Classifications

• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07D—HETEROCYCLIC COMPOUNDS
  • C07D401/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom
  • C07D401/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings
  • C07D401/12—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings linked by a chain containing hetero atoms as chain links
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
  • C07C257/00—Compounds containing carboxyl groups, the doubly-bound oxygen atom of a carboxyl group being replaced by a doubly-bound nitrogen atom, this nitrogen atom not being further bound to an oxygen atom, e.g. imino-ethers, amidines
  • C07C257/10—Compounds containing carboxyl groups, the doubly-bound oxygen atom of a carboxyl group being replaced by a doubly-bound nitrogen atom, this nitrogen atom not being further bound to an oxygen atom, e.g. imino-ethers, amidines with replacement of the other oxygen atom of the carboxyl group by nitrogen atoms, e.g. amidines
  • C07C257/18—Compounds containing carboxyl groups, the doubly-bound oxygen atom of a carboxyl group being replaced by a doubly-bound nitrogen atom, this nitrogen atom not being further bound to an oxygen atom, e.g. imino-ethers, amidines with replacement of the other oxygen atom of the carboxyl group by nitrogen atoms, e.g. amidines having carbon atoms of amidino groups bound to carbon atoms of six-membered aromatic rings
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07D—HETEROCYCLIC COMPOUNDS
  • C07D209/00—Heterocyclic compounds containing five-membered rings, condensed with other rings, with one nitrogen atom as the only ring hetero atom
  • C07D209/02—Heterocyclic compounds containing five-membered rings, condensed with other rings, with one nitrogen atom as the only ring hetero atom condensed with one carbocyclic ring
  • C07D209/04—Indoles; Hydrogenated indoles
  • C07D209/08—Indoles; Hydrogenated indoles with only hydrogen atoms or radicals containing only hydrogen and carbon atoms, directly attached to carbon atoms of the hetero ring
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07D—HETEROCYCLIC COMPOUNDS
  • C07D235/00—Heterocyclic compounds containing 1,3-diazole or hydrogenated 1,3-diazole rings, condensed with other rings
  • C07D235/02—Heterocyclic compounds containing 1,3-diazole or hydrogenated 1,3-diazole rings, condensed with other rings condensed with carbocyclic rings or ring systems
  • C07D235/04—Benzimidazoles; Hydrogenated benzimidazoles
  • C07D235/06—Benzimidazoles; Hydrogenated benzimidazoles with only hydrogen atoms, hydrocarbon or substituted hydrocarbon radicals, directly attached in position 2
  • C07D235/16—Radicals substituted by carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07D—HETEROCYCLIC COMPOUNDS
  • C07D235/00—Heterocyclic compounds containing 1,3-diazole or hydrogenated 1,3-diazole rings, condensed with other rings
  • C07D235/02—Heterocyclic compounds containing 1,3-diazole or hydrogenated 1,3-diazole rings, condensed with other rings condensed with carbocyclic rings or ring systems
  • C07D235/04—Benzimidazoles; Hydrogenated benzimidazoles
  • C07D235/20—Two benzimidazolyl-2 radicals linked together directly or via a hydrocarbon or substituted hydrocarbon radical
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07D—HETEROCYCLIC COMPOUNDS
  • C07D333/00—Heterocyclic compounds containing five-membered rings having one sulfur atom as the only ring hetero atom
  • C07D333/02—Heterocyclic compounds containing five-membered rings having one sulfur atom as the only ring hetero atom not condensed with other rings
  • C07D333/04—Heterocyclic compounds containing five-membered rings having one sulfur atom as the only ring hetero atom not condensed with other rings not substituted on the ring sulphur atom
  • C07D333/06—Heterocyclic compounds containing five-membered rings having one sulfur atom as the only ring hetero atom not condensed with other rings not substituted on the ring sulphur atom with only hydrogen atoms, hydrocarbon or substituted hydrocarbon radicals, directly attached to the ring carbon atoms
  • C07D333/08—Hydrogen atoms or radicals containing only hydrogen and carbon atoms
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07D—HETEROCYCLIC COMPOUNDS
  • C07D409/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having sulfur atoms as the only ring hetero atoms
  • C07D409/02—Heterocyclic compounds containing two or more hetero rings, at least one ring having sulfur atoms as the only ring hetero atoms containing two hetero rings
  • C07D409/12—Heterocyclic compounds containing two or more hetero rings, at least one ring having sulfur atoms as the only ring hetero atoms containing two hetero rings linked by a chain containing hetero atoms as chain links

Definitions

• This invention relates to anti -inflammatory and anti -allergy agents and, more particularly, relates to novel compounds, formulations and methods for the prophylaxis and treatment of inflammation, allergy and pulmonary disorders.
• the invention particularly relates to compositions and methods that are efficacious for the treatment of tryptase-related and mast cell mediated inflammatory disorders.
• the disorders noted above include, among others, asthma and other inflammatory diseases of the pulmonary system like allergic rhinitis, chronic obstructive pulmonary disease, respiratory syncytial virus and smoker's emphysema where the methods and compositions described herein are useful. Furthermore, the compositions and methods are particularly useful in treating the underlying pathological changes in the airways associated with these diseases such as basement membrane thickening, cell hypertrophy and hyperplasia, inflammatory cell influx, and other tissue remodeling. Other inflammatory conditions, including, for example, rheumatoid arthritis, multiple sclerosis, inflammatory bowel disease, conjunctivitis, psoriasis, scleroderma, and related diseases can be treated with the compounds and methods described herein.
• mast cell mediated disease particularly asthma
• Human asthma is a complex inflammatory disease. Genetic susceptibility and repeated allergen exposure from a variety of sources lead to allergen sensitization that, via IL-4 production from T-cells and mast cells, can ultimately induce B-cell derived IgE levels that are significantly elevated over normal levels. Subsequent exposure to allergen coupled with these newly elevated IgE levels can activate the Fc ⁇ RI high affinity IgE receptor on the surface of mast cells and other pro- inflammatory cells in the lung to induce degranulation/activation and thus trigger a cascade of inflammatory responses. This early phase of the response is characterized by severe bronchoconstriction that reaches its peak at about 15 minutes followed by a recovery of several hours.
• cytokines including, for example, IL-3, IL-4, IL-5, IL-6, and TNF-
• proteases including, for example, cathepsin G, chymase, carboxy peptidase A, tryptase
• tryptase is released in very large amounts - up to 35 pg per cell (see Caughey, Am- J. Phvsiol . ,
• tryptase is long lived, and has been shown to have a myriad of significant effects as a peptidase, protease and cytokine that intensify the inflammatory response.
• tryptase can cause further mast cell degranulation to amplify the allergen response (see Molinari et al., -J. APPI . Phvsiol. , 79 (6) . 1966-70 (1995) ) and induce eosinophil and neutrophil migration into the lung (see Walls et . al. , Int. Arch.
• tryptase can inactivate fibrinogen to act as a local anti -coagulant and promotes plasma extravasation bringing more circulating cells and mediators into the lung (see Schwartz et al, J. Immunol .. 135, 2762-7 (1985)). Further, tryptase can process high and low molecular weight kininogen to bradykinin and activates kallikrein to produce neurogenic inflammation (see Proud et aJL • , Biochem. Pharm.. 37 (8) . 1473-80 (1988); Walls et al., Biochem. Soc.
• Tryptase is a mitogen/activator of fibroblast (see Ruoss et a_l. , J. Clin. Invest.. 88, 493-9 (1991); Gruber et al., J. Immunology. 158. 2310-17 (1997)) and bronchial smooth muscle cells which can contribute to airway hyperresponsiveness to the lung as seen in a variety of pulmonary disorders (see Brown et al. , Chest. 107 (3) .
• tryptase is a mitogen for airway epithelial cells and induces IL-8 and ICAM-1 expression (see Cairns and Walls J. Immunology. 156. 275-83 (1996)) and recently tryptase has been shown to activate cellular receptors (see Molino et al. , J. Biol. Chem.. 272 (7) , 4043-49 (1997) ) .
• lipid mediators such as the leukotrienes (LTD4, LTC4 , LTE4 , LTB4) , the prostaglandins (PGD2) and platelet activating factor (PAF) .
• LTD4 leukotrienes
• PPD2 prostaglandins
• PAF platelet activating factor
• This late phase response is associated with a significant influx of inflammatory cells, most notably eosinophils, neutrophils, and lymphocytes, into the lung tissue and airway space. These cells are activated and release even more mediators which can contribute to the significant tissue damage and development of hyperresponsiveness seen in chronic asthma.
• inflammatory cells most notably eosinophils, neutrophils, and lymphocytes
• tryptase contributes to the early and late phase bronchoconstriction as well as to the development of airway hyperresponsiveness, a hallmark of asthma. Furthermore, in chronic asthma and other long term respiratory diseases, these activities cause profound changes to the airway such as desquamation of the epithelial lining, fibrosis and thickening of the underlying tissues. These changes are not treated by present therapeutics. Tryptase can be detected in a variety of biological fluids and recently tryptase 's relatively long biological half -life (vis a vis histamine) has become appreciated and clinicians now use circulating levels of tryptase as a marker of anaphylaxis (see Schwartz et al., N. En ⁇ l . J. Med., 316., 1622-26
• Tryptase can process prostromelysin to mature stromelysin (MMP-3) which can further activate collagenase (MMP-1) .
• MMP-3 mature stromelysin
• tryptase could play a significant role in the tissue remodeling of various pulmonary disorders (most notably asthma) but also in rheumatoid and osteo- arthritis.
• Tryptase is stored in the mature form as a homotetramer within the secretory granules of the mast cell and probably is held in an inactivated form by the low pH of this intracellular media. When released it is stabilized by interactions with heparin.
• This unique assembly of 4 catalytically active subunits could also be considered to be a dimer of dimers because computational models indicate that two adjacent active sites may face one another.
• the present invention provides novel compounds which inhibit tryptase activity. Also provided are formulations containing the novel compounds and methods of using the compounds to treat a patient in need thereof. More specifically, there are provided methods for the treatment of a patient suffering from a mast cell mediated disorder, including for example, asthma, allergic rhinitis, rheumatoid arthritis, dermatological diseases, multiple sclerosis, conjunctivitis, inflammatory bowel disease, anaphylaxis, osteoarthritis, peptic ulcers, cardiovascular disease, or other disease state in which mast cells and, in particular, tryptase activation is involved. In addition, there are described processes for preparing the inhibitory compounds of the invention.
• a mast cell mediated disorder including for example, asthma, allergic rhinitis, rheumatoid arthritis, dermatological diseases, multiple sclerosis, conjunctivitis, inflammatory bowel disease, anaphylaxis, osteoarthritis, peptic ulcers, cardiovascular disease, or other disease state in which
• the present invention relates to tryptase inhibitors, pharmaceutically acceptable salts and prodrugs thereof useful in the treatment or prophylaxis of inflammatory diseases, particularly asthma and other related inflammatory diseases.
• the invention also encompasses pharmaceutical compositions and methods for prophylaxis and treatment of asthma, pulmonary disorders and related inflammatory, mast-cell mediated diseases, particularly those which involve activation of tryptase. Also provided are processes for making such compounds as well as intermediates useful in such processes.
• the present invention provides compounds useful for the treatment or prophylaxis of inflammatory diseases.
• a compound of Formula (I) is particularly useful for the treatment or prophylaxis of inflammatory diseases.
• a compound of Formula (I) is particularly useful for the treatment or prophylaxis of inflammatory diseases.
• a compound of Formula (I) is particularly useful for the treatment or prophylaxis of inflammatory diseases.
• a compound of Formula (I) is a compound of Formula (I) :
• X is -C(O)-, -(CH 2 ) n - or -SO.-;
• R is -H, straight or branched chain (C.-C 6 ) -alkyl , - (C.-C ⁇ ) -cycloalkyi or - (CH 2 ) m -Ar' - (Y) t ;
• Ar or Ar' is aryl, heteroaryl, or a 5-membered to 7-membered carbocyclic or heterocyclic ring;
• alkyl refers to a univalent saturated, straight- or branched- chain alkyl group containing the designated number of carbon atoms.
• C. -C 6 alkyl refers to a univalent saturated, straight- or branched- chain alkyl group which can contain from one to six carbon atoms, for example, methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, t-butyl, n-pentyl, 2 -methylbutyl , 3 -methyl - butyl, n-hexyl, 2 -methylpentyl , 3 -methylpentyl , 4 -methylpentyl , 2,2- dimethylbutyl , 2,3- dimethylbutyl , and the like.
• alkoxy refers to an alkyl group bonded through an oxygen atom to another substituent.
• C.-C. alkoxy refers to a C--C 4 alkyl group bonded through an oxygen atom to another substituent and includes, for example, methoxy, ethoxy, n-propoxy, n-butoxy, t-butoxy and isobutoxy.
• Carbocyclic refers to an organic cyclic moiety in which the cyclic skeleton is comprised of only carbon atoms whereas the term “heterocyclic” refers to an organic cyclic moiety in which the cyclic skeleton contains one or more heteroatoms selected from nitrogen, oxygen, or sulfur and which may or may not include carbon atoms.
• cycloalkyi refers to a carbocyclic moiety containing the indicated number of carbon atoms.
• C 3 -C 8 cycloalkyi therefore, refers to an organic cyclic substituent in which three to eight carbon atoms form a three, four, five, or six, seven, or eight -membered ring, including preferably, for example, a cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl ring .
• Aryl refers to an aromatic carbocyclic group having a single ring, for example, a phenyl ring, multiple rings, for example, biphenyl , or multiple condensed rings in which at least one ring is aromatic, for example, naphthyl , 1,2,3,4,- tetrahydronaphthyl , anthryl , or phenanthryl , which can be unsubstituted or substituted with one or more substituents selected from halogen, lower (C.-C 4 ) alkyl, lower (C.-C alkoxy, lower (C.-C alkylthio, trifluoromethyl, lower (C.-C acyloxy, aryl, heteroaryl and hydroxy.
• substituents selected from halogen, lower (C.-C 4 ) alkyl, lower (C.-C alkoxy, lower (C.-C alkylthio, trifluoromethyl, lower (C.-C acyloxy, ary
• the substituents attached to a phenyl ring portion of an aryl moiety may be configured in the ortho-, meta- or para- orientations, with the meta- and para- orientations being preferred.
• Heterocycle or “heterocyclic” refers to a saturated, unsaturated or aromatic carbocyclic group having a single ring, multiple rings or multiple condensed rings, and having at least one hetero atom such as nitrogen, oxygen or sulfur within at least one of the rings.
• Heteroaryl refers to a heterocycle in which at least one ring is aromatic. Any of the heterocyclic or heteroaryl groups can be unsubstituted or optionally substituted with one or more groups selected from halogen, lower (C.-C alkyl, lower (C.-C 4 ) alkoxy, lower (C.-C alkythio, trifluoromethyl, lower (C.-C.) acyloxy, and hydroxy.
• heterocyclic moieties may exist in several isomeric forms, all of which are to be encompassed by the present invention.
• a 1, 3 , 5- triazine moiety is isomeric to a 1, 2 , 4- triazine group.
• Such positional isomers are to be considered within the scope of the present invention.
• the heterocyclic or heteroaryl groups can be bonded to other moieties in the compounds of the invention. The point (s) of attachment to these other moieties is not to be construed as limiting on the scope of the invention.
• a pyridyl moiety may be bound to other groups through the 2-, 3-, or 4 - position of the pyridyl group. All such configurations are to be construed as within the scope of the present invention.
• heterocyclic or heteroaryl moieties included in the scope of the present invention may include, but are not limited to, the following:
• halo refers to a halogen atom which may include fluoro, chloro, bromo and iodo.
• Preferred halo groups include chloro, bromo and fluoro with chloro and fluoro being especially preferred.
• “Pharmaceutically acceptable salt” refers to an organic or inorganic salt which is useful in the treatment of a warm-blooded animal. Such salts can be acid or basic addition salts, depending on the nature of the compound of Formula (I) .
• “warm blooded animal” includes a mammal, including a member of the human, equine, porcine, bovine, murine, canine or feline species.
• a salt may be formed by treatment of a compound of Formula (I) with a basic compound, particularly an inorganic base.
• Preferred inorganic salts are those formed with alkali and alkaline earth metals such as lithium, sodium, potassium, barium and calcium.
• Preferred organic base salts include, for example, ammonium, dibenzylammonium, benzylammonium, 2- hydroxyethylammonium, bis (2 -hydroxyethyl) ammonium, phenylethylbenzylamine, dibenzyl - ethylenediamine, and the like salts.
• salts of acidic moieties may include, for example, those salts formed with procaine, quinine and N-methylglusoamine, plus salts formed with basic amino acids such as glycine, ornithine, histidine, phenylglycine, lysine and arginine.
• An especially preferred salt is a sodium or potassium salt of a compound of Formula (I) .
• a salt is formed by the treatment of a compound of Formula (I) with an acidic compound, particularly an inorganic acid.
• Preferred inorganic salts of this type may include, for example, the hydrochloric, hydrobromic, hydroiodic, sulf ric, phosphoric or the like salts.
• Preferred organic salts of this type may include, for example, salts formed with formic, acetic, succinic, citric, lactic, maleic, fumaric, palmitic, cholic, pamoic, mucic, d-glutamic, d-camphoric, glutaric, glycolic, phthalic, tartaric, lauric, stearic, salicyclic, methanesulfonic, benzenesulfonic, para- toluenesulfonic, sorbic, puric, benzoic, cinnamic and the like organic acids.
• An especially preferred salt of this type is a hydrochloride or sulfate salt of a compound of Formula (I) .
• esters of a carboxylic acid or hydroxyl containing group including a metabolically labile ester or a prodrug form of a compound of Formula (I) .
• a metabolically labile ester is one which may produce, for example, an increase in blood levels and prolong the efficacy of the corresponding non- esterified form of the compound.
• a prodrug form is one which is not in an active form of the molecule as administered but which becomes therapeutically active after some in vivo activity or biotransformation, such as metabolism, for example, enzymatic or hydrolytic cleavage.
• Esters of a compound of Formula (I) may include, for example, the methyl, ethyl, propyl, and butyl esters, as well as other suitable esters formed between an acidic moiety and a hydroxyl containing moiety.
• Metabolically labile esters may include, for example, methoxymethyl , ethoxymethyl, iso-propoxymethyl, ⁇ -methoxyethyl, groups such as ⁇ - ( (C.-C 4 ) alkyloxy) ethyl; for example, methoxyethyl , ethoxyethyl , propoxyethyl , iso- propoxyethyl , etc.; 2 -oxo- 1, 3 -dioxolen-4 -ylmethyl groups, such as 5-methyl-2 -oxo- 1, 3 , dioxolen-4 -ylmethyl , etc.; C.-C 3 alkylthiomethyl groups, for example,
• stereoisomer or “stereoisomeric” refers to a compound which has the same molecular weight, chemical composition, and constitution as another, but with the atoms grouped such that their orientation in three- dimensional space is different.
• stereoisomers may exist as enantiomeric mixtures, diastereomers or may be resolved into individual stereoisomeric components (e.g. specific enantiomers) by methods familiar to one skilled in the art.
• the compounds of the invention may exist as crystalline solids which can be crystallized from common solvents such as ethanol, N, N- dimethyl - formamide, water, or the like.
• crystalline forms of the compounds of the invention may exist as solvates and/or hydrates of the parent compounds or their pharmaceutically acceptable salts. All of such forms likewise are to be construed as falling within the scope of the invention.
• the compounds of the invention are useful for the therapeutic or prophylactic treatment of an inflammatory disease state in warm-blooded animals.
• the compounds of the invention may be used as anti- inflammatory agents in an inflammatory disease, especially a mast-cell mediated disease, for example, asthma, allergy or pulmonary disorders.
• a formulation comprising a compound of Formula (I) in combination, admixture, or associated with a pharmaceutically acceptable carrier, diluent or excipient therefor.
• composition used in the noted therapeutic methods can be in a variety of forms. These include, for example, solid, semi -solid and liquid dosage forms, such as tablets, pills, powders, liquid solutions or suspensions, liposomes, injectable and infusible solutions.
• solid, semi -solid and liquid dosage forms such as tablets, pills, powders, liquid solutions or suspensions, liposomes, injectable and infusible solutions.
• the preferred form depends on the intended mode of administration and therapeutic application. Considerations for preparing appropriate formulations will be familiar to one skilled in the art and are described, for example, in Goodman and Gilman's: "The Pharmacological Basis of Therapeutics", 8th Ed., Pergamon Press, Gilman et. al. eds. (1990); and
• Typical carriers, diluents, and excipients may include water (for example, water for injection) , buffers, lactose, starch, sucrose, and the like.
• a compound of the invention can be administered orally, topically or parenterally (e.g.
• Such forms of the compounds of the invention may be administered by conventional means for creating aerosols or administering dry powder medications using devices such as for example, metered dose inhalers, nasal sprayers, dry powder inhaler, jet nebulizers, or ultrasonic nebulizers.
• devices such as for example, metered dose inhalers, nasal sprayers, dry powder inhaler, jet nebulizers, or ultrasonic nebulizers.
• Such devices optionally may be include a mouthpiece fitted around an orifice.
• aerosol typically includes any gas -borne suspended phase of a compound of the invention which is capable of being inhaled into the bronchioles or nasal passages.
• aerosol includes a gas -borne suspension of droplets of the desired compound, as may be produced in a metered dose inhaler or nebulizer, or in a mist sprayer.
• Aerosol also includes a dry powder composition of a compound of the instant invention suspended in air or other carrier gas, which may be delivered by insufflation from an inhaler device, for example.
• the preferred range of concentration of the compounds of the invention is 0.1-100 milligrams (mg) /milliliter (mL) , more preferably 0.1-30 mg/mL, and most preferably 1-10 mg/mL.
• a physiologically compatible buffer such as phosphate or bicarbonate.
• the usual pH range is from about 5 to about 9, preferably from about 6.5 to about 7.8, and more preferably from about 7.0 to about 7.6.
• sodium chloride is added to adjust the osmolarity to the physiological range, preferably within 10% of isotonic.
• such methods comprise pressurizing or providing a means of pressurizing a container of the solution, usually with an inert carrier gas, and passing the pressurized gas through a small orifice, thereby pulling 'roplets of the solution into the mouth and ⁇ .rachea of. the animal to which the drug is to be administered.
• a mouthpiece is fitted to the outlet of the orifice 'to facilitate delivery into the mouth and trachea.
• revices of the present invention comprise solutions of the compounds of the invention connected to or contained within any of the conventional means for creating aerosols in asthma medication, such as metered dose inhalers, jet nebulizers, or ultrasonic nebulizers.
• such devices may include a mouthpiece fitted around the orifice.
• a device may comprise a solution of a compound of the instant invention in a nasal sprayer.
• a dry powdei comprising a compound of the invention, optionally with an excipient is another embodiment. This may be administered by a drug powder inhaler containing the described powder.
• ⁇ -Adrenergic agonists are especially useful in these combinations, because they provide symptomatic relief of the initial asthmatic response, whereas the compounds of the present invention may provide relief and be better suited to treating the late asthmatic response.
• Preferred ⁇ -adrenergic agonists in these solutions include any of the usual ⁇ -agonists employed for the relief of asthma, for example, albuterol, terbutaline, bitolterol mesylate, or the like.
• agents useful in combination with the compounds of the invention include anticholinergics, such as ipratropium bromide, and antiinflammatory corticosteroids (adrenocortical steroids) such as beclomethasone, triamcinolone, flurisolide, or dexamethasone.
• a compound of the invention may be used in the treatment of immunomediated inflammatory skin conditions, such as urticaria and angioedema, eczematous dermatitis, and hyperproliferative skin disease, for example, psoriasis. In such cases, a compound of the invention could be administered topically so as treat the condition involved.
• a topical preparation comprising a compound of the invention
• the dosage of medicament and the length of time required for treating each patient may vary, but one skilled in the art will recognize that variations may occur from patient to patient and adjust the treatment regimen accordingly.
• a pharmaceutical preparation for topical application comprising a compound of the invention, typically in concentrations in the range of from about 0.001% to about 10%, in combination with a pharmaceutically acceptable carrier, excipient, or diluent therefor.
• Such topical preparations can be prepared by combining the compound of the invention with conventional pharmaceutical diluents and carriers commonly used in topical dry, liquid, cream and aerosol formulations.
• Ointment and creams may be formulated, for example, with an aqueous or oily base with the addition of suitable thickening and/or gelling agents.
• bases may include water and/or an oil such as a liquid paraffin or a vegetable oil such as peanut oil or castor oil.
• Thickening agents which may be used according to the characteristics of the base may include, for example, soft paraffin, aluminum stearate, cetostearyl alcohol, propylene glycol, polyethylene glycols, woolfat, hydrogenated lanolin, beeswax, and the like.
• Lotions may be formulated with an aqueous or oily base and will include also, in general, one or more of the following: stabilizing agents emulsifying agents, dispersing agents, suspending agents, thickening agents, coloring agents, perf mes, and the like.
• Powders may be formed with the aid of any suitable powder bases, for example, talc, lactose, starch and the like. Drops may be formulated with an aqueous base or non- aqueous base also comprising one or more dispersing agents, suspending agents solubilizing agents, and the like.
• any of the formulations of the invention may also include one or more preservatives or bacteriostatic agents, for example, methyl hydroxybenzoate, ethyl hydroxybenzoate, propyl hydroxybenzoate, chlorocresol , benzalkonium chlorides, and the like. Additionally, the formulations may contain other active ingredients such as antimicrobial agents, particularly antibiotics, anesthetics, analgesics and antipruritic agents.
• the pharmaceutical formulations of the invention may be administered by parenteral or oral administration for prophylactic and/or therapeutic treatment.
• the pharmaceutical compositions can be administered in a variety of unit dosage forms depending on the method of administration.
• unit dosage forms suitable for oral administration may include, powders, tablets, pills, capsules and dragees.
• the pharmaceutical formulations can be administered intravenously. Therefore, the invention further provides formulations for intravenous administration which comprise a compound of the invention dissolved or suspended in a pharmaceutically acceptable carrier or diluent therefor.
• aqueous carriers can be used, for example, water, buffered water or other buffer solutions, saline, and the like.
• the resulting aqueous solutions can be packaged for use as is, or lyophilized, the lyophilized preparation being combined with a sterile aqueous solution prior to administration.
• the sterile aqueous solution for the lyophilized product can be packaged as a kit for use with the lyophilized formulation.
• the compositions can contain pharmaceutically acceptable substances to aid in administration and more closely mimic physiological conditions. Such substances, can include, for example, pH adjusting substances such as acids, bases or buffering agents, tonicity adjusting agents, wetting agents and the like.
• Such substances may include but are not limited to, for example, sodium hydroxide, hydrochloric acid, sulfuric acid, sodium acetate, sodium lactate, sodium chloride, potassium chloride, calcium chloride, sorbitan monolaurate, triethanolamine oleate, and the like or any other means familiar to one skilled in the art for maintaining pH at a desired level.
• carriers, diluents, and excipients known to one skilled in the art may be used.
• Such carriers, diluents and excipients may include, for example, mannitol, lactose, starch magnesium stearate, sodium saccharin, talcum, cellulose, glucose, sucrose, or other solid polyol sugar, magnesium carbonate, and the like.
• a pharmaceutically acceptable formulation is prepared by admixing any of the usual carrier, diluents, and excipients, such as those noted, with from about 0.1 to about 95% of a compound of the invention.
• the compounds of Formula (I) may be prepared by a variety of methods familiar to one skilled in the art. For example, to prepare a compound of the formula:
• the product of the reaction may be salified to prepare a pharmaceutically acceptable salt of the invention.
• the product of the reaction may be esterified to prepare a pharmaceutically acceptable ester of the invention as previously defined.
• the reactions used to prepare the compounds of Formula (I) may be carried out in any number of solvents in which the reactants may be mutually soluble, including, for example, tetrahydrofuran, benzene, toluene, chloroform, dichloromethane, N, N- dimethylformamide, ethyl ether, dioxane, acetonitrile, or the like.
• the reaction is carried out at a temperature of between -80° and 150°C, preferably, however, at room tempature.
• the product and intermediates may be isolated or purified using one or more standard purification techniques, including, for example, one or more of simple solvent evaporation, recrystallization, distillation, sublimation, filtration, chromatography, including thin- layer chromatography, HPLC (e.g. reverse phase HPLC using, for example, dilute trifluoroacetic acid in water, acetonitrile, or methanol mixtures as eluent) , column chromatography, flash chromatography, radial chromatography, trituration, and the like.
• HPLC e.g. reverse phase HPLC using, for example, dilute trifluoroacetic acid in water, acetonitrile, or methanol mixtures as eluent
• Protease inhibition was assayed according to published procedures with minor modifications using various proteases and specific chromogenic peptide p-nitroanilide substrates. Assays were performed in Costar ultra- low cluster 96 -well microtiter plate (Costar Corning Corp., Cambridge MA) . Each protease was incubated with various concentrations of the test compound for 15 min. at 37 °C or as otherwise indicated, in specific assay buffer, and the residual activity was then measured by addition of the substrate. p-Nitroaniline produced by the proteolysis was determined by measuring the change in absorbance at 405 nm on a SpectraMAX 340 plate reader (Molecular devices, Sunnyvale, CA) .
• K. The inhibition constant, K. is calculated from individual data points using the equation for a tight- binding inhibitor (See Beith, "Proteinase Inhibitors - Proceedings of 2nd Int. Res. Conference", Fritz, et. al . eds., New York, p.4463-4469 (1974)):
• K.' is apparent inhibition constant
• v, and v o are the inhibited and uninhibited rates, respectively
• [I] o and [E] o are the total concentrations of inhibitor and enzyme, respectively.
• IC 50 is determined by fitting the individual inhibition data point to Sigmoid or four-parameter curve- fit equations.
• GPR-pNA Tosyl-Gly-Pro-Arg-p- nitroanilide
• the reaction was performed in 50 mM Tris -HCI, pH 8.0, containing 150 mM NaCl and 0.02% Triton X-100 at 37°C in Costar ultra- low cluster 96 -well microtiter plates (Costar Corning Corp., Cambridge, MA).
• the amount of pNA produced by tryptase was determined by measuring the change in absorbance at 405 nm on a SpectraMAX 340 plate reader (Molecular devices, Sunnyvale, CA) .
• the Km for the substrate was determined by-* Lineweaver-Burk analysis from initial velocities of substrate hydrolysis.
• the inhibition assay was carried out in a total volume of 200 ⁇ L. Tryptase (30 ⁇ L- final concentration 1 nM) was incubated with various concentrations of sample compound (50 ⁇ L) to be tested in the above assay buffer for 5 min. The reaction was started by the addition of substrate GPR-pNA (40 ⁇ L- final concentration 320 ⁇ M) , and the residual activity was measured after 15 min. of incubation.
• the inhibition constant, K ⁇ was determined by fitting the inhibition data to a two- site competitive binding equation using data analysis program GraphPad PRISM (GraphPad Software, Inc., San Diego, CA) .
• Human Neutrophil elastase activity was determined by using pyroGlu-Pro-Val-pNA in 100 mM Tris -HCI, pH
• Bovine pancreatic Trypsin Bovine pancreatic Trypsin (TPCK- treated) activity was determined by using N- ⁇ -Benzoyl -L-Arg-pNA in 50 mM Tris -HCI, pH 8.2, 20 mM CaCl 2 (See Somorin, et al. , J. Biochem. 85, 157-162 (1979)).
• Bovine Pancreatic Chymotrypsin activity was determined by using N-Suc-Ala-Ala-Pro-Phe-pNA in 100 mM Tris -HCI, pH 7.8, 10 mM CaCl 2 (See Delmar, et al. , J. Biochem. j35_, 157-162 (1979)).
• Human plasma factor Xa activity was determined by using N-Benzoyl - Ile-Glu-Gly-Arg-pNA in 50 mM Tris-HCl, pH 7.8, 200 mM NaCl, 0.05% BSA (See Lottenberg, et al . Meth. Enzvmol. __0_, 341-361 (1981)).
• Human plasma thrombin activity was determined by using H-D-Phe-Pip-Arg-pNA in 50 mM Tris-HCl, pH 8.3, 100 mM NaCl, 1% BSA (See Lottenberg, et al. , Meth. Enzvmol . 80, 341-361 (1981)).
• Human plasma and r- tissue kallikrein activity were determined in 50 mM Tris-HCl, pH 7.8 , 200 mM NaCl, 0.05% BSA by using H-D-Prolyl -Phe-Arg-pNA and DL-Val- Leu-Arg-pNA, respectively (See Lottenberg, e al . , Meth. Enzvmol . M, 341-361 (1981)).
• the inhibition constant K of the test compounds against each proteolytic enzyme was determined according to Zitnik et. al. , Biochem. Biophvs . Res . Commun. 232. 687-697 (1997)). The results are provided below.

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