Arylbicyclooctanes as inhibitors of leukotriene biosynthesis

BACKGROUND OF THE INVENTION
The leukotrienes constitute a group of locally acting hormones, produced in living systems from arachidonic acid. The major leukotrienes are Leukotfiene B.sub.4 (abbreviated at LTB.sub.4), LTC.sub.4, LTD.sub.4, and LTE.sub.4. The biosynthesis of these leukotrienes begins with the action of the enzyme 5-lipoxygenase on arachidonic acid to produce the epoxide known as Leukotfiene A.sub.4 (LTA.sub.4), which is converted to the other leukotrienes by subsequent enzymatic steps. Further details of the biosynthesis as well as the metabolism of the leukotfienes are to be found in the book Leukotrienes and Lipoxygenases, ed. J. Rokach, Elsevier, Amsterdam (1989). The actions of the leukotrienes in living systems and their contribution to various diseases states are also discussed in the book by Rokach.
EP 385,662 describes a series of heterocycles differing notably from the present invention in not containing the bicyclic structure characterized by the oxygen and X.sup.1 bridges. EP 462,812 and 462,813 contain 5-membered heterocyclenes (Ar.sup.2) and a substituted phenylene (Ar), respectively. EP 462,830 and 462,831 lack and contain an oxygen attached to R.sup.1, respectively. However, none of the above compounds contains the bicyclic bridged structure of the present invention, characterized by the oxygen and X.sup.1 bridges.
Hence the compounds of the present invention are completely novel and unexpectedly have biological activity as leukotriene biosynthesis inhibitors. ##STR2##
SUMMARY OF THE INVENTION
The present invention relates to arylbicyclooctanes having activity as leukotriene biosynthesis inhibitors, to methods for their preparation, and to methods and pharmaceutical formulations for using these compounds in mammals (especially humans).
Because of their activity as leukotriene biosynthesis inhibitors, the compounds of the present invention are useful as anti-asthmatic, anti-allergic, anti-inflammatory, and cytoprotective agents. They are also useful in treating angina, cerebral spasm, glomerular nephritis, hepatitis, endotoxemia, uveitis and allograft rejection and in preventing the formation of atherosclerotic plaques.

DETAILED DESCRIPTION OF THE INVENTION
The compounds of the present invention may be represented by the following formula I: ##STR3## wherein:
Ar.sup.1 is arylene-(R.sup.2).sub.2, wherein arylene is a 5-membered aromatic ring containing one O or S and in which 0-2 carbon atoms are replaced by N; a 5-membered aromatic ring containing 1-3 nitrogen atoms; a 6-membered aromatic ring wherein 0-3 carbon atoms are replaced by N; 2- or 4-pyranone; or 2- or 4-pyridinone;
Ar.sup.2 is aryl-(R.sup.3).sub.2, wherein aryl is a 9 or 10-membered bicyclic heterocyclic moiety containing 1 or 2 N's and optionally a further N, O or S heteroatom;
Ar.sup.3 is aryl-(R.sup.4).sub.2 wherein aryl is a 5-membered aromatic ring containing one O, S, or N and in which 0-3 carbon atoms are replaced by N; a dihydro derivative of the aforesaid 5-membered aromatic ring; a 6-membered aromatic ring wherein 0-3 carbon atoms are replaced by N; 2- or 4-pyranone; 2- or 4-pyridinone; or a bicyclic 8-, 9-, or 10-membered aromatic ring wherein 0-2 carbon atoms are replaced by either O or S or a combination thereof and 0-3 carbon atoms are replaced by N;
A is a direct link to X.sup.3 or [C(R.sup.5).sub.2 ].sub.n ;
X.sup.1 is --OCH.sub.2 --, --CH.sub.2 O--, --CH.sub.2 CH.sub.2 -- or --CH=CH--;
X.sup.2 is O or S or a bond;
X.sup.3 is O, S, S(O), or S(O).sub.2 ;
R.sup.1 is H, lower alkyl, or lower alkylcarbonyl;
R.sup.2 is H, lower alkyl, lower alkoxy, lower alkylthio, CN, CF.sub.3, or halogen;
R.sup.3 is R.sup.2, oxo, thioxo, hydroxy lower alkyl, lower alkoxy lower alkyl, lower alkylthio lower alkyl, (R.sup.4).sub.2 -phenyl lower alkyl, (R.sup.4).sub.2 -phenylthio lower alkyl, lower alkylthio lower alkylcarbonyl, NO.sub.2, N.sub.3, N(R.sup.6).sub.2, NR.sup.6 COR.sup.7, NR.sup.6 CON(R.sup.7).sub.2, SR.sup.8, S(O)R.sup.8, S(O).sub.2 R.sup.8, S(O).sub.2 N(R.sup.6).sub.2, COR.sup.7, CON(R.sup.6).sub.2, CO.sub.2 R.sup.7, C(R.sup.7).sub.2 OC(CR.sup.7).sub.2 --CO.sub.2 R.sup.7, C(R.sup.7).sub.2 CN, Ar.sup.3, or COAr.sup.3 ;
R.sup.4 is H, lower alkyl, lower alkoxy, lower alkylthio, halogen, CF.sub.3 or CN;
R.sup.5 is H or lower alkyl, or two R.sup.5 groups attached to the same carbon can form a ring of 3 to 8 members;
R.sup.6 is H or lower alkyl, or two R.sup.6 groups attached to the same nitrogen may form a saturated ring of 5 or 6 members, optionally containing a second heteroatom chosen from O, S or NR.sup.5 ;
R.sup.7 is H or lower alkyl;
R.sup.8 is lower alkyl, CF.sub.3, or phenyl-(R.sup.4).sub.2 ;
n is 1 to 3;
or a pharmaceutically acceptable salt thereof.
A preferred embodiment of the present invention is represented by Formula Ia: ##STR4## wherein:
X.sup.1 is --OCH.sub.2 --or --CH.sub.2 O--;
R.sup.1 is H or lower alkyl;
R.sup.2 is H or F; and
R.sup.3 is H or lower alkyl, or one R.sup.3 is lower alkyl attached to position 1 and the other R.sup.3 is oxo or thioxo attached to position 2.
Another preferred embodiment of the present invention is represented by Formula Ib: ##STR5## wherein:
X.sup.1 is --OCH.sub.2 -- or --CH.sub.2 O--;
R.sup.1 is H or lower alkyl;
R.sup.2 is H, F, CF.sub.3 or CN; and
Ar.sup.3 is furan or thiophene.
Definitions
The following abbreviations have the indicated meanings:
______________________________________Ac = acetylBn = benzyli-Pr = isopropyln-Pr = normal propyln-Bu = normal butyli-Bu = isobutyls-Bu = secondary butylt-Bu = tertiary butylEt = ethylFu = 2- or 3- furylMe = methylPh = phenylPy = 2-, 3-, or 4- pyridylTh = 2- or 3- thienylTz = 2- or 5- thiazolylTf = trifluoromethanesulfonylAIBN = azoisobutylronitrileCH.sub.2 N.sub.2 = diazomethaneCSA = camphor sulfonic acidDCC = 1,3-dicyclohexylcarbodiimideDDQ = 2,3-dichloro-5,6-dicyano-1,4-benzoquinoneDHP = 3,4-dihydro-2H-pyranDIBAL-H = diisobutylaluminum hydrideDIPHOS = 1,2-bis(diphenylphosphino)ethaneDMAP = 4-dimethylaminopyridineDMF = N,N-dimethylformamideDMSO = dimethylsulfoxideEt.sub.3 N = triethylamineLDA = lithium diisopropylamideMs = methanesulfonyl = mesylPhe = benzenediylPye = pyridindiylPCC = pyridinium chlorochromateRIA = radioimmuno assayr.t. = room temperatureSuper-Hydride = lithium triethylborohydridet-BOC = tertiary butyloxy carbonylTFA = trifluoroacetic acidTFAA = trifluoroacetic anhydrideTHF = tetrahydrofuranTMSCl = chlorotrimethylsilaneTMSCN = trimethylsilylcyanideTs = p-toluenesulfonyl = tosylTsA = trifluoromethanesulfonic anhydrideTze = thiazoldiyl______________________________________
Alkyl is intended to include linear and branched structures and combinations thereof.
The term "alkyl" includes "lower alkyl" and extends to cover carbon fragments having up to 20 carbon atoms. Examples of alkyl groups include octyl, nonyl, undecyl, dodecyl, tridecyl, tetradecyl, pentadecyl, eicosyl, 3,7-diethyl-2,2-dimethyl-4-propylnonyl, and the like.
The term "lower alkyl" includes those alkyl groups of from 1 to 7 carbon atoms. Examples of lower alkyl groups include methyl, ethyl, propyl, isopropyl, butyl, s- and t-butyl, pentyl, hexyl, heptyl, and the like.
The term "lower alkoxy" includes those alkoxy groups of from 1 to 7 carbon atoms of a straight, branched, or cyclic configuration. Examples of lower alkoxy groups include methoxy, ethoxy, propoxy, isopropoxy, cyclopropyloxy, cyclohexyloxy, and the like.
"Lower alkylcarbonyl" means alkylcarbonyl groups of from 1 to 8 carbon atoms of a straight, branched, or cyclic configuration. Examples of lower alkylcarbonyl groups are formyl, 2-methylbutanoyl, cyclohexylacetyl, etc. By way of illustration, the 2-methylbutanoyl group signifies --C(O)CH(CH.sub.3)CH.sub.2 CH.sub.3.
Examples of saturated tings which may be formed by two R.sup.5 groups attached to the same nitrogen are pyrrolidine, piperidine, morpholine, thiamorpholine, piperazine, or N-lower alkyl piperazine.
Halogen includes F, Cl, Br, and I.
It is intended that the definitions of any substituent (e.g., R.sup.4, R.sup.8, etc.) in a particular molecule be independent of its definitions elsewhere in the molecule.
The aromatic rings herein are comprised of carbon atoms and, where applicable, specifically named heteroatoms. Thus, "a 5-membered aromatic ting containing one 0" means furan.
Examples of Ar.sup.1 are furan, thiophene, oxazole, thiazole, 1,3,4-oxadiazole, 1,3,4-thiadiazole, 1,2,5-oxadiazole, 1,2,5-thiadiazole, pyrrole, imidazole, 1,3,4-triazole, benzene, pyridine, pyrazine, pyrimidine, pyridazine, 1,2,3-triazine, 1,2,4-triazine, and 1,3,5-triazine.
Examples of Ar.sup.2 are a 9- or 10-membered benzo-fused heterocyclic moiety such as indolyl, isoindolyl, benzimidazolyl, 1H-indazolyl, benzoxazolyl, benzothiazolyl, quinolyl, isoquinolyl, cinnolinyl, quinazolinyl, quinoxalinyl, 4H-1,4-benzoxazinyl, or 4H-1,4-benzothiazinyl, or a hydrogenated derivative thereof such as indolinyl, 2,3-dihydrobenzimidazolyl, 2,3-dihydrobenzoxazolyl, 2,3-dihydrobenzothiazolyl, 1,2-dihydroquinolyl, 1,2,3,4-tetrahydroquinolyl, 1,2-dihydroisoquinolyl, 2,3-dihydro-4H-1,4-benzoxazinyl or 2,3-dihydro-4H-1,4-benzothiazinyl, 1,2-dihydro-2-oxoquinolyl, 1,2-dihydro-2-thioxoquinolyl, or 2,3-dihydro-2-oxoindolyl, additional examples of Ar.sup.2 are a 9- or 10-membered pyrido-fused heterocyclic moiety such as 1H-pyrrolo[2,3-b]pyridyl, imidazo[[4,5-b]pyridyl, 1,7-naphthyridinyl, 1,8-naphthyridinyl, pyrido[2,3-d]pyrimidinyl, pyrido[2,3-b]pyrazinyl, 4H-pyrido[3,2-b][1,4]oxazinyl and 4H-pyrido[3,2-b][1,4]thiazinyl, or a hydrogenated derivative thereof.
The heterocyclic moiety may be attached through any available position including from either of the two rings of the bicyclic heterocyclic moiety and including through an available nitrogen atom. The heterocyclic moiety may bear a suitable substituent such as, for example, a (C.sub.1-4)alkyl, phenyl, benzoyl, or phenyl-(C.sub.1-4)alkyl substituent on an available nitrogen atom.
Examples of Ar.sup.3 are furan, thiophene, oxazole, thiazole, isoxazole, isothiazole, 1,3,4-oxadiazole, 1,3,4-thiadiazole, 1,2,5-oxadiazole, 1,2,5-thiadiazole, pyrrole, pyrazole, imidazole, 1,3,4-triazole, tetrazole, benzene, pyridine, pyrazine, pyrimidine, pyridazine, 1,2,3-triazine, 1,2,4-triazine, 1,3,5-triazine, thieno[2,3-b]furan, thieno[3,2-b]pyrrole, indole, benzofuran, benzothiophene, benzimidazole, benzoxazole, benzothiazole, benzo[2,1,3]thiadiazole, furano[3,2-b]pyridine, naphthalene, quinoline, isoquinoline, quinoxaline, quinazoline, cinnoline, phthalazine, 1,8-naphthyridine, and the like.
Optical Isomers--Diatereomers--Geometric lsomers
Some of the compounds described herein contain one or more asymmetric centers and may thus give rise to diastereomers and optical isomers. The present invention is meant to comprehend such possible diastereomers as well as their racemic and resolved, enantiomerically pure forms and pharmaceutically acceptable salts thereof.
Some of the compounds described herein contain olefinic double bonds, and unless specified otherwise, are meant to include both E and Z geometric isomers.
Salts
The pharmaceutical compositions of the present invention comprise a compound of Formula I as an active ingredient or a pharmaceutically acceptable salt, thereof, and may also contain a pharmaceutically acceptable carrier and optionally other therapeutic ingredients. The term "pharmaceutically acceptable salts" refers to salts prepared from pharmaceutically acceptable non-toxic bases including inorganic bases and organic bases. Salts derived from inorganic bases include aluminum, ammonium, calcium, copper, ferric, ferrous, lithium, magnesium, manganic salts, manganous, potassium, sodium, zinc and the like. Particularly preferred are the ammonium, calcium, magnesium, potassium and sodium salts. Salts derived from pharmaceutically acceptable organic non-toxic bases include salts of primary, secondary, and tertiary amines, substituted amines including naturally occurring substituted amines, cyclic amines and basic ion exchange resins, such as arginine, betaine, caffeine, choline, N,N'-dibenzylethylenediamine, diethylamine, 2-diethylaminoethanol, 2-dimethylaminoethanol, ethanolamine, ethylenediamine, N-ethylmorpholine, N-ethylpiperidine, glucamine, glucosamine, histidine, hydrabamine, isopropylamine, lysine, methylglucamine, morpholine, piperazine, piperidine, polyamine resins, procaine, purines, theobromine, triethylamine, trimethylamine, tripropylamine, tromethamine, and the like.
When the compound of the present invention is basic, salts may be prepared from pharmaceutically acceptable non-toxic acids, including inorganic and organic acids. Such acids include acetic, benzenesulfonic, benzoic, camphorsulfonic, citric, ethanesulfonic, fumaric, gluconic, glutamic, hydrobromic, hydrochloric, isethionic, lactic, maleic, rnalic, mandelic, methanesulfonic, mucic, nitric, pamoic, pantothenic, phosphoric, succinic, sulfuric, tartaric, p-toluenesulfonic acid and the like. Particularly preferred are citric, hydrobromic, hydrochloric, maleic, phosphoric, sulfuric, and tartaric acids.
It will be understood that in the discussion of methods of treatment which follows, references to the compounds of Formula I are meant to also include the pharmaceutically acceptable salts.
Utilities
The ability of the compounds of Formula I to inhibit biosynthesis of the leukotrienes makes them useful for preventing or reversing the symptoms induced by the leukotrienes in a human subject. This inhibition of the mammalian biosynthesis of leukotrienes indicates that the compounds and pharmaceutical compositions thereof are useful to treat, prevent, or ameliorate in mammals and especially in humans: 1) pulmonary disorders including diseases such as asthma, chronic bronchitis, and related obstructive airway diseases, 2) allergies and allergic reactions such as allergic rhinitis, contact dermatitis, allergic conjunctivitis, and the like, 3) inflammation such as arthritis or inflammatory bowel disease, 4) pain, 5) skin disorders such as atopic eczema, and the like, 6) cardiovascular disorders such as angina, formation of atherosclerotic plaques, myocardial ischemia, hypertension, platelet aggregation and the like, 7) renal insufficiency arising from ischaemia induced by immunological or chemical (cyclosporin) etiology, 8) migraine or cluster headache, 9) ocular conditions such as uveitis, 10) hepatitis resulting from chemical, immunological or infectious stimuli, 11 ) trauma or shock states such as burn injuries, endotoxemia, and the like, 12) allograft rejection, 13) prevention of side effects associated with therapeutic administration of cytokines such as Interleukin II and tumor necrosis factor, 14) chronic lung diseases such as cystic fibrosis, bronchitis and other small- and large-airway diseases, 15) cholecystitis, and 16) multiple sclerosis.
Thus, the compounds of the present invention may also be used to treat or prevent mammalian (especially, human) disease states such as erosive gastritis; erosive esophagitis; diarrhea; cerebral spasm; premature labor; spontaneous abortion; dysmenorrhea; ischemia; noxious agent-induced damage or necrosis of hepatic, pancreatic, renal, or myocardial tissue; liver parenchymal damage caused by hepatoxic agents such as CCl.sub.4 and D-galactosamine; ischemic renal failure; disease-induced hepatic damage; bile salt induced pancreatic or gastric damage; trauma- or stress-induced cell damage; and glycerol-induced renal failure. The compounds also act as inhibitors of tumor metastasis and exhibit cytoprotective action.
The cytoprotective activity of a compound may be observed in both animals and man by noting the increased resistance of the gastrointestinal mucosa to the noxious effects of strong irritants, for example, the ulcerogenic effects of aspirin or indomethacin. In addition to lessening the effect of non-steroidal anti-inflammatory drugs on the gastrointestinal tract, animal studies show that cytoprotective compounds will prevent gastric lesions induced by oral administration of strong acids, strong bases, ethanol, hypertonic saline solutions, and the like.
Two assays can be used to measure cytoprotective ability. These assays are; (A) an ethanol-induced lesion assay and (B) an indomethacin-induced ulcer assay and are described in EP 140,684.
Dose Ranges
The magnitude of prophylactic or therapeutic dose of a compound of Formula I will, of course, vary with the nature of the severity of the condition to be treated and with the particular compound of Formula I and its route of administration. It will also vary according to the age, weight and response of the individual patient. In general, the daily dose range for anti-asthmatic, anti-allergic or anti-inflammatory use and generally, uses other than cytoprotection, lie within the range of from about 0.001 mg to about 100 mg per kg body weight of a mammal, preferably 0.01 mg to about 10 mg per kg, and most preferably 0.1 to 1 mg per kg, in single or divided doses. On the other hand, it may be necessary to use dosages outside these limits in some cases.
For use where a composition for intravenous administration is employed, a suitable dosage range for anti-asthmatic, anti-inflammatory or anti-allergic use is from about 0.001 mg to about 25 mg (preferably from 0.01 mg to about 1 mg) of a compound of Formula I per kg of body weight per day and for cytoprotective use from about 0.1 mg to about 100 mg (preferably from about 1 mg to about 100 mg and more preferably from about 1 mg to about 10 mg) of a compound of Formula I per kg of body weight per day.
In the case where an oral composition is employed, a s suitable dosage range for anti-asthmatic, anti-inflammatory or anti-allergic use is, e.g. from about 0.01 mg to about 100 mg of a compound of Formula I per kg of body weight per day, preferably from about 0.1 mg to about 10 mg per kg and for cytoprotective use from 0.1 mg to about 100 mg (preferably from about 1 mg to about 100 mg and more preferably from about 10 mg to about 100 mg) of a compound of Formula I per kg of body weight per day.
For the treatment of diseases of the eye, ophthalmic preparations for ocular administration comprising 0.001-1% by weight solutions or suspensions of the compounds of Formula I in an acceptable ophthalmic formulation may be used.
The exact amount of a compound of the Formula I to be used as a cytoprotective agent will depend on, inter alia, whether it is being administered to heal damaged cells or to avoid future damage, on the nature of the damaged cells (e.g., gastrointestinal ulcerations vs. nephrotic necrosis), and on the nature of the causative agent. An example of the use of a compound of the Formula I in avoiding future damage would be co-administration of a compound of the Formula I with a non-steroidal anti-inflammatory drug that might otherwise cause such damage (for example, indomethacin). For such use, the compound of Formula I is administered from 30 minutes prior up to 30 minutes after administration of the NSAID. Preferably it is administered prior to or simultaneously with the NSAID, (for example, in a combination dosage form).
Pharmaceutical Compositions
Any suitable route of administration may be employed for providing a mammal, especially a human with an effective dosage of a compound of the present invention. For example, oral, rectal, topical, parenteral, ocular, pulmonary, nasal, and the like may be employed. Dosage forms include tablets, troches, dispersions, suspensions, solutions, capsules, creams, ointments, aerosols, and the like.
The pharmaceutical compositions of the present invention comprise a compound of Formula I as an active ingredient or a pharmaceutically acceptable salt thereof, and may also contain a pharmaceutically acceptable carrier and optionally other therapeutic ingredients. The term "pharmaceutically acceptable salts" refers to salts prepared from pharmaceutically acceptable non-toxic bases or acids including inorganic bases or acids and organic bases or acids.
The compositions include compositions suitable for oral, rectal, topical, parenteral (including subcutaneous, intramuscular, and intravenous), ocular (ophthalmic), pulmonary (nasal or buccal inhalation), or nasal administration, although the most suitable route in any given case will depend on the nature and severity of the conditions being treated and on the nature of the active ingredient. They may be conveniently presented in unit dosage form and prepared by any of the methods well-known in the an of pharmacy.
For administration by inhalation, the compounds of the present invention are conveniently delivered in the form of an aerosol spray presentation from pressurized packs or nebulisers. The compounds may also be delivered as powders which may be formulated and the powder composition may be inhaled with the aid of an insufflation powder inhaler device. The preferred delivery system for inhalation is a metered dose inhalation (MDI) aerosol, which may be formulated as a suspension or solution of compound I in suitable propellants, such as fluorocarbons or hydrocarbons.
Suitable topical formulations of Compound I include transdermal devices, aerosols, creams, ointments, lotions, dusting powders, and the like.
In practical use, the compounds of Formula I can be combined as the active ingredient in intimate admixture with a pharmaceutical carder according to conventional pharmaceutical compounding techniques. The carder may take a wide variety of forms depending on the form of preparation desired for administration, e.g., oral or parenteral (including intravenous). In preparing the compositions for oral dosage form, any of the usual pharmaceutical media may be employed, such as, for example, water, glycols, oils, alcohols, flavoring agents, preservatives, coloring agents and the like in the case of oral liquid preparations, such as, for example, suspensions, elixirs and solutions; or carders such as starches, sugars, microcrystalline cellulose, diluents, granulating agents, lubricants, binders, disintegrating agents and the like in the case of oral solid preparations such as, for example, powders, capsules and tablets, with the solid oral preparations being preferred over the liquid preparations. Because of their ease of administration, tablets and capsules represent the most advantageous oral dosage unit form in which case solid pharmaceutical carriers are obviously employed. If desired, tablets may be coated by standard aqueous or nonaqueous techniques.
In addition to the common dosage forms set out above, the compounds of Formula I may also be administered by controlled release means and/or delivery devices such as those described in U.S. Pat. Nos. 3,845,770; 3,916,899; 3,536,809; 3,598,123; 3,630,200 and 4,008,719, the disclosures of which are hereby incorporated herein by reference.
Pharmaceutical compositions of the present invention suitable for oral administration may be presented as discrete units such as capsules, cachets or tablets each containing a predetermined amount of the active ingredient, as a powder or granules or as a solution or a suspension in an aqueous liquid, a non-aqueous liquid, an oil-in-water emulsion or a water-in-oil liquid emulsion. Such compositions may be prepared by any of the methods of pharmacy but all methods include the step of bringing into association the active ingredient with the carder which constitutes one or more necessary ingredients. In general, the compositions are prepared by uniformly and intimately admixing the active ingredient with liquid carders or finely divided solid carders or both, and then, if necessary, shaping the product into the desired presentation. For example, a tablet may be prepared by compression or molding, optionally with one or more accessory ingredients. Compressed tablets may be prepared by compressing in a suitable machine, the active ingredient in a free-flowing form such as powder or granules, optionally mixed with a binder, lubricant, inert diluent, surface active or dispersing agent. Molded tablets may be made by molding in a suitable machine, a mixture of the powdered compound moistened with an inert liquid diluent. Desirably, each tablet contains from about 1 mg to about 500 mg of the active ingredient and each cachet or capsule contains from about 1 to about 500 mg of the active ingredient.
The following are examples of representative pharmaceutical dosage forms for the compounds of Formula I:
______________________________________Injectable Suspension (I.M.) mg/ml______________________________________Compound of Formula I 10Methylcellulose 5.0Tween 80 0.5Benzyl alcohol 9.0Benzalkonium chloride 1.0Water for injection to a total volume of 1 mL______________________________________Tablet mg/tablet______________________________________Compound of Formula I 25Microcrystalline Cellulose 415Povidone 14.0Pregelatinized Starch 43.5Magnesium Stearate 2.5 500______________________________________Capsule mg/capsule______________________________________Compound of Formula I 25Lactose Powder 573.5Magnesium Stearate 1.5 600______________________________________ PerAerosol canister______________________________________Compound of Formula I 24 mgLecithin, NF Liquid Concentrate 1.2 mgTrichlorofluoromethane, NF 4.025 gDichlorodifluoromethane, NF 12.15 g______________________________________
Combinations with Other Drugs
In addition to the compounds of Formula I, the pharmaceutical compositions of the present invention can also contain other active ingredients, such as cyclooxygenase inhibitors, non-steroidal anti-inflammatory drugs (NSAIDs), peripheral analgesic agents such as zomepirac diflunisal and the like. The weight ratio of the compound of the Formula I to the second active ingredient may be varied and will depend upon the effective dose of each ingredient. Generally, an effective dose of each will be used. Thus, for example, when a compound of the Formula I is combined with an NSAID the weight ratio of the compound of the Formula I to the NSAID will generally range from about 1000:1 to about 1:1000, preferably about 200:1 to about 1:200. Combinations of a compound of the Formula I and other active ingredients will generally also be within the aforementioned range, but in each case, an effective dose of each active ingredient should be used.
NSAIDs can be characterized into five groups:
(1) the propionic acid derivatives;
(2) the acetic acid derivatives;
(3) the fenamic acid derivatives;
(4) the oxicams; and
(5) the biphenylcarboxylic acid derivatives;
or a pharmaceutically acceptable salt thereof.
The propionic acid derivatives which may be used comprise: alminoprofen, benoxaprofen, bucloxic acid, carprofen, fenbufen, fenoprofen, fluprofen, flurbiprofen, ibuprofen, indoprofen, ketoprofen, miroprofen, naproxen, oxaprozin, pirprofen, prano-profen, suprofen, tiaprofenic acid, and tioxaprofen. Structurally related propionic acid derivatives having similar analgesic and anti-inflammatory properties are also intended to be included in this group.
Thus, "propionic acid derivatives" as defined herein are non-narcotic analgesics/non-steroidal anti-inflammatory drugs having a free --CH(CH3)COOH or --CH2CH2COOH group (which optionally can be in the form of a pharmaceutically acceptable salt group, e.g., --CH(CH.sub.3)COO.sup.- Na.sup.+ or --CH.sub.2 CH.sub.2 COO.sup.- Na.sup.+), typically attached directly or via a carbonyl function to a ring system, preferably to an aromatic ring system.
The acetic acid derivatives which may be used comprise: indomethacin, which is a preferred NSAID, acemetacin, alclofenac, clidanac, diclofenac, fenclofenac, fenclozic acid, fentiazac, furofenac, ibufenac, isoxepac, oxpinac, sulindac, tiopinac, tolmetin, zidometacin and zomepirac. Structually related acetic acid derivatives having similar analgesic and anti-inflammatory properties are also intended to be encompassed by this group.
Thus, "acetic acid derivatives" as defined herein are non-narcotic analgesics/non-steroidal anti-inflammatory drugs having a free --CH.sub.2 COOH group (which optionally can be in the form of a pharmaceutically acceptable salt group, e.g. --CH.sub.2 COO.sup.- Na.sup.+), typically attached directly to a ring system, preferably to an aromatic or heteroaromatic ring system.
The fenamic acid derivatives which may be used comprise: flufenamic acid, meclofenamic acid, mefenamic acid, niflumic acid and tolfenamic acid. Structurally related fenamic acid derivatives having similar analgesic and anti-inflammatory properties are also intended to be encompassed by this group.
Thus, "fenamic acid derivatives" as defined herein are non-narcotic analgesics/non-steroidal anti-inflammatory drugs which contain the basic structure: ##STR6## which can bear a variety of substituents and in which the free --COOH group can be in the form of a pharmaceutically acceptable salt group, e.g., --COO.sup.- Na.sup.+.
The biphenylcarboxylic acid derivatives which can be used comprise: diflunisal and flufenisal. Structurally related biphenylcarboxylic acid derivatives having similar analgesic and anti-inflammatory properties are also intended to be encompassed by this group.
Thus, "biphenylcarboxylic acid derivatives" as defined herein are non-narcotic analgesics/non-steroidal anti-inflammatory drugs which contain the basic structure: ##STR7## which can bear a variety of substituents and in which the free --COOH group can be in the form of a pharmaceutically acceptable salt group, e.g., --COO.sup.- Na.sup.+.
The oxicams which can be used in the present invention comprise: isoxicam, piroxicam, sudoxicam and tenoxican. Structurally related oxicams having similar analgesic and anti-inflammatory properties are also intended to be encompassed by this group.
Thus, "oxicams" as defined herein are non-narcotic analgesics/non-steroidal anti-inflammatory drugs which have the general formula: ##STR8## wherein R is an aryl or heteroaryl ring system.
The following NSAIDs may also be used: amfenac sodium, aminoprofen, anitrazafen, antrafenine, auranofin, bendazac lysinate, benzydanine, beprozin, broperamole, bufezolac, cinmetacin, ciproquazone, cloximate, dazidamine, deboxamet, delmetacin, detomidine, dexindoprofen, diacerein, di-fisalamine, difenpyramide, emorfazone, enfenamic acid, enolicam, epirizole, etersalate, etodolac, etofenamate, fanetizole mesylate, fenclorac, fendosal, fenflumizole, feprazone, floctafenine, flunixin, flunoxaprofen, fluproquazone, fopirtoline, fosfosal, furcloprofen, glucametacin, guaimesal, ibuproxam, isofezolac, isonixim, isoprofen, isoxicam, lefetamine HCl, leflunomide, lofemizole, lonazolac calcium, lotifazole, loxoprofen, lysin clonixinate, meclofenamate sodium, meseclazone, nabumetone, nictindole, nimesulide, orpanoxin, oxametacin, oxapadol, perisoxai citrate, pimeprofen, pimetacin, piproxen, pirazolac, pirfenidone, proglumetacin maleate, proquazone, pyridoxiprofen, sudoxicam, talmetacin, talniflumate, tenoxicam, thiazolinobutazone, thielavin B, tiaramide HCl, tiflamizole, timegadine, tolpadol, tryptamid and ufenamate.
The following NSAIDs, designated by company code number (see e.g., Pharmaprojects), may also be used: 480156S, AA861, AD 1590, AFP802, AFP860, AI77B, AP504, AU8001, BPPC, BW540C, CHINOIN 127, CN100, EB382, EL508, F1044, GV3658, ITF182, KCNTEI6090, KME4, LA2851, MR714, MR897, MY309, ONO3144, PR823, PV102, PV108, R830, RS2131, SCR152, SH440, SIR133, SPAS510, SQ27239, ST281, SY6001, TA60, TAI-901 (4-benzoyl-1- indancarboxylic acid), TVX2706, U60257, UR2301, and WY41770.
Finally, NSAIDs which may also be used include the salicylates, specifically acetyl salicylic acid and the phenylbutazones, and pharmaceutically acceptable salts thereof.
In addition to indomethacin, other preferred NS AIDS are acetyl salicylic acid, diclofenac, fenbufen, fenoprofen, flurbiprofen, ibuprofen, ketoprofen, naproxen, phenylbutazone, piroxicam, sulindac and tolmetin.
Pharmaceutical compositions comprising the Formula I compounds may also contain inhibitors of the biosynthesis of the leukotrienes such as are disclosed in EP 138,481 (Apr. 24,1985), EP 115,394 (Aug. 8, 1984), EP 136,893 (Apr. 10, 1985), and EP 140,709 (May 8, 1985), which are hereby incorporated herein by reference.
The compounds of the Formula I may also be used in combination with leukotriene antagonists such as those disclosed in EP 106,565 (Apr. 25, 1984) and EP 104,885 (Apr. 4, 1984) which are hereby incorporated herein by reference and others known in the art such as those disclosed in EP Application Nos. 56,172 (Jul. 21, 1982) and 61,800 (Jun. 10, 1982); and in U.K. Patent Specification No. 2,058,785 (Apr. 15, 1981), which are hereby incorporated herein by reference.
Pharmaceutical compositions comprising the Formula I compounds may also contain as the second active ingredient, prostaglandin antagonists such as those disclosed in EP 11,067 (May 28, 1980) or thromboxane antagonists such as those disclosed in U.S. Pat. No. 4,237,160. They may also contain histidine decarboxylase inhibitors such as .alpha.-fluoromethylhistidine, described in U.S. Pat. No. 4,325,961. The compounds of the Formula I may also be advantageously combined with an H.sub.1 - or H.sub.2 -receptor antagonist, such as for instance acetamazole, aminothiadiazoles disclosed in EP 40,696 (Dec. 2, 1981), benadryl, cimetidine, famotidine, framamine, histadyl, phenergan, ranitidine, terfenadine and like compounds, such as those disclosed in U.S. Pat. Nos. 4,283,408; 4,362,736; and 4,394,508. The pharmaceutical compositions may also contain a K.sup.+ /H.sup.+ ATPase inhibitor such as omeprazole, disclosed in U.S. Pat. No. 4,255,431, and the like. Compounds of Formula I may also be usefully combined with most cell stabilizing agents, such as 1,3-bis(2-carboxychromon-5-yloxy)-2-hydroxypropane and related compounds described in British Patent Specifications 1,144,905 and 1,144,906. Another useful pharmaceutical composition comprises the Formula I compounds in combination with serotonin antagonists such as methysergide, the serotonin antagonists described in Nature, Vol. 316, pages 126-131, 1985, and the like. Each of the references referred to in this paragraph is hereby incorporated herein by reference.
Other advantageous pharmaceutical compositions comprise the Formula I compounds in combination with anti-cholinergics such as ipratropium bromide, bronchodilators such as the beta agonist salbutamol, metaproterenol, terbutaline, fenoterol, and the like, and the anti-asthmatic drugs theophylline, choline theophyllinate and enprofylline, the calcium antagonists nifedipine, diltiazem, nitrendipine, verapamil, nimodipine, felodipine, etc. and the corticosteroids, hydrocortisone, methylprednisolone, betamethasone, dexamethasone, beclomethasone, and the like.
Methods of Synthesis
Compounds of the present invention can be prepared according to the following method. Temperatures are in degree Celsius. The substituents are the same as in Formula I except where defined otherwise.
Scheme I
The route which is used to prepare compounds of Formula Ic and Id is outlined in Scheme I. The bicyclic ketone II is condensed with the aryl halide III in the presence of a base such as n-BuLi in an organic solvent such as THF. The benzyloxy derivative IV is then reduced using H.sub.2 and Pd on charcoal, in an organic solvent such as EtOH providing phenol V. Coupling of phenol V with the appropriate halide or activated alcohol of type VI (wherein X=Cl, Br, I, OMs, OTs) in a polar solvent such as DMF in presence of an inorganic base such as K.sub.2 CO.sub.3, provides compounds of Formula Ic. Alternatively, coupling of phenol V with the appropriate alcohol VI (wherein X=OH) by treatment with a phosphine and a diazodicarboxylate in an organic solvent such as THF also provides compounds of Formula Ic. Alkylation of compounds of Formula Ic is achieved by treatment with a hydride such as KH with an alkyl halide in an organic solvent such as THF providing compounds of Formula Id. ##STR9##
Representative Compounds
Tables 1-5 illustrate compounds representative of the present invention.
TABLE 1______________________________________ ##STR10## IeEx. X.sup.1 R.sup.1______________________________________1 OCH.sub.2 H2 OCH.sub.2 Me3 CH.sub.2 O H4 CH.sub.2 O Me5 CH.sub.2 CH.sub.2 H6 CH.sub.2 CH.sub.2 Me______________________________________
TABLE 2______________________________________ ##STR11## IfEx. R.sup.1 Ar.sup.2______________________________________ 7 H ##STR12## 8 Me ##STR13## 9 Me ##STR14##10 Me ##STR15##11 Me ##STR16##12 Me ##STR17##13 Me ##STR18##14 Me ##STR19##______________________________________
TABLE 3______________________________________ ##STR20## IgEx. R.sup.1 Ar.sup.2______________________________________15 H ##STR21##16 Me ##STR22##17 Me ##STR23##18 Me ##STR24##19 Me ##STR25##20 Me ##STR26##21 Me ##STR27##22 H ##STR28##______________________________________
TABLE 4______________________________________ ##STR29## IhEx. R.sup.1 R.sup.3 Ar.sup.3 Ar.sup.1______________________________________23 H CN 3-Fu 5-(3-FPhe)24 H CN 3-Th 5-(3-FPhe)25 H CN 3-Fu 6,2-Pye26 H CN 3-Th 6,2-Pye27 H C.sub.3 H.sub.4 NS* 3-Fu 5-(3-FPhe)28 H C.sub.3 H.sub.4 NS* 3-Th 5-(3-FPhe)29 H C.sub.3 H.sub.2 NS** 3-Fu 5-(3-FPhe)30 H C.sub.3 H.sub.2 NS** 3-Th 5-(3-FPhe)31 H COC.sub.3 H.sub.2 NS** 3-Fu 5-(3-FPhe)32 H COC.sub.3 H.sub.2 NS** 3-Th 5-(3-FPhe)33 H CN H 5-(3-FPhe)34 H CN Ph 5-(3-FPhe)35 H CN C.sub.3 H.sub.2 NS** 5-(3-FPhe)36 Me CN 3-Fu 5-(3-FPhe)37 Me CN 3-Th 5-(3-F Phe)______________________________________ *C.sub.3 H.sub.4 NS = 2(4,5-dihydrothiazolyl) **C.sub.3 H.sub.2 NS = 2thiazolyl
TABLE 5______________________________________ ##STR30## IiEx. R.sup.1 R.sup.2 Ar.sup.1______________________________________38 H 3-Fu 5-(3-FPhe)39 H 3-Th 5-(3-FPhe)40 Me 3-Fu 5-(3-FPhe)41 Me 3-Th 5-(3-FPhe)42 H 3-Fu 6,2-Pye43 H 3-Th 6,2-Pye______________________________________
Assays for Determining Biological Activity
Compounds of Formula I can be tested using the following assays to determine their mammalian leukotriene biosynthesis inhibiting activity.
Human 5-Lipoxygenase Inhibitor Screen
Objective of the Assay: The objective of the assay is to select agents which specifically inhibit the activity of human 5-lipoxygenase using a 100,000x g supernatant fraction prepared from insect cells infected with recombinant baculovirus containing the coding sequence for human 5-lipoxygenase. Enzyme activity is measured spectrophotometrically from the optimal rate of conjugated diene formation (A.sub.234) measured after the incubation of the enzyme with arachidonic acid in the presence of ATP, calcium ions, and phosphatidylcholine.
Description of Procedure: The activity of 5-lipoxygenase is measured using a spectrophotometric assay and recombinant human 5-lipoxygenase as a source of enzyme. The 100,000x g fraction from S19 cells infected with the recombinant baculovirus rvH5LO(8-1) containing the coding region sequence for human 5-lipoxygenase is prepared as described by Denis et al. (J. Biol. Chem., 266, 5072-5079 (1991)). The enzymatic activity is measured, using a spectrophotometric assay from the optimal rate of conjugated diene formation (A.sub.234) using the procedure described by Riendeau et al. (Biochem. Pharmacol. 38, 2323-2321, (1989)) with minor modifications. The incubation mixture contains 50 mM sodium phosphate pH 7.4, 0.2 mM ATP, 0.2 mM CaCl.sub.2, 20 .mu.M arachidonic acid (5 .mu.L from a 100-fold concentrated solution in ethanol), 12 .mu.g/mL phosphatidylcholine, an aliquot of the 100,000x g fraction (2-10 .mu.L) and inhibitor (0.5 mL final volume). Inhibitors are added as 500-fold concentrated solutions in DMSO. Reactions are initiated by the addition of an aliquot of the enzyme preparation and the rate of conjugated diene formation is followed for 2 minutes at room temperature. The reactions are performed in semi-micro cuvettes (0.7 mL capacity, 10 mm path length and 4 mm internal width) and the absorbance changes are recorded with a Hewlett-Packard diode array spectrophotometer (HP 8452A) connected to the ChemStation using UV/VIS Kinetics Software (Hewlett-Packard). Enzymatic activity is calculated from the optimal rate of the reaction by a linear fit of the variation of A.sub.234 during the first twenty seconds using the least square method for the equation A.sub.234 =V.sub.o t+A.sub.o where V.sub.o is the rate, t is the time, and A.sub.o is the absorbance at zero time. The results are expressed as percentages of inhibition of the reaction rate relative to controls (typically between 0.15-0.21 AU/min) containing the DMSO vehicle.
Rat Peritoneal Polymorphonuclear (PMN) Leukocyte Assay
Rats under ether anesthesia are injected (i.p.) with 8 mL of a suspension of sodium caseinate (6 grams in ca. 50 mL water). After 15-24 hr. the rats are sacrificed (CO.sub.2) and the cells from the peritoneal cavity are recovered by lavage with 20 mL of buffer (Eagles MEM containing 30 mM HEPES adjusted to pH 7.4 with NaOH). The cells are pelleted (350x g, 5 min.), resuspended in buffer with vigorous shaking, filtered through lens paper, recentrifuged and finally suspended in buffer at a concentration of 10 cells/mL. A 500 .mu.L aliquot of PMN suspension and test compound are preincubated for 2 minutes at 37.degree. C., followed by the addition of 10 .mu.M calcium ionophore A-23187. The suspension is stirred for an additional 4 minutes then bioassayed for LTB.sub.4 content by adding an aliquot to a second 500 .mu.L portion of the PMN at 37.degree. C. The LTB.sub.4 produced in the first incubation causes aggregation of the second PMN, which is measured as a change in light transmission. The size of the assay aliquot is chosen to give a submaximal transmission change (usually -70%) for the untreated control. The percentage inhibition of LTB.sub.4 formation is calculated from the ratio of transmission change in the sample to the transmission change in the compound-free control.
Human Polymorphonuclear (PMN) Leukocyte LTB.sub.4 Assay
A. Preparation of Human PMN. Human blood is obtained by antecubital venepuncture from consenting volunteers who have not taken medication within the previous 7 days. The blood is immediately added to 10% (v/v) trisodium citrate (0.13M) or 5% (v/v) sodium heparin (1000 IU/mL). PMNs are isolated from anticoagulated blood by dextran sedimentation of erythrocytes followed by centrifugation through Ficoll-Hypaque (specific gravity 1.077), as described by Boyurn (Scand, J. Clin. Lab. Invest., 21 (Supp 97), 77 (1968)). Contaminating erythrocytes are removed by lysis following exposure to ammonium chloride (0.16M) in Tris buffer (pH 7.65), and the PMNs are resuspended at 5.times.10.sup.5 cells/mL in HEPES (15 mM)-buffered Hanks balanced salt solution containing Ca.sup.2+ (1.4 mM) and Mg.sup.2+ (0.7 mM), pH 7.4.
B. Generation and Radioimmunoassay of LTB.sub.4. PMNs (0.5 mL; 2.5.times.10.sup.5 cells) are placed in plastic tubes and incubated (37.degree. C., 2 min) with test compounds at the desired concentration or vehicle (DMSO, final concentration 0.2%) as control. The synthesis of LTB.sub.4 is initiated by the addition of calcium ionophore A23187 (final concentration 10 .mu.M) or vehicle in control samples and allowed to. proceed for 5 minutes at 37.degree. C. The reactions are then terminated by the addition of cold methanol (0.25 mL) and samples of the entire PMN reaction mixture are removed for radioimmunoassay of LTB.sub.4.
Samples (50 .mu.L) of authentic LTB.sub.4 of known concentration in radioimmunoassay buffer (RIA) buffer (potassium phosphate 1 mM; disodium EDTA 0.1 mM; Thimerosal 0,025 mM; gelatin 0.1%, pH 7.3) or PMN reaction mixture diluted 1:1 with RIA buffer are added to reaction tubes. Thereafter [.sup.3 H]-LTB.sub.4 (10 nCi in 100 .mu.L RIA buffer) and LTB.sub.4 -antiserum (100 .mu.L of a 1:3000 dilution in RIA buffer) are added and the tubes vortexed. Reactants are allowed to equilibrate by incubation overnight at 4.degree. C. To separate antibody-bound from free LTB.sub.4, aliquots (50 .mu.L) of activated charcoal (3% activated charcoal in RIA buffer containing 0.25% Dextran T-70) are added, the tubes vortexed, and allowed to stand at room temperature for 10 minutes prior to centrifugation (1500x g; 10 min; 4.degree. C.). The supernatants containing antibody-bound LTB.sub.4 are decanted into vials and Aquasol 2 (4 mL) is added. Radioactivity is quantified by liquid scintillation spectrometry. The specificity of the antiserum and the sensitivity of the procedure have been described by Rokach et al, Prostaglandins Leukotrienes and Medicine, 13, 21 (1984). The amount of LTB.sub.4 produced in test and control samples is calculated. Inhibitory dose-response curves are constructed using a four-parameter algorithm and from these the IC.sub.50 values are determined.
Human Whole Blood Assay In Vitro for LTB.sub.4 Production
Fresh blood is collected in heparinized tubes by venipuncture from human volunteers. A 500 .mu.L aliquot is incubated with one of the test compounds at final concentrations varying from 3 nM to 3 mM at 37.degree. C. for 15 min. Drug stock solutions are made up in DMSO and 1 .mu.L of the stock solution is added to each assay tube. The blood is then incubated with A23187 (in 5 .mu.L autologous plasma, 25 .mu.M final concentration) at 37.degree. C. for 30 min. At the end of incubation, plasma is obtained (12,000x g, 15 min) and a 100 .mu.L aliquot is added to 400 .mu.L methanol for protein precipitation. The mixture is vortexed, centrifuged and the supematant stored at -70.degree. C. until assayed for LTB.sub.4 by standard RIA.
Asthmatic Rat Assay
Rats are obtained from an inbred line of asthmatic rats. Both female (190-250 g) and male (260-400 g) rats are used.
Egg albumin (EA), grade V, crystallized and lyophilized, is obtained from Sigma Chemical Co., St. Louis. Aluminum hydroxide is obtained from the Regis Chemical Company, Chicago. Methysergide bimaleate is supplied by Sandoz Ltd., Basel.
The challenge and subsequent respiratory recordings are carried out in a clear plastic box with internal dimensions 10.times.6.times.4 inches. The top of the box is removable; in use, it is held firmly in place by four clamps and an airtight seal is maintained by a soft rubber gasket. Through the center of each end of the chamber a DeVilbiss nebulizer (No. 40) is inserted via an airtight seal and each end of the box also has an outlet. A Fleisch No. 0000 pneumotachograph is inserted into one end of the box and coupled to a Grass volumetric pressure transducer (PT5-A) which is then connected to a Buxco Electronics preamplifier (Buxco Electronics Inc., Sharon, Conn.). The preamplifier is connected to a Beckman Type R Dynograph and to a Buxco computer consisting of waveform analyser, Data Acquisition Logger with special software. While aerosolizing the antigen, the outlets are open and the pneumotachograph is isolated from the chamber. The outlets are closed and the pneumotachograph and the chamber are connected during the recording of the respiratory patterns. For challenge, 2 mL of a 3% solution of antigen in saline is placed into each nebulizer and the aerosol is generated with air from a small Potter diaphragm pump operating at 10 psi and a flow of 8 liters/minute.
Rats are sensitized by injecting (subcutaneously) 1 mL of a suspension containing 1 mg EA and 200 mg aluminum hydroxide in saline. They are used between days 12 and 24 post sensitization. In order to eliminate the serotonin component of the response, rats are pretreated intravenously 5 minutes prior to aerosol challenge with 3.0 .mu.g/kg of methysergide. Rats are then exposed to an aerosol of 3% EA in saline for exactly 1 minute, then their respiratory profiles are recorded for a further 30 minutes. The duration of continuous dyspnea is measured by the Buxco computer.
Compounds are generally administered either orally 2-4 hours prior to challenge or intravenously 2 minutes prior to challenge. They are either dissolved in saline or 1% methocel or suspended in 1% methocel. The volume injected is 1 mL/kg (intravenously) or 10 mL/kg (orally). Prior to oral treatment rats are starved overnight. The activity of compounds is determined in terms of their ability to decrease the duration of antigen-induced dyspnea in comparison with a group of vehicle-treated controls. Usually, a compound is evaluated at a series of doses and an ED.sub.50 is determined. This is defined as the dose (mg/kg) which would inhibit the duration of symptoms by 50%.
Pulmonary Mechanics in Trained Conscious Squirrel Monkeys
The test procedure involves placing trained squirrel monkeys in chairs in aerosol exposure chambers. For control purposes, pulmonary mechanics measurements of respiratory parameters are recorded for a period of about 30 minutes to establish each monkey's normal control values for that day. For oral administration, compounds are dissolved or suspended in a 1% methocel solution (methylcellulose, 65HG, 400 cps) and given in a volume of 1 mL/kg body weight. For aerosol administration of compounds, a DeVilbiss ultrasonic nebulizer is utilized. Pretreatment periods vary from 5 minutes to 4 hours before s the monkeys are challenged with aerosol doses of either leukotriene D.sub.4 (LTD.sub.4) or Ascaris summ antigen, 1:25 dilution.
Following challenge, each minute of data is calculated by computer as a percent change from control values for each respiratory parameter including airway resistance (R.sub.L) and dynamic compliance (C.sub.dyn). The results for each test compound are subsequently obtained for a minimum period of 60 minutes post challenge which are then compared to previously obtained historical baseline control values for that monkey. In addition, the overall values for 60 minutes post-challenge for each monkey (historical baseline values and test values) are averaged separately and are used to calculate the overall percent inhibition of LTD.sub.4 or Ascaris antigen response by the test compound. For statistical analysis, paired t-test is used. (References: McFarlane, C. S. et al., Prostaglandins, 28, 173-182 (1984) and McFarlane, C. S. et al., Agents Actions, 22, 63-68 (1987).)
Prevention of Induced Bronchoconstriction in Allergic Sheep
A. Rationale. Certain allergic sheep with known sensitivity to a specific antigen (Ascaris summ) respond to inhalation challenge with acute and late bronchial responses. The time course of both the acute and the late bronchial responses approximates the time course observed in asthmatics and the pharmacological modification of both responses is similar to that found in man. The effects of antigen in these sheep are largely observed in the large airways and are conveniently monitored as changes in lung resistance or specific lung resistance.
B. Methods. Animal Preparation: Adult sheep with a mean weight of 35 kg (range, 18 to 50 kg) are used. All animals used meet two criteria: a) they have a natural cutaneous reaction to 1:1,000 or 1:10,000 dilutions of Ascaris summ extract (Greer Diagnostics, Lenois, N.C.) and b) they have previously responded to inhalation challenge with Ascaris summ with both an acute bronchoconstriction and a late bronchial obstruction (W. M. Abraham, et al., Am. Rev. Resp. Dis., 128, 839-44 (1983)).
Measurement of Airway Mechanics: The unsedated sheep are restrained in a cart in the prone position with their heads immobilized. After topical anesthesia of the nasal passages with 2% lidocaine solution, a balloon catheter is advanced through one nostril into the lower esophagus. The animals are then intubated with a cuffed endotracheal tube through the other nostril using a flexible fiberoptic bronchoscope as a guide. Pleural pressure is estimated with the esophageal balloon catheter (filled with one mL of air), which is positioned such that inspiration produces a negative pressure deflection with clearly discernible cardiogenic oscillations. Lateral pressure in the trachea is measured with a sidehole catheter (inner dimension, 2.5 mm) advanced through and positioned distal to the tip of the nasotracheal tube. Transpulmonary pressure, the difference between tracheal pressure and pleural pressure, is measured with a differential pressure transducer (DP45; Validyne Corp., Northridge, Calif.). For the measurement of pulmonary resistance (R.sub.L), the maximal end of the nasotrachel tube is connected to a pneumotachograph (Fleisch, Dyna Sciences, Blue Bell, Pa.). The signals of flow and transpulmonary pressure are recorded on an oscilloscope (Model DR-12; Electronics for Medicine, White Plains, N.Y.) which is linked to a PDP-11 Digital computer (Digital Equipment Corp., Maynard, Mass.) for on-line calculation of R.sub.L from transpulmonary pressure, respiratory volume obtained by integration and flow. Analysis of 10-15 breaths is used for the determination of R.sub.L. Thoracic gas volume (V.sub.tg) is measured in a body plethysmograph, to obtain specific pulmonary resistance (SR.sub.L =R.sub.L .cndot.V.sub.tg).
Aerosol Delivery Systems: Aerosols of Ascaris summ extract (1:20) are generated using a disposable medical nebulizer (Raindrop.RTM., Puritan Bennett), which produces an aerosol with a mass median aerodynamic diameter of 6.2 .mu.M (geometric standard deviation, 2.1 ) as determined by an electric size analyzer (Model 3030; Thermal Systems, St. Paul, Minn.). The output from the nebulizer is directed into a plastic t-piece, one end of which is attached to the nasotracheal tube, the other end of which is connected to the inspiratory part of a Harvard respirator. The aerosol is delivered at a tidal volume of 500 mL of a rate of 20 per minute. Thus, each sheep receives an equivalent dose of antigen in both placebo and drug trials.
Experimental Protocol: Prior to antigen challenge baseline measurements of SR.sub.L are obtained, infusion of the test compound is started 1 hr prior to challenge, the measurement of SR.sub.L repeated and then the sheep undergoes inhalation challenge with Ascaris summ antigen. Measurements of SR.sub.L are obtained immediately after antigen challenge and at 1, 2, 3, 4, 5, 6, 6.5, 7, 7.5, and 8 hrs after antigen challange. Placebo and drug tests are separated by at least 14 days. In a further study, sheep are given a bolus dose of the test compound followed by an infusion of the test compound for 0.5-1 hr prior to Ascaris challenge and for 8 hrs after Ascaris as described above.
Statistical Analysis: A Kruskal-Wallis one way ANOVA test is used to compare the acute immediate responses to antigen and the peak late response in the controls and the drug-treated animals.
The invention is further defined by reference to the following examples, which are intended to be illustrative and not limiting. All temperatures are in degrees Celsius. Unless stated otherwise:
(i) all operations were carried out at room or ambient temperature, that is, at a temperature in the range 18.degree.-25.degree. C.;
(ii) evaporation of solvent was carried out using a rotary evaporator under reduced pressure (600-4000 pascals: 4.5-30 mm. hg) with a bath temperature of up to 60.degree. C.;
(iii) the course of reactions was followed by thin layer chromatography (TLC) and reaction times are given for illustration only;
(iv) melting points are uncorrected and `d` indicates decomposition; the melting points given are those obtained for the materials prepared as described; polymorphism may result in isolation of materials with different melting points in some preparations;
(v) the structure and purity of all final products were assured by at least one of the following techniques: TLC, mass spectrometry, nuclear magnetic resonance (NMR) spectrometry or elemental analysis;
(vi) yields are given for illustration only;
(vii) when given, NMR data are in the form of delta (.delta.) values for major diagnostic protons, given in parts per million (ppm) relative to tetramethylsilane (TMS) as internal standard, determined at 300 MHz or 400 MHz using the indicated solvent; conventional abbreviations used for signal shape are: s. singlet; d. doublet; t. triplet; m. multiplet; br. broad; etc.; in addition, "Ar" signifies an aromatic signal;
(viii) chemical symbols have their usual meanings; the following abbreviations have also been used v (volume), w (weight), b.p. (boiling point), m.p. (melting point), L (liter(s)), mL (milliliters), g (gram(s)), mg (milligrams(s)), mol (moles), mmol (millimoles), and eq (equivalent(s)).
EXAMPLE 1
[1S,5R]1,2-Dihydro-1-methyl-6-{5-[3-(3.alpha.-hydroxy-6,8-dioxabicyclo[3.2.1]-octanyl)]-3-fluoro phenoxymethyl}quinolin-2-one
Step 1: 2,4-Di-O-p-toluenesulfonyl-1,6-anhydro-.beta.-D-glucose
To a solution of 1,6-anhydro-.beta.-D-glucose (50 g, 308 mmoL) in dry pyridine (100 mL) at 0.degree. C. was added dropwise a solution of p-toluenesulfonyl chloride (123 g, 647 mmoL) dissolved in CHCl.sub.3 (350 mL) and pyridine (200 mL). The reaction mixture was stirred at r.t. for at least 2 days. Water was added and the reaction mixture was stirred for .about.1 hr, then the organic layer was decanted and the aqueous phase was reextracted with CHCl.sub.3. The combined organic layers were washed with H.sub.2 SO.sub.4 (10%) until the pH remains acidic, then finally washed with a saturated NH.sub.4 OAc solution. The resulting organic layer was dried over MgSO.sub.4 and the solvent evaporated. The syrup obtained was flash chromatographed on silica gel eluting with hexane:EtOAc (1:1) to give the title compound an oil.
Step 2 [1S,3S,5R]6,8-Dioxabicyclo[3.2.1]octan-3-ol
The ditosylate derivative from Step 1 (107 g, 0.228 mmoL) was dissolved in THF (1.6 L) at -40.degree. C. and Super-Hydride in THF (800 mL, 1M, 0.8 mmoL) was slowly added. The resulting reaction mixture was stirred at r.t. overnight. The reaction was cannulated into cold H.sub.2 O(226 mL) using external cooling, then NaOH 3N (640 mL, 1.92 mmol) and H.sub.2 O.sub.2 (30%) (490 mL, 4.3 mmol) were successively added. The reaction mixture was stirred at r.t. for 1 hr. Then the supernatant (THF layer) was separated from the aqueous layer and concentrated. The resulting residue was combined with the aqueous layer and extracted with CH.sub.2 Cl.sub.2 using a continuous extractor. The organic layer was dried (MgSO.sub.4) and evaporated to dryness. The oily residue was dissolved in hot Et.sub.2 O, filtered and evaporated to dryness affording the title compound contaminated with the 2-octanol isomer. The crude product was used as such for the next step.
Step 3: [1S,5R]6,8-Dioxabicyclo[3.2.1octan-3-one
The crude alcohol from Step 2 (16.6 g, 89 mmoL) in CH.sub.2 Cl.sub.2 (200 mL) was added slowly to a suspension of PCC (38.4 g, 178 mmoL) and celite (22 g) in CH.sub.2 Cl.sub.2 (400 mL) and stirred for 1 hr. The reaction mixture was diluted with Et.sub.2 O (600 mL) and filtered over celite. The filtrate was evaporated and the residue distilled with a Kugelrohr apparatus (10.degree. C., 1.8 mm/Hg) affording the title product as an oil.
Step 4: [1S,5R] O-Benzyl-5-[3-(3.alpha.-hydroxy-6,8-dioxabicyclo[3.2.1]octanyl)]-3-fluorophenol
To a solution of O-benzyl-3-bromo-5-fluorophenol (1.03 g, EP 385,662) in THF (15 mL) stirred at -78.degree. C. was added a solution of n-BuLi in hexane (2.5M, 1.62 mL). After 1 hr., a solution of [1S,5R] 6,8-dioxabicyclo[3.2.1]octan-3-one (471 mg) from Step 3 in THF (2 mL) was added dropwise to the resulting mixture. After 45 min. at -78.degree. C., the reaction mixture was stirred at 0.degree. C. for 1 hr. Saturated aqueous NH.sub.4 Cl was then added and the organic phase separated. The aqueous phase was extracted was EtOAc (3x) and the combined organic phases were washed with brine, dried (MgSO.sub.4), and evaporated to afford the title product as an oil.
Step 5: [1S,5R]5-[3-(3.alpha.-Hydroxy-6,8-dioxabicyclo[3.2.1]octanyl)]-3-fluorophenol
A mixture of O-benzylphenol derivative (1.1 g) from Step 4, Pd/C (10%) (100 mg) in EtOH (20 mL) was stirred under H.sub.2 (1 atm.) for 1 hr. Then CH.sub.2 Cl.sub.2 (20 mL) was added and the resulting mixture was filtered over celite. The flitrate was evaporated and the crude product was flash chromatographed on silica gel eluting with EtOAc/hexane (3:2) to afford the title product as a white solid.
Step 6: [1S,5R]1,2-Dihydro-1-methyl-6-{5-[3-(3.alpha.-hydroxy-6,8-dioxabicyclo[3.2.1]octanyl)]-3-fluorophenoxymethyl}-quinolin-2-one
A mixture of phenol (290 mg) from Step 5, 6-bromo-methyl-1,2-dihydro-1-methylquinolin-2-one (335 mg, EP 385,662) and K.sub.2 CO.sub.3 (250 mg) in DMF (6 mL) was stirred overnight. To the reaction mixture was added H.sub.2 O(15 mL) followed by CHCl.sub.3 (30 mL) and the organic layer was decanted. The aqueous layer was extracted with CHCl.sub.3 and the combined organic layers were washed with H.sub.2 O, brine, dried over MgSO.sub.4 and evaporated. The residue was chromatographed on silica gel, eluting with toluene/acetone (65:35) to afford the title compound as a white solid; m.p. 164.degree.-165.degree..
EXAMPLE 2
[1S,5R]1,2-Dihydro-1-methyl-6-{5-[3-(3.alpha.-methoxy-6,8-dioxabicyclo[3.2.1]octanyl)]-3-fluorophenoxymethyl quinolin-2-one
To a suspension of KH (35% in mineral oil, 423 mg) in THF (10 mL) was added [1S,5R] 1,2-dihydro-1-methyl-6-{5-[3-(3.alpha.-hydroxy-6,8-dioxabicyclo[3.2.1]octanyl)]-3-fluorophenoxymethyl}-quinolin-2-one (303 mg) as a solid. After 10 min., methyl iodide (0.23 mL) was added and the resulting mixture was stirred at r.t. for 30 min. Water was slowly added to the reaction mixture and extracted with EtOAc (3x). The combined organic layers were washed with brine, dried (MgSO.sub.4), and evaporated. The crude product was flash chromatographed on silica gel eluting with EtOAc/toluene (9:1) to afford the title product as a pale yellow foam.
.sup.1 H NMR (250 MHz, acetone - d.sub.6). .delta.1.98-2.14 (m, 3H), .sub.2.34 (dt, 1H, J=14.9, 1.8 Hz), 3.00 (s, 3H), 3.59 (t, 1H, J=5.5 Hz), 3.68 (s, 3H), 4.34 (dd, 1H, J=0.6, 6.2 Hz), 4.54 (m, 1H), 5.24 (s, 2H), 5.54 (t, 1H, J=1.8 Hz), 6.60 (d, 1H, J=9.5 Hz), 6.71-6.77 (m, 2H), 6.89 (t, 1H, J=1.7 Hz), 7.55 (d, 1H, J=8.7 Hz), 7.73 (dd, 1H, J=2.1, 8.7 Hz), 7.79 (d, 1H, J=1.9 Hz), 7.84 (d, 1H, J=9.5 Hz).
EXAMPLE 3
[1R,5S]1,2-Dihydro-1-methyl-6-{ 5-[3-(3.alpha.-hydroxy-6,8-dioxabicyclo[3,2.1]-octanyl)]-3-florophenoxymethyl}quinolin-2-one
Step 1: (R)3-tert-Butyldimethylsilyloxy-1.2-epoxypropane
To a 0.degree. C. mixture of tert-butyldimethylsilyl chloride (12.2 g) and imidazole (11.5 g) in CH.sub.2 Cl.sub.2 (100 mL) was slowly added (S)-glycidol (5.0 g; Aldrich). After 2.5 hr at 0.degree. C., the reaction mixture was treated with saturated aqueous NH.sub.4 Cl and the organic phase was separated. The aqueous phase was extracted with CH.sub.2 Cl.sub.2 (100 mL) and the combined organic phases were washed with H.sub.2 O and brine and then dried (MgSO.sub.4) and evaporated. Flash chromatography of the residue (silica gel; hexane/EtOAc (9:1)) afforded the title epoxide as a colorless liquid.
Step 2: [2R]6,6-Dimethoxy-4-(1,3-propanediyldithio)-3-tert-butyldimethylsilyloxyhexan-1-ol
To a solution of 2-(2,2-dimethoxyethyl)-1,3-dithiane (10.4 g; Liebigs Ann. Chem. 1989, 1045) in THF (50 mL) stirred at -30.degree. C. was added a solution of n-BuLl in hexane (1.3M; 38.5 mL; Aldrich). After 1 hr., the resulting purple solution was treated with a solution of the epoxide (9.4 g) from Step 1 in THF (10 mL). After 3 hr at -30.degree. C., the reaction mixture was quenched with saturated aqueous NH.sub.4 Cl. Water was then added and the aqueous phase was extracted with Et.sub.2 O (2x). The combined organic phases were washed with H.sub.2 O and brine, dried (MgSO.sub.4) and evaporated. Flash chromatography of the residue (silica gel; hexane/EtOAc (4:1)) afforded the title alcohol as a yellow liquid.
Step 3: [2R]6,6-Dimethoxy-4-(1,3-propanediyldithio)hexan-1,2-diol
To a solution of the silyl ether from Step 2 (9.94 g) in THF (50 mL) was added a solution of Bu.sub.4 NF in THF (1.0M; 26.5 mL; Aldrich). After 90 min at r.t., the volatiles were evaporated. Flash chromatography of the residue (silica gel; hexane/EtOAc (1:3)) afforded the title diol as an amber oil.
Step 4: [2R]6-Hydroxy-2-hydroxymethyl-3-(1,3-propanediyldithio)tetrahydropyran
A solution of the diol from Step 3 (3.11 g) in THF (40 mL) and aqueous HCl (5% v/v; 25 mL) was stirred at 60.degree. C. for 18 hr. Water was then added and the aqueous phase was extracted with EtOAc (3x). The combined organic phases were washed with brine, dried (Na.sub.2 SO.sub.4) and evaporated to afford the crude title compound as an amber gum.
Step 5: [2R]6-Hydroxy-3-(1,3-propanediyldithio)-p-toluene-sulfonylmethyltetrahydropyran
p-Toluenesulfonyl chloride (1.98 g; Aldrich) was added to a 0.degree. C. solution of the alcohol from Step 4 (2.05 g) in pyridine (10 mL). The mixture was stirred at r.t. for 90 min. before MeOH (1 mL) and CHCl.sub.3 (50 mL) were added. The organic phase was washed with H.sub.2 O (2x), dried (MgSO.sub.4) and evaporated. Flash chromatography of the residue (silica gel; hexane/EtOAc (3:2)) yielded the title tosylate as an amber semi-solid.
Step 6: [1R,5S]3-(1,3-Propanediyldithio)-6,8-dioxabicyclo[3.2.1]octane
1,8-Diazabicyclo[5.4.0] undec-7-ene (1.5 mL; Aldrich) was added to a solution of the tosylate from Step 5 (1.95 g) in CH.sub.2 Cl.sub.2 (30 mL). The mixture was stirred at r.t. for 16 hr. before the volatiles were evaporated. Flash chromatography of the residue (silica gel; hexane/EtOAc (3:1)) gave the title compound as a white solid.
Step 7: [1R,5S]6,8-Dioxabicyclo[3.2.1]octan-3-one
To a vigorously stirred mixture of the dithioketal from Step 6 (0.76 g) and CaCO.sub.3 (0.42 g) in THF (12 mL) and H.sub.2 O (2.3 mL) was added in 10 min. a solution of mercury (II) perchlorate trihydrate (1.73 g; Aldrich) in H.sub.2 O (1 mL). The mixture was stirred for 40 min. at r.t. before another portion of mercury salt solution (0.56 g in 0.2 mL of H.sub.2 O) was slowly added. After 15 min., Et.sub.2 O (50 mL) was added and the reaction mixture was filtered. The organic tiltrate was dried (MgSO.sub.4) and evaporated to afford the title ketone as an amber liquid.
Step 8: [1R,5S]1,2-dihydro-1-methyl-6-{5-[3-(3.alpha.-hydroxy-6,8-dioxabicyclo[3.2.1]octanyl)]-3-fluorophenoxymethyl}-quinolin-2-one
Following the procedure described in Example 1, Steps 4-6, but substituting [1R,5S]6,8-dioxabicyclo[3.2.1]octan-3-one (from Step 7) for [1S,5R] 6,8-dioxabicyclo[3.2.1]octan-3-one, the title product is obtained.
EXAMPLE 4
[1R,5S]1,2-Dihydro-1-methyl-6-{5-[3-(3.alpha.-methoxy-6,8-dioxabicyclo-3.2.1]-octanyl)]-3-fluorophenoxymethyl}quinolin-2-one
Following the procedure described in Example 2, but substituting [1R,5S]1,2-dihydro-1-methyl-6-{5-[3-(3.alpha.-hydroxy-6,8dioxabicyclo[3.2.1]octanyl)]-3-fluorophenoxymethyl}quinolin-2-one (Example 3) for [1S,5R]1,2-dihydro-1-methyl-6-{3-[3-(3.alpha.-hydroxy-6,8-dioxabicyclo[3.2.1]octanyl)]-5-fluoro phenoxymethyl}quinolin-2-one, the title product is obtained.
EXAMPLE 5
1,2-Dihydro-1-methyl-6-{5-[3-(3.alpha.-hydroxy-8-oxabicyclo[3.2.1]-octanyl)]-3-fluorophenoxymethyl}quinolin-2-one
Following the procedure described in Example 1, Steps 4-6, but substituting 8-oxabicyclo[3.2.1]oct-6-en-3-one (J.A.C.S. 100, 1765 (1978)) for [1S,5R]6,8-dioxabicyclo[3.2.1]octan-3-one, the title product is obtained.
EXAMPLE 6
1,2-Dihydro-1-methyl-6-{5-[3-(3.alpha.-methoxy-8-oxabicyclo[3.2.1]-octanyl)]-3-fluorophenoxymethyl}quinolin-2-one
Following the procedure described in Example 2, but substituting 1,2-dihydro-1-methyl-6-{5-[3-(3.alpha.-hydroxy-8-oxabicyclo[3.2.1]octanyl)]-3-fluorophenoxymethyl}quinolin-2-one (Example 5) for 1,2-dihydro-1-methyl-6-{5-[3-(3.alpha.-hydroxy-6,8-dioxabicyclo[3.2.1]octanyl)]-3-fluorophenoxymethyl}quinolin-2-one, the title product is obtained.
EXAMPLE 23
[1S,5R]2-Cyano-4-(3-furyl)-7-{5-[3-(3.alpha.-hydroxy-6,8-dioxabicyclo[3.2.1]octanyl]-3-fluorophenoxymethyl}quinoline
Step 1: 4-(3-Furyl)-7-methoxy-2-quinolinone
A solution of m-anisidine (6.2 mL), ethyl .beta.-oxo-3-furan-propionate (8.4 g) and pyridine (10 drops) in xylene (15 mL) was heated at reflux for 15 hr. The mixture was cooled to r.t. and concentrated. The residual material was subjected to chromatography (silica gel; hexane/EtOAc (2:1)), affording the .beta.-keto amide as an oil. A portion of this material (2.5 g), O-phosphoric acid (30 mL of 85% acid), and water (30 mL) was heated at 100.degree.-110.degree. C. for 5 hr. After cooling to room temperature, H.sub.2 O (100 mL) was added and the precipitate that formed was collected by filtration. The title compound was obtained as a mixture with the regioisomeric product, 4-(3-furyl)-5-methoxy-2-quinolinone.
Step 2: 4-(3-Furyl)-7-hydroxyquinoline
To a solution of 4-(3-furyl)-7-methoxy-2-quinolinone (4.1 g) from Step 1 in THF (250 mL) at reflux was added LiAlH.sub.4 (1M in THF; 68 mL). The mixture was refluxed for 50 hr., cooled to 0.degree. C., and then treated successively with H.sub.2 O (2.6 mL), 15% aq. NaOH (2.6 mL), and H.sub.2 O (7.8 mL). The precipitate that formed was removed by filtration, washing with THF, and the tiltrate was concentrated and redissolved in MeCN (100 mL). To this solution at r.t. was added a solution of Ce(NH.sub.4).sub.2 (NO.sub.3).sub.6 (8.7 g) in H.sub.2 O (30 mL). After the addition was complete, H.sub.2 O (500 mL) was added and the resulting mixture extracted with EtOAc (3x). The combined organics were washed with H.sub.2 O (2x) and brine, dried (MgSO.sub.4), and concentrated. Flash chromatography of the residue (silica gel; EtOAc/hexane (3:1)) provided 4-(3-furyl)-7-methoxyquinoline as a solid. A portion of this material (1.0 g) and pyridine hydrochloride (4 g) were heated at 175.degree.-180.degree. C. for 5 hr. While still hot, H.sub.2 O (50 mL) and ice was added, causing the formation of a precipitate. Filtration and drying of this material provided the title compound as a solid.
Step 3: 4-(3-Furyl)-7-carboethoxyquinoline
To a solution/suspension of 4-(3-furyl)-7-hydroxyquinoline (740 mg) from Step 2 in CH.sub.2 Cl.sub.2 (50 mL) at -78.degree. C. was added pyridine (1.3 mL) and TSA (1.2 mL). The mixture was stirred at ice bath temperature for 20 min and then H.sub.2 O was added. The organics were removed and the aqueous phase was extracted with CH.sub.2 Cl.sub.2 (3x). The combined organics were dried (MgSO.sub.4) and concentrated. Flash chromatography of the residue (silica gel; hexane/EtOAc (2:1 )) provided the triflate as a solid. A portion of this material (940 mg), Et.sub.3 N (0.8 mL) and bis(triphenylphosphine)palladium dibromide (217 mg) in absolute ethanol (85 mL) was stirred at 70.degree. C. under an atmosphere of CO (1 atm) for 18 hr. The mixture was cooled to r.t., filtered, and concentrated. Flash chromatography of the residue (silica gel; chloroform/EtOAc (4:1) provided the title compound as a solid.
Step 4: 4-(3-Furyl)-7-t-butyldimethylsiloxymethylquinoline
To a solution of 4-(3-furyl)-7-carboethoxyquinoline (583 mg) from Step 3 in THF (50 mL) at 0.degree. C. was added LiAlH.sub.4 (2.18 mL of a 1M solution in THF). After 15 min., H.sub.2 O (0.08 mL), 15% NaOH (0.08 mL), and H.sub.2 O (0.24 mL) were added successively. The mixture was filtered through Celite, washed with THF, and then concentrated. The residue was dissolved in DMF (10 mL) and treated with t-butyldimethylsilyl chloride (362 mg) and imidazole (327 mg). After 1 hr, H.sub.2 O was added and the mixture extracted with EtOAc (3x). The combined organics were washed with H.sub.2 O (3x), dried (MgSO.sub.4), and concentrated. Flash chromatography of the residue (silica gel; hexane/EtOAc (2:1)) provided the title compound as an oil.
Step 5: 2-Cyano-4-(3-furyl)-7-hydroxymethylquinoline
To a solution of 4-(3-furyl)-7-t-butyldimethylsiloxy methylquinoline from Step 4 (690 mg) in CHCl.sub.3 (40 mL) was added mCPBA (570 mg, 80-85% pure). After 1 hr, saturated aqueous NaHCO.sub.3 (50 mL) was added followed by H.sub.2 O (50 mL). The organics were removed and the aqueous phase was extracted with CHCl.sub.3 (2x). The combined organics were washed with H.sub.2 O, brine, dried (MgSO.sub.4), and concentrated. The residue was dissolved in CH.sub.2 Cl.sub.2 (15 mL) and then TMSCN (0.27 mL) was added. After 5 min, dimethylcarbamoyl chloride (0.19 mL) was added and the mixture stirred at r.t. for 18 hr. Additional TMSCN (0.27 mL) and dimethylcarbamoyl chloride (0.19 mL) were added and the mixture was refluxed for 2 hr. After cooling to r.t., 10% K.sub.2 CO.sub.3 (25 mL) was added followed by CH.sub.2 Cl.sub.2 (50 mL). The organics were removed and the aqueous layer neutralized with 25% NH.sub.4 OAc buffer. The aqueous was extracted with CH.sub.2 Cl.sub.2 (3x) and then the combined organics were washed with water, brine, dried (MgSO.sub.4), and concentrated. Flash chromatography of the residue (hexane/EtOAc (4:1)) provided the silyl ether of the title compound as a solid. A portion of this material (690 mg) dissolved in THF (30 mL) was treated with (n-Bu).sub.4 NF (2.1 mL) at 0.degree. C. After 30 min, 25% NH.sub.4 OAc buffer (50 mL) was added and the mixture was extracted with EtOAc (3x). The combined organics were washed with H.sub.2 O brine (MgSO.sub.4) and concentrated. The residue was triturated with hexane/Et.sub.2 O (1:1) to provide the title compound as a pale beige solid.
Step 6: [1S, 5R]2-Cyano-4-(3-furyl)-7-{5-[3-(3.alpha.-hydroxy-6,8-dioxabicyclo[3.2.1]octanyl]-3-fluorophenoxymethyl}-quinoline
A mixture of 2-cyano-4-(3-furyl)-7-hydroxymethylquinoline from Step 5 (72 mg), [1S, 5R]5-[3-(3.alpha.-hydroxy-6,8-dioxabicyclo[3.2.1]octanyl)]-3-fluorophenol from Example 1, Step 5 (76 mg), triphenylphosphine (98 mg) and di-t-butylazodicarboxylate (86 mg) in THF (5 mL) was stirred at r.t. for 30 min. The mixture was concentrated and the residue subjected to flash chromatography (silica gel; toluene/acetone (5:1), then hexane/EtOAc (1:1)). The title compound was obtained as a white solid, m.p. 158.5.degree.-159.5.degree. C.
EXAMPLE 24
[1S,5R]2-Cyano-4-(3-thienyl)-7-{5-[3-(3.alpha.-hydroxy-6,8-dioxabicyclo[3.2.1]octany]-3-fluorophenoxymethyl}quinoline
Following the procedures described in Example 23, but substituting ethyl .beta.-oxo-3-thiophenepropionate for ethyl .beta.-oxo-3-furanpropionate in Step 1, the title compound was obtained as a white solid, m.p. 189.degree.-190.degree. C.
EXAMPLE 27
[1S,5R]2-[2-(4,5-Dihydrothiazolyl)]-4-(3-furyl)-7-{3-[3-(3.alpha.-hydroxy-6,8-dioxabicyclo[3.2.1]octanyl]-5-fluorophenoxymethyl]quinoline
Step 1: 2-[2-(4,5-Dihydrothiazolyl)]-4-(3-furyl)-7-hydroxymethylquinoline
A mixture of 2-cyano-4-(3-furyl)-7-hydroxymethylquinoline (168 mg) (Example 23, Step 5) and 2-aminoethanethiol hydrochloride (380 mg) in chlorobenzene (8 mL) was refluxed for 22 hr. The mixture was cooled to r.t., diluted with EtOAc (50 mL) and washed with saturated NaHCO.sub.3. The aqueous phase was neutralized with 25% NH.sub.4 OAc buffer and extracted with EtOAc (3x). The combined organics were washed with H.sub.2 O (2x), brine, dried (MgSO.sub.4) and concentrated. The residue was triturated with hexane/EtOAc (1:1) to provide the title compound as a pale yellow solid.
Step 2: [1S,5R]2-[2-(4,5-Dihydrothiazolyl)]-4-(3-furyl)-7-{5-[3(3.alpha.-hydroxy-6,8-dioxabicyclo[3.2.1]octanyl]-3-fluorophenoxymethyl]quinoline
Following the procedure described in Example 23, Step 6, but substituting 2-[2-(4,5-dihydrothiazolyl)]-4-(3-furyl)-7-hydroxymethylquinoline for 2-cyano-4-(3-furyl)-7-hydroxymethylquinoline, the title compound was obtained as a colorless foam.
.sup.1 H NMR (300 MHz, acetone d.sub.6): .delta.1.90-2.05 (m, 2H), 2.16 (dd, 1H), 2.32 (dd, 1H), 3.40 (t, 2H), 3.64 (t, 1H), 4.31 (s, 1H), 4.49 (d, 1H), 4.56 (t, 2H), 4.62 (t, 1H), 5.42 (s, 2H), 5.61 (s, 1H), 6.75 (dt, 1H), 6.88 (ddd, 1H), 6.96 (dd, 1H), 7.09 (1H), 7.75-7.84 (m, 2H), 8.13-8.14 (m, 2H), 8.21-8.28 (m, 2H).
EXAMPLES 7-22, 25, 26, and 28-41
Using the procedures described above and the procedures described in EP 462,812, 462,813,462,830, and 462,831, the compounds of Examples 7-22, 25, 26, and 28-41 are prepared.

Number	Date	Country
0385662	Sep 1990	EPX
420511	Apr 1991	EPX
0462812	Dec 1991	EPX
0462813	Dec 1991	EPX
0462830	Dec 1991	EPX
0462831	Dec 1991	EPX
466452	Jan 1992	EPX
501578	Sep 1992	EPX
501579	Sep 1992	EPX

Arylbicyclooctanes as inhibitors of leukotriene biosynthesis

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