HUT64750A - Process for the production of substituted benzoyl-benyole, biphenyl- and 2-oxazole-alcanic acid derivatives and of medical preparatives containing them - Google Patents

Abstract

There are disclosed compounds of the formula: A(CH2)nO-B, wherein A is phenoxyethyl, phenoxyphenyl or a group having formula (a), wherein X is -N- or (b); Z is (c), (d), (e), (f), -S- or -O-; R<1> is hydrogen, lower alkyl or phenyl; R<2> is hydrogen or lower alkyl; or R<1> and R<2> taken together form a benzene ring; R<3> is hydrogen or lower alkyl; n is 1 -2; B is (g), (h), (i) or (j), wherein R<4> and R<5> are each, independently, hydrogen or lower alkyl; R<6> is halo or nitro; R<7> is (k) or (l); R<8> is lower alkyl; m is 0-3; and the pharmacologically acceptable salts thereof, and their use in the treatment of inflammatory conditions, such as rheumatoid arthritis, ulcerative colitis, psoriasis and other immediate hypersensitivity reactions; in the treatment of leukotriene-mediated naso-bronchial obstructive air-passageway conditions, such as allergic rhinitis, allergic bronchial asthma and the like; and as gastric cytoprotective agents.

Description

The present invention relates to novel substituted benzoylbenzene, biphenyl and 2-oxazole alkanoic acid derivatives having lipoxygenase inhibiting, phospholipase A ₂ inhibiting and leukotriene antagonist activity, useful as anti-inflammatory, antiallergic and cell protective agents.

It is well known that arachidonic acid (AA) is metabolised in mammals by two different pathways. The cyclooxygenase enzymatic pathway of arachidonic acid metabolism results in the production of prostaglandins and thromboxanes. The physiological activity of prostaglandins has been extensively demonstrated in recent years. Prostaglandins are known to be formed from the PGG ₂ and PGH ₂ endoperoxides by the cyclooxygenase pathway of arachidonic acid metabolism. These endoperoxides are also A ₂ and B ₂ precursors of thromboxane (Tx). Tx A _{2 is a} vasoconstrictor that stimulates platelet aggregation. Under normal circumstances, the vasoconstrictor and platelet aggregating properties of thromboxanes are counterbalanced by another product derived from endoperoxides via the cyclooxygenase pathway, prostacyclin (PGI ₂ ), with PGI ₂ vasodilator and platelet aggregation inhibitory activity. A situation in which prostacyclin synthesis is impaired and / or platelet activation is elevated favors thrombosis and vasoconstriction. The role of prostanoids in hemostasis and thrombosis is described by RJ Gryglewski, [CRC Crit. Rev. Biochem. 7, 291 (1980)] and JB Smith (Am. J. Pathol 99, 743 (1980)]. Cyclooxygenase meta bits are known to be directly involved in the inflammatory response (see Higgs et al., 1984, Annals of Clinical Research, 16, 287-299). This is accomplished through their antihypertensive activity, their involvement in causing pain and fever, their peptide mediator permeability, and their edema-forming property. Finally, various aspects of cell-mediated immunity are influenced by cyclooxygenase products.

Lipoxygenase enzymes are involved in another pathway of arachidonic acid metabolism, a product of several pathways known as leukotrienes. These are designated by the LT nomenclature, the most important products of the lipoxygenase metabolic pathway are B ₄ , C ₄ and D ₄ leukotrienes. The substance, which is referred to as the primary product of the slow-acting anaphylaxis (SRS-A), has been shown to contain a mixture of LTC ₄ and LTD ₄ leukotrienes, and contains various amounts of other leukotriene metabolites [see Bach et al., J. Immun. ., 215. 115-118 (1980); Biochem. Biophys. Gap. Commun. 93: 1121-1126 (1980).

The importance of these leukotrienes is that, according to a large body of evidence, leukotrienes are involved in inflammatory reactions, exert their chemotactic activity, stimulate the release of lysosomal enzymes, and play an important role in the immediate hypersensitivity reaction. LTC ₄ and LTD ₄ have been shown to be potent constrictors of human bronchi (Dahlen et al., 1980, Natur. 288, 484-486) and Piper, Int. Arch. Appl. Immunol., 76, burns. 1, 43 (1985)] stimulate the release of mucus from the airways in vitro [Marom et al., Am. Rev. Resp. Dis. 126, 449 (1982)], skin-active vasodilators (Bisgaard et al., Prostaglandins, 23, 797 (1982)), and induce a redness and bladder response (Camp et al., Br. J. Pharmacol. 497 (1983)]. The non-peptide leukotriene LTB ₄ is a highly potent chemotactic factor for leukocytes [AW Ford-Hutchinson, J. Roy. Soc. Med., 74, 831-833 (1981)], which stimulates cell accumulation and affects vascular smooth muscle (Bray, Br. Med. Bull., 39, 249 (1983)). The activity of leukotrienes as mediators of inflammation and hypersensitivity is described in detail in Bailey and Casey, Ann. Reports. Chem., 17, 203-217 (1982), and Bray, Agents and Actions 19, 87 (1986).

Phospholipase A2 (PLA ₂ ) is a critical rate-limiting enzyme in the arachidonic acid (AA) cascade as it is responsible for the hydrolysis of esterified AA at the C2 position of membrane phospholipids. This reaction produces two products: (1) free arachidonic acid, which is available for subsequent metabolism by any of the cyclooxygenase or lipoxygenase enzymes, and (2) lysophospholipid. When PLA2 is exposed to alkyl arachidonoyl glycerophosphatidylcholine, platelet activating factor (PAF) formation begins; PAF is a precursor of inflammation (pro-inflammatory agent) due to its effect (see Wedmore et al., 1981, Br. J. Pharmacol. 74, 916-917). In this regard, it should be noted that anti-inflammatory steroids are believed to inhibit eicosanoid synthesis by inducing the synthesis of a PLA2 inhibitory protein, termed macrocortin or lipomodulin [See Flower et al., Nat. (1979) and Hirata et al., Proc. Natn. Acad. Sci. USA, 77, 2533 (1980).

• · · ·

- 5 As a first step in the subsequent conversion of arachidonic acid by cyclooxygenase or lipoxygenase to various eicosanoids, ΡΙΑ ₂ -mediated release of arachidonic acid from membrane phospholipids is a critical event in attempting to address the various physiological manifestations that based. Thus, while PLA2 is required for platelet aggregation (Pickett et al., 1976, Biochem. J. 160, 405), in terms of cardiac contraction and pacing, Geisler et al., Pharm. Gap. Commun., 9, 117 (1977)] and in prostaglandin synthesis [Vogt. Adv. Prostagl. Thromb. Gap. 3, 89 (1978)], inhibition of PLA2 can be labeled for the treatment of physiological conditions mediated by both PAF and products of the cyclooxygenase and / or lipoxygenase pathway.

It has also been shown that the cyclooxygenase / lipoxygenase pathway plays a key role in both the pathogenesis and the cellular pathogenesis of gastric mucosal damage due to extracellular (gastrointestinal contents, microorganisms, etc.) and intracellular (ischemia, viruses, etc.) factors. . Thus, prostaglandins, on the one hand, exert a cell protective effect on the gastric mucosa (see Robert, Gastroenterology, 77, 761-767 (1979)), and this role of prostaglandins, especially of the E series, is considered important in the treatment of gastrointestinal ulcers (see Isselbachher, Drugs, 33, (burn), 38-46 (1987)). On the other hand, ex vivo experiments prove that the gastric mucosal tissue of rats pretreated with ethanol is able to · · · ·

LTC ₄ production, and that this LTC ₄ production is quantitatively dependent on the severity of the ethanol damage [see Lángé et al., Naunyn-Schmiedeberg's Arch. Pharmacol. Suppl., 330, R27 (1985)]. It has also been shown that LTC ₄ may cause vasoconstriction in rat mucosal tissue during both the venous and arterial blood vessels [see Whittle, IUPHAR Ninth Int. Cong. ofPharm., pp. 30-32, London, England (1984). This is significant because ethanol-induced damage to the gastric mucosa is multifactorial, such as stomach congestion (Guth et al., 1984, Gastroenterology 87: 1083-90) contributing significantly to the development of a bleeding necrosis form of tissue injury. In addition, in anesthetized cats, exogenous LTD _{4 results in} both increased pepsin secretion and decreased transgastric potential (Pendleton et al., 1986, Eur. J. Pharmacol. 125: 297-99). In this regard, a significant recent discovery is that 5-lipoxygenase inhibitors and certain leukotriene antagonists protect the gastric mucosa from damage caused by oral or parenteral administration of most non-steroidal anti-inflammatory drugs [Rainsford, Agents and Actions, 21 , 316-319 (1987)]. Platelet activating factor (PAF) is also involved as a mediator in gastrointestinal damage, and recently 5lipoxygenase inhibitors have been shown to inhibit PAF-induced gastric mucosal damage [Gastroenterology, 96, A55, 1989]. As mentioned above, there is strong evidence that lipoxygenase products are involved in the development of pathological conditions associated with gastric mucosal injury, such as those caused by exposure to ethanol and non-steroidal anti-inflammatory drugs. Thus, compounds that inhibit the biological activity of leukotrienes and PAF and / or regulate the biosynthesis of these substances, for example by inhibiting 5-lipoxygenase, are considered valuable cytoprotective agents.

Accordingly, the biological activity of the enzyme lipoxygenase, which leads to the metabolism of leukotrienes and SRSs and arachidonic acid to leukotrienes, indicates that the prevention, elimination or amelioration of allergies, anaphylaxis, asthma and inflammation, and gastric mucosal cell protection medication. its approach should be to inhibit the release of mediators of these conditions or to antagonize their effects. Thus, compounds that inhibit the biological effects of leukotrienes and SRSs and / or which regulate the biosynthesis of these substances, for example, by inhibiting Pd-mediated release of arachidonic acid from membrane phospholipids or by inhibiting lipoxygenase, are useful in the treatment of conditions such as allergic bronchial asthma, allergic rhinitis, and other immediate hypersensitivity reactions, and are valuable in cell protection of the gastric mucosa.

We have discovered that certain novel substituted benzoylbenzene, biphenyl, and 2-oxazole alkanoic acid derivatives inhibit PLA ₂ and lipoxygenase, antagonists of the lipoxygenase pathway products, and thus useful as anti-inflammatory, anti-<

allergic and cell protective agents. The novel compounds of the invention are

They are represented by the formula (CH ₂ ) _n ° -B, where

A is phenoxyethyl, phenoxyphenyl or a group of formula (a) wherein

R ³

I

X is -N = or -C-;

R ³ R ³ R ³ R ³ R ³

I I I

Z is -C = C-, -C = N-, -N = C-, -N-, -S- or -O-,

R ¹ is hydrogen, lower alkyl or phenyl,

R ² is hydrogen or lower alkyl, or

R ¹ and R ² together form a benzene ring,

R ³ is hydrogen or lower alkyl, n is 1 or 2,

B is a group of formula (g), (h), (i) or (j) wherein

R ⁴ and R ⁵ are independently hydrogen or lower alkyl,

R ⁶ is halogen or nitro,

R ⁴

R ⁷ is -CR ⁸ or -CHCOOR ⁵

^R8 is lower alkyl, and m is an integer from 0 to 3,

• · · ·

And pharmaceutically acceptable salts of the above compounds.

The term lower alkyl refers to chains of 1-6 carbon atoms. The term halogen means fluorine, chlorine or bromine.

Substituent A includes, inter alia, 5- or 6-membered unsaturated hetero rings fused to mono- or benzene containing nitrogen, sulfur or oxygen, which may be optionally substituted with lower alkyl or phenyl. The above definition includes the following heterocyclic moieties: furyl, pyrrolyl, thienyl, oxazolyl, thiazolyl, imidazolyl, pyridyl, pyrazinyl, pyrimidinyl, benzofuranyl, benzothienyl, benzothiazolyl, indolyl, benzoxazolyl , quinolyl, quinazolyl, benzimidazolyl and quinoxalyl. Quinolyl, benzothiazolyl and benzimidazolyl groups are particularly preferred.

The compounds of the present invention may form pharmaceutically acceptable salts with pharmaceutically acceptable organic or inorganic acids such as hydrochloric acid, hydrobromic acid, sulfonic acid, sulfuric acid, phosphoric acid, nitric acid, maleic acid, fumaric acid, benzoic acid, ascorbic acid, pamoic acid, Propionic acid, tartaric acid, citric acid, lactic acid, malic acid, mandelic acid, cinnamic acid, palmitic acid, itaconic acid and benzenesulfonic acid, Compounds containing carboxylic acid groups, alkali metal and alkaline earth metal carboxylates or «

They can also form carboxylates with pharmaceutically acceptable cations derived from amines. These include, but are not limited to, cations such as ammonium, monor, di- and thymethylammonium, mono-, di- and triethylammonium, mono-, di- and tripropylammonium (iso- and normal). ), ethyldimethylammonium, benzyldimethylammonium, cyclohexylammonium, benzylammonium, dibenzylammonium, piperidinium, morpholinium, pyrrolidinium, piperazinium, 1-methylpiperidinium, 4-ethylmorpholinium, 1-isopropylpyrrolidinium, 1,4-dimethylpiperazinium, 1- (n-butyl) piperidinium, 2-methylpiperidinium, 1-ethyl-2-methylpiperidinium -, mono-, di- and triethanolammonium, ethyl diethanolammonium, n-butylmonoethanolammonium, tris (hydroxyethyl) methylammonium and phenyl monoethanolammonium carboxylates.

The following describes the preparation of the compounds of the invention. The preparation of compounds of formula (I) is illustrated in Schemes 1, 2 and 3. For the preparation of compounds of formula (1), as shown in Scheme 1, 4-methoxybenzonitrile is reacted with, for example, 3-bromo-toluene, and the resulting compound is then reacted with bromine in ethylene bromide to give 3-bromomethyl- [ 4'-methoxy] benzophenone. This bromine intermediate is reacted with sodium cyanide as shown in Scheme 2 to give the cyano intermediate, which is hydrolyzed in the presence of a base to give the carboxylic acid intermediate, which is demethylated to give the hydroxycarboxylic acid intermediate.

As shown in Scheme 3, the hydroxy-11-carboxylic acid intermediate is converted to the methyl ester with methanol in the presence of p-toluenesulfonic acid and then treated with a suitable halogenated alkyl-A compound, wherein A is as defined above, Hal is halogen, to give the desired final product. is obtained in the form of its methyl ester.

Hydrolysis of the ester in known manner yields the desired final product in the form of the free carboxylic acid.

The compounds of formula (2) may be prepared in several different ways. Compounds in which R ⁶ is a nitro group and R 'is -CR ⁶ may be prepared, for example, by reacting 4-bromo-3-nitroacetophenone with 4-iodoanisole in the presence of copper biphenyl intermediate which is demethylated with aluminum bromide to give the hydroxyl intermediate. The latter is reacted with the appropriate halogenated alkyl-A compound - where A is as defined above and Hal is halogen to give the final product. This reaction sequence is illustrated in Scheme 4.

Compounds of formula (2) wherein R ⁶ is halogen and R ⁷ is -CHCOOR ^{5 may be} prepared by a process employing the 4-methoxybiphenyl intermediate of Scheme 4. Thus, as shown in Scheme 5, the 4-acetyl-4-methoxy-2-nitrobiphenyl intermediate is reduced with tin (II) chloride to the amino derivative and this amino group is replaced with halogen. For example, the amino group • · ···. »• · ♦ · · ** ♦. · »... ...

- 12 fluorine can be exchanged for diazonium fluoroborate via an intermediate prepared from the amino intermediate with sodium nitrite and tetrafluoroboric acid. The resulting acetyl fluoromethoxybiphenyl intermediate is converted to the corresponding carboxylic acid and then demethylated with hydrogen bromide. The intermediate 2-fluoro-4'-hydroxy [1,1'-biphenyl] -4-acetic acid is thus obtained. Further steps of the reaction are shown in Scheme 6. The carboxylic acid intermediate obtained in Scheme 5 is esterified with methanol in the presence of p-toluenesulfonic acid and then reacted with the appropriate halogenated alkyl A derivative, where A is as defined above, Hal is halogen, to give the final product as the methyl ester. The ester can be hydrolyzed in a known manner to give the desired end product in the form of the free carboxylic acid.

The preparation of compounds of formula (3) or (4) according to the invention is carried out as shown in Schemes 7 and 8. Benzaldehyde and 4-methoxybenzaldehyde are reacted with each other and the resulting 4-methoxybenzene is reacted with succinic anhydride to form hemisuccinate. The latter compound is reacted with urea and acetic acid to give 4- (4-methoxyphenyl) -5-phenyl-2-oxazole-propionic acid intermediate.

This intermediate is demethylated with hydrogen bromide and then esterified with methanol to give the corresponding hydroxymethyl ester intermediate. This intermediate is reacted with the corresponding halogenated alkyl A derivative - A is as defined above, Hal is halogen - to give the desired final product as the methyl ester. The ester is known by a known procedure. _the "_#

13 can be hydrolyzed to give the desired end product as the free carboxylic acid.

The starting materials conventionally used in the above schemes are commercially available, commercially available, or can be prepared by anyone skilled in the art. For example, the 2-bromomethylquinoline intermediate can be prepared as shown in Scheme 9.

The benzene-fused heterocyclic compounds used in the reactions of the above schemes are also commercially available or may be prepared by methods known to those skilled in the art. For example, intermediates such as 1-methyl-2-chloromethylbenzimidazole, 2-chloromethylbenzthiazole, and 2-chloromethylbenzoxazole can be prepared as shown in Scheme 11 in the following formulas: X is O or S, or -NCH 3. This reaction is preferably carried out at a controlled low temperature in an organic solvent such as methylene chloride.

The compounds of the present invention, by virtue of their ability to inhibit the activity of PLA ₂ and lipoxygenase enzymes, are antagonists of enzymatic pathway mediators and are useful in the treatment of conditions mediated by arachidonic acid oxidation products. Accordingly, the compounds of the present invention are useful in the treatment of diseases such as rheumatoid arthritis, inflamed bowel, osteoarthritis, tendinitis, mucosal inflammation, psoriasis (and related dermatitis), and similar inflammatory conditions. In addition, the compounds of the invention have the ability to,

14, which are antagonists of the effects of LTC4, LTD4 and LTE4, which are components of SRS-A, are useful in inhibiting the symptoms induced by these leukotrienes. Therefore, the compounds of this invention are capable of ko ^ os, the prevention and treatment of a condition which triggers for LTC4, _LTD4, and LTE4, such conditions as allergic rhinitis, allergic bronchial asthma and other induced by leukotrienes nose-bronchial obstructive airways conditions, and other immediate hypersensitivity reactions, such as allergic conjunctivitis. The compounds of the invention are particularly valuable for the prevention and treatment of allergic bronchial asthma.

The compounds of the present invention are cell protective agents, particularly useful when administered in conjunction with conventional non-steroidal anti-inflammatory drugs, the majority of which have the side effect of irritating the gastrointestinal tract. The cell protective effect of the compounds of the invention significantly reduces the gastric irritating effect of conventional anti-inflammatory drugs. This effect is based not only on the ability of the compounds of the invention to inhibit the biological activity of leukotrienes and / or to regulate the biosynthesis of these substances by inhibiting lipoxygenase, but also plays a role in inhibiting the oxidation of arachidonic acid by the lipoxygenase pathway. cyclooxygenase, thereby increasing the production of cellular protective prostaglandins. These biological effects make the compounds of the invention particularly useful in the treatment of conditions such as erosive esophagitis, inflammatory bowel syndrome, induced bleeding injuries, e.g.

- 15 triggered by alcohol or non-steroidal anti-inflammatory drugs (NSAIDs), liver ischemia, liver-induced necrosis caused by harmful substances, pancreatic, renal or ipi-cerebral palsy, hepatotoxic agents such as carbon tetrachloride or D-galactosamine-induced parenchymal injury, ischemic renal disease, disease-induced liver injury, pancreatic or gastric damage caused by bile salts, cell injury caused by trauma or stress, and glycerol-induced renal disease.

When used as anti-inflammatory and / or cell protective agents for the treatment of allergic airway disorders, the compounds of the present invention may be formulated into oral dosage forms such as tablets or capsules. The compounds of the invention may be administered alone or in combination with suitable carriers such as magnesium carbonate, magnesium stearate, talc, sugar, lactose, pectin, dextrin, starch, gelatin, tragacanth, methylcellulose, sodium carboxylic acid, with melting point waxes or cocoa butter. Diluents, flavors, dissolution and glidants, suspending agents, binders, tablet disintegrating agents and the like may be used. The compounds of the invention may also be encapsulated with or without a carrier. The ratio of active ingredient in the above formulations, whether solid or liquid, is at least sufficient to provide the desired activity when administered orally. The compounds of the invention may also be injected parenterally, in this case in the form of sterile solutions containing additional solutes, for example, sufficient saline or glucose to make the solution isotonic. The compounds of the invention may be formulated into aqueous or partially aqueous solutions for aerosol or inhalation administration.

The dosage employed will depend upon the particular formulation employed, the route of administration, the symptoms, and the subject being treated. Treatment is generally initiated at a low dose, below the optimum dose of the compound. The dosage is then increased until the optimum is achieved under the circumstances. In general, the compounds of the present invention are most preferably administered in a concentration that generally produces an effective result without causing any adverse or deleterious side effects, this amount may be administered in a single unit dose or, if desired, may be subdivided into expedient sub-units at appropriate intervals throughout the day. administration.

The compounds of the present invention exhibit PLA ₂ and lipoxygenase inhibitory and leukotriene antagonist activity, as well as anti-inflammatory and eventual gastric irritant activity, as described in more detail in the following examples.

These methods determine, inter alia, the activity specificity of the compounds of the present invention as PLA ₂ inhibitors by measuring their ability to inhibit the synthesis of LTB4 and PGE ₂ polymorphonuclear leukocytes from rat glycogen and by measuring their ability to inhibit the release of arachidopsic acid mediated by the human PLA ₂ source. In addition, pharmacological studies demonstrate the in vivo effect of the compounds of the present invention on inhibiting the lipoxygenase and cyclooxygenase pathways of arachidonic acid metabolism.

The following examples illustrate the preparation of the compounds of the present invention and the pharmacological testing of the compounds.

Example 1 Preparation of 1- {2-Nitro-4 '- (2-quinolylmethoxy) [1,1'-biphenyl] -4-yl} ethanone

A. Preparation of 4-Acetyl-4'-methoxy-2-nitrobiphenyl

A mixture of 43.65 g, 0.187 mole of 4-iodoanisole, 40.6 g, 0.166 mole of 4-bromo-3-nitroacetophenone and 36 g, 0.567 mole of copper powder (copper bronze) was stirred under a nitrogen atmosphere in an oil bath at 80 ° C. place. The temperature is slowly raised to 110 ° C and the mixture is maintained at this temperature for 5 days (TLC by hexane: ethyl acetate = 8: 2). The mixture was then cooled, dissolved in dichloromethane and filtered on a celite pad. The filtrate and washings were evaporated, and the remaining dark brown oil (58.4 g) was purified by flash chromatography on dichloromethane pre-adsorbed on silica (Merek 60), impurities eluting with 9: 1 hexane: ethyl acetate. hexane: ethyl acetate (8: 2). This gave 16.2 g (32%) of the title compound as a yellow solid. Product

mp 124-126 ° C. NMR (CDCl 3, 400 MHz): δ = 2.67 (s, 3H, ΟΟΟΗ ₃ ), 3.85 (s, 3H, OCH 3), 6.97 (d, 2H, J = 8.74 Hz, ArH), 7.27 (d, 2H, J = 8.74 Hz, ArH), 7.56 (d, 1H, J = 8Hz, ArH), 8.15 (d, 1H, J = 8Hz,

ArH), 8.34 (S, 1H, ArH).

MS (EI, m / z): 271 (M) &lt; + &gt;.

B. Preparation of 4-Acetyl-4'-hydroxy-2-nitrobiphenyl

To a solution of aluminum tribromide (12.6 g, 47.4 mmol) in benzene (45 mL) was added dropwise a solution of methyl ether (Step A) (12 g) in benzene (5 g, 18.45 mmol) dropwise over 30 minutes under nitrogen. The resulting solution was stirred at room temperature for 3.5 hours (TLC (8: 2 hexane: ethyl acetate)). The mixture was then cooled in an ice bath, and the complex was decomposed by dropwise addition of about 37 mL of 6N hydrochloric acid. The organic layer was separated and the aqueous layer was extracted three times with ether. The combined ethereal extracts were concentrated to a small volume and extracted with 2.5N sodium hydroxide (2 x 50 mL + 1 x 10 mL). The alkaline extracts were cooled and then acidified to pH 2 with concentrated hydrochloric acid. The solid was collected and dried. 4.27 g of product are obtained in 90% yield. This product was used in the next step without purification.

NMR (CDCl ₃ , 400 MHz): δ = 2.67 (s, 3H, COCH 3), 5.03 (broad, 1H, OH), 6.91 (d, 2H, J = 8.56 Hz, ArH). , 7.23 (d, 2H, J = 8.57 Hz, · · ·

- 19 ArH), 7.55 (d, 1H, J = 7.9 Hz, ArH), 8.15 (d, 1H, J = 8.1 Hz, ArH), 8.34 (s, 1H, ArH) ).

MS (EI, m / z): 257 (M) ⁺

C. Preparation of 1- [2-Nitro-4 '- (2-quinolyl) - [1,1'-biphenyl-4-yl] ethanone

A mixture of 4.4 g, 17.12 mmol of phenol from Step B, 2.37 g, 17.12 mmol of powdered anhydrous potassium carbonate, 0.453 g, 1.71 mmol of 18-crown-6 and 38 mL of acetonitrile was added at room temperature. , stirring under nitrogen for 15 minutes. To the mixture was then added 3.34 g (18.83 mmol) of 2-chloromethylquinoline which was freshly prepared from its hydrochloride salt and refluxed for 10 hours (TLC eluting with hexane and hexanes). ethyl acetate (7: 3). An excess of 10% potassium carbonate, 18-crown-6 and chloromethylquinoline was added and refluxing continued for a further 4 hours. The solvent was removed and the residue was diluted with water and extracted three times with ethyl acetate. The extracts were washed and dried over magnesium sulfate. The residue was purified by flash chromatography after chromatography on pre-absorbed silica gel with dichloromethane (Merek 60), eluting with increasing polarity, hexane: ethyl acetate = 7: 3, 1: 1 and 1: 3, then pure. ethyl acetate. 2.59 g of pure title compound are obtained. This compound was recrystallized from toluene to give 2.05 g (30%) of a yellow solid, m.p. 160-162 ° C.

NMR (CDCl 3, 400 MHz): δ = 2.66 (s, 3H, COCH 3), 5.43 (s, 2H, OCH ₂ Ar), 7.10 (d, 2H, J = 8.7 Hz, ArH ), 7.27 (d, 2H, J = 8.7 Hz, ArH), 7.56 (m, 2H, J = 8.49 Hz, ArH), 7 , 75 (dt, 1H, ArH), 7.84 (d, 1H, J = 8.1 Hz, ArH), 8.09 (d, 1H, J = 8.5 Hz, ArH), 8.14 ( dd, 1H, ArH), 8.22 (d, 1H, J = 8.49 Hz, ArH), 8.34 (s, 1H, ArH).

MS (EI, m / z): 398 (M) ⁺ , 256, 158, 142

Analysis: Calculated for C24H38N2O4: C, 72.35; H, 4.55; N, 7.03; Found: C, 71.96; H, 4.75; N, 6.80.

Example 2

2-Fluoro-4 '- (2-quinolinylmethoxy) - [1,1 ¹ -biphenyl] -4-acetic acid

A. 4-acetyl-4 ^{1-methoxy-2-aminobiphenyl} Preparation

To a warm solution of 49.4 g (218.9 mmol) of tin (II) chloride in 72 mL of concentrated hydrochloric acid and 99 mL of ethanol was added 10.7 g (39.5 mmol) of Example 1A under stirring over 45 min. nitro derivative prepared according to example. The resulting yellow solution was refluxed for 3.5 hours (TLC checking, eluting with hexane: ethyl acetate = 1: 1). The ethanol was removed from the mixture and the residue was poured into a mixture of 360 ml of 50% sodium hydroxide and ice. The resulting solid was extracted three times with dichloromethane, and the extracts were washed with water and dried over sodium sulfate. Removal of the solvent gave 9.31 g (97.8%) of a yellow solid. Melting point 152-154 ° C.

NMR (CDCl 3, 400 MHz): δ = 2.59 (s, 3H, COCH 3), 3.80 (s, 3H, OCH 3), 6.97 (d, 2H, J = 8.7 Hz, ArH). , 7.23 (d, 1H, J = 7.4 Hz, ArH), 7.40 (d, 2H, J = 8.7 Hz, ArH), 7.48 (d, 1H, J = 7, 3 Hz, ArH), 7.49 (s, 1H, ArH).

MS (+, m / z): 241 (M) ⁺ , 226 (M-CH 3) ⁺ , 198 (M-CO 2 CH 3) ⁺ , 83.

B. Preparation of 4-Acetyl-4'-methoxy-2-fluorobiphenyl by stirring in an ice-cold mixture of 9.2 g, 38.2 mmol of the above aniline, 9.8 ml of water and 35.1 ml of 48% HBF4 in tetrahydrofuran a solution of sodium nitrite (2.82 g, 40.85 mmol) in water (5 mL) was added slowly. The internal temperature of the mixture was maintained below 5 ° C during the addition. The mixture was then stirred at 0 to 5 ° C for a further 20 minutes. The diazonium fluoroborate was filtered off and washed with ether containing 10% HBF 4 and 10% methanol and dried in vacuo. The resulting salt was decomposed by heating in 95 ml xylene at 70 ° C. After this time, the mixture was refluxed for a further 2.5 hours (TLC checking with 1: 1 hexane: ethyl acetate, UV detection). The xylene was removed from the mixture and the residue was extracted three times with ethyl acetate and once with ether. The combined extracts were washed with 10% sodium carbonate followed by brine and dried over magnesium sulfate. After removal of the solvent, an amber oil (6.03 g) was purified by flash chromatography followed by Merek 60 silica gel pre-absorption in dichloromethane eluting with 95: 5 hexane: ethyl acetate. The yield of the title compound is between 33% and 51% (depending on the particular procedure being performed), 3.12-4.75 g, m.p. 100-101 ° C.

NMR (CDCl ₃ , 400 MHz): δ = 2.62 (s, 3H, COCH 3), 3.86 (s, 3H, OCH 3), 700 (d, 2H, J = 8.9 Hz, ArH), δ , 50-7.80 (m, 5H, ArH). MS (EI, m / z): 244 (M) ⁺ , 229 (M-CH ₃ ) ⁺ .

C. Preparation of 2-Fluoro-4'-ethoxy- [1,1'-biphenyl] -4-acetic acid

A mixture of 0.468 g (14.6 mmol) of sulfur, 2.57 ml of morpholine and 3.95 g (16.2 mmol) of the ketone prepared in Step A was heated at reflux for 17 hours (thin layer chromatography, acid-treated silica plate). and hexane: ethyl acetate (8: 2). After cooling, glacial acetic acid (9.9 ml), sulfuric acid (1.6 ml) and water (4 ml) were added and the mixture was refluxed for 30 hours. Water was added and the mixture was extracted three times with ether. The combined extracts were concentrated to a smaller volume and extracted with 10% sodium carbonate. The alkaline extracts are carefully acidified to pH 2 with concentrated hydrochloric acid. The title acid was extracted three times with ether, the extracts washed and dried over magnesium sulfate. Removal of solvent from the solution afforded 2.37 g of tan-brown solid (56.3%), m.p. 140-142 ° C.

NMR (CDCl 3, 400 MHz): δ = 3.68 (s, 2H, CH ₂ COO), 3.85 (s, 3H, OCH 3), 6.96-7.50 (m, 7H, ArH).

MS (EI, m / z): 260 (M) ⁺ , 215 (M-COOH) ⁺ .

D. Preparation of 2-Fluoro-4'-hydroxy [1,1'-biphenyl] -4-acetic acid

To a solution of the methyl ester from Step C (1.31 g, 5.04 mmol) in glacial acetic acid (17 mL) was added dropwise hydrobromic acid (48%, 48% eq.) And refluxed for 4.5 hours ( TLC (hexane: ethyl acetate = 7: 3). A little water was added and the mixture was extracted three times with ether. The extracts were washed and dried over magnesium sulfate. The solvent was removed from the solution to afford the title compound (1.13 g, 92%) as a tan solid. M.p. 208-210 ° C.

NMR (DMSO-dg, 400 MHz): δ = 3.61 (s, 2H, _CH2 COOH), 6.83 (d, 2H, J = 8.64 Hz, Ar H), 7.1 to 7.42 (m, 5H, ArH), 9.61 (s, 1H, COOH). MS (Cl, m / z): 246 (M) ⁺ , 201 (M-COOH) ⁺ .

E. Preparation of 2-Fluoro-4H-hydroxy- [1,1'-biphenyl] -4-acetic acid methyl ester

A solution of 1.1 g (4.47 mmol) of the acid obtained in Step D in 10 ml of methanol containing 0.159 g of p-toluenesulfonic acid * H ₂ O is refluxed for 1.5 hours (thin layer chromatography checked with acid treated silica). plate, eluting with 7: 3 hexane: ethyl acetate). The solvent was removed from the solution and the residue was dissolved in ethyl acetate, washed with brine and dried over magnesium sulfate. 1.16 g of a tan solid are obtained in quantitative yield which is used as such in the next step. Mp 115-118 ° C.

NMR (CDCl ₃ , 400 MHz): δ = 3.65 (s, 2H, CH ₂ CO 3), 3.73 (s, 3H, COOCH 3), 6.88 (d, 2H, J = 8.8 Hz), ArH), 7.10 (m, 2H, ArH), 7,327.44 (Itl, 3H, ArH).

MS (EI, m / z): 260 (M) ⁺ , 201 (M-COOCH ₃ ) ⁺ .

F 2 ^4-Fluoro-1 - (2-quinolinylmethoxy] - [l, ^l, -biphenyl] -4-acetic acid methyl ester

1.16 g (4.46 mmol) of phenol from Step E,

A mixture of 0.616 g (4.46 mmol) of powdered anhydrous potassium carbonate, 0.118 g, 0.445 mmol of 18-crown-6 and 10 mL of acetonitrile was stirred at room temperature under nitrogen for 15 minutes. Subsequently, 0.871 g (4.9 mmol) of 2-chloromethylquinoline, freshly prepared from its hydrochloride salt, was added and the mixture was placed in an oil bath and heated at 65 ° C for 5 hours. An excess of 10% potassium carbonate, 18-crown-6 and chloromethylquinoline was added and heating continued for a further 6 hours (TLC checking with dichloromethane: methanol = 19: 1). hexane: ethyl acetate = 7: 3). The solvent was removed from the mixture, the residue diluted with water and extracted three times with ethyl acetate. The extracts were washed and dried over magnesium sulfate. The resulting solution was removed to afford a tan solid which was purified by flash chromatography on Merek 60 silica gel pre-absorbed with dichloromethane (7: 3 hexane: ethyl acetate).

Yield: 1.55 g (87%), which was recrystallized from methanol. The resulting off-white solid had a melting point of 99-101 ° C.

NMR (CDCl ₃ ,, 400 MHz): δ = 3.64 (s, 2H, CH ₂ COO), 3.72 (s, 3H, COOCH 3), 5.43 (S, 2H, OCH ₂ Ar), δ , 1 (m, 4H, ArH), 7.35 (t, 1H,

ArH), 7.47 (d, 2H, ArH), 7.55 (t, 1H, ArH). 7.69 (d, 1H, ArH), 7.74 (t, 1H, ArH), 7.84 (d, 1H, ArH), 8.09 (d, 1H, ArH), 8.20 (d, 1H, ArH). MS (EI, m / z): 401 (M) ⁺ , 142, 114.

Analysis calculated for C ₂₅ H ₂₀ OFNO ₃ : C, 74.80; H, 5.02; N, 3.49; Found: C, 74.68; H, 4.65; N, 3.49.

G. Preparation of 2-Fluoro-4- (2-quinolylmethoxy) - [1,1'-biphenyl] -4-acetic acid

To a solution of the ester from Step F (1.69 g, 4.21 mmol) in dry tetrahydrofuran (20 mL) was added dropwise 1 N LiOH (12.6 mL) under nitrogen, and the mixture was stirred at room temperature for 3 hours (TLC, dichloromethane as eluent). methane: methanol: 19: 1 or hexane: ethyl acetate = 1: 1). The solvent was removed from the mixture, water was added to the residue, and the mixture was neutralized to pH 6.5 with 10% acetic acid. The acid was extracted with ethyl acetate (high volume required) and the extracts were dried over magnesium sulfate and evaporated to dryness. 1.65 g of an off-white solid are obtained in quantitative yield. M.p. 190-193 ° C (dec.). This product was recrystallized from ethyl acetate in a yield of 1.32% (80%)

4 g of a white solid are obtained. The purified material has a melting point of 195-196 ° C (dec.).

The analytical sample was dried under vacuum at 40 ° C.

NMR (DMSO-d6,

400 MHz): S

3.62 (s, 2H, _CH2 COO), 5.41 (s, 2H,

CH ₂ OAr), 7.15 (m, 4H, ArH),

7.41 (t, 1H, J = 8Hz, ArH), 7.48 (d,

2H, ArH),

7.61 (t, 1H, ArH),

7.69 (d, 1H, ArH), 7.78 (dt,

1H,

ArH), 8.01 (m, 2H, ARH), 8.42 (d, 1H, ArH), 12.42 (s, COOH).

MS (+ FAB, m / z): 388 (M) &lt; ^{+ &gt};.

Calcd for C ₂₄ HigFNO3 formula:

Found: C, 74.41; H, 4.68; N, 3.62.

found:

Example 3

Preparation of 3- [4- (2-quinolylmethoxy) benzoyl] -benzole acetic acid

A. Preparation of 3-Methyl- [4'-methoxy] -benzophenone

A three-necked flask fitted with a condenser, a mechanical stirrer and a dropping funnel was charged with 1.925 g (79.19 grams) of magnesium shavings under nitrogen atmosphere and enough ether to cover the shavings. A solution of 15.79 g (92.28 mmol) of 3-bromo-toluene in ml of ether is then added with a few drops of iodine crystal to initiate the reaction. The remainder of the solution is added dropwise to the mixture and the mixture is refluxed until most of the magnesium disappears. The mixture was cooled and 10 g (75.1 mmol) of 4-methoxybenzonitrile solution dried in vacuo over phosphorus pentoxide was added in portions. The mixture is heated under reflux for 2 hours (thin layer chromatography is used to check the starting material is consumed by 4 ° C). · · ·

After cooling to ice bath, 130 ml of cold water and then 25 ml of 1: 1 dilute sulfuric acid are added slowly. To complete the decomposition of the complex, the mixture was heated under reflux for 4 hours (TLC checking with 8: 2 ether: ethyl acetate). After stirring overnight at room temperature, the phases were separated and extracted three times with ether. The extracts were washed with 5% sodium bicarbonate, dried over magnesium sulfate and evaporated to dryness. The resulting crude amber oil (13.93 g) was purified by flash chromatography on Merck-60 silica, petroleum ether / ethyl acetate 8: 2 as eluent. Yield: 12.5 g (73.5%) as a pale yellow oil.

NMR (CDCl ₃ , 400 MHz): δ = 2.4 (s, 3H, CH 3), 3.9 (s, 3H, OCH 3), 6.96 (d, J = 8.8 Hz, 2H, ArH) , 7.38 (m, 2H, ArH), 7.53 (d, J = 6.9 Hz, 1H, ArH), 7.57 (s, 1H, ArH), 7.82 (d, J = 8 , 7 Hz, 2H, ArH).

MS (EI, m / z): 226 (M) &lt; ^{+ &gt};, 135 (91).

B. Preparation of 3-Bromomethyl- [4'-methoxy] -benzophenone

A solution of 17.5 g (77.4 mmol) of benzophenone from Step A in 26.5 mL of ethylene bromide containing a small amount of benzoyl peroxide is heated to reflux. A solution of 12.7 g (79.6 mmol) of bromine in 15 ml of ethylene bromide is then added over a period of about 30 minutes while irradiating with a 300 W lamp (Photolamp). Refluxing for 17 hours • ♦ • • · J J J 4 4 4 4 4 4 4 4 4 4 a

- 28 ta •

continued (TLC checking with 9: 1 petroleum ether / ethyl acetate as eluent, traces of starting material still present). The solvent was removed in vacuo, and the remaining 39.22 g of a brown oily substance was purified by flash chromatography, followed by chromatography on Merck-60 silica, pre-absorbed in dichloromethane, eluting with 9: 1 petroleum ether / ethyl acetate. . Chromatography gave about 15%, 2.59 g of unreacted starting material and 61.3 or 71.5% yield (taking into account the recovered unreacted starting material), 14.36 g of the desired product, and about 5.20 g of a mixed fraction is obtained. The resulting pale yellow solid had a melting point of 58-61 ° C. This material is used as such in the next step. NMR (CDCl ₃ , 400 MHz): δ = 3.88 (s, 3H, OCH 3), 4.52 (s, 2H, CH ₂ Br), 6.96 (d, J = 8.8 Hz, 2H, ArH), 7.44 (t, J = 7.6Hz, 1H, ArH), 7.58 (d, J = 7.8Hz, 1H, ArH), 7.66 (d, J = 7.6). Hz, 1H, ARH), 7.76 (s, 1H, ArH), 7.81 (d, J = 8.8 Hz, 1H, ArH).

MS (EI, m / z): 306/304 (1 bromo, M) +, 225, 135. Dibroma trace at 386/384/382 (possibly 5).

C. Preparation of 3-cyanomethyl- [4'-methoxy] -benzophenone g, 45.9 mmol of the bromine compound obtained in Step B is dissolved in 30 ml of dioxane and a solution of 7 g of sodium cyanide in 28.5 ml of water is added. . The mixture was refluxed for 6 hours (TLC checking with petroleum ether / ethyl acetate 8: 2). If necessary, the Μ · · * 4 4 «« 4 · «• • a a a a a a a a a

- 29 was treated with charcoal and extracted three times with ether. The extracts were dried over magnesium sulfate and evaporated to dryness. The resulting crude oily crude product (13.44 g) was purified by flash chromatography (Merck-60 silica eluting with hexane: ethyl acetate = 6: 4) after pre-adsorption on dichloromethane. This gave 10.69 g (92%) of pure product as a pale yellow oil which solidified on standing. The nearly colorless solid had a melting point of 70-71 ° C.

NMR (CDCl ₃ , 400 MHz): δ = 3.80 (s, 2H, CH ₂ CN), 3.87 (s, 3H, OCH 3), 6.95 (d, J = 8.6 Hz, 2H, ArH), 7.48 (t, J = 7.7 Hz, 1H, ArH), 7.54 (d, J = 7.6 Hz, 1H, ArH), 7.68 (s + d, J = 7 , 6 Hz, 2H, ArH), 7.79 (d, J = 8.6 Hz, 2H, ArH).

MS (EI, m / z): 251 (M) ⁺ .

D. Preparation of 3- [4-methoxybenzoyl) phenylacetic acid The nitrile prepared in step C (15.9 mmol) was dissolved in 40% sodium hydroxide (40 mL) and refluxed under nitrogen for 7 hours. Thin layer chromatography (9: 1 toluene / methanol) was added as eluent, water was added to the mixture under ice-cooling, and the solution was washed with ethyl acetate and acidified to pH 2 with cold concentrated hydrochloric acid. After extraction three times, the extracts were dried over magnesium sulfate and evaporated to dryness to give the crude product (3.56 g, 82%) as a yellow solid, m.p. 138-140 ° C NMR (CDCl 3, 400 MHZ): δ = 3.72 (s, 2H, CH ₂ COO), 3.88 (s, 3H,?

A ·· * · · · · ··· c · ··· «·

OCH ₃ ), 6.95 (d, J = 8.8 Hz, 2H, ArH), 7.43 (t, 1H, ArH), 7.48 (d, 1H, ArH), 7.65 (d, 1 H, ArH), 7.68 (s, 1H, ArH), 7.81 (d, J = 8.6 H 2, 2H, ArH).

MS (EI, m / Z): 270 (M) ⁺ , 211 (M-CH ₂ COOH) ⁺ , 135, 107.

E. Preparation of 3- [4-Hydroxybenzoyl] phenylacetic acid

8.1 g (0.030 mol) of the acid from Step D and

Thoroughly stirred mixture of pyridine hydrochloride (13.87 g, 0.120 mol) was stirred under nitrogen in an oil bath at 200-210 ° C for 7 hours (TLC: toluene / methanol 9: 1, dichloromethane: methanol 9: 1). 1 ratio). The mixture was cooled and then dissolved in dichloromethane. The solution was extracted with 1N sodium hydroxide, the extract was acidified with concentrated hydrochloric acid under cooling and extracted three times with ethyl acetate. After drying over magnesium sulfate, the solvent is removed from the solution. 7.61 g of the title compound are obtained in quantitative yield in the form of a tan solid. 147-149 ° C.

NMR (DMSO-dg, 400 MHz): 8 = 3.67 (s, 2H, _CH2 COO), 6.88 (d, J = 8.84 Hz, 2H, Ar-H), ca. 7.5 (m, 4H, ArH), 7.65 (d, J = 8.8 Hz, 2H, ArH), 10.4 (s, 1H, OH), ca. 12.3 (s, 1H, COOH).

MS (m / z): 257 (M + H) ⁺ , 217, 131.91.

F. Preparation of 3- [4-Hydroxy-benzoyl] -phenyl-acetic acid methyl ester

8.56 g (33.4 mmol) of the acid from Step E and

A mixture of p-toluenesulfonic acid monohydrate (1.05 g, 5.6 mmol) in methanol (70 mL) was refluxed for 2.5 hours (TLC control, 1: 9 methanol: toluene). Methanol was evaporated from the solution, the residue was dissolved in ethyl acetate and washed with brine. The solution was dried over magnesium sulfate and the solvent was evaporated. Yield: 8.68 g (96.2%) of a tan solid. Melting point 111-113 ° C. The crude product obtained is used in this state in the next step.

NMR (CDCl ₃ , 400 MHz): δ = 3.69 (s, 2H, CH ₂ COO), 3.69 (s, 3H, COOCH 3), 6.86 (d, J = 8.4 Hz, 2H, ArH), 7.41 (t, 1H, 7.58 Hz, 1H, ArH), 7.47 (d, J = 7.56 Hz, 1H, ARH), 7.62 (d, J = 7.4). Hz, 1H, ArH), 7.64 (s, 1H, ArH), 7.74 (d, 2H, J = 8.4 Hz, ArH). MS (m / z): 271 (M + H) ⁺ , 217, 131.91.

G. Preparation of 3- [4- (2-Quinolylmethoxy) benzoyl] -benzolacetic Acid Methyl Ester g, 14.8 mmol, phenol prepared in Step F, 2.05 g, 14.8 mmol, powdered, anhydrous. of potassium carbonate and 0.4 g (1.48 mmol) of 18-crown-6 in 35 mL of acetonitrile was stirred at room temperature under nitrogen for 15 minutes. 2-Chloromethyl-quinoline (2.9 g, 16.28 mmol) was added at one time and the free base was prepared fresh from its hydrochloride salt and kept in an oil bath at 65-70 ° C for 8 hours. TLC (toluene: methanol = 9: 1). An excess of 10% potassium carbonate, crown ether and chloromethylquinoline was added and heating continued for a further 8 hours. Acetonitrile was evaporated and the residue partitioned between water and ethyl acetate. The organic layer was dried over magnesium sulfate and evaporated. This gives 6.57 g of a tan solid crude product which is purified by flash chromatography on dichloromethane pre-adsorbed on Merck-60 silica eluting with petroleum ether / ethyl acetate 7: 3 to give 82.7% yield. 5.03 g of the title compound are obtained in the form of a pale yellow solid. M.p. 93-95 ° C.

NMR (CDCl ₃ , 400 MHZ): δ = 3.67 (s, 5H, CH ₂ COO + OCH ₃ ), 5.45 (s, 2H, ArCH ₂ O), 7.08 (d, J = 8, 8 Hz, 2H, ArH), 7.40 (t, J = 7.8 Hz,

1H, ArH), 7.46 (d, J = 7.7 Hz, 1H, ARH), 7.55 (T, J = 7.3 Hz 1H, ArH), 7.6 to 7.66 (M, 3H, Ar-H) , 7 , 7-7.82 (m, 4H, ArH), 8.07 (d, 1H, ArH), 8.20 (d, J = 8.4 Hz, 1H, ArH).

MS (EI, m / z): 411 (M) ⁺ , 142, 121.

H. Preparation of 3- [4- (2-quinolylmethoxy) benzoyl] -benzole acetic acid To a solution of the ester prepared in Step G (12.16 mmol) in dry tetrahydrofuran (66 mL) was added lithium hydroxide (37 mL, 37 mmol). and the mixture was stirred under nitrogen at room temperature for 2.5 hours (TLC checking with toluene / methanol 9: 1 as eluent). The tetrahydrofuran was evaporated and the residue was diluted with water, acidified to pH 6.5 with 10% acetic acid and extracted three times with ethyl acetate. The extracts were washed with brine, dried over magnesium sulfate and evaporated to dryness. The resulting crude yellow solid (4.94 g) was recrystallized from ethyl acetate to give 3.65 g (75%) of the title pure product as a white solid. 146-147 ° C.

NMR (DMSO-dg, 400 MHz): δ = 3.68 (s, 2H, _CH2 COO), 5.48 (s, 2H,

ArCH ₂ O), 7.22 (d, 2H, J = 8.8 Hz, ArH), 7.47 (m, 1H, ArH), 7.53 (m, 2H, ARH), 7.62 (m , 2H, ArH), 7.69 (d, J = 8.4 Hz, 1H, ArH),

7.74-7.82 (m, 3H, ArH), 8.2 (m, 2H, ArH), 8.43 (d, J = 8.5 Hz,

1H, ArH), 12.39 (1H, COOH).

MS (EI, m / z): 397 (M) ⁺ , 380 (M-OH) ⁺ , 142.

Analysis: Calculated for C25H19NO4: C, 75.57; H, 4.78; N, 3.53; Found: C, 75.22; H, 4.76; N, 3.39.

Example 4

Preparation of 3- [4- (2-Naphthylmethoxy) benzoyl] -benzole acetic acid

A. Preparation of 3- [4- (2-Naphthylmethoxy) -benzoyl] -benzolacetic acid methyl ester Ig, 3.7 mmol; A mixture of phenol (0.48 g), powdered potassium carbonate (0.48 g, 3.7 mmol), 8-crown-6 (0.098 g, 0.37 mmol) and acetonitrile (10 mL) was stirred for 15 minutes under nitrogen. 2-Bromomethyl-naphthalene (0.496 g, 4.07 mmol) was added and the oil was placed in an oil bath for 10 hours at 6570 ° C (TLC checking with dichloromethane / ethyl acetate 8: 2). An excess of 10% potassium carbonate, crown ether and bromomethylnaphthalene was added and heating continued for a further 4 hours, then acetonitrile was evaporated, the residue was dissolved in water and extracted three times with ethyl acetate. After washing with sodium hydroxide, brine, drying over magnesium sulfate and evaporation to dryness, 1.49 g of crude product are obtained in the form of a waxy solid which is used in the next step.

NMR (CDCl ₃ , 400 MHz): δ = 3.7 (s, 5H, OCH ₃ + CH ₂ COO), 5.32 (s, 2H, ArCH ₂ O), 7.08 (d, J = 8, 7Hz, 2H, ArH), 7.4-7.56 (m, 5H, ArH), 7.63-7.68 (m, 2H, ArH), 7.82-7.92 (m, 6H, Ar-H).

MS (m / z): 410 (M) ⁺ , 141.

B. Preparation of 3- [4- (2-Naphthylmethoxy) benzoyl] -benzolacetic acid

To a solution of 1.29 g (3.15 mmol) of the ester prepared in Step A is added dropwise 1 N lithium hydroxide and the mixture is stirred overnight under a nitrogen atmosphere. The solvent was evaporated and the residue was dissolved in water, acidified to pH 3 with 10% acetic acid under cooling and extracted three times with ethyl acetate. The extracts were dried over magnesium sulfate and evaporated to dryness. The residue (1.24 g, quantitative yield) was recrystallized by dissolving in a relatively large volume of warm ethyl acetate in dichloromethane and evaporating to half volume. The precipitate was collected and dried in vacuo at 45 ° C. Yield: 0.610 g (48.8%). Melting point: 150-152 ° C.

NMR (DMSO-dg, 400 MHz): δ = 3.70 (s, 2H, _CH2 COO), 5.40 (s, 2H, _ArCH2), 7.20 (d, 2H, Ar H), 7. 45-7.60 (m, 7H, ArH), 7.75 (d, 2H, ArH), 7.95 (m, 3H, ArH), 8.02 (s, 1H, ArH), 12.47 ( wide s, 1H, *

COOH).

MS (+ FAB, m / z): 397 (M + H) ⁺ , 217, 141.

Calcd for C 6 ^H ₂ 20 O 4 Calculated:% C = 78.78, H% = 5.09; Found: C, 78.12; H, 5.13.

Example 5

Preparation of 5-Phenyl-4- [4- (2-quinolin-1-methoxy) -phenyl] -2-oxa-2-propionic acid

A. Preparation of 4-methoxybenzoin To a solution of g of potassium cyanide in 35 ml of water was added 4-methoxybenzaldehyde (27.2 g, 0.2 mol), benzaldehyde (21.2 g, 0.2 mol) and 95% ethanol. The mixture was refluxed under nitrogen for 4.5 hours and then the ethanol was evaporated in vacuo. Water (200 mL) was added to the residue and the water was evaporated in vacuo to remove any unreacted benzaldehyde. The procedure was repeated twice and the remaining water was removed in the form of an azeotrope with ethanol. 56.3 g of an orange semi-solid crude product are obtained by flash chromatography, after pre-absorption with dichloromethane / ethyl acetate, chromatography on Merck-60 silica, eluting with 8: 2 hexane: ethyl acetate. Yield: 20.1 g (41.5%) of a pale yellow solid. 99-101 ° C.

NMR (CDCl ₃ , 400 MHz): δ = 3.82 (s, 3H, OCH 3), 4.62 (broad s, 1H, OH), 5.88 (s, 1H, CHOH), 6.86 (d , 2H, J = 8.94 Hz, ArH), 7.22-7.38 (m, 5H, ArH), 7.91 (d, 2H, J = 8.94 Hz, ArH).

MS (Cl, m / z): 243 (M + H) &lt; ⁺ & gt ^; , 225, 197, 137 (M-PhCO) &lt; ^{+ &gt};.

B. Preparation of 4-methoxybenzoin hemisuccinate A mixture of g, 0.083 mol of 4-methoxybenzoin and 9.1 g, 0.091 mol of succinic anhydride in 6 ml of toluene was heated at 135 ° C (internal temperature) for 7 hours under nitrogen. The solution was then poured into 0.5N sodium bicarbonate, the organic phase was separated and extracted with 0.5N sodium bicarbonate. The combined extracts were washed with ether and then acidified with concentrated hydrochloric acid under cooling. The resulting oil was extracted three times with ethyl acetate, and the extracts were washed with water and dried over magnesium sulfate. The solvent was removed from the solution. Yield: 20.89 g (73.8%) as a yellow solid. 104-108 ° C. This material was used in the next step without further purification.

NMR (CDCl ₃ , 400 MHZ): δ = 2.72-2.82 (mm, 4H, CH ₂ CH ₂ COO), 3.82 (S, 3H, OCH 3), 6.86 (d, 2H, J = 9.1 Hz, ArH), 7.34-7.46 (m, 5H, ArH), 7.92 (d, 2H, J = 9.1 Hz, ArH).

MS (EI, m / z): 342 (M) &lt; ^{+ &gt};, 135.

C. Preparation of 4- (4-Methoxyphenyl) -5-phenyl-2-oxazole propionic acid

A mixture of crude 4-methoxybenzoine hemisuccinate (20.8 g, 0.061 mol) prepared in Step B, 8.7 g (0.146 mol) of urea and 60 ml of acetic acid was refluxed under nitrogen for 5.5 hours. The mixture was cooled and poured into ice water. The resulting oil was extracted three times with ethyl acetate. The extracts were washed neutral with water and then extracted with saturated sodium carbonate. The combined aqueous extracts were carefully acidified with concentrated hydrochloric acid under cooling and then extracted with ethyl acetate. The organic extract was dried over magnesium sulfate and evaporated to dryness. 19.6 g of a waxy yellow oil are obtained. This material was purified by flash chromatography on Merck-60 silica, eluting with 8: 2 dichloromethane: ethyl acetate. 14.3 g (72.7%) of a pale yellow solid are obtained. Melting point 100-101 ° C.

NMR (CDCl ₃ , 400 MHz): δ = 2.96 (t, 2H, CH ₂ CO), 3.20 (t, 2H, CH ₂ COO), 3.83 (s, 3H, OCH ₃ ), δ , 90 (d, 2H, ArH), 7.28-7.38 (m, 3H, ArH), 7.54-7.62 (m, 4H, ArH).

MS (EI, m / z): 323 (M) ⁺ , 278 (M-COOH) ⁺ , 152, 77.

D. Preparation of 4- (4-Hydroxy-phenyl) -5-phenyl-2-oxazole-propionic acid

To a solution of methoxyacid (5.6 g, 17.3 mmol) in acetic acid (55 ml) was added 48% hydrobromic acid (84 ml) and the mixture was refluxed under nitrogen for 8 hours (TLC checking 1: 1 hexane: ethyl acetate). After cooling, water was added and the solution was extracted three times with ethyl acetate. The extract was dried over magnesium sulfate and evaporated to dryness. The residual brown waxy oil (5.25 g, 99%) was used in the next step without purification. For analytical characterization, a small sample is purified by flash chromatography on Merck-60 silica, eluting with dichloromethane: methanol (98: 2 to 95: 5).

NMR (DMSO-dg, 400 MHz): δ = 2.75 (t, 2H, J = 7.14 Hz, CH ₂ C);

3.02 (t, 2H, J = 7.1 Hz, CH ₂ COO), 6.77 (d, 2H, J = 8.7 Hz, ArH),

7.35 (d, 2H, J = 8.55 Hz, ArH), 7.41 t, 3H, J = 7.12 Hz, ArH),

7.50 (d, 2H, J = 7Hz, ArH), 9.64 (s broad, interchangeable).

MS (EI, m / z): 309 (M) +, 264 (M-COOH) ⁺ , 121, 105.77.

E. 5-Phenyl-4- (4-hydroxy-phenyl) -2-oxazole-propionic acid methyl-

Preparation of the ester g (16.18 mmol), the crude acid prepared in Step D

A solution of 0.58 g of p-toluenesulfonic acid monohydrate in 40 ml of methanol is refluxed for 2.5 hours. The methanol was evaporated and the residue was partitioned between ethyl acetate and 20% sodium chloride. The extracts were washed, dried over magnesium sulfate and evaporated. 4.8 g of a thick oil are thus obtained. This material was purified by flash chromatography eluting with dichloromethane on silica after chromatography on Merck-60 silica gel, eluting with a gradient of dichloromethane / ethyl acetate 90:10 to 75:25. 3.56 g of a white solid are obtained in 68% yield. 115-116 ° C.

NMR (CDCl ₃ , 400 MHz): δ = 2.92 (t, 2H, J = 7.4 Hz, CH ₂ C), 3.20 (t, 2H, J = 7.4 Hz, CH ₂ COO) , 3.71 (s, 3H, OCH ₃ ), 6.74 (d, 2H, J = 8.59 Hz, ArH), 7.26-7.36 (m, 3H, ARH), 7.40 ( d, 2H, J = 8.7 Hz, ArH), 7.54 (d, 2H, J = 7.56 Hz, ArH).

• · »· · · · ················ · · · · · · ·

MS (EI, m / z): 323 (M) ^+, 264 _(M-COOCH3) ^+, 105.77.

F. Preparation of 5-Phenyl-4- [4- (2-quinolylmethoxy) phenyl] -2-oxazole-propionic acid methyl ester

2.46 g (7.61 mmol) of the ester from Step E,

A mixture of 1.05 g (7.60 mmol) of powdered anhydrous potassium carbonate, 0.223 g, 0.843 mmol of 18-crown-6 and 33 mL of acetonitrile (molecular sieve dried) is stirred at room temperature under nitrogen for 15 minutes. To the mixture was added 1.35 g (7.60 mmol) of 2-chloromethylquinoline, freshly prepared from its hydrochloride salt and kept at 65 ° C for 10 hours. After 6 hours, 10% excess chloromethylquinoline, 18-crown-6 and potassium carbonate were added. After 10 hours, the solvent was removed and the residue was partitioned between ethyl acetate and water. The extracts were washed with brine, dried over magnesium sulfate and evaporated. The crude yellow solid was purified by flash chromatography using Merck-60 silica eluting with toluene followed by toluene / methanol (97.5: 2.5). 3.5 g of the title compound are obtained in quantitative yield.

NMR (CDCl ₃ , 400 MHz): δ = 2.90 (t, 2H, J = 7.2 Hz, CH ₂ C), 3.16 (t, 2H, J = 7.2 Hz, CH ₂ COO ), 3.72 (s, 3H, OCH 3), 5.41 (s, 2H, ArCH ₂ O), 7.02 (d, 2H, J = 8.8 Hz, ArH), 7.29-7, 36 (m, ca. 4H, ArH), 7.54-7.84 (m, ca. 6H, ArH), 7.83 (d, 1H, J = 8.1 Hz, ArH), 8.08 (t, 1H, J = 8.5 Hz, ArH), 8.19 (d, 1H, J = 8.5 Hz, ArH). MS (+ FAB, m / z): 487 (M + Na) +, 465 (M + H) ⁺ .

• · · ·

G. Preparation of 5-Phenyl-4- [4- (2-quinolylmethoxy) phenyl] -2-oxazole propionic acid

To a solution of the ester prepared in Step F (3.4 g, 7.32 mmol) in dry tetrahydrofuran (37 mL) under nitrogen was added dropwise 21.98 mL (3 equivalents) of 1 N lithium hydroxide and the mixture was stirred at room temperature for 3 hours (TLC check). Elution with dichloromethane: methanol (97: 3) or toluene: methanol (95: 5) was carried out, the solvent was evaporated and the residue was dissolved in water under cooling with 10% acetic acid to pH 5.5-6. The extracts were washed with brine, dried over magnesium sulfate and evaporated to give a pale yellow solid (3.18 g) which was recrystallized from warm ethyl acetate with sufficient dichloromethane. to obtain a clear solution, a first crop of crystals (2.63 g), m.p. 192194 ° C (dec.) A second crop is obtained by evaporation of the mother liquor, 0.327 g, m.p. 192-193 ° C (dec.). The combined yield was 85.8%.

IR (KBr, cm ^-1 ): 1720 (CO).

NMR (DMSO-dg, 400 MHz): δ = 2.76 (t, 2H, J = 7Hz, _CH2 C), 3.03 (t, 2H, J = 7Hz, CH ₂ COO), 5, 38 (s, 2H, _ArCH2 O), 7.11 (d, 2H, J = 8.8 Hz, Ar H), 7.36-7.56 (m, 7H, Ar H), 7.61 (t, 1H, ArH), 7.69 (d, 1H, J = 8.5 HZ, ArH), 7.78 (t, 1H, ArH), 8.00 (t, J = 7.9 Hz, 2H, ArH) ), 8.42 (d, 1H, J = 8.5 Hz, ArH).

MS (EI or Cl, m / z): 451 (M + H) &lt; ⁺ & gt ^; , 310.

· • «· ·

44 4 * 44 444 · ♦ »

41 - Analysis for C 28 ^H 22 ^N 2 O 4 Calculated:% C = 74.65, H% = 4.92, N% = 6.22; Found: C, 74.20; H, 4.86; N, 6.00.

Example 6

Preparation of 4- [4- (2-Naphthylmethoxy) phenyl] -5-phenyl-2-oxazole propionic acid

A. Preparation of 4- [4- (2-Naphthylmethoxy) phenyl] -5-phenyl-2-oxazole-propionic acid methyl ester

A mixture of 1.5 g, 4.6 mmol of the hydroxy ester prepared in Example 5E, 0.636 g, 4.6 mmol of powdered anhydrous potassium carbonate, 0.123 g, 0.46 mmol of 18-crown-6 and 18 ml of acetonitrile was added at room temperature. under nitrogen atmosphere for 15 minutes. 2-Bromomethyl-naphthalene (1.13 g, 5.1 mmol) was then added and the mixture was placed in an oil bath at 70 ° C for 8-9 hours (TLC checking with hexane / ethyl acetate 9: 1 or dichloromethane). methane / methanol 9: 1). The solvent was evaporated and the residue was dissolved in water and extracted with ethyl acetate. The extracts were washed and dried over magnesium sulfate. The solvent was evaporated to yield 2.17 g of a tan solid. The sample is recrystallized from methanol (methanol must contain sufficient dichloromethane to give a clear solution), the methanolic solution is concentrated to a small volume and cooled in an ice bath. The resulting white solid was collected and dried under vacuum overnight. 134-135 ° C.

IR (KBr, cm ^-1 ): 1740 (CO).

• · · ·

- 42 NMR (CDCl3, 400 MHz): δ = 2.89 (t, 2H, J = 7.5 Hz, _CH2 C), 3.16 (t, 2H, J = 7.5 Hz, _CH2 COO ), 3.71 (S, 3H, OCH 3), 5.2 (s, 2H, ArCH ₂ O), 7.00 (d, 2H, J = 8.6 Hz, ArH), 7.25-7, 35 (m, 3H, ArH), 7.46-7.58 (m, 7H, ArH), 7.8-7.9 ((m, 4H, ArH).

MS (Cl, m / z): 464 (M + H) &lt; ⁺ &gt;, 324.

Analysis calculated for C 30 H ₂₅ NO ₄ : C, 77.73; H, 5.44; N, 3.02; Found: C, 77.44; H, 5.36; N, 3.03.

B. Preparation of 4- [4- (2-Naphthylmethoxy) phenyl] -5-phenyl-2-oxazole propionic acid

The ester from Step A (1.49 g, 3.21 mmol) was obtained

A solution of 9.6 ml of 1 N lithium hydroxide in 18 ml of dry tetrahydrofuran was stirred under nitrogen at room temperature overnight (TLC control, eluting with 75:25 hexane: ethyl acetate). The solvent was evaporated and the residue was dissolved in water and then acidified to pH 5 with dilute hydrochloric acid. The mixture was extracted with ethyl acetate, and the extracts were dried over magnesium sulfate and evaporated to give 1.39 g of crude product, m.p. 145-150 ° C. The crude product is purified by purification in hot ethyl acetate containing sufficient dichloromethane to give a clear solution, then the solution is concentrated to half volume and precipitated with ether. 1.07 g of a white solid are obtained in 58% yield. M.p. 151-152 ^° C.

IR (KBr, cm ^-1 ): 1720 (CO).

• * • a «· ··« · ”« 4 ·· 4 4 «*

- 43 NMR (DMSO-dg, 400 MHz): 8 = 2.78 (t, 2H, _CH2 C), 3.03 (t, 2H, J = 7Hz, CH ₂ COO), 5.29 (s , 2H, ArCH ₂ O), 7.10 (d, 2H, J = 8.9 Hz, ArH), 7.34-7.60 (m, 10H, ArH), 7.90-8.00 (m , 4H, ArH), 12.28 (s, 1H, COOH).

MS (EI, m / z): 450 (M + H) ⁺ 310 Analysis for C ₂ 3NO4 gH ₂ O: Calculated: C% = 77.48, H% = 5.15, N% = 3, 11; Found: C, 76.40; H, 5.16; N, 3.04.

Example 7

Preparation of 4- [4- (1-Methyl-1H-benzoimidazol-2-yl) -methoxy-phenyl] -5-phenyl-2-oxazole-propionic acid

A. Preparation of 4- [4- (1-Methyl-1H-benzoimidazol-2-yl) -methoxy-phenyl] -5-phenyl-2-oxazole-propionic acid methyl ester

A mixture of 0.5 g, 1.55 mmol of the ester prepared in Example 5E, 0.214 g, 1.55 mmol of powdered anhydrous potassium carbonate, 0.0416 g, 0.155 mmol of 18-crown-6 and 6 mL of acetonitrile was added under nitrogen atmosphere at room temperature. Stir for 15 minutes. Then, 2-chloromethyl-1-methylbenzimidazole (0.307 g, 1.7 mmol) was added and the mixture was placed in an oil bath at 65-70 ° C for 4 hours (TLC checking with dichloromethane and ethyl acetate). acetate (9: 1), blotting with iodine). An excess of 10% potassium carbonate, 2-chloromethyl-1-methylbenzimidazole and 18-crown-6 was added and heating continued for a further 10 hours. The solvent was evaporated and the residue was dissolved in water and extracted with ethyl acetate. The extracts were washed, dried over magnesium sulfate, and dried over magnesium sulfate. , then evaporate to dryness. The residue (1.64 g) was purified by flash chromatography using Merck-60 (8: 2 dichloromethane / ethyl acetate) as the eluent on a small amount of methanol pre-absorbed in dichloromethane. 1.03 g (71.2%) of a pale yellow solid are obtained. 142-144 ° C (dec.).

NMR (CDCl ₃ , 400 MHz): δ = 2.87 (t, 2H, J = 7.1 Hz, CH ₂ C), 3.16 (t, 2H, J = 7.8 Hz, CH ₂ COO) 3.72 (s, 3H, COOCH3), 3.90 (s, 3H, NCH3), 5.41 (s, 2H, _ArCH2 0), 7.07 (d, 2H, J = 8.8 Hz , ArH), 7.25-7.40 (m, 7H, ArH), 7.5-7.6 (m, 3H, ArH), 7.78 (d, 1H, ArH). MS (+ CI, m / z): 468 (M + H) ⁺ , 324, 293, 147.

B. Preparation of 4- [4- (1-Methyl-1H-benzoimidazol-2-yl) -methoxy] -5-phenyl-2-oxazole-propionic acid lg, 2.14 mmol of the ester from Step A, 6.42 A solution of 1 N lithium hydroxide (1 ml) in THF (13 ml) was stirred under nitrogen at room temperature for 1 hour (TLC checking with dichloromethane: ethanol 9: 1). The solvent was evaporated, water was added to the residue and the pH was adjusted to 6.5 with 10% acetic acid. The resulting pale yellow precipitate was collected, washed with water and dried in vacuo. The solid was redissolved in hot ethyl acetate, the ethyl acetate should contain enough methanol to give a clear solution, evaporated to a smaller volume and cooled in an ice bath. The crystals were collected and dried. Yield: 0.642 g (66.2%). M.p. 222-224 ° C. The product obtained has a melting point of 222-224 ° C.

NMR (DMSO-dg, 400 MHz): S = 2.76 (t, 2H, J = 7Hz, _CH2 C), 3.03 (t, 2H, J = 7Hz, CH ₂ COO) 3, 86 (s, 3H, NCH ₃ )), 5.43 (s, 2H, ArCH ₂ O), 7.14-7.66 (m, 13H, ArH).

MS (Cl, m / z): 454 (M + H) &lt; ⁺ & gt ^; , 147.

Analysis: Calculated for C27H23N3O4: C, 71.51; H, 5.11; N, 9.27; Found: C, 71.62; H, 5.17; N, 9.40.

1. Test Example

5- and 12-hydroxy-eicosetraenoic acid (5-HETE and 12-HETE) and LTB4 are the early arachidonic acid oxidation products of the lipoxygenase cascade, which are known to mediate the allergic response and some aspects of inflammation. This is especially true for 5,12-diHETE, also known as LTB4 (see Ford-Hitchinson, J. Roy. Soc. Med. 74, 831 (1981)]. Consequently, compounds that inhibit arachidonic acid PLA2-mediated release effectively prevent the oxidation of arachidonic acid to various leukotriene products via the lipoxygenase cascade. Accordingly, the specificity of the effect of PLA ₂ inhibitors can be determined by measuring the activity of the test compounds in a study that measures the ability of the compounds to inhibit LTB4 synthesis by rat glycogen-induced polymorphonuclear leukocytes (PMN).

Carry out the test in the following way:

Rat polymorphonuclear leukocytes (PMNs) were obtained from female Wistar rats weighing 150 to 200 g, which were <4 · 4 · 10 ml of 6% glycogen was injected intraperitoneally. The rats were quenched with carbon dioxide 18 to 24 hours after glycogen injection and the glycogen-induced cells were harvested by peritoneal wash in physiological saline (0.9% sodium chloride solution). This material was centrifuged at 400 xg for 10 minutes. The supernatant fluid was discarded and the cell pellet suspended in 2.0 x 10 ⁷ cells / ml in HBSS containing Ca ⁺⁺ , Mg ⁺⁺ and 10 mM L-cysteine.

To a 1 ml aliquot of the above cell suspension is added test substance or vehicle and the sample is preincubated at 37 ° C for 10 minutes. Samples were then added with 1 g A23187, [ ³ H] -AA (3.0 gCi / ml) and 1 g unlabelled arachidonic acid (AA), and incubated for a further 10 minutes. The reaction is stopped by settling the cells by centrifugation. The supernatant was then analyzed by high performance liquid chromatography on a 15 cm x 4.6 mm ID supelcosil LC-18 (Supelco) (3M) column using a two solvent solvent system having a flow rate of 1.4 mL, given below:

The solvent was 70:30 17.4 mmol ΗβΗΡΟΟΗΟΗΟΗ

Solvent B: CH 3 CN

Gradient: (system equilibrated with solvent)

Time THE % B% 0 100 0 15.0 100 0 20.0 65 35

Time THE % B% 40.0 65 35 42.0 10 90 50.0 10 90 50.1 100 0

The percentage change in the solvents is linear.

Injection: 140 μΐ of each supernatant was injected directly onto the column and the ³ H arachidonic acid metabolites were monitored by an on-line radioactivity detector (Ramona, IN / US, Fairfield, NJ).

Standards: 10 ⁴ -2.0 x 10 ⁴ dpm of the desired eicosanoid was injected in a 90 μΐ ethanol cocktail.

Percent inhibition was determined by comparison with co-chromatography of standard [ ³ H] leukotriene B ₄ (LTB ₄ ) on stimulated PMN medium exposed to test substance and medium on non-exposed stimulated cells.

Our results are expressed as percent inhibition at a given compound dose or in the form of an IC 50.

The results of testing the compounds of the invention are shown in Table I below.

Table I

Example Compound% Inhibition Ketoprofen

First

-50 * (at 10 μιπόΙ / 1) (0.5 μιπόΙ / 1)

Table I (continued)

Example rinti error %-ancestor Inhibition Second 91 (0.5 Mmol / l) Third 87 (10 μιηόΙ / than 1) 38 (0.5 μιπόΙ / than 1) 4th 8 (10 μιπόΙ / than 1) 5th 96 (10 μιηόΙ / than 1) 6th 95 (10 μιηόΐ / l) 81 (0.5 μιηόΐ / l) 6A. 94 (10 μιπόΐ / l) 63 (0.5 μιηόΐ / l) 7th 85 (10 μιηόΐ / l)

* Negative value indicates potentiation of cyclooxygenase (PGE ₂ synthesis).

2. Test Example

Test Example 1 was used to determine the extent to which compounds of the invention inhibit the synthesis of the arachidonic acid cyclooxygenase oxidation product, PGE ₂ .

In this assay, the procedure of Assay Example 1 is followed, however, for determination of cyclooxygenase activity, the samples are co-chromatographed with authentic reference [ ³ H] -PGE ₂ .

The results are calculated as in Test Example 1. Our results are shown in Table II. in Table.

II. Spreadsheet

Example Compound% Inhibition

ketoprofen 87 (10 μχηόΐ / 1) First -13 * (0.5 gmol / 1 in) Second -22 * (0.5 gmol / l) Third 8 (10 gmol / 1 in) -8 * (0.5 μπιόΙ / than 1) 4th -31 * (10 μπιόΐ / 1) 5th -275 * (10 μπιόΐ / 1) 6th -191 * (10 μιηόΙ / than 1) -12 * (0.5 μτηόΐ / 1) 6A. -79 * (10 μπιόΐ / 1) -29 * (0.5 μιηόΐ / 1) 7th -268 * (10 μιηόΐ / ΐ-ηέΐ)

* Negative value indicates potentiation of cyclooxygenase (PGE ₂ synthesis).

3. Test Example

The compounds of the present invention were tested in vitro in isolated phospholipase A ₂ assay to determine their ability to inhibit the release of arachidonic acid from a substrate containing arachidonic acid by the enzyme human and non-human phospholipase A ₂ .

Carry out the test in the following way:

Add the following ingredients to a 1 ml polypropylene tube:

Material Volume, u.1 Final concentration

³ H-AA E. coli substrate ¹ CaCl ₂ (0.1 M) ² 25 5 5 nM / L PL 5 mmol / l Tris-HCl (0.5 M) pH 7.5 ³ 20 100 mM Water ⁴ 25 Drug / carrier ⁵ 1 50 μιηόΐ / ΐ PLA ₂ 25 12% hydrolysis over 10 minutes

Volume resulting in 100 zist * Preincubate at room temperature for 30 minutes before adding substrate.

¹ The substrate is prepared by adding 2 ml deionized and distilled water to 2 ml of ³ H-arachidonate-labeled E. coli (low count) and then to the mixture is added 1 ml of ³ H-arachidonate-labeled E. coli (higher count) ) to give a total of 5 ml of substrate containing 1000 nmol of phospholipid.

² 0.1 M CaCl ₂ stock solution, required for enzyme activity.

³ 0.5 M Trisma-base stock solution.

0.5 M Trisma-HCl stock solution to adjust pH to 7.5 (optimal pH for the enzyme) ⁴ Non-ionic and distilled water.

⁵ 10 mM stock solution in dimethylsulfoxide. This is diluted 1: 2 with dimethylsulfoxide and 1 μΐ is added to a 100 μΐ test tube.

⁶ Two types of human PLA ₂ enzymes are used:

a) Semi-purified human platelet acid extract in PLA ₂ (10 mM sodium acetate pH 4.5 buffer). The protein precipitate was removed by centrifugation at about 2200 rpm for 10 minutes.

(b) Purified human synovial fluid.

The 100 μΐ reaction mixture was incubated for 10 minutes in a water bath at 37 ° C. The reaction was quenched with 2 mL of tetrahydrofuran and vigorously stirred. NH ₂ columns (100 mg / ml - Analytichem International) were conditioned with 0.5 ml of tetrahydrofuran followed by 0.5 ml of tetrahydrofuran-water (2 ml: 0.1 ml).

The samples are placed on the columns and slowly passed through them. The hydrolyzed arachidonic acid remains on the column and is eluted from the column with 1 ml of glacial acetic acid in 2% tetrahydrofuran. Arachidonic acid was transferred to scintillation tubes and quantified by B-count analysis. Total counts were prepared by pipetting 25 μΐ of ³ H-arachidonate E. coli directly into a scintillation tube and adding 1 ml of tetrahydrofuran. To each sample is added 10 ml of aquazole (scintillation cocktail).

Calculations:

pH] AA dpm (sample) - pHJAA dpm (non-specific hydrolysis)% hydrolysis = ------------------------------- ------------------------ x 100 total counts dpm carrier dpm - agent dpm% change = _______________________________________ x 100 carrier dpm

Standard active ingredient activity:

IC ₅₀ (μπιό1 / 1)

agent human platelet pla ₂ human synovial fluid pla ₂ arachidonic acid 8.6 3.2 Monoalid 25.2 0.14 Above by the invention for compounds of formula

the following results were obtained:

III. Spreadsheet

Example% Inhibition at 10 μιηόΙ / 1 compound ______________________________ HSF * ________________ ketoprofen -16.9 **

3. 25.5

4. 15.1 * human synovial fluid ** the negative value represents the potentiation of HSF.

4. Test Example

The ability of the compounds of the present invention to inhibit the lipidoxygenase and / or cyclooxygenase pathways of arachidonic acid metabolism is measured in vivo in a mouse zymosan peritonitis assay.

Carry out the test in the following way:

7-week-old male CD-1 mice are housed in plastic cages in groups of six. Animals were intraperitoneally administered with 1 ml of a 1% solution of zymosan in 0.9% pyrogen-free saline or saline (unstimulated control). Test compounds are administered orally 1 hour prior to zymosan injection. 20 minutes after the zymosan injection, the mice were strangled with carbon dioxide inhalation and the abdomen was flushed with 2 mL of ice-cold Hanks Balanced Salt Solution (HBSS) without CaCl ₂ , MgSO ₄ · 7Η 2θ and MgCl ₂ · 6H ₂ O. The peritoneal lavage fluid is aspirated from each mouse using a syringe and placed in a 5 mL ice-cooled test tube, and the volume of the fluid is recorded. For evaluation by ELISA, the samples are prepared by centrifuging the sample at 800 xg for 15 minutes, adding 1 mL of supernatant to 8 mL of ice-cold methanol and maintaining overnight at -70 ° C to precipitate proteins at 800 xg. Centrifuge for 15 minutes and dry in a Savant speed vac concentrator. Samples were reconstituted in 1 ml ice-cold ELISA buffer and stored at -70 ° C until assay. _Xa assays for eicosanoids (LTC ₄ and 6-keto-PGF _Xa) were performed by standard ELISA.

Compounds for oral administration were suspended in 0.5% Tween 80. Substances for intraperitoneal examination were suspended in 0.5% methylcellulose in 0.9% saline.

The total metabolite level per mouse in washing fluid was calculated and significance was determined by one-way analysis of variance by comparison with control LSD (p <0.05). The effect of the drugs is given as the percentage change from the control value. The activity of some standard drugs in this study is as follows:

ED5o-fflg / p-G ^ - = - g L

Compound

ltc ₄ 6-ketO-PGF1 _a / TxB2 BW755C <10 22.0 phenidone 24.0 <30.0 indomethacin inactive 0.126 ibuprofen inactive 7.0 That's it procedure one according to the invention congregation and anti-inflammatory etodolac we obtained the following results: ARC. Spreadsheet Example Dose % Inhibition of mg / kg ltc ₄ 6-keto-PGF compound

5th

ve

-27 ** (i.p.) * 86 * intraperitoneally ** negative value indicates potentiation.

The results show that the compound of the present invention is effective in inhibiting the lipoxygenase pathway but has no inhibitory effect on the cyclooxygenase pathway.

Claims (11)

Claims CLAIMS 1. A compound of formula (CH2) n ^O-B and pharmaceutically acceptable salts thereof A is phenoxyethyl, phenoxyphenyl or a group of formula (a) wherein R ³ I X is -N = or -C-; R ³ R ³ R ³ R ³ R ³ II II Z is -C-C-, -ON-, -N = C-, -N-, -S- or -O-, R ¹ is hydrogen, lower alkyl or phenyl, R ² is hydrogen or lower alkyl, or R ¹ and R ² together form a benzene ring, R ³ is hydrogen or lower alkyl, n is 1 or 2, B is a group of formula (g), (h), (i) or (j) wherein R ⁴ and R ⁵ are independently hydrogen or lower alkyl, R ⁶ is halogen or nitro, 0 R ⁴ R ⁷ is -CR ⁸ or -CHCOOR ⁵ R ⁸ is lower alkyl and m is from 0 to 3. »· • ·« «· · · * * * · · * ··« · · ··· · «· - 56 Á The compound of claim 1, wherein the compound is 1- [2-nitro-4 '- (2-quinolylmethoxy) - [1,1'-biphenyl] -4-yl] ethanone. 3. A compound according to claim 1 wherein the compound is 2-fluoro-4- (2-quinolylmethoxy- [1,1'-biphenyl] -4-acetic acid. 4. The compound of claim 1, wherein the compound is 3- [4- (2-quinolylmethoxy) benzoyl] -benzole acetic acid. 5. The compound of claim 1, wherein the compound is 3- [4- (2-naphthylmethoxy) benzoyl] -benzole acetic acid. The compound of claim 1, wherein the compound is 5-phenyl-4- [4- (2-quinolylmethoxy) phenyl] -2-oxazole-propionic acid. The compound of claim 1, wherein the compound is 4- {4- [2-naphthylmethoxy] phenyl} -5-phenyl-2-oxazole propionic acid. 8. The compound of claim 1, wherein the compound is 4- {4- [2-naphthylmethoxy] phenyl} -5-phenyl-2-oxazole-propionic acid methyl ester. The compound of claim 1, wherein the compound is 4- {4 - [(1-methyl-1H-benzimidazol-2-yl) methoxy] phenyl} -5-phenyl-2-oxazole propionic acid . 10. A process for the preparation of the compounds of the formula A- (CH 2) _n -OB and their pharmaceutically acceptable salts as defined in claim 1, characterized in that: a) ^a compound of formula (6), (7), (8) or (9) wherein R ⁴ , R ⁶ and n are as defined in claim 1; OR ⁴ R ⁷ is -CR ⁸ or -CHCOOR ⁵ , and -COOR ^{5 is} an ester function, etherified with a compound of formula A (CH 2) _n " ^x - wherein A and n are as defined in claim 1, X is a leaving group -, and if desired the resulting compound is hydrolyzed with a compound selected from hydrogen or lower alkyl R ⁵ position; obsession b) a compound of formula (I), (II), (III) or (IV) having -COOR ⁵ units wherein R ⁵ is lower carbon. hydrolyzing an alkoxy group, the corresponding compound wherein R ⁵ is hydrogen, or a pharmaceutically acceptable salt; c) converting a compound of formula (I), (II), (III) or (IV) into a pharmaceutically acceptable salt. 11. A process for the preparation of a pharmaceutical composition comprising a compound of formula A (CH ₂ ) _n -OB, or a pharmaceutically acceptable salt thereof, wherein the substituents are as defined in claim 1, with pharmaceutically acceptable carriers and / or other excipients. formulated as a pharmaceutical composition.