=negative, 2=positive, 4=gross bleeding
 Weight Change: 0=0-1%, 1=1 to &lt;5%, 2=5% to &lt;10%, 3=10 to &lt;20%, 4=&gt;20%
 MPO (Day 7 Only)
 Two strips of colonic tissue/mouse
 MPO activity by spectrophotometric assay
 Bioassay Example 3
 Establishment of the Dose-response Characteristics in the Mouse Dextran
 Sulfate Model
 To determine the dose-response relationship of a test compound in the DSS
 Mouse Model, the following procedure is used.
 Experimental conditions and statistical analyses are the same as the Mouse
 Dextran Sulfate IBD Model, except four groups of mice (n=8-10/group) are
 used. Animals are dosed orally with either test compound (3, 10 or 30
 mg/kg) or vehicle alone. In addition, the following procedure is
 introduced to evaluate the histology in the animals:
 Histology Score
 5-6 slices/segment with 15-18 total pieces/colon
 Score for extent of damage: 0=1-25% involvement, 1=26-50%
 involvement, 2=51-75% involvement, 3=76-100% involvement
 Score for Grade:
 0=intact crypt, 1=loss of 1/3 crypt, 2=loss of 2/3 crypt, 3=loss of entire
 crypt with surface epithelium intact, 4=loss of entire crypt and erosion
 of surface epithelium
 Score for Severity:
 0=normal, 1=focal inflammatory cell infiltrate including PMNs,
 2=inflammatory cell infiltration, gland dropout and crypt abscess,
 3=mucosal ulceration
 Single, evaluator (qualified pathologist) blinded to the treatment
 conditions.
 Bioassay Example 4
 Effect of Test Compounds on Flux of Reactive Oxygen Species Induced by
 TNF-.alpha.
 Oxidative stress agents (OSA) are thought to be involved in cell death in
 IBD and are key initiator in the cascade of events leading to apoptosis.
 The purpose of this study is to evaluate the effect of a test compound on
 cytokine-induced OSA flux.
 To visualize OSA, the dye dihydrodichlorofluorescein diacetate is used.
 This non-fluorescent dye is taken up by cells and deacetylated to its
 non-fluorescent congener dihydrodichlorofluorescein (H.sub.2 DCF), which
 is trapped within cells. Reactive oxygen species ("ROS") react with
 H.sub.2 DCF, converting it to the highly fluorescent DCF. DCF fluorescence
 can be measured spectrofluorometrically and can also be visualized in
 intact cells using fluorescent microscopy.
 SK-N-MC cells (American Type Culture Collection, Rockville, Md.) are plated
 at 250,000 cells/well in 24-well Corning plates. Following plating, the
 cells are maintained in retinoic acid medium (5 .mu.M) for five days and
 then treated with a test compound at 100 .mu.M for 1 hour prior to
 TNF-.alpha. (3.0 ng/mL) treatment. TNF-.alpha. and H.sub.2 DCF are added
 simultaneously and cultures are incubated for an additional 4 hours.
 Following incubation, cultures are read in a cytofluorometer at 485-530 nm
 wavelength to detect increased DCF formation. Relative fluorescence units
 (RFU) values for the respective treatment conditions are compared. In this
 assay, higher fluorescence readings indicate ROS production. Thus,
 reductions in fluorescence indicates reduction in ROS production.
 Bioassay Example 5
 Effect of Compound A on TNF-.alpha. Induced Apoptosis in a Human Cell Model
 This test is used to evaluate the potential of a test compound to prevent
 TNF-.alpha. induced apoptosis.
 A test compound is evaluated in an in vitro model of TNF-.alpha. induced
 toxicity (see Pulliam et al. J. Neurosci. Res. 21:521-530 (1998)). In this
 model, human brain cell aggregates from fetal tissue are treated with
 TNF-.alpha. which caused an apoptotic cell death. Brain cell aggregates
 prepared from 1 brain were incubated for 10-12 days before
 experimentation. Aggregates are weighed out (100 mg/flask) and aliquoted
 into 10 mL flasks. TNF-.alpha. is used at a concentration of 1 ng. The
 test compound is added 1 hour prior to the TNF-.alpha.. Experiments
 include untreated brain aggregates, TNF-.alpha.-treated brain aggregates,
 TNF-.alpha.-+test compound treated aggregates and test compound treated
 aggregates. After TNF-.alpha. is added, aggregates are incubated for an
 additional 48 h. After this time, brain aggregates are centrifuged for 5
 min at 500 rpm. The supernatant is removed and the pellet is lysed for
 determination of programmed cell death (Boeringer Mannheim Cell Death Kit
 ELISA).
 Bioassay Example 6
 Effect of Test Compound on TNF-.alpha. Induced Reduction in bcl-2
 Cytokine-mediated apoptosis or programmed cell death is believed to be
 involved in a number of diseases including IBD. Reductions in bcl-2 are a
 major signal in initiation of the apoptotic cascade (see Jourd'heuil et
 al., J. Clin Gastroenterol. 25(Suppl):S61-S72 (1997)). The purpose of this
 study is to investigate the effects of a test compound on bcl-2 protein
 levels in a cellular model of cytokine mediated apoptosis.
 SK-N-MC cells (American Type Culture Collection, Rockville, Md.) are plated
 at 500,000 cells/plate and treated with retinoic acid ("RA") (5 .mu.M) for
 5 days. Following RA treatment, the cells are incubated with a test
 compound (100 .mu.M) for 1 hour. Cells are then treated with increasing
 concentrations of TNF-.alpha. (0, 0.3 and 3 ng/mL) for 6 h. The cells are
 harvested and lysed and bcl-2 is measured in the lysate using an ELISA
 assay (Boehringer Manheim). Quantification of bcl-2 is based on a standard
 curve and results are expressed as units/mL of bcl-2 in the sample.