WO1993022285A1 - Leukotriene antagonists - Google Patents

Abstract

This invention relates to a compound of formula (I) where the several groups are defined herein. These compounds are leukotriene antagonists and as such can be used in treating various diseases associated with leuktrienes.

Description

LEUKOTRIENE ANTAGONISTS

The field of this invention is that of certain mono and diacids which have been found to be useful for treating diseases arising from or related to leukotrienes, particularly leukotriene B₄. As such there utility lies in antagonizing the affects of leukotrienes.

Background of the Invention

The family of bioactive lipids known as the leukotrienes exert

pharmacological effects on respiratory, cardiovascular and gastrointestinal systems. The leukotrienes are generally divided into two sub-classes, the peptidoleukotrienes

(leukotrienes C₄, D₄ and E₄) and the dihydroxyleukotrienes (leukotriene B₄). This invention is primarily concerned with the hydroxy leukotrienes (LTB) but is not limited to this specific group of leukotrienes.

The peptidoleukotrienes are implicated in the biological response associated with the "Slow Reacting Substance of Anaphylaxis" (SRS-A). This response is expressed in vivo as prolonged bronchoconstriction, in cardiovascular effects such as coronary artery vasoconstriction and numerous other biological responses. The pharmacology of the peptidoleukotrienes include smooth muscle contractions, myocardial depression, increased vascular permeability and increased mucous production.

By comparison, LTB₄ exerts its biological effects through stimulation of leukocyte and lymphocyte functions. It stimulates chemotaxis, chemokinesis and aggregation of polymorphonuclear leukocytes (PMNs).

Leukotrienes are critically involved in mediating many types of

cardiovascular, pulmonary, dermatological, renal, allergic, and inflammatory diseases including asthma, adult respiratory distress syndrome, cystic fibrosis, psoriasis, and inflammatory bowel disease.

Leukotriene B₄ ( LTB₄) was first described by Borgeat and Samuelsson in

1979, and later shown by Corey and co-workers to be 5(S),12(R)-dihydroxy-(Z,E,E,Z)-6,8,10,14-eicosatetraenoic acid.

It is a product of the arachidonic acid cascade that results from the enzymatic hydrolysis of LTA₄. It has been found to be produced by mast cells,

polymorphonuclear leukocytes, monocytes and macrophages. LTB₄ has been shown to be a potent stimulus in vivo for PMN leukocytes, causing increased chemotactic and chemokinetic migration, adherence, aggregation, degranulation, superoxide production and cytotoxicity. The effects of LTB₄ are mediated through distinct receptor sites on the leukocyte cell surface that exhibit a high degree of stereospecificity. Pharmacological studies on human blood PMN leukocytes indicate the presence of two classes of LTB₄-specific receptors that are separate from receptors specific for the peptide chemotactic factors. Each of the sets of receptors appear to be coupled to a separate set of PMN leukocyte functions.

Calcium mobilization is involved in both mechanisms.

LTB₄ has been established as an inflammatory mediator in vivo. It has also been associated with airway hyper-responsiveness in the dog as well as being found in increased levels in lung lavages from humans with severe pulmonary dysfunction.

By antagonizing the effects of LTB₄, or other pharmacologically active mediators at the end organ, for example airway smooth muscle, the compounds and pharmaceutical compositions of this invention are valuable in the treatment of diseases in subjects, including human or animals, in which leukotrienes are a factor.

Summary of the Invention

In a first aspect, this invention covers a compound of formula I

or an N-oxide, or a pharmaceutically acceptable salt, where

m is 1 - 8;

R is C₁ to C₂₀-aliphatic, unsubstituted or substituted phenyl- C₁ to C₁₀-aliphatic where substituted phenyl has one or more radicals selected from the group consisting of lower alkoxy, lower alkyl, trihalomethyl, and halo, or R is C₁ to C₂₀-aliphatic-O-, or R is unsubstituted or substituted phenyl-C₁ to C₁₀-aliphatic-O- where substituted phenyl has one or more radicals selected from the group consisting of lower alkoxy, lower alkyl, trihalomethyl, and halo;

R₁ is R₃, -(C₁ to C₅ aliphatic)R₃, -(C₁ to C₅ aliphatic)CHO, -(C₁ to C₅ aliphatic)CH₂OR₄;

R₂ is H, R₃, -(C₁ to C₅ aliphatic)R₃ or tetrazol-5-yl;

R₃ is tetrazol-5-yl or COOH or an ester or amide thereof; and

R₄ is hydrogen, C₁ to C₅ alkyl, or C₁ to C₆ acyl. In a further aspect, this invention relates to compositions comprising a compound of formula I, or a salt thereof, in admixture with a carrier. Included in these compositions are those suitable for pharmaceutical use and comprising a pharmaceutically acceptable excipient or carrier and a compound of formula I which may be in the form of a pharmaceutically acceptable salt.

Processes for making these compounds are also included in the scope of this invention, which processes comprise:

a) forming a salt, or

b) forming an ester;

c) oxidizing a thio ether to the sulfoxide or sulfone;

d) forming a compound of formula I by treating a 6-halomethylpyridyl compound with the appropriate mercaptan, or hydroxy compound.

General Embodiments

The following definitions are used in describing this invention.

"Aliphatic" is intended to include saturated and unsaturated radicals. This includes normal and branched chains, saturated or mono or poly unsaturated chains where both double and triple bonds may be present in any combination. The phrase "lower alkyl" means an alkyl group of 1 to 6 carbon atoms in any isomeric form, but particularly the normal or linear form. "Lower alkoxy" means the group lower alkyl-O-. "Acyl-lower alkyl" refers to the group (O)C-lower alkyl where the carbonyl carbon is counted as one of the carbons of the 1 to 6 carbons noted under the definition of lower alkyl. "Halo" refers to and means fluoro, chloro, bromo or iodo. The phenyl ring may be substituted with one or more of these radicals.

Multiple substituents may be the same or different, such as where there are three chloro groups, or a combination of chloro and alkyl groups and further where this latter combination may have different alkyl radicals in the chloro/alkyl pattern.

The phrase "a pharmaceutically acceptable ester-forming group" covers all esters which can be made from the acid function(s) which may be present in these compounds. The resultant esters will be ones which are acceptable in their application to a pharmaceutical use. By that it is meant that the mono or diesters will retain the biological activity of the parent compound and will not have an untoward or deleterious effect in their application and use in treating diseases.

Amides may be formed from acid groups. The most preferred amides are those where the nitrogen is substituted by hydrogen or alkyl of 1 to 6 carbons. The diethylamide is particularly preferred.

Pharmaceutically acceptable salts of the instant compounds are also intended to be covered by this invention. These salts will be ones which are acceptable in their application to a pharmaceutical use. By that it is meant that the salt will retain the biological activity of the parent compound and the salt will not have untoward or deleterious effects in its application and use in treating diseases.

Pharmaceutically acceptable salts are prepared in a standard manner. The parent compound, dissolved in a suitable solvent, is treated with an excess of an organic or inorganic acid, in the case of acid addition salts of a base, or an excess of organic or inorganic base where R₂ is COOH for example.

Oxides of the pyridyl ring nitrogen may be prepared by means known in the an and as illustrated herein. These are to be considered pan of the invention.

If by some combination of substituents, a chiral center is created or another form of an isomeric center is created in a compound of this invention, all forms of such isomer(s) are intended to be covered herein. Compounds with a chiral center may be administered as a racemic mixture or the racemates may be separated and the individual enantiomer used alone.

As leukotriene antagonists, these compounds can be used in treating a variety of diseases associated with or attributing their origin or affect to

leukotrienes, particularly LTB₄. Inflammatory diseases such as psoriasis and inflammatory bowel disease may be treated by applying or administering the compounds described herein. It is also expected that these compounds can be used to treat allergic diseases including those of a pulmonary and non-pulmonary nature. For example these compounds will be useful in antigen-induced anaphylaxis. They are useful in treating asthma and allergic rhinitis. Ocular diseases such as uveitis, and allergic conjunctivitis can also be treated by these compounds.

Preferred compounds are those where R is C₈ to C₂₀ alkoxy, phenyl-C₄ to

C₁₀ alkoxy or substituted-phenylC₄ to C₁₀ alkoxy; R₁ -(C₁-C₃alkyl)R₃, or -(C₂- C₃alkenyl)R₃ and R₂ is R₃, -(C₁ to C₅ aliphatic)_R3 or tetrazol-5-yl. The more preferred compounds are those where R is substituted phenyl-C₄ to C₁₀ alkoxy, particularly the substituted-phenyl(CH₂)_4-8-O- group or CH₃(CH₂)_7-9-O-; m is 2 -6, most preferably 2, 3 or 4; R₁ is HO₂C-CH=CH-, or HO₂C-CH₂CH₂- or a salt, ester or amide derivative thereof and R₂ is tetrazol-5-yl or COOH or an ester or amide thereof. The most preferred compounds are:

3-[6-(carboxyethanyl)-5-[8-(4-methoxyphenyl)octyloxy]pyrid-2-yl]ethanyl beπzoic acid, dilithium salt;

(E)-3-[3-[4-(4-methoxyphenyl)butyioxy]-6-[(E)-3-oxo-3-phenyl-1-propenyl]-2-pyridinyl]-2-propenoic acid, lithium salt; 3-[3-[4-(4-methoxyphenyl)butyloxy]-6-(3-phenylpropyl)-2-pyridinyljpropanoic acid, lithium salt;

(E)-3-[3-[4-(4-methoxyphenyl)butyloxy]-6-(2-phenylethyl)-2-pyridinyl]-2-propenoic acid, lithium salt;

3-[3-[4-(4-methoxyphenyl)butyloxy]-6-(2-phenylethyl)-2-pyridinyl]propanoic acid, lithium salt; or

another pharmaceutically acceptable salt.

Synthesis

Several methods, variations on the same process, have been used for

preparing these compounds. In general, the approach taken was to first make the intermediates needed to make the R group, then to prepare the phenyl intermediate needed for forming the core structure of formula I; the pyridyl intermediate was then prepared and reacted with the phenyl intermediate to form the basic skeletal core structure. Hydrogenation procedures were used to saturate the alkene chain to make the -(CH₂)_m group. Salts, free acids, amides, alternative esters and the like were then prepared.

As noted, the first step was to make the intermediates needed for forming those R groups where the intermediates were not available commercially. This

chemistry is illustrated for the case of the substituted phenyl-C₁ to C₁₀-aliphatic-O- groups. The same or similar chemistry has been disclosed in published patent

applications, for example PCT international application numbers PCT/US91/03772, PCT/US91/03940, and PCT/US91/03399. The chemistries set out in those

documents can be used in place of or in conjunction with those given here to prepare the R groups of formula I.

A second aspect of the synthesis is preparing the substituted phenyl

intermediates; this is in reference to the phenyl group on the right hand side of

formula I. For the case where m is 2, this involves treating a bromobenzoate with trimethylsilyl acetylene and Pd(OAc)₂ and Ph3P with heating for up to a day in

order to prepare a ethynyl benzoate. This chemistry can be used to prepare

positional isomers and analogs of the acid where the aliphatic chain contains two or more carbons or where phenyl is substituted by one of the other radicals which fall within the defintion of R₂. An inert atmosphere should be used when carrying out this reaction. The TMS group is removed with a weak base, eg potassium carbonate using a alcoholic solvent, and the resulting ethynyl compound is ready for use in the reaction which creates the unsaturated analogs of formula I.

Making the substituted pyridyl triflate can begin with the starting compound and the chemistry disclosed in the PCT application PCT/US91/03940 and the other PCT cases cited above. The chemistry set out in the '03940 case can be use to convert the starting material, 3-hydroxy-2(hydroxymethyl)pyridine HCl, through to the 6-(E-2-carboxymethylethenyl)-5-[8-(4-methoxyphenyl)octyloxy]-2-pyridone, the synthesis outlined in Scheme II below. This compound is then treated with trifluoromethanesulfonic anhydride to make the triflate. The ethynyl benzoate is then added to the 6 position by combining the triflate and the ethynyl benzoate in the presence of Pd(Ph₃P)₂Cl₂ using conventional conditions. Catalytic

hydrogenation is then used to reduce the triple bond (the double bond R₁ at the 2-position is also reduced). Base, or acid, can then be used to hydrolyze the ester groups, if so desired. The free acid can be obtained from the salt by acidifying a solution of the salt. Esters and amide can be prepared using standard reaction conditions and reagents. Tetrazoles are prepared from the corresponding acid halide, e.g., the acid chloride, by literature methods.

Scheme I illustrates one way to prepare an intermediate which is useful for making the R group in formula I.

Scheme I

The starting alcohol, represented here as the 3-octyn-1-ol, is commercially available (Lancaster Synthesis). To migrate the triple bond to the co-carbon, KH and 1,3-diaminopropane are combined and stirred to a homogeneous mix. This can be done at ambient temperature or thereabouts. This mix is then cooled, preferably to about 0°C or thereabouts, whereupon the alcohol is added. Stirring is then commenced at about room temperature for 15 to 20 hours or so. Water is added to quench the reaction and the product is recovered.

Protecting the alcohol is accomplished by forming a silyl ether illustrated here as the t-butyldiphenylsilyl ether. Other silyl ethers could be used. The alcohol is dissolved in a polar solvent, for example dimethylformamide, and imidazole is added followed by the desired silane. All this is carried out under an inert atmosphere such as argon. Ambient temperature is acceptable for effecting the reaction.

Adding the phenyl group is done in a dry environment using an amine for a solvent and an inert atmosphere. To a flask containing a solvent such as

triethylamine under argon is added the silylether followed by a halophenyl compound, eg. iodoanisole, a palladium catalyst (Ph₃P)₂PdCl₂ and CuI, both of the latter in catalytic amounts. Heat is used to effect the reaction, usually a temperature of up to about 50°C will be sufficient. Two or more hours, up to six but often about four at the elevated temperature will usually cause the reaction to go to completion.

The triple bond is then hydrogenated, preferably by catalytic hydrogenation.

For example, the silyl ether can be dissolved in a saturated solvent such as an alcohol, a heavy metal catalyst added (Pd-C) and the mixture put under H₂ for a time sufficient to reduce the triple bond. Stirring for 2 to 6 hours will usually effect the reaction.

Recovering the alcohol is done by treating the silyl ether with a fluoride source such as tetrabutylammonium fluoride. Reactants are combined at a mildly reduced temperature, eg. 0°C, then the reaction is allowed to run its course at ambient temperature or there about. Several hours may be needed for the reaction to go to completion. Product was recovered by extraction means. Methods for making these compounds are illustrated in the following reaction schemes.

A method for preparing the alkynyl phenyl precursor is illustrated in Scheme II.

Scheme II

The reagents illustrated in this Scheme can be varied to suit that which is needed to make the intended form of formula I.

When making the TMS-ethynylbenzene compound, the reaction vessel should be dry and the reaction should be carried out under an inert atmosphere, argon is preferred. A solvent such as diethylamine can be used. The bromobenzene adduct is first dissovled in the solvent, then an excess of the trimethylsilyl acetylene (about a 50% excess) is added along with a catalytic amount of Pd(OAc)₂ and Ph₃P. An elevated temperature, about 50 to 150°C depending on the solvent, is used to effect the reaction. About 10 to 20 hours is required for the reaction to go to completion. After the product is recovered, a weak base is used to hydrolyze the TMS group. Potassium carbonate is preferred using a simple alcohol as the solvent.

Using the precursors prepared as per Schemes I and II, or compounds which have been purchased from a commercial source, and following the steps outlined in Scheme III which follows, one can prepare compounds of formula I where m is 2.

Scheme III

A general description of the conditions and reagents which can be used for converting the diol to the pyrid-6-one compound can be found in PCT application number PCT/US91/03940. The triflate is prepared by dissolving the pyridone in an inert solvent such as dichloromethane under conditions which exclude water and employs an inert atmosphere. This solution is best cooled to around 0 °C or thereabouts. Pyridineis added followed by trifluoromethanesulfonic anhydride after which the pot is stirred for a few minutes; 10 to 20 minutes should be sufficient to effect the reaction. Product is recovered using standard procedures.

Triflate and the ethynylbenzene compound are coupled by first dissolving the triflate in a solvent such as dimethylformamide (DMF) under conditions excluding water and which has an inert atmosphere, then adding the ethynyl compound and an amine such as diethylamine, Cul and Pd(Ph₃P)₂Cl₂. This mixture is heated to 50 to 100 °C or thereabouts for 1 to 10 hours, usually about 5. About a two-fold excess of the ocetylinic compound is used and about one equivalent of the amine, relative to the triflate is employed. The Cul and

Pd(Ph₃P)₂Cl₂ are used in catalytic amounts. Normal sepratory and

chromatographic means are used to isolate the product. Catalyitc hydrogenation is used to saturate the acetylinic group; this hydrogenates the double bond in the 2-position group as well. Base or acid can be used to hydrolyze the esters, which gives the salt or the free acid. Amides, esters, and the like can be prepared by conventional means as noted above.

Compounds where m is 3 can be prepared by following the steps outlined in Scheme IV. The first two compounds in this Scheme illustrate the specific case of where R is a substituted phenylbutyl group and is made, for all intents and purposes, by the same means set out in Scheme I above.

The general case for the chemistry for converting the starting diol to the aldehyde, and so on down throught the fully substituted carboxaldehyde has been previously described in the above cited PCT published patent applications. That description of the generalized case for each step is incorporated herein by reference.

Forming the structure labeled 2 is accomplished by first dissolving a compound such as phenylacyl triphenylphosphonium bromide in a dry solvent such as toluene under an inert atmosphere and then adding sodium hydride. This mixture is heated to an intermediate temperature, eg 45 °C for several hours, 2 to 6 hours is preferable. Thereafter the mixture is cooled and the carboxaldehyde is added in the same solvent. The reaction is then allowed to proceed at about room temperature for up to several days, 1 to 2 days is preferred. In this procedure, an excess of the phenacyl compound relative to the carboxaldehyde is used. The product is worked up using standard techniques. In this case the ester was hydrolyzed, or the double bonds were hydrogenated and the ketone reduced prior to hydrolysis as illustrated. A heavy metal catalyst is useful for reducing these functional groups.

Pharmaceutical compositions of the present invention comprise a

pharmaceutical carrier or diluent and some amount of a compound of the formula (I). The compound may be present in an amount to effect a physiological response, or it may be present in a lesser amount such that the user will need to take two or more units of the composition to effect the treatment intended. These compositions may be made up as a solid, liquid or in a gaseous form. Or one of these three forms may be transformed to another at the time of being administered such as when a solid is delivered by aerosol means, or when a liquid is delivered as a spray or aerosol.

Included within the scope of this disclosure is the method of treating a disease mediated by LTB₄ which comprises administering to a subject a

therapeutically effective amount of a compound of formula I, preferably in the form of a pharmaceutical composition. For example, inhibiting the symptoms of an allergic response resulting from a mediator release by administration of an effective amount of a compound of formula I is included within the scope of this disclosure. The administration may be carried out in dosage units at suitable intervals or in single doses as needed. Usually this method will be practiced when relief of symptoms is specifically required. However, the method is also usefully carried out as continuous or prophylactic treatment. It is within the skill of the an to determine by routine experimentation the effective dosage to be administered from the dose range set forth above, taking into consideration such factors as the degree of severity of the condition or disease being treated, and so forth.

The nature of the composition and the pharmaceutical carrier or diluent will, of course, depend upon the intended route of administration, for example parenterally, topically, orally or by inhalation.

For topical administration the pharmaceutical composition will be in the form of a cream, ointment, liniment, lotion, pastes, aerosols, and drops suitable for administration to the skin, eye, ear, or nose.

For parenteral administration the pharmaceutical composition will be in the form of a sterile injectable liquid such as an ampule or an aqueous or non-aqueous liquid suspension.

For oral administration the pharmaceutical composition will be in the form of a tablet, capsule, powder, pellet, atroche, lozenge, syrup, liquid, or emulsion.

When the pharmaceutical composition is employed in the form of a solution or suspension, examples of appropriate pharmaceutical carriers or diluents include: for aqueous systems, water; for non-aqueous systems, ethanol, glycerin, propylene glycol, com oil, cottonseed oil, peanut oil, sesame oil, liquid parafins and mixtures thereof with water; for solid systems, lactose, kaolin and mannitol; and for aerosol systems, dichlorodifluoromethane, chlorotrifiuoroethane and compressed carbon dioxide. Also, in addition to the pharmaceutical carrier or diluent, the instant compositions may include other ingredients such as stabilizers, antioxidants, preservatives, lubricants, suspending agents, viscosity modifiers and the like, provided that the additional ingredients do not have a detrimental effect on the therapeutic action of the instant compositions.

The pharmaceutical preparations thus described are made following the conventional techniques of the pharmaceutical chemist as appropriate to the desired end product.

In these compositions, the amount of carrier or diluent will vary but preferably will be the major proportion of a suspension or solution of the active ingredient. When the diluent is a solid it may be present in lesser, equal or greater amounts than the solid active ingredient.

Usually a compound of formula I is administered to a subject in a

composition comprising a nontoxic amount sufficient to produce an inhibition of the symptoms of a disease in which leukotrienes are a factor. Topical formulations will contain between about 0.01 to 5.0% by weight of the active ingredient and will be applied as required as a preventative or curative agent to the affected area. When employed as an oral, or other ingested or injected regimen, the dosage of the composition is selected from the range of from 50 mg to 1000 mg of active ingredient for each administration. For convenience, equal doses will be

administered 1 to 5 times daily with the daily dosage regimen being selected from about 50 mg to about 5000 mg.

No unacceptable toxicological effects are expected when these compounds are administered in accordance with the present invention.

Bioassays

The specificity of the antagonist activity of a number of the compounds of this invention is demonstrated by relatively low levels of antagonism toward agonists such as potassium chloride, carbachol, histamine and PGF₂.

The receptor binding affinity of the compounds used in the method of this invention is measured by the ability of the compounds to bind to [³H]-LTB₄ binding sites on human U937 cell membranes. The LTB₄ antagonist activity of the compounds used in the method of this invention is measured by their ability to antagonize in a dose dependent manner the LTB₄ elicited calcium transient measured with fura-2, the fluorescent calcium probe. The methods employed were as follows:

U937 Cell Culture Conditions

U937 cells were obtained from Dr. John Bomalaski (Medical College of PA) and Dr. John Lee (SmithKline Beecham Corp., Dept. of Immunology) and grown in RPMI-1640 medium supplemented with 10% (v/v) heat inactivated fetal calf serum, in a humidified environment of 5% CO_2, 95% air at 37°C. Cells were grown both in T-flasks and in Spinner culture. For differentiation of the U937 cells with DMSO to macrophage-like cells, the cells were seeded at a concentration of 1 × 10⁵ cells/ml in the above medium with 1.3% DMSO and the incubation continued for 4 days. The cells were generally at a density of 0.75-1.25 × 10⁶ cells/ml and were harvested by centrifugation at 800 x g for 10 min.

Preparation of U937 Cell Membrane Enriched Fraction

Harvested U937 cells were washed with 50 mM Tris-HCl, pH 7.4 at 25° C containing 1 mM EDTA (buffer A). Cells were resuspended in buffer A at a concentration of 5 × 10⁷ cells/ml and disrupted by nitrogen cavitation with a Parr bomb at 750 psi for 10 min at 0° C. The broken cell preparation was centrifuged at 1,000 x g for 10 min. The supernatant was centrifuged at 50,000 x g for 30 min. The pellet was washed twice with buffer A. The pellet was resuspended at about 3 mg membrane protein/ml with 50mM Tris-HCl, pH 7.4 at 25°C and aliquots were rapidly frozen and stored at -70°C.

Binding of [³H]-LTB₄ to U937 Membrane Receptors

[³H]-LTB₄ binding assays were performed at 25° C, in 50 mM Tris-HCl (pH 7.5) buffer containing 10 mM CaCl₂, 10 mM MgCl₂, [³H]-LTB₄, U937 cell membrane protein (standard conditions) in the presence or absence of varying concentrations of LTB₄, or test compounds. Each experimental point represents the means of triplicate determinations. Total and non-specific binding of [³H]-LTB₄ were determined in the absence or presence of 2 μM of unlabeled LTB₄,

respectively. Specific binding was calculated as the difference between total and non-specific binding. The radioligand competition experiments were performed, under standard conditions, using approximately 0.2 nM [³H]-LTB₄, 20-40 μg of U937 cell membrane protein, increasing concentrations of LTB₄ (0.1 μM to 10 μM) or other competing ligands (0.1 μM to 30 μM) in a reaction volume of 0.2 ml and incubated for 30 minutes at 25° C. The unbound radioligand and competing drugs were separated from the membrane bound ligand by a vacuum filtration technique. The membrane bound radioactivity on the filters was determined by liquid scintillation spectrometry.

Saturation binding experiments for U937 cells were performed, under standard conditions, using approximately 15-50 mg of U937 membrane protein and increasing concentrations of [³H]-LTB₄ (0.02-2.0 nM) in a reaction volume of 0.2 ml and incubation at 22°C, for 30 minutes. LTB₄ (2 mM) was included in a separate set of incubation tubes to determine non-specific binding. The data from the saturation binding experiments was subjected to computer assisted non-linear least square curve fitting analysis and further analyzed by the method of Scatcnard. Loading by Differentiated U937 Cells with Fura-2

Harvested cells were resuspended at 2 × 10⁶ cells/ml in Krebs Ringer Hensilet buffer containing 0.1% BSA (RIA grade), 1.1 mM MgSO₄, 1.0 mM CaCl₂ and 5 mM HEPES (pH 7.4, buffer B). The diacetomethoxy ester of fura-2 ((ura-2/AM) was added to a final concentration of 2 μM and cells incubated in the dark for 30 minutes at 37° C. The cells were centrifuged at 800 x g for 10 minutes and resuspended at 2 × 10⁶ cells/ml in fresh buffer B and incubated at 37°C for 20 minutes to allow for complete hydrolysis of entrapped ester. The cells were centrifuged at 800 x g for 10 minutes and resuspended in cold fresh buffer B at 5 × 106 cells/ml. Cells were maintained on ice in the dark until used for fluorescent measurements.

Fluorescent Measurements-Calcium Mobilization

The fluorescence of fura-2-containing U937 cells was measured with a fluorometer designed by the Johnson Foundation Biomedical Instrumentation

Group. A fluorometer was equipped with temperature control and a magnetic stirrer under the cuvette holder. The wave lengths are set at 339 nm for excitation and 499 nm for emission. All experiments were performed at 37°C with constant mixing.

U937 cells were diluted with fresh buffer to a concentration of 1 × 10⁶ cells/ml and maintained in the dark on ice. Aliquots (2 ml) of the cell suspension were put into 4 ml cuvettes and the temperature brought up to 37°C, (maintained in 37°C, water bath for 10 min). Cuvettes were transferred to the fluorometer and fluorescence measured for about one minute before addition of stimulants or antagonists and followed for about 2 minutes post stimulus. Agonists and antagonists were added as 2 μl aliquots.

Antagonists were added first to the cells in the fluorometer in order to detect potential agonist activity. Then after about one minute 10 nM LTB₄ (a near maximal effective concentration) was added and the maximal Ca²⁺ mobilization [Ca²⁺]_i was calculated using the following formula:

F was the maximum relative fluorescence measurement of the sample. Fmax was determined by lysing the cells with 10 μl of 10% Triton X-100 (final Concentration 0.02%). After ¥max was determined 67 μl of 100 mM EDTA solution (pH 10) was added to totally chelate the Ca²⁺ and quench the fura-2 signal and obtain the Fmin. The [Ca²⁺]_i level for 10 nM LTB₄ in the absence of an antagonist was 100% and basal [Ca²⁺]_i was 0%. The IC₅₀ concentration is the concentration of antagonist which blocks 50% of the 10nM LTB₄ induced [Ca²⁺]_i mobilization. The EC₅₀ for LTB₄ induced increase in [Ca²⁺]_i mobilization was the concentration for half maximal increase. The K_i for calcium mobilization was determined using the formula:

With the experiments described, the LTB₄ concentration was 10 nM and the EC₅₀ was 2 nM.

Specific Embodiments

The following examples are given to illustrate how to make and use the compounds of this invention. These Examples are just that, examples, and are not intended to circumscribe or otherwise limit the scope of this invention. Reference is made to the claims for defining what is reserved to the inventors.

Example 1

3-[6-(Carboxyethanyl)-5-[8-(4-methoxyphenyl)octyloxylpyrid-2-yl]ethanyl benzoic acid, dilithium saltSB 201044

1A 7-Octyn-1-ol. 35% KH in mineral oil (27g, 240mmol) under an argon atmosphere was washed with hexane and treated dropwise with 1,3-diaminopropane.

The mixture was stirred at room temperature until it became homogeneous. The flask was cooled to 0 °C and 3-octyn-l-ol (10g, 79mmol, Lancaster Synthesis) was slowly added. The reaction was then stirred at room temperature for 18h. The reaction was quenched with H₂O (50mL) and the product was extracted into ether.

The organic layer was washed with 10% HCl and brine and dried (MgSO₄).

Evaporation gave 9.73g (97%) of product as a colorless oil which was used without further purification: ¹H NMR (90MHz, CDCI₃) 63.65 (t, J=5Hz, 2H, O-CH₂), 2.23 (m, 2H, CH₂), 2.0 (m, 1H, acetylenic), 1.7-1.2 (m, 8H, (CH₂)₄); IR (neat) v_max 3350, 2930, 2125 cm^{- 1}.

1B 7-Octyn-1-¹butyldiphenylsilyl ether. To a cooled (0 °C) solution of 7-octyn- 1-ol (9.3g, 73.7mmol) in DMF (70mL) under an argon atmosphere was added imidazole (7.5g, 110mmol) followed by the dropwise addition of ¹butylcniorodtpnenyisttane (21mL, 81mmol). The reaction was tnen stirrea at room temperature for 2 hours. The reaction solution was diluted with Et₂O and washed with H₂O and brine and dried (MgSO₄). Purification by flash column

chromatography (silica, 3% EtOAc in hexane ) provided 24.9g (93%) as a colorless oil: ¹H NMR (250MHz, CDCl₃) δ 7.7 (d, 4H, phenyl), 7.4 (m, 6H, phenyl), 3.63 (t, 2H, O-CH₂), 2.23 (m, 2H, CH₂), 1.97 (t, 1H, acetylenic), 1.6-1.3 (m, 8H, (CH₂)₄), 1.05 (s, 9H, ¹butyl); IR (film) v_max 3321, 2940, 2125 cm^-1.

1C 8-(4-MethoxyphenyI)-7-octyn-1-¹butyldiphenylsilyl ether. To a flame dried flask containing triethylamine (140mL) under an argon atmosphere was added 4-iodoanisole (13.3g, 56.9mmol), 7-octyn-1-^tbutyldiphenylsilyl ether (24.9g, 68.3mmol), (Ph₃P)₂PdCl₂ catalyst (793mg, 1.13mmol), and CuI (431mg,

2.27mmol). The resulting mixture was heated at 50 °C for 4 hours. Upon cooling to room temperature the reaction mixture was filtered, the solids were washed with Et₂O and the solvent was evaporated. The residue was diluted with Et₂O and washed with 5% HCl, H₂O, NaHCO₃, and brine and dried (MgSO₄). Purification by flash column chromatography (silica, 2% EtOAc in hexane) gave 30g (93%) as an orange oil: ¹HNMR (250MHz, CDCl₃) δ 7.7 (d, 4H, phenyl), 7.4 (m, 6H, phenyl), 7.35 (d, 2H, phenyl), 6.8 (d, 2H, phenyl), 3.8 (s, 3H, OMe), 3.7 (t, 2H, O-CH₂), 2.4 (t, 2H, CH₂), 1.7-1.3 (m, 8H, (CH₂)₄), L05 (s, 9H, ^¹butyl). 1D 8-(4-Methoxyphenyl)octan-1-¹hutyldiphenylsilyl ether.

8-(4-Methoxyphenyl)-7-octyn-1-¹butyldiphenylsilyl ether (30g, 63.7mmol) was dissolved in EtOH (125mL) and EtOAc (125mL) and treated with 5% Pd-C catalyst (3g). The reaction was vigorously stirred under an H₂ atmosphere (balloon pressure) for 4 hours. The reaction mixture was filtered through a pad of Celite and the solvent was evaporated. The resulting pale yellow oil was pure by nmr analysis and was used directly for the next step: -B. NMR (250MHz, CDCl₃) δ 7.7 (d, 4H, phenyl), 7.4 (m, 6H,phenyl), 7.05 (d, 2H, phenyl), 6.8 (d, 2H, phenyl), 3.8 (s, 3H, OMe), 3.6 (t, 2H, O-CH₂), 2.5 (t, 2H, benzylic), 1.75-1.3 (m, 12H, (CH₂)6), 1.0 (s, 9H, ¹butyl).

1E 8-(4-Methoxyphenyl)octan-1-ol. To a cooled (0 °C) solution of

8-(4-methoxyphenyl)octan-1-¹butyldiphenylsilyl ether (63mmol) was added tetrabutylammonium fluoride (70mL, 70mmol; IM solution in THF). The cooling bath was removed and the reaction was stirred at room temperature for 4.5 hours. The solvent was evaporated and the residue was dissolved in Et₂O. This was washed with H₂O, 5% HCl, NaHCO₃, and brine and dried (MgSO₄). Purification by flash column chromatography (silica, 30% EtOAc in hexane) gave 12.6g (85%; two steps) as a colorless solid: ¹H NMR (250MHz, CDCl₃) δ 7.15 (d, 2H, phenyl), 6.86 (d, 2H, phenyl), 3.85 (s, 3H, OMe), 3.68 (t, 2H, O-CH₂), 2.62 (t, 2H, benzylic), 1.75-1.3 (m, 12H, (CH₂)₆); MS (CI): 254.2 (M+NH₄); mp 47-49 °C.

1F 1 -Iodo-8-(4-methoxyphenyl)octane. To a stirred solution of 8-(4-methoxyphenyl)octan-1-ol (12.3g, 52mmol) in dry toluene (200mL) under an argon atmosphere was added triphenylphosphine (17.8g, 67.6mmol) and imidazole (10.6g, 156mmol). After the imidazole had dissolved I₂ (17.1g, 67.6mmol) was added. The reaction was then heated at 65 °C for 30 minutes. Upon cooling to room temperature the reaction was concentrated to 1/4 volume. The remaining solution was diluted with Et₂O and washed with H₂O and brine and dried (MgSO₄). The solvent was removed and the resulting residue was dissolved in CH₂Cl₂ and applied to a flash chromatography column (silica). Elution with 2% EtOAc in hexane provided the captioned 16.3g (90%) of product as a colorless oil (slight

contamination with triphenylphosphine): ¹H NMR (250MHz, CDCl₃) δ 7.08 (d, J=8.6Hz, 2H, phenyl), 6.82 (d, J=8.6Hz, 2H, phenyl), 3.78 (s, 3H, OMe), 3.17 (t, J=7.4Hz, 2H, I-CH₂), 2.54 (t, J=7.6Hz, 2H, benzylic), 1.85 (m, 2H, CH₂), 1.60 (m, 2H, CH₂), 1.31 (m, 8H, aliphatic); MS (CI): 364.2 (M+NH₄).

1G 3-Hydroxy-2-pyridine carboxaldehyde.

3-Hydroxy-2-(hydroxymethyl)pyridine hydrochloride (3.56g, 19mmol, Aldrich, 85%) was suspended in CH₂Cl₂ (40mL) under an argon atmosphere and

triethylamine (4.0mL, 30mmol) was added; the solution was stirred for 10 minutes. MnO₂ (11.3g, 133mmol) was added and the reaction was stirred at room

temperature for 18 hours. The reaction mixture was filtered through Celite and concentrated in vacuo. The crude aldehyde was used directly in the next step.

1H 3-[8-(4-Methoxyphenyl)octyloxyl-2-pyridine carboxaldehyde. The crude aldehyde obtained above was dissolved in dry DMF (25mL) and treated with 1-iodo-8-(4-methoxyphenyl)octane (6.6g, 19mmol) and anhydrous K₂CO₃ (13.1g, 95mmol). The reaction was vigorously stirred at 90 °C under an argon atmosphere for 4 hours. Upon cooling to room temperature the reaction mixture was diluted with EtOAc and washed with H₂O and brine and dried (MgSO₄). After removing solvent, the product was used directiy in next step.

1I 2-(E-2-CarboxymethyIethenyl)-3-[8-(4-methoxyphenyl)octyloxyIpyridine. The crude aldehyde obtained above and methyl (triphenylphosphoranylidene) acetate (6.3g, 19mmol) were dissolved in toluene (20mL) and heated at 50 °C under an argon atmosphere for 30 minutes. Upon cooling to room temperature the reaction was diluted with EtOAc and washed with H₂O and brine and dried (MgSO₄). The product was purified by flash column chromatography (silica, 10: 5: 85, EtOAc: CH₂Cl₂: hexane) gave 3.75g (49%, three steps) as a pale yellow waxy solid: ¹H NMR (250MHz, CDCl₃) δ 8.28 (d, J=6.5Hz, 1H, 6-pyridyl), 8.17 (d, J=15.8Hz, 1H, olefin), 7.28 (m, 2H, 4-pyridyl, 5-pyridyl), 7.12 (d, J=8.6Hz, 2H, phenyl), 7.02 (d, J=15.8Hz, 1H, olefin), 6.89 (d, J=8.6Hz, 2H, phenyl), 4.08 (t, J=6.5Hz, 2H, OCH₂), 3.87 (s, 3H, OMe), 3.85 (s, 3H, methyl ester), 2.61 (t, J=7.5Hz, 2H, benzylic), 1.85 (m, 2H, CH₂), 1.60 (m, 2H, CH₂), 1.30 (m, 8H, aliphatic).

1J 2-(E-2-Carboxymethylethenyl)-3-[8-(4-methoxyphenyl)-octyloxylpyridine N-oxide. 2-(E-2-Carboxymethylethenyl)-3-[8-(4-rnethoxyphenyl)octyloxy]pyridine (620mg, L56mmol) was dissolved in dry CH₂Cl₂ (15mL) under an argon atmosphere, cooled to 0 °C, and treated with 80% MCPBA (378mg, 1.71mmol) in three portions. Following the addition, the cooling bath was removed and the reaction was stirred at room temperature for 24 hours. The reaction solution was diluted with CH₂Cl₂ and poured into saturated aqueous NaHCO₃. The aqueous phase was extracted with CH₂Cl₂ and the combined CH₂Cl₂ extracts were washed with brine and dried (MgSO₄). The crude product was used directly in the next step: ¹H NMR (250MHz, CDCl₃) δ 8.02 (d, J=15.8Hz, 1H, olefin),

7.80 (d, J=6.5Hz, 1H, 6- pyridyl), 7.39 (d, J=15.8Hz, 1H, olefin), 7.00 (m, 2H, 5-pyridyl, 4-pyridyl), 6.85 (d, J=8.6Hz, 2H, phenyl), 6.65 (d, J=8.6Hz, 2H, phenyl), 3.91 (t, J=6.5Hz, 2H, OCH₂), 3.68 (s, 3H, OMe), 3.62 (s, 3H, methyl ester), 2.37 (t, J=7.5Hz, 2H, benzylic), 1.85 (m, 2H, CH₂), 1.60 (m, 2H, CH₂), 1.30 (m, 8H, aliphatic).

1K 6-(E-2-Carboxymethylethenyl)-5-[8-(4-methoxyphenyl)octyloxy]-2-pyridone. The crude product obtained above was dissolved in dry DMF (10mL) under an argon atmosphere and cooled to 0 °C. To this was slowly added trifluoroacetic anhydride (2.2mL, 15.6mmol) followed by removal of the cooling bath. The reaction was stirred at room temperature for 18h. The reaction solution was diluted with EtOAc and slowly poured into saturated aqueous NaHCO₃. The organic layer was washed with NaHCO₃ and brine and dried (MgSO₄). The product was obtained as a yellow solid and was used without further purification: ¹H NMR (250MHz, CDCl₃) δ 7.75 (d, J=15.8Hz, 1H, olefin), 7.40 (d, J=9.8Hz, 1H,

3-pyridyl), 7.10 (d, J=8.6Hz, 2H, phenyl), 7.00 (d, J=15.8Hz, 1H, olefin), 6.82 (d, J=8.6Hz, 2H, phenyl), 6.70 (d, J=9.8Hz, 1H, 4-pyridyl), 3.95 (t, J=6.5Hz, 2H, OCH₂), 3.85 (s, 3H, OMe), 3.82 (s, 3H, methyl ester), 2.57 (t, J=7.5Hz, 2H, benzylic), 1.85 (m, 2H, CH₂), 1.60 (m, 2H, CH₂), 1.30 (m, 8H, aliphatic). 1L 6-(Ε-2-Carboxymethylethenyl)-5-[8-(4-methoxyphenyl)octyIoxy]-2-pyridine trifluoromethylsulfonate. To a cooled (0 °C) solution of the crude pyridone obtained above in dry CH₂Cl₂ (10mL) under an argon atmosphere was added dry pyridine (0.38mL, 4.65mmol) and trifluoromethanesulfonic anhydride (0.35mL, 2.08mmol). The reaction was stirred at 0 °C for 15 minutes. The reaction was diluted with EtOAc and washed with H₂O, 2% HCl, saturated NaHCO₃, and brine and dried (MgSO₄). Purification by flash column chromatography (silica, 20% EtOAc in hexane) gave 490mg (58%, three steps) as a colorless oil: ¹H NMR (250MHz, CDCl₃) δ 7.96 (d, J=15.8Hz, 1H, olefin), 7.33 (d, J=9.8Hz, 1H, 3-pyridyl), 7.10 (d, J=9.8Hz, 1H, 4-pyridyl), 7.06 (d, J=8.6Hz, 2H, phenyl), 6.94 (d, J=15.8Hz, 1H, olefin), 6.89 (d, J=8.6Hz, 2H, phenyl), 4.02 (t, J=6.5Hz, 2H, OCH₂), 3.82 (s, 3H, OMe), 3.76 (s, 3H, methyl ester), 2.54 (t, J=7.5Hz, 2H, benzylic), 1.85 (m, 2H, CH₂), 1.60 (m, 2H, CH₂), 1.30 (m, 8H, aliphatic).

1M Methyl (3-trimethylsiIyl)ethynyl benzoate. To a flame dried flask containing triethylamine (150mL) under an argon atmosphere was added methyl 3-bromobenzoate (13.2g, 6L3mmol), trimethylsilyl acetylene (10g, 101mmol), Pd(OAc)2 (0.15g, 2mmol), and Ph₃P (0.3g, 4mmol). The resulting mixture was heated at 100 °C for 16 hours. Upon cooling to room temperature the reaction mixture was filtered and the solvent was evaporated. The residue was diluted with EtOAc and washed with NH₄Cl, H₂O, brine and dried (MgSO₄). Purification by flash column chromatography (silica, 5% EtOAc: hexane) gave 8g (60%) as an amber oil: ¹H NMR (250MHz,CDCl₃) δ 8.07 (d, J=1Hz, 1H, aromatic), 7.85 (dd, J=3Hz, 1H, aromatic), 7.48 (dd, J=3Hz, 1H, aromatic), 7.21 (t, 1H, aromatic), 3.8 (s, 3H, methyl ester), 0.2 (s, 9H, Me₃Si). 1N Methyl 3-ethynyI benzoate. To a solution of methyl (3-trimethylsilyl)ethynyl benzoate (30mmol) in methanol (100mL) was added K₂CO₃ (1g). The reaction was stirred at room temperature for 3 hours. The solvent was evaporated and the residue was dissolved in CH₂Cl₂. This was washed with

NaHCO₃, H₂O, brine and dried (MgSO₄). Purification by flash column

chromatography (silica, 5% EtOAc: hexane) gave 1.13g (70%) as a solid: ¹H NMR (90MHz, CDCl₃) δ 7.9 (m, 1H, aromatic), 7.75 (dt, 1H, aromatic), 7.4 (dt, 1H, aromatic), 7.1 (t, 1H, aromatic), 3.7 (s, 3H, methyl ester), 2.9 (s, 1H, acetylenic). 1O Methyl 3-[6-(E-2-carboxyrnethyIethenyl)-5-[8-(4-methoxyphenyl)octyloxy]pyrid-2-yl]ethynyI benzoate. To a solution of 6- (E-2-carboxymethylethenyl)-5-[8-(4-methoxyphenyl)octyloxy]-2-pyridine

trifluoromethylsulfonate (0.1g, 0.18mmol) in dry DMF (3mL) under an argon atmosphere was added the acetylene prepared above (0.043g, 0.27mmol) along with triethylamine (0.3mL, 1.8mmol), CuI (1.2mg, 4%mol), and Pd(Ph₃P)₂Cl₂ (4mg, 2% mol). The resulting mixture was heated at 75 °C for 5 hours. Upon cooling to room temperature the reaction πύxture was diluted with EtOAc and washed with H₂O, brine and dried (Na₂SO₄). Purification by flash column chromatography (silica, 0-10% EtOAc: hexane) gave 0.03g (16%) as an oil: ¹H NMR (250MHz, CDCl₃) δ 8.25 (s, 1H, aromatic), 8.06 (d, J=15.8Hz, 1H, olefin), 8.01 (m, 1H, aromatic), 7.75 (d, 1H, aromatic), 7.5 (m, 2H, aromatic, 4-pyridyl), 7.27 (d, 1H, 5-pyridyl), 7.18 (d, J=15.8Hz, 1H, olefin), 7.08 (d, 2H, phenyl), 6.8 (d,2H, phenyl), 4.05 (t, 2H, OCH₂), 3.9 (s, 3H, methyl ester), 3.7 (s, 3H, methyl ester), 3.68 (s, 3H, OCH₃), 2.52 (t, 2H, benzylic), 1.9- 1.3 (m, 12H, aliphatic).

1P Methyl 3-[6-(carboxymethylemanyl)-5-[8-(4-methoxyphenyl)-octyloxylpyrid-2-yl]ethanyl benzoate. The solution of ester prepared above (0.03g, 0.3mmol) in ethanol (5mL) and 5% Pd/C catalyst (2mg) was shaken in a Parr hydrogenation apparatus under 50 psi of H₂ for 4 hours. The reaction mixture was filtered through Celite and the solvent evaporated to give the captioned 0.013g (65%) of product as an oil: ¹H NMR (250MHz , CDCl₃) δ 7.8 (s, 1H, aromatic), 7.75 (m, 1H, aromatic), 7.25 (m, 2H, aromatic, 4-pyridyl), 7.01 (d, 2H, phenyl), 6.89 (d, 1H, 5-pyridyl), 6.75 (m, 3H, aromatic, phenyl), 3.85 (m, 5H, OCH₂, methyl ester), 3.75 (s, 3H, methyl ester), 3.65 (s, 3H, OCH₃), 3.1 (t, 2H, CH₂-methyl ester), 3.0 (m, 4H, benzylic), 2.7 (t, 2H, benzylic), 2.5 (t, 2H, benzylic), 1.8-1.2 (m, 12H, aliphatic). 1O 3-[6-(Carboxyethanyl)-5-[8-(4-methoxyphenyl)octyloxy]pyrid-2-yl]ethanyl . benzoic acid, dilithium salt. The diester prepared above (0.013g, 0.2mmol) was dissolved in tetrahydrofuran (0.5mL) and MeOH (0.5mL) and treated with 1.0M LiOH (1mL, 1mmol). The reaction was stirred under an argon atmosphere for 24 hours. The solvent was evaporated and the resulting residue was purified by

Reverse Phase MPLC (RP-18 silica, 0-40% MeOH: H₂O). Lyophilization yielded a colorless amorphous solid: ¹H NMR (250MHz, CD₃OD) δ 8.87 (s, 1H, aromatic), 7.75 (dd, 1H, aromatic), 7.2 (m, 3H, aromatic, 4-pyridyl), 7.08 (d, 2H, phenyl), 6.95 (d, 1H, 5-pyridyl), 6.81 (d, 2H, phenyl), 3.95 (t, 2H, OCH₂), 3.75 (s, 3H, OCH₃), 3.12 (m, 2H, CH₂-CO₂Li), 3.0 (s, 4H, benzylic), 2.5 (m, 4H, benzylic), 1.8-1.3 (m, 12H, aliphatic); MS (FAB): 546.4 (M+H), 552.4 (M+Li).

Example 2

(Ε)-3-[3-[4-(4-Methoxyphenyl)butyloxyl-6-[(E)-3-oxo-3-phenyl-1-propenyl]-2- pvridinyl]-2-propenoic acid, lithium salt

2A. 1-Iodo-4-(4-methoxyphenyl)butane. To a stirred solution of 4-(4- methoxyphenyl)butan-1-ol (9.37g, 52mmol, Aldrich) in dry toluene (185mL) under an argon atmosphere was added triphenylphosphine (17.8g, 67.6mmol) and

imidazole (10.6g, 156mmol). After ten minutes I₂ (17.1g, 67.6mmol) was added. The reaction was then heated at 65 °C for 30 minutes. Upon cooling to room temperature the reaction was concentrated to 1/4 volume. The remaining solution was diluted with Et₂O and washed with H₂O and brine and dried (MgSO₄). The solvent was removed and the resulting residue was dissolved in CH₂Cl₂ and applied to a flash chromatography column (silica). Elution with 2% EtOAc in hexane provided 12.7g (84%) of product as a colorless oil: ¹H NMR (250MHz, CDCl₃) δ 7.08 (d, J=8.6Hz, 2H, phenyl), 6.82 (d, J=8.6Hz, 2H, phenyl), 3.78 (s, 3H, OMe), 3.17 (t, J=7.4Hz, 2H, I-CH₂), 2.54 (t, J=7.6Hz, 2H, benzylic), 1.85 (m, 2H, CH₂), 1.60 (m, 2H, CH₂). 2B 3-Hydroxy-6-methyl-2-pyridine carboxaldehyde. 2,6-Lutidine-α²,3-diol (15g, 107.8mmol; Aldrich) was suspended in dry CH₂Cl₂ (200mL) and treated with MnO₂ (47g, 539mmol). The reaction was stirred at room temperature for 6 hours. The reaction mixture was filtered through a pad of Celiteand the solvent was evaporated. The crude aldehyde was obtained as a tan solid and was used directly for the next step: ¹H NMR (250MHz, CDCl₃) δ 10.65 (s, 1H, OH), 10.30 (s, 1H, aldehyde), 7.30 (m, 2H, 4,5-pyridyl), 2.55 (s, 3H, methyl). 2C 3-[4-(4-MethoxyphenyI)butyloxyl-6-methyl-2-pyridine carboxaldehyde. a solution of 1-iodo-4-(4-methoxyphenyl)butane (12.6g, 43.4mmol) in dry dimethylformamide (45mL) under an argon atmosphere was added 3-hydroxy-6-methyl-2-pyridine carboxaldehyde (7.2g, 52.5mmol) and anhydrous K₂CO₃ (30g, 217mmol). The reaction was vigorously stirred at 90 °C for 2.5 hours. Upon cooling to room temperature the reaction was diluted with EtOAc and washed with H₂O, aq NH₄CI, and brine and dried (MgSO₄). Evaporation provided crude aldehyde as a dark oil that was used without further purification. 2D 2-(E-2-Carboxymethylethenyl)-3-[4-(4-methoxyphenyl)butyloxy]-6-methylpyridine. 3-[4-(4-Methoxyphenyl)butyloxy]-6-methyl-2-pyridine

carboxaldehyde obtained above was dissolved in dry toluene (100mL) under an argon atmosphere and treated with methyl (triphenylphosphoranylidene)acetate (14.5g, 43.4mmol). The reaction was heated for 1 hour at 50 °C. Upon cooling to room temperature the reaction was diluted with EtOAc and washed with H₂O and brine and dried (MgSO₄). Purification by flash column chromatography (silica, 20% EtOAc in hexane) gave 13.6g (88%; from iodide) as a pale yellow oil: ¹H NMR (250MHz, CDCl₃) δ 8.07 (d, J=15.7Hz, 1H, olefin), 7.10 (m, 4H, phenyl, 4,5-pyridyl), 7.07 (d, J=15.7Hz, 1H, olefin), 6.81 (d, J=8.6Hz, 2H, phenyl), 3.97 (t, J=6.5Hz, 2H, O-CH₂), 3.79 (s, 3H, methyl ester), 3.78 (s, 3H, OMe), 2.54 (t, J=7.6Hz, 2H, benzylic), 2.48 (s, 3H, methyl), 1.85 (m, 2H, CH₂), 1.60 (m, 2H, CH₂); MS (ES): 356.4 (M+H).

2E 2-(E-2-Carboxymethylethenyl)-3-[4-(4-methoxvphenyl)butyloxy]-6-methylpyridine N-oxide. 2-(E-2-Carboxymethylethenyl)-3-[4-(4- methoxyphenyl)butyloxy]-6-methylpyridine (13.6g, 38.2mmol) was dissolved in dry CH₂Cl₂ (100mL) and cooled to 0 °C; 50% m-chloroperoxybenzoic acid (13.2g, 38.3mmol) was added in three portions over 10 minutes. The cooling bath was removed and the reaction was stirred for 15 hours at room temperature. The reaction was poured into aqueous NaHCO₃ and the product extracted into CH₂Cl₂. The organic extract was washed with H₂O and brine and dried (MgSO₄). The crude product was obtained as a yellow solid and was used without further purification.

2F 2-(E-2-Carboxymethylethenyl)-3-[4-(4-methoxyphenyl)butyloxy]-6-hydroxymethylpyridine. 2-(E-2-Carboxymethylethenyl)-3-[4-(4- methoxyphenyl)butyloxy]-6-methylpyridine N-oxide obtained above was suspended in dry dimethylformamide (100mL) and cooled to 0 °C under an argon atmosphere. To this was slowly added trifluoroacetic anhydride (54mL, 380mmol). The reaction was maintained at 0 °C for 20 minutes followed by 18 hours at room temperature. The reaction solution was slowly added to a solution of saturated aqueous Na₂CO₃ and stirred for 1 hour. The product was then extracted into EtOAc; the combined organic extracts were washed with H₂O and brine and dried (MgSO₄). Purification by flash column chromatography (silica, EtOAc:hexane:CH₂Cl₂, 25:25:50) gave 9.0g (64%; two steps) as a waxy solid: ¹H NMR (250MHz, CDCl₃) δ 8.08 (d, J=15.7Hz, 1H, olefin), 7.23 (d, J=8.6Hz, 1H, 5-pyridyl), 7.16 (d, J=8.6Hz, 1H, 4-pyridyl), 7.09 (d, J=8.6Hz, 2H, phenyl), 7.03 (d, J=15.7Hz, 1H, olefin), 6.82 (d, J=8.6Hz, 2H, phenyl), 4.69 (d, J=4.1Hz, 2H, CH₂-OH), 4.01 (t, J=6.5Hz, 2H, O-CH₂), 3.82 (s, 3H, methyl ester), 3.78 (s, 3H, OMe), 3.62 (t, J=4.1Hz, 1H, OH), 2.55 (t, J=7.6Hz, 2H, benzylic), 1.85 (m, 2H, CH₂), 1.58 (m, 2H, CH₂); MS (CI): 374.3 (M+H).

2G 2-(E-2-Carboxymethyiethenyl)-3-[4-(4-methoxyphenyl)butyloxy]-6-pyridine carboxaldehyde. 2-(E-2-Carboxymethylethenyl)-3-[4-(4-methoxyphenyl)butyloxy]- 6-hydroxymethylpyridine obtained above (0.5g, 1.3mmol) was dissolved in dry CH₂Cl₂ (20mL). Manganese Oxide (1.21g, 13mmol) was added and the reaction was stirred at room temperature for 24 hours. The reaction was filtered through

Celite; evaporation provided 0.33g (66%) the product as an oil: ¹H NMR (250MHz, CDCl₃) δ 9.90 (s, 1H, aldehyde), 8.01 (d, J=15.8Hz, 1H, olefin), 7.89 (d, J=8.5Hz, 1H, 5-pyridyl), 7.20 (d, J=8.5Hz, 1H, 4-pyridyl), 7.11 (d, J=15.8Hz, 1H, olefin), 7.01 (d, 2H, phenyl), 6.75 (d, 2H, phenyl), 4.09 (t, 2H, O-CH₂), 3.81 (s, 3H, methyl ester), 3.78 (s, 3H, OMe), 2.60 (t, 2H, benzylic), 1.85 (m, 4H, CH₂-CH₂).

2H (E)-Methyl 3-[3-[4-(4-methoxyphenyl)butyloxy]-6-[(E)-3-oxo-3-phenyl-1-propenyl]-2-pyridinyl]-2-propenoate. To a solution of phenacyl

triphenylphosphonium bromide (0.51g, 1.1mmol) in dry toluene (5mL) under an argon atmosphere was added sodium hydride (0.04g, 1.1mmol). The reaction was heated at 45 °C for 4 hours. The reaction was cooled to room temperature and the carboxaldehyde prepared above (0.33g, 0.9mmol) in dry toluene (5mL) was added. The reaction was stirred at room temperature for 48 hours. The reaction was diluted with EtOAc and washed with H₂O, brine and dried (Na₂SO₄). The solvent was removed and the resulting residue was dissolved in CH₂Cl₂ and applied directly to a flash chromatography column (silica). Elution with 5-40% EtOAc in hexane provided 0.18g (43%) as an oil: ¹H NMR (250MHz, CDCI₃) δ 8.11 (d, J=15.8Hz, 1H, olefin), 8.08 (m, 2H, aromatic), 8.01 (d, J=16.1Hz, 1H, olefin), 7.68 (d,

J=16.1Hz, 1H, olefin), 7.55 (m, 3H, aromatic), 7.39 (d, J=8.6Hz, 1H, 5-pyridyl), 7.21 (d, J=15.8Hz, 1H, olefin), 7.18 (d, J=8.6Hz, 1H, 4-pyridyl), 7.11 (d, 2H, phenyl), 6.8 (d, 2H, phenyl), 4.1 (t, 2H, O-CH₂), 3.85 (s, 3H, methyl ester), 3.78 (s, 3H, OMe), 2.7 (t, 2H, benzylic), 1.85 (m, 4H, CH₂-CH₂); MS (ES): 472 (M+H).

2I (E)-3-[3-[4-(4-Methoxyphenyl)butyloxy]-6-[(E)-3-oxo-3-phenyl-1-propenyl]-2-pyridinyl]-2-propenoic acid, lithium salt. (E)-Methyl 3-[3-[4-(4-methoxyphenyl)butyloxy]-6-[(E)-3-oxo-3-phenyI-1-propenyl]-2-pyridinyl]-2-propenoate (0.03g, 0.06mmol) was dissolved in tetrahydrofuran (2mL) and MeOH (2mL). Lithium hydroxide (0.1mL, 0.12mmol) 1M solution in H₂O was added and the reaction was stirred at room temperature for 48 hours. The mixture was evaporated and redissolved in MeOH. Purification by Reversed Phase MPLC (RP-18 silica, 0-65% MeOH: H₂O) gave 0.014g (63%) a solid after lyophilization: ¹H NMR (250MHz, CD₃OD) δ 8.11 (m, 3H, olefin, aromatic), 7.98 (d, J=16.1Hz, 1H, olefin), 7.71 (d, J=16.1Hz, 1H, olefin), 7.55 (m, 4H, 5-pyridyl, aromatic), 7.35 (d, J=8.6Hz, 1H, 4-pyridyl), 7.21(d, J=15.8Hz, 1H, olefin), 7.18 (d, 2H, phenyl), 6.8 (d, 2H, phenyl), 4.1 (t, 2H, O-CH₂), 3.78 (s, 3H, OMe), 2.7 (t, 2H, benzylic), 1.85 (m, 4H, CH₂-CH₂); MS (ES): 458.2 (M+H-Li); C, H, N calculated for C₂₈H₂₆NO₅Li-⁷/₄H₂O: C, 67.94; H, 6.01; N, 2.83; found: C, 67.63; H, 5.57; N, 2.56.

Example 3

3-[3-(4-(4-Methoxyphenyl)butyloxy]-6-(3-phenylpropyl)-2-pyridinyl]propanoic acid, lithium salt

3A. Methyl 3-r3-[4-(4-methoxyphenyl)butyloxy]-6-(3-phenylpropyl)-2-pyridinyl]propanoate. (E)-Methyl 3-[3-[4-(4-methoxyphenyl)butyloxy]-6-[(E)-3-oxo-3-phenyl-1-propenyl]-2-pyridinyl]-2-propenoate (0.15g, 0.3mmol) was dissolved in absolute EtOH (20mL) and 15mg of catalyst (5% Pd/C) was added. The reaction was shaken in a Parr hydrogenation apparatus under 80 psi of H₂ for 16 hours. The reaction mixture was filtered through a pad of Celite and the solvent was evaporated. Purification by flash column chromatography (silica, 10% EtOAc: hexane) gave 0.04g (65%) of the captioned product as an oil: ¹H NMR (250MHz, CDCl₃) δ 7.32-6.75 (m, 11H, aromatic, 4,5-pyridyl), 3.91 (t, 2H, O-CH₂), 3.81 (s, 3H, methyl ester), 3.65 (s, 3H, OCH₃), 3.1 (t, 2H, CH₂-ester), 2.8-2.5 (m, 8H, benzylic), 2.01 (m, 2H, aliphatic), 1.8 (m, 4H, aliphatic); MS (ES): 462.4 (M+H). 3B 3-[3-[4-(4-Methoxyphenyl)butyloxy]-6-(3-phenylpropyl)-2-pyridinyl]propanoic acid, lithium salt. The methyl ester (0.037g, 0.1mmol) prepared above was dissolved in tetrahydrofuran (1mL) and MeOH (1mL). Lithium hydroxide (0.2mL, 0.02mmol) 1M solution in H₂O was added and the reaction was stirred at room temperature for 24 hours. The solvent was evaporated and the residue was dissolved in H₂O. Purification by Reversed Phase MPLC (RP-18 silica, 0-65% MeOH: H₂O) gave 0.022g (65%) as a lyophilized solid: ¹H NMR (250MHz, CD₃OD) δ 7.4-7.1 (m, 8H, aromatic, 5-pyridyl), 7.01 (d, 1H, 4-pyridyI), 6.81 (d, 2H, aromatic), 4.01 (t, 2H, O-CH₂), 3.81 (s, 3H, OCH₃), 3.1 (m, 2H, CH₂-CO₂Li), 2.75-2.5 (m, 8H, benzylic), 2.1-1.85 (m, 6H, aliphatic); MS (ES): 448.2 (M+H-Li); C, H, N calculated for C₂₈H₃₂NO₄Li· ³/4H₂O: C, 72.01; H, 7.23; N, 3.00; found: C, 71.82; H, 6.87; N 2.89.

Example 4

Preparation of Free Acids

The acid form of any of the foregoing salts may be prepared by dissolving the salt in water if it is not already in solution, then acidifying that solution with an acid such as a mineral acid eg. dilute (6N) HCl. The acid is recovered by filtering out the precipitate.

Example 5

Formulations for pharmaceutical use incorporating compounds of the present invention can be prepared in various forms and with numerous excipients. Means for making various formulations can be found in standard texts such as Remington's Pharmaceutical Sciences, and similar publications and compendia. Specific examples of formulations are given below.

OINTMENTS

Hydrophyllic Petrolatum

Ingredients Amount (% Weight/weight)

Cholesterol 30.0g

Stearyl Alcohol 30.0g

White Wax 78.0g

Active Ingredient 2.0g

White Petrolatum 860.0g The stearyl alcohol, white wax and white petrolatum are melted together (steam bath for example) and cholesterol and the active ingredient are added.

Stirring is commenced and continued until the solids disappear. The source of heat is removed and the mix allowed to congeal and packaged in metal or plastic tubes.

Emulsion Ointment

Ingredients Amount (% W/W)

Methylparaben 0.25g

Propylparaben 0.15

Sodium Lauryl Sulfate 10.0g

Active Ingredient 5.0g

Propylene Glycol 120.0g

Stearyl Alcohol 250.0g

White Petrolatum 250.0g

Purified Water QS to 1000.0g

The stearyl alcohol and white petrolatum are combined over heat. Other ingredients are dissolved in water, then this solution is added to the warm (ca 50 to 100° C) alcohol/petrolatum mixture and stirred until the mixture congeals. It can then be packed in tubes or another appropriate package form.

Example 6

Inhalation Formulation

A compound of formula 1, 1 to 10 mg/ml, is dissolved in isotonic saline and aerosolized from a nebulizer operating at an air flow adjusted to deliver the desired amount of drug per use.

Claims

What is claimed is:

1. A compound of formula I

or an N-oxide, or a pharmaceutically acceptable salt, where

m is 1 - 8;

R is C₁ to C₂₀-atiphatic, unsubstituted or substituted phenyl-C₁ to C₁₀-aliphatic where substituted phenyl has one or more radicals selected from the group consisting of

(a) lower alkoxy,

(b) lower alkyl,

(c) trihalomethyl, and

(d) halo; or

R is C₁ to C₂₀-aliphatic-O-; or

R is unsubstituted or substituted phenyl-C₁ to C₁₀-aliphatic-O- where substituted phenyl has one or more radicals selected from the group consisting of

(a) lower alkoxy,

(b) lower alkyl,

(c) trihalomethyl, and

(d) halo;

R₁ is R₃, -(C₁ to C₅ aliphatic)R₃, -(C₁ to C₅ aliphatic)CHO, -(C₁ to C₅ aliphatic)CH₂OR₄;

R₂ is H, R₃, -(C₁ to C₅ aliphatic)R₃ or tetrazol-5-yl;

R₃ is tetrazol-5-yl or COOH or an ester or amide thereof; and

R₄ is hydrogen, C₁ to C₆ alkyl, or C₁ to C₆ acyl.

2. A compound of claim 1 where R is C₈ to C₂₀ alkoxy, phenyl-C₄ to C₁₀ alkoxy or substituted-phenylC₄ to C₁₀ alkoxy; R₁ is -(C₁-C₃alkyl)R₃, or -(C₂-C₃alkenyl)R₃ and R₂ is hydrogen.

3. A compound of claim 2 where R is substituted phen_yl-C₄ to C₁₀ alkoxy, particularly the substituted-phenyl(CH₂)_4-8-O- group or CH₃(CH₂)_7-9-O-; m is 2 - 6; R₁ is HO₂C-CH=CH-, or HO₂C-CH₂CH₂- or a salt, ester or amide derivative thereof and R₂ is tetrazol-5-yl or COOH or an ester or amide thereof.

4. A compound of claim 3 where m is 2, 3 or 4.

5. A compound of claim 4 which is (E)-3-[3-[4-(4-methoxyphenyl)butyloxy]-6-[(E)-3-oxo-3-phenyl-1-propenyI]-2-pyridinyl]-2-propenoic acid, lithium salt, the free acid or another pharmaceutically acceptable salt.

6. A compound of claim 4 which is 3-[3-[4-(4-methoxyphenyl)butyloxy]-6-(3-phenylpropyl)-2-pyridinyl]propanoic acid, lithium salt, the free acid or another pharmaceutically acceptable salt.

7. A compound of claim 4 which is (E)-3-[3-[4-(4-methoxyphenyl)butyloxy]-6-(2-phenylethyl)-2-pyridinyl]-2-propenoic acid, lithium salt, the free acid or another pharmaceutically acceptable salt.

8. A compound of claim 4 which is 3-[3-[4-(4-methoxyphenyl)butyloxy]-6-(2-phenylethyl)-2-pyridinyl]propanoic acid, lithium salt, the free acid or another pharmaceutically acceptable salt.

9. A compound of claim 1 where R₂ is R₃, -(C₁ to C₅ aliphatic)R₃ or terrazol-5-yl.

10. A compound of claim 9 where m is 2, 3 or 4.

11. A compound of claim 10 which is 3-[6-(carboxyethanyl)-5-[8-(4-methoxyphenyl)octyloxy]pyrid-2-yl]ethanyl benzoic acid, dilithium salt, or another pharmaceutically acceptable salt

12. A composition comprising a carrier or excipient and a compound of formula I according to claim 1.

13. A method for treating psoriasis, which method comprises administering to a patient in need thereof, an effective amount of a compound of formula I according to claim 1 either alone in combination with a pharmaceutically acceptable excipient.