CA2067138A1 - Substituted benzoylbenzene-, biphenyl-and 2-oxazole-alkanoic acid derivatives as inhibitors of pla2 and lipoxygenase - 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-;
R1 is hydrogen, lower alkyl or phenyl; R2 is hydrogen or lower alkyl; or R1 and R2 taken together form a benzene ring;
R3 is hydrogen or lower alkyl; n is 1 -2; B is (g), (h), (i) or (j), wherein R4 and R5 are each, independently, hydrogen or lower alkyl; R6 is halo or nitro; R7 is (k) or (l); R8 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

2~7138 ~,.'0 91/06538 PCT/US90/062~2 I
SUBSTITUTED BENZOYLBENZENE-, BIPHENYL-AND 2-OXAZOLE- ALKANOIC A~ID DERIVATIVES
AS INHIBITORS OF PLA2 AND LIPOXl~'GENASE

This invention relates to novel substituted benzoylben~ene-, biphenyi- and 2-oxazole- alkanoic acid derivatives possessing lipoxygenase inhibitory, phospholipase A2 inhibitory and leukotriene antagonist activity, which are useful as anti-infla~mmatory, anti-allergic and cytoprotective agents.
It is now well-established that arachidonic acid (AA) is metabolized in mammals 10by two distinct pathways. The metabolism of arachidonic acid by cyclooxygenase enzymes results in the production of prostaglandins and thromboxanes. The physiological activity of the prostaglandins has already been amply elucidated in recent years. It is now known that prostaglandins arise from the endoperoxides PGG2 and PGH2 by the cyclooxygenase pathway of arachidonic acid metabolism. These endoperoxides are also the precursors of the throm-15boxanes (Txj A2 and B2. TxA2 is a vasoconstrictor which stimulates platelet aggregation. In the normal situation, the vasoconstrictive and platelet aggregating praperties of the thrombox-anes are balanced by another product arising from the endoperoxides in the cyclooxygenase pathway, prostacyclin (PGI2), which is a vasodilator with platelet aggregation inhibitory activity. In the event prostacyclin synthesis is impaired and/or platelet activation is enhanced, 20then thrombosis and vasoconstriction is favored. The role of prostanoids in haemostasis and thrombosis are reviewed by R.J. Gryglewski, CRC Crit. Rev. Biochem., 7. 291 (1980) and J.
B. Smith, Am. J. Pathol., 99, 743 (1980). Cyclooxygenase metabolites are known to partici-pate directly in thé inflammatory response [see Higgs et al., Annals of Clinical Research, 16, 287-299 (1984)]. This is through their vasodepressor activities, participation in pain and ~ever 25augmentation of peptide mediator vascular permeability and edema forming propenies.
Finally, various aspects of cell mediated immunity are influenced by cyclooxygenase products.
The other pathway of AA metabolism involves lipoxygenase enzymes and results in the production of a number of oxidative products called leukotrienes. The latter are designated by the LT nomenclature system, and the most significant ptoducts of the lipoxy-; 30genase metabolic pathway are the leukotrienes B4, C4 and D4. The substance denominated slow-reacting substance of anaphylaxis (SRS-A) has been shown to consist of a mixture of leuko~rienes, with LTC4 and LTD4 as the primary products and having varying amounts of other leukotriene metabolites [see Bach et al., J. Immun., 215, 115-118 (1980); Biochem.
Biophvs. Res. Commun., 2~, 1121-1126 (1980)].
35The significance of these leukotrienes is that a great deal of evidence has been accumulated showing that leukotrienes participate in inflammatory teactions, exhibit chemo-.

,. . . .

.

WO 91/06538 ~ ~1 6 7 1 3 8 PCr/US90/06252 ~

!~actic activities, s~imulate Iysosomal enzyme release and act as important factors in the immedi-ate hypetsensitivity reaction. It has been shown that LTC4 and LTD4 are potent bronchocon-strictors of the human bronchi [see Dahlen et al., Nature, 288, 484-486 (1980) and Piper, Int.
~Arch. Appl. Immunol., 76, suppl. 1, 43 (1985)] which stimulale the release of mucus from S airways in vitro [Marom et al., Am. Re~. Resp. Dis., ~, 449 (1982)], are potent vaso-dilators in skin [see Bisgaard et al., Prostaglandins~ 23. 797 (1982)], and produce a wheal and flare response [Camp et al., Br. J. Pharmacol., 80, 497 (1983)]. The nonpeptide leukotriene, LTB4, is a powerful chemotactic factor for leukocytes [see A. W. Ford-Hutchinson, J. Rov.
Soc. Med., 74, 831-833 (1981), which stimulates cell accumulation and affects vascular smooth muscle [see Bray, Br. Med. Bull., 39, 249 (1983)]. The activity of leukotrienes as mediators of inflammation and hypersensitivity is extensively reviewed in Bailey and Casey, Ann Reports Med. Chem., 19, 87 (1986).
Phospholipase A2 (PLA2) is the critical rate limiting enzyme in the arachidonic acid (AA) cascade since it is responsible for the hydrolysis of esterified AA from the C-2 position of membrane phospholipids. This reaction generates two products (1) free AA which is then available for subsequent metabolism by either the cyclooxygenase or lipoxygenase enzymes and (2) Iysophospholipid. When alkyl-arachidonoyl-glycerophosphatidylcholine is acted upon by the PLA2 the géneration of platelet activating faclor (PAF) is initiated; PAF is pro-inflammatory in its own right [see Wedmore et al" Br. J. Pharmacol., 74, 916-917 (1981)] In this regard it may be noted that the anti-inflammatory steroids are thought to inhibit eicosanoid synthesis by inducing the synthesis of a PLA2 inhibitory protein denominated macrocortin or lipomodlllin [sée Flower et al., Nature, London, 278, 456 (1979) and Hirata et al., Proc. Natn. Acad. Sci. U.S.A., 77, 2533 (1980)].
As the initial step leading to subsequent conversion of AA to the various eicosanoids by the cyclooxygenase and lipoxygenase pathways, the PLA2-mediated release of AA from membrane phospholipids is a critical event in attempting to deal with the various physiological manifestations which are based on the activity of the eicosanoids andlor PAF.
Thus, while PLA2 has been shown to be required for platelet aggregation lPickett et al., Biochem. J., 160, 405 (1976)], cardiac contraction and excitation [Geisler et al., Pharm. Res.
Commun., ~, 117 (1977)], as well as prostaglandin synthesis [Vogt, Adv. Prosta~el. Thromb.
Res., 3, 89 ~1978)~, the inhibition of PLA2 is indicated in the therapeutic treatment of both PAF induced or cyclooxygenase and/or lipoxygenase pathway product-mediated physiological conditions.
There is also evidence that products of the cyclooxygenase/lipoxygenase pathways play key roles in both the pathogenesis of gastri~ mucosal damage due to extra-cellular (gastric and intestinal contents, rnicroorganisms, and the like) or intracellular (ischemia, ' : :

! :') 91/06~38 2 ~ ~ 713 8 PCI/US~0/06252 ~iruses, etc.) agents, as well as in cytoprotection against such damage. Thus, on the one hand prostaglandins exert a cytoprotective effect on the gastric mucosa [see Robert, Gastro-nterolo~v, Z~, 761-767 (1979)] and this action of the prostaglandins, especially of the E
series, is considered lo be of importance in the treatment of gastro-intestinal ulceration [see Isselbacher, Dru,~s, 33 (suppl.), 38-46 (1987)]. On the other hand, ex vivo experiments have shown that gastric mucosal tissue from ethanol-pretreated rats is capable of LTC4 generation and that this LTC4 production is quantitatively related to the severity of the ethanol damage [see Lange et al., Naunvn-Schmiedeber~'s Arch. Pharmacol. Suppl., ~Q, R27, (1985)]. It has also been demonstrated that LTC4 can induce vasoconstriction in both venous and arteriolar vessels in the rat submucosa [see Whittle, IUPHAR Ninth Int. Con,~. of Pharrn., S30-2.
London, England (1984)]. This is significant since ethanol-induced lesion formation in gastric mucosa may be multifactorial with, for example, stasis of gastric blood flow contributing significantly to the development of the hemorrhagic necrotic aspects of the tissue injury [see Guth et al., Gastroenterolo~y, 87, 1083-90 (1984)]. Moreover, in the anesthetized cat, exogenous LTD4 evokes both increased pepsin secretion and decreased transgastric potential [Pendleton et al., Eur. J. Pharrnacol., 125, 297-99 (1986)]. A particularly significant recent finding in this regard is that 5-lipoxygenase inhibitors and some leukotriene antagonists protect the gastric mucosa against lesions induced by the oral or parenteral administration of most nonsteroidal anti-inflammatory drugs lsee Rainsford, A~ents and Actions, 21, 316-319 (1987)]. Platelet activating factor (PAF) is also implicated as a mediator of gastrointestinal damage, and it has been recently shown that 5-lipoxygenase inhibitors inhibit PAF-induced gastric mucosal damage (Gastroenterolo~y, ~, A55, A434, 1989). Accordingly, a significant body of evidence implicates the involvement of lipoxygenase products in the development of pathological features associated with gastric mucosal lesions, such as for example, those induced by ethanol exposure and administration of non-steroidal anti-inflammatory drugs.
Thus, compounds which inhibit the biological effects of leukotrienes and PAF and/or which control the biosynthesis of these substances, as by inhibiting 5-lipoxygenase, are considered IO
be of value as cytoprotective agents.
Accordingly, the biological activity of the leukotrienes and SRS's, and of lipoxygenase as the enzyme leading to the metabolism of AA to leukotrienes, indicates that a rational approach to drug therapy to prevent, remove or ameliorate the symptoms of allergies, anaphylaxis, asthma and inflammation and for gastric cytoprotection must focus on either blocking the release of mediators of these conditions or antagonizing their effects. Thus, compounds which inhibit the biological effects of the leukotrienes and SRS's and/or which control the biosynthesis of these substances, as by inhibiting the PLA2-mediated release of arachidonic acid from membrane phospholipids, or by inhibiting lipoxygenase, are considered Wo 91/06538 ~ ~ ~) 7 ~ 3 8 PCr/US90/06252 to be of value in treating such conditions as allergic bronchial asthma, allergic rhinitis, as weil as in other in~nediale hypersensitivity reactions and in providing gastJic cytoprotection.
It has now been found that certain novel substituted benzoylbenzene-, biphenyl-and 2-oxazole- alkanoic acid derivatives inhibit PLA2 and lipoxygenase, and antagonize S products of the lipoxygenase pathway, and so are useful as anti-inflarnmatory, anti-allergic and cytoprotective agents. The present invention provides novel compounds having the following formula:

A(CH2)nO-B
wherein A is phenoxyethyl, phenoxyphenyl or a group having the forrnula Rl~ X~

wherein X is -N- or-C-;

I
Z is -C= C-, -C=N-, -N=C-, -N-, -S- or -O-;
Rl is hydrogen, lower alkyl or phenyl;
R2 is hydrogen or lower alkyl; or -Rl and R2 talcen together form a benzene ring;
R3 is hydrogen or lower allcyl;
nis 1 -2;
B is 0~CJ~CHCOOR5 e~R7 Il R6 ~0~ ~

; ~, .. .
.. ~

91/06538 2 0 6 713 8 Pcr/US90/0625~

wherein R4 and R5 are each, independen~ly, hydrogen or lower alkyl;
R6 is halo or nitro;

R7 is -C-R8 or -CHCOORs ;
S R8 is lower alkyl;
misO-3;
and the pharmacologically acceptable salts thereof.
The terrn "lower alkyl" refers to moieties having 1 - 6 carbon atoms in the carbon chain. The terrn "halo" refers to fluoro, chloro or bromo.
The grouping A embraces, inter alia, 5- or 6- membered unsaturated nitrogen, sulfur or oxygen containing mono- or benzofused-heterocycles, optionally substituted with Iower alkyl or phenyl. The foregoing definition embraces the following heterocyclic moieties:
furyl, pyrrolyl, thienyl, oxazolyl, thiazolyl, imidazolyl, pyridyl, pyrazinyl, pyrimidinyl, benzo-furanyl, benzothieriyl, benzothiazolyl, indolyl, benzoxazolyl, quinolinyl, quinazolinyl, benz-15 imidazolyl, quinoxalinyl, quinazolinyl and the like. Especially preferred are qui~olinyl, benzothiazolyl, and benzimidazolyl.
The compounds of the invention can forrn pharrnacologically acceptable salts from pharnnacologically acceptable organic and inorganic acids such as hydrochloric, hydro-bromic, sulfonic, sulfuric, phosphoric, nitric, maleic, fumaric, benzoic, ascorbic, pamoic, 20 succinic, methanesulfonic, acetic, propionic, tartaric, citric, lactic, malic, mandelic, cinnamic, palmitic, itaconic and benzenesulfonic. The compounds which are carboxylic acids are capable of forming alkali metal and alkaline earth carboxylates and carboxylates of pharmacologically acceptable cations derived from ammonia or a basic amine Examples of the latter include but are not limited to cations such as ammonium, mono-, di-, and trimethylammonium, mono-, di-25 and triethylammonium, mono-, di- and tripropylammonium (iso and norrnal), ethyldimethyl-ammonium, benzyldimethylammonium, cyclohexylammonium, benzylammonium, dibenzyl-ammonium, piperidinium, morpholinium, pyrrolidinium, piperazinium, 1-methylpiperidinium, 4-ethylmorpholinium, 1-isopropylpyrrolidinium,- 1,4-dimethylpiperazinium, 1-n-butyl-piperi-diniumj 2-methylpiperidinium, 1-ethyl-2-methylpiperidiMum, mono-, di- and triethanolammo-30 nium, ethyl diethanolammonium, n-butylmonoethanolammonium, tris(hydroxymethyl)methyl-an~moniurn, phenylmonoethanolammonium, and the like, The compounds of the invention can be prepared by the following reaction schemes. When it is desired to prepare compounds having the forrnula :~.~: .

WO 91/06538 2 ~ 6 7 1 3 8 PCrtUS90/06252 ~;;

A(CH2)nO~ LcHcooR5 Il R4 O
4-methoxybenzonitrile, for example, is reacted wi~h 3-bromotoluene, followed by reaction with bromine in ethylene bromide to yield the intermediate 3-bromomethyl-[4'-methoxy]benzo-phenone.

CH30~3~ 3-bromotoluene ~
CN Mg C~CH3 S O
bromine /
. BrCH2CH2B,~
/benzoyl peroxide ~/

O
The bromo interrnediate is reacted with sodium cyanide to yield the cyano intermediate, which is hydrolyzed in the presence of base to yield the carboxylic acid, which in turn is demethylated - 10to yield the hydroxy carboxylic acid intermediate:
CH30~LCH2Br dioxane / H20 ~ O
CH30~ LCH2CN KOH

o .

,.

., .:
"~ , .

.
. :

91/06538 2 ~ 6 7 1 3 8 P~/US90/~6252 ,. -,.~

CH30~ LCH2COOH ~ HCI/N2 Ho~cJ~--CH2COOH
O ', The hydroxy carboxylic acid intermediate is converted to the methyl ester with methanol in the presence of p-toluenesulfonic acid followed by reaction with an appropriate haloalkyl-A
S compound, where A is as deflned hereinbefore and hal is halo, to yield the desired final product as the methyl ester.
H~ ~ CH30H
)~LcH2cooH p-toluenesulfonic acid o H0~ A(CH2)nhal !LCH2COOCH3 K2C03 / 1 8-crown-6 Il O

A(CH2)nO~ ~
~C/~CH2COOCH3 O
10 The ester can be hydrolyzed by conventional methods to yield the desired final product in free carboxylic acid form.

~' i .
- . .
-.: ~ . .
;: . .

- : , .: ,. ~ . . ..
~ ,. .. . .

WO91/06538 2~ 7138 PCr/US90/0625Z

Compounds of the invention having the formula A(CH2)n~ O

can be prepared by several routes. Compounds in which R6 is nitro and R7 is the -C-R8 moiety can be prepared as follows; for example: 4-bromo-3-nitroacetophenone is reacted with 5 4-iodoanisole in the presence of copper bronze, to yield the intermediate methoxy-containing biphenyl, which is demethylated with aluminum bromide to yield the hydroxy intermediate The latter is then reacted with an appropriate haloaL~cyl-A compound, where A is as defined hereinbefore and hal is halo, to yield the desired final product.

\~c, , C~ /~ \~OC~3 AlBr3 benzene .
O O
CH3C~ CH3C~

~ K2CO3/18-crown-6 ~OH

Compounds in which R6 is halo and R7 is the -CHCOOR5 moiety can be prepared by aprocess which utilizes the 4-methoxy-biphenyl intermediate of the preceding scheme. Thus, the 4-acetyl-4-methoxy-2-nitrobiphenyl intermediate of the previous scheme is subjected to - 15 reduction with stannous chloride to yield the intermediate amino derivative, which is then subjected to replacement of the amino group with a halo group. For example, the amino group can be replaced with fluorine via a diazonium fluoroborate transitory intermediate prepared from the atnino intermediate using sodium nitrite and tetrafluoroboric acid. The resulting .

~: -, . .

";,? 91/06538 - 2 (~ 6 7 ~ ~ 8 PCI/US9OtO62~2 acetyl-fluoro-methoxy biphenyl interrnediate is converted to the corresponding carboxylic acid followed by demethylation with hydrogen bromide to yield the 2-fluoro-4'-hydroxy-[1,1'-biphenyl]-~acetic acid intermediate:
O O

CH3~ ~ ~OCH3 NaNO2 / HBF4 / THF-H~

CH3C~ HO2C-CH2 . I S /morpholine/a ~
F ~ H2SO4 / HOAc / ~ F ~ OCH3 HBr / HOA,~/
/~
~/
HO2C-CH2 ~

~OH

'~ . . ~ ' .~ . .

wo gl/06538 2 0 ~ 7 1 3 g P(~/US90/06252 ~;

The latter carboxylic acid intermediate is esterified with methanol in the presence of p-toluene-sulfonic acid and lhe latter is reacted with an appropriate haloallcyl-A eompound, where A is as clefined hereinbefore and hal is halo, to yield the desired final product as the methyl ester.
HO2C-CH2 ,e~ CH3O2CCH2 ,~

F 1 J~ p-toluenesulfonie aeid F W~OH

A(CH2)nh~
~K2CO3 / 1 ~-crown-6 s CH3O2CCH2 ~

F ~--J~o(cH2)nA
The ester ean be hydrolyzed by conventional methods to yield the desired final product in its free carboxylic acid fonn.
Compounds of the invention havirrg the formula CH(CHz) CoOR5 or ~ ~CH(CH2)mCooR5 o ~ 0 can be prepared as follows. Ber.zaldehyde and 4-methoxybenzaldehyde are reacted to yield 4-methoxybenzoin, which is convertecl to the hemisuecinate by reaction with succinic anhydride.
The latter is reacted with urea and acetic acid to yield the interrnediate 4-(4-methoxyphenyl)-5-phenyl-2-oxazole-propionic acid.
CH30~ CHO CH30~ OH
J--C-CH--~\
CHO EtOH / ~ ~ ll ~J

.

.. . . . . . .
.
' ' ,. ~ ' . : ' ., - . ' ~67~ 3~
-s? 91/06538 PCl/US90/062~2 o O=C--CH2CH2COOH

CH3~LC CH NH2CNH2 , O ~ HOAc /
toluene / ~ ~

o The latter intermediate is demethylated with hydrogen brornide and esterified with methanol to yield the corresponding hydroxy methyl ester intermediate, which is then reacted with an S appropriate haloalky-A compound, where A is as defined hereinbefore and hal is halo, to yield the desired final product as the methyl ester.
CH30~

~CH2CH2COOH Br ~~0 . H;~

HO

-toluenesulfonic acid /

HO

~CH2CH2CCH3 ~H2)Ah21 o - K2C03 / 1 8-crown-6\
' ~ - . -- ~ ..... . . .

WO 91/06538 2 0 ~ 713 8 Pcr/US90/06252 A(CH2)n \~N

o The ester can be hydrolyzed by conventional methods to yield the desired final product in its free carboxylic acid form.
The conventional starting materials used in the reaction sequences outlined 5 above are available commercially or can be prepared by methods known in the art. Thus, for example, the intennediate compound 2-bromomethylquinoline can be prepared by thefollowing reaction sequence:

N-Br (C6H5C0)202 ~CH2Br 10 The benzo-fused heterocyclic compounds used in the above reaction sequences are also either commercially available or can be prepared by methods conventional in the art. Thus, for example, such intermediates as 1-methyl-2-chloromethylbenzimidazole, 2-chloromethylbenz-thiazole and 2-chloromethylbenzoxazole can be prepared by the following reaction scheme ~NH2 I Rl~ CH2CI

15 wherein X is O, S or NCH3. The reaction is preferably carried out at a controlled low temperature in an organic solvent, such as methylene chloride.
The compounds of the invention, by virtue of their ability to inhibit the activity of PLA2 enzyme, as well as that of lipoxygenase enzyme and to antagonize mediators arising ' from the enzymatic pathway, are useful in the treatment of conditions mediated by products of , .; ~ .. . . ~ . . :
.; . . - .. - -., ... . . , - :.
- ,....

:
., ~ . . .. . -.: - i . , . , , . :
. . .. ~ ... - - 1 -`~O 91/06538 2 ~ 6 7 1 3 8 Pcr/US9~/06252 the oxidaion of arachidonic acid. Accordingly, the compounds are indicated in the treatrnent of such diseases as rheumatoid arthritis, inflammatory bowel disease, osteoarthritis, tendinitis, bursitis, psoriasis (and related skin inflammation) and similar conditions involving inflarnma-tion. Moreover, by virtue of their ability to antagonize the effect of LTC4, LTD4 and LTE4, 5 which are the constituents of SRS-A, they a~e useful for the inhibition of symptoms induced by these leukotrienes. Accordingly, the compounds are indicated in the prevention and treatment of those disease states in which LTC4, LTD4 and LTE4 are causative fact~rs, for example allergic rhinitis, allergic bronchial asthma and other leukotriene mediated naso-bronchial obstructive air-passageway condiions, as well as in other imrnediate hypersensitivity reactions, 10 such as allergic conjunctivitis. The compounds are especially valuable in the prevention and treatment of allergic bronchial asthma.
The compounds of the invention are cytoprotective agents and are considered especially useful when adrninistered with conventional non-steroidal anti-inflammatory drugs, whose major side effect is gastrointestinal irritation. The cytoprotective effect of the 15 compounds of the invention significantly reduces the gastroirritant impact of conventional anti-inflammatory drugs. This effect is based not only on the ability of the compounds of the invention to inhibit the biological effects of leukotrienes and/or control the biosynthesis of these substances, as by inhibiting lipoxygenase, but also by a shunting effect, whereby the control of the lipoxygenase pathway "shunts" the oxidadon of arachidonic acid into the cyclooxygenase 20 pathway, giving rise to an increase in the formation of cytoprotective prostaglandins. These biological effects make the compounds of the invention especially useful in treating such conditions as erosive esophagitis, inflammatory bowel disease and induced hemorrhagic lesions such as those induced by alcohol or non-steroidal anti-inflamrnatory drugs (NSAID's), hepatic ischemia, noxious agent induced damage or necrosis of hepatic, pancreatic, renal or 25 myocardial tissue; liver parenchymal darnage caused by hepatotoxic agents such as carbon tetrachloride and D-galactosamine; ischemic renal failure; disease-induced hepatic damage; bile salt-induced pancreatic or gastric damage; trauma or stress-induced cell damage; and glycerol-induced renal failure.
When the compounds of the invention are employed in the treatment of allergic 30 airway disorders, as anti-inflammatory agents and/or as cytoprotective agents, they can be formulated into oral dosage forms such as tablets, capsules and the like. The compounds can be administered alone or by combining them with conventional carriers, such as magnesium carbonate, magnçsium stearate, talc, sugar, lactose, pectin, dextrin, starch, gelatin, tragacanth, methylcellulose, sodium carboxymethylcellulose, low melting wax, cocoa butter and the like.
35 Diluents, flavoring agents, solubilizers, lubricants, suspending agents, binders, tablet-disintegrating agents and the like may be employed. The compounds may be encapsulated with ... .

, WO 9l/06538 2 ~ 6 7 1 ~ ~ PCr/US90/062~2 or without other carriers. In all cases, the proportion of active ingredients in said compositions both solid and liquid will be at least to impart the desired activity thereto on oral administration.
The compounds may also be injected parenterally, in which case they are used in the form of a sterile solution containing other solutes, for example, enough saline or glucose to make the 5 solution isotonic. For administration by inhalation or insufflation, the compounds may be formulated into an aqueous or partially aqueous solution, which can then be utilized in the form of an aerosol.
The dosage requirements vary with the particular compositions employed, the route of administration, the severity of the symptoms presented and the particular subject being 10 treated. Treatment will generaily be initiated with small dosages less than the optimum dose of the compound. Thereafter the dosage is increased until the optimum effect under the circumstances is reached. In general, the compounds of the invention are most desirably administered at a concentration that will generally afford effective results without causing any harmful or deleterious side effects, and can be administered either as a single unit dose, or if 15 desired, the dosage may be divided into convenient subunits administered at suitable times throughout the day.
The PLA2 and lipoxygenase inhibitory and leukotriene antagonist effects, as well as the anti-inflammatory and potential gastroirritant effects of the compounds of the invention, may be demonstrated by standard pharmacological procedures which are described 20 more full in the examples given hereinafter.
These procedures, inter alia, determine the specificity of action of the compounds of the invention as PLA2 inhibitors as measured by their ability to inhibit the synthesis of LTB4 and PGE2 by rat glycogen-elicited polymorphonuclear leukocytes, as well as measure their ability to inhibit arachidonic acid release mediated by human source PLA2.
25 The pharmacological testing additionally demonstrates the ability of the compounds of the invention to inhibit, in vivo, the lipoxygenase and cyclooxygenase pathways of arachidonic acid metabolism.
` The following examples show the preparation and pharmacological testing of compounds within the invention.

.
, :

, ~~ o gl/06~38 2 Q ~ 7 1 3 ~ PCr/US90/06252 ~m~
1 -r2-Nitro-4'-(2-allinolinvlmetlm~v)-r1.l '-bi~hellY114-vllethanone A. 4-Acetvl-4'-methoxv-2-nitro biphenvl A st~lTed mixture of 4-iodoanisole (43.65 g, 0.187 mole), 4-bromo-3-nitTo acetophenone (40.6 g, 0.166 mole) and copper powder (copper bronze, 36 g, 0.567 mole) kept under nitrogen is placed in an oil bath heated at 80C. The temperature is slowly raised to 110C and the mixture is kept at this temperature for S days (lLC,8:2 hexane-ethyl acetate). Upon cooling the rnixture is dissolved in dichloromethane and filtered through a Celite pad. The filtrate and washings are evaporated and the residual thick, dark brown oil (58.4 g) is flash chromatographed (on silica Merck 60, preabsorbed in dichloromethane, eluted with 9: 1 hexane-ethyl acetate to remove the impurities and 8:2 hexane-ethyl acetate to recover the main product) to provide 16.2 g (32%) of the title compound (yellow solid, m.p. 124-126C).

NMR (CDCl3, 400 MHz): o 2.67 (s, 3H, COCH3), 3.85 (s, 3H, OCH3), 6.97 (d, 2H, J 8.74 Hz, ArH), 7.27 (d, 2H, J 8.74 Hz, ArH), 7.56 (d, lH, J 8 Hz, ArH), 8.15 ( d, 1 H, J 8 Hz, ArH), 8.34 (s, lH, ArH) MS (EI, m/z): 271 (M) ' - B. 4-Acetvl-4'-hydroxv-2-nitro bi~henvl To a stirred solution of AlBr3 (12.6 g, 47.4 mmole) in benzene (A5 mL) is added dropwise under nitrogen a solution of the methylether (5 g, 18.45 mmole) of Step -~ A in benzene (12 mL) over 30 minutes. The resulting solution is stilTed at room temperature for 3.5 hours. (TLC, 8:2 hexane-ethyl acetate). The mixture is cooled in an ice bath and the complex is decomposed by the dropwise addition of 6N-HCl (ca. 37 mL). The organic layer is separated and the aqueous phase is reextracted with ether (3x). The combined ~5 extracts are concentrated to a small volume and extracted again with 2.5N-NaOH (2xS0 mL
+ lxlO mL). The basic extracts are cooled and acidified with concentrated HCI (to pH 2).
The solid is collected and dried (4.27 g, 90%). It is used in the next step without further purification.

NMR (CDC13, 400 MHz): o 2.67 (s, 3Hj COCH3), 5.03 (broad, lH, OH), 6.91 (d, 2H, J 8.56 Hz, ArH), 7.23 (d, 2H, J 8.57 Hz, ArO, 7.55 (d, IH, J 7.9 Hz, ArH), 8.15 (d, lH, J 8.1 Hz, ArH), 8.34 (s, lH, ArH).
MS (EI, m/z): 257 (b.p., M)+

. . .

. .: .

. ' ~ , 2067~38 C. I - ~ 2-Nitro-4'-(2-quinolinvl)~ 1 ~ l '-biphenYl-4-vllethanone A rnixture of the phenol (4.4 g, 17.12 rnmole) of Step B, powdered anhy-drous potassium carbonate (2.37 g, 17.12 mmole), 18-crown-6 (0.453 g, 1.71 mrnole) and acetonitrile (38 mL) is stirred at room temperature under nitrogen for 15 minutes. 2-Chlor~
methylquinoline (3.34 g, 18.83 mmole, fr~e base freshly prepared from the hydrochloride salt) is added and the rnixture is refluxed for 10 hours. (TLC, 7:3 hexane-ethyl acetate). A 10%
excess of potassium carbonate, 18-crown-6 and the chloromethylquinoline is added and reflux continued for another 4 hours. The solvent is removed and the residue is diluled with water and extracted with ethyl acetate (3x). The extracts are washed and dried (MgS04). The residue 10 is flash chromatographed (on silica Merck 60, preabsorbed in dichloromethane and eluted in order of increasing polarity with 7:3, 1:1 and 1:3 hexane-ethyl acetate followed by pure ethyl acetate) to provide the pure title compound (2.59 g). Recrystallization from toluene yields a yellow solid, m.p. 160-162C (2.05 g, 30%).

NMR (CDCI3, 400 MHz): ~ 2.66 (s, 3H, COCH3), 5.43 (s, 2H, OCH2Ar), 7.10 (d, 2H, J 8.7 Hz, ArH), 7.27 (d, 2H, J 8.7 Hz, ArH), 7.56 (m, 2H, ArH), 7.68 (d, IH, J 8.49 Hz, ArH), 7.75 (dt, lH, ArH), 7.84 (d, lH, J 8.1 Hz, ArH), 8.09 (d, lH, J 8.5 Hz, ArH), 8.14 (dd, lH, ArH), 8.22 (d, lH, J 8.49 Hz, ArH), 8.34 (s, lH, ArH) MS (EI, m/z): 398 (M)+, 256, 158, 142 (b.p.) 20 Analvsis for: C24HI8N2o4 Calculated: C, 72.35; H, 4.55; N, 7.03 Found: C, 71.96; H, 4.75; N, 6.80.

: . , . , . .. :: . . . - . . . , -',: . . - ~:- : . .

~ .,0 91/06538 2 ~ 6 7 1 3 8 PCr/US90/0625t ~aml~le 2 2-Fluoro-4'-(2-~uinolinvlmethu2~Y)-~1.1 '-biphenvll-4-acetic acid A. 4-Acetvl-4'-methoxy-2-arnino bipheml To a stirred, warm solution of tin (II) chloride (49.4 g, 218.9 mmole) in a mixture of concentrated HCI (72 rnL) and ethanol (99 mL) is added over a period of 45 minutes the ni~o derivative (10.7 g, 39.5 rnmole) of Exarnple lA. The resulting yellow solution is refluxed for 3.5 hours (TLC, 1: 1 hexane-ethyl acetate). The ethanol is removed and the residue is poured into a mixture of 50% NaOH (360 mL) and ice. The resulting solid is extracted (dichloromethane, 3x), the extracts are washed with water and dried 10 (Na2SO4). Removal of the solvent provides a yellow solid (9.31 g, 97.8%), m.p. lS2-NMR (CDCI3, 400 MHz): o 2.59 (s, 3H, COCH3), 3.80 (s, 3H, OCH3), 6.97 (d, 2H, J 8.7 Hz, ArH), 7.23 (d, lH, J 7.4 Hz, ArH), 7.40 (d, 2H, J 8.7 Hz, ArH), 7.48 (d, lH, J 7.3 Hz, ArH), 7.49 (s, lH, ArH).
1~ MS (EI, mlz): 241 (b.p., M)+, 226 (M-CH3)+, 198 (M-COCH3)+, 83 B. 4-Acetvl-4'-methoxY-2-fluoro biphenvl To a stiITed, ice cold mixture of the aniline (9.2 g, 38.2 mmole) in tetrahydro-furan (26 mL), water (9.8 mL) and HB~i4 (48%, 35.1 mL) is slowly added a solution of sodium nitrite (2.82 g, 40.85 mmole) in water (5 mL). The internal temperature is kept 20 below 5C during the addition. The mixture is then stirred for an additional 20 minutes at ' 0-5C. The diazonium fluoroborate is filtered off and washed with 10% HBF4 and 10%
methanol in ether and dried in vacuo. The sal~ is decomposed by heating at 70C in xylene (95 mL). When the decomposition subsides, the mixture is refluxed for another 2.5 hours (TLC, 1: 1 hexane-ethyl acetate, UV). The xylene is removed and the residue is extracted with 2s ethyl acetate (3x) and ether. The combined extracts are washed with 10% sodium carbonate and brine and dried (MgSO4). Removal of the solvent provides an a nber oil (6.03 g) which is purified by flash chromatography (on silica Merck 60, preabsorbed in dichloromethane and eluted with 95:5 hexane-ethyl acetate). The title compound is obtained as a yellow solid 3.12-4.75 g, (33-51% depending on the run); m.p. 100-101C.

30 NMR (CDC13, 400 MHz): ~ 2.62 (s, 3H, COCH3), 3.86 (s, 3H, OCH3), 7.00 (d, 2H, J 8.9 Hz, ArH), 7.50-7.80 (m, SH, ArH).
MS (EI, m/z): 244 (M)+, 229 (b.p., M-CH3)+
,~

. -. . .

. , ~, . ............ . . .
. - . - . : ~ .
;. . .. . . .. . . .... ..

WO 91/06538 2 0 6 7 1 3 8 PCr/US90/06252 C. 2-Fluoro-4'-methoxv-~1,1'-biphenvll-4-acetic acid A mixture of sulfur (0.468 g, 14.6 mmole), morpholine t2.57 mL) and the ketone (3.95 g, 16.2 mmole) of Step A is refluxed for 17 hours (TLC, acid treated silica plate;
8:2 hexane-ethyl acetate). Upon cooling, glacial acetic acid (9.9 mL), sulfuric acid (1.6 rnL) and water (4 rnL) are added and the reflux resumed for 30 hours. Water is then added and the mixnlre is extracted with ether (3x). The combined extTacts are concentrated to a smaller volume and extracted with 10% sodium carbonate. The basic extracts are carefully acidified in the cold with concentrated HCI (to pH 2). The title acid is extracted w~th ether (3x) and the extracts are washed and dried (MgS04). Removal of the solvent provides a tan to brown 10 solid (2.37 g, 56.3%) melting at 140-142C.

NMR (CDCI3, 400 MHz): o 3.68 (s, 2H, CH2COO), 3.85 (s, 3H, OCH3), 6.96-7.50 (m, 7H, ArH).
MS (EI, rn/z): 260 (M)+, 215 (b.p., M-COOH)+ .
D. 2-Fluoro-4'-hvdroxv-~ 1. I '-biphenvll-4-acetic acid To a solution of the methylether (1.31 g, 5.04 mmole) of Step C in glacial acetic acid ~17 rnL) is added dropwise 48% HBr in acetic acid (25 mL) and the mixture is refluxed for 4.5 hours (TLC, 7:3 hexane-ethyl acetate). A little water is added and the mLx-ture is extracted with ether (3x). The extracts are washed and dried (MgSO4). Removal of the solvent provides the title compound as a tan solid (1.13 g, 92%), m.p. 208-210C.

20 NMR (DMSO-d6, 400 MHz): ~ 3.61 (s, 2H, CH2COO), 6.83 (d, 2H, J 8.64 Hz, ArH), 7.1-7.42 (m, SH, ArH), 9.61 (s, lH, COOH).
MS ( C I, m/z): 246 (M)+, 201 (b.p., M-COOH)+
E. 2-Fluoro-4'-hvdroxv-rl.l'-biphenvll-4-acetic acid methvlester A solution of the acid (1.1 g, 4.47 rnmole) of Step D in methanol (10 mL) 25 containing p-toluenesul~onic acid H20 (0.159 g) is refluxed for 1.5 hours (TLC, acid treated silica plate, hexane-ethyl acetate 7:3). The solvent is removed~ the residue is dissolved in ethyl acetate, washed with brine and dried (MgSO4). The tan solid (I .16 g, m.p. 115-118C, quantitative yield) is used as such in the next step.

NMR (CDCl3, 400 MHz): ~- 3.65 (s, 2H, CH2COO), 3.73 (s, 3H, COOCH3), 6.88 (d, 2H, J 8.8 Hz, ArH), 7.10 (m, 2H, ArH), 7.32-7.44 (m, 3H, ArH).
MS (EI, IllJz): 260 (M)+, 201 (b.p., M-COOCH3)+ .

.

, ,~'vo 9l/0653X 2 0 ~ 71 3 8 PCr/US90/06252 F. 2-Fluoro-4'-(2-quinolinvlmethoxv)~ biphenvll-4-acetic acid methYlester A stirred mixture of the phenol (1.16 g, 4.46 mmole) of Step E, powdered anhydrous potassium carbonate (0.616 g, 4.46 mmole), 18-crown-6 (0.118 g, 0.445 mmole) and acelonitrile (10 mL) is stirred at room temperature under nitrogen for 15 minutes.
5 2-Chloromethylquinoline (0.871 g, 4.9 mmole, free base freshly prepared from the hydro-chloride salt) is then added and the mixture is placed in an oil bath heated at 65C for 5 h~urs.
A 10% excess of potassium carbonate, 18-crown-6 and the chloromethylquinoline is added and the heating continued for another 6 hours (TLC, 19: 1 dichloromethane-methanol or 7:3 hexane-ethyl acetate). The solvent is removed and the residue is diluted with water and extracted with ethyl acetate (3x). The extracts are washed and dried (MgSO4). Removal of the solvent provides a tan solid which is purified by flash chromatography (on silica Merck 60, preabsorbed with dichloromethane, eluted with 7:3 hexane-ethyl acetate). The title com-pound thus obtained (1.55 g, 87%) is recrystallized from methanol. The off-white solid melts at 99-101C.

13 NMR (CDC13, 400 MHz): ~ 3.64 (s, 2H, CH2COO), 3.72 (s, 3H, COOCH3), 5.43 (s, 2H, OCH2Ar), 7.1 (m, 4H, ArH), 7.35 (t, lH, ArH), 7.47 (d, 2H, ArH), 7.55 (t, lH, ArH), 7.69 (d, lH, ArH), 7.74 (t, lH, ArH), 7.84 (d, lH, ArH), 8.09 ~d, lH, ArH), 8.20 (d, lH, ArH~.
MS (EI, nlJz): 401 (M)~, 142, 114 (b.p.) Anal~,~sis for: C25H20FNO3 Calculated: C, 74.80; H, 5.02; N, 3.49.
Found: C, 74.68; H, 4.65; N, 3.49.

,,' ,, ''` '' ' ' -, .
`" ~ '', .

W O 91/06538 2 0 ~ 713 8 PC~r/US90/062~2 G. 2-Fluor~-4'-(2-quinolinvlmethoxv)-~ 1.1'-biphenYll-4-acetic acid A solution of the ester (1.69 g, 4.21 mmole) of Step F, in dry tetrahydrofuran (20 rnL) is treated dropwise under nitrogen with lN-LiOH (12.6 rnL) and ~he mixture is s,tirred for 3 hours at room temperature (TLC, 19: 1 dichloromethane-methanol or 1: 1 hexane-5 ethyl acetate). The solvent is removed, the residue is treated with water and neutralized (topH 6.5) with 10% acetic acid. The acid is extracled with ethyl acetate (large volume needed ) and the extracts are dried (MgSO4) and evaporated to dryness to yield an off-white solid (1.65 g, quantitative yield, m.p. 190-193C, dec.). Recrystallization from ethyl acetate pro-vides a white solid (1.32 g, 80%, m p. 195-196C dec.). The analytical sample is dried in 10 vacuo at 40C.

NMR (DMSO-d6, 400 MHz): ~ 3.62 (s, 2H, CH2COO), 5.41 (s, 2H, CH20Ar), 7.15 (m, 4H, ArH), 7.41 (t, IH, J 8 Hz, ArH), 7.48 (d, 2H, ArH), 7.61 (t, IH, ArH), 7.69 (d, lH, ArH), 7 78 (dt, lH, ArH), 8.01 (m, 2H, ArH), 8.42 (d, lH, ArH), 12.42 (s, COOH).
15 MS (+ FAB, rn/z): 388 (M)+

Analvsis for: C24HI8FNO3 Calculated: C, 74.41; H, ~.68; N, 3,62.
EQ~: C, 74.28; H, 4 48; H, 3.69 : . : .
. . . . ~ . -, - . . . .. -: : ~ . . ~ : -,. . ; , : ' . . . . .
. .

1'~) 91/06538 2 ~ ~ 7 1 3 8 PCr/US90/06252 ,l~xamDle 3 3-r4-(2-Ouinolinvlmethoxv)henzovllbenzene acetic acid A. 3-Melhvl-~4'-methoxvl-benzoDhenone A 3-neck flask equipped with a condenser, mechanical stirrer and dropping funnel is charged under nitrogen with 1.925 g (79.19 g.a.) of magnesium turnings and enough ether to cover the turnings. A few drops of a solution of 3-bromotoluene (15.79 g, 92.28 mmole) in ether (40 mL) is then added along with a crystal of iodine to initiate the reaction. The remainder of the solution is then added dropwise and the mixture is refluxed until most of the magnesium has disappeared. After cooling, a solution of 4-methoxybenzo-10 nitrile (10 g,75.1 mmole, dried in vacuo over P2O5) is added in one portion. The tnixture is refluxed for 2 hours ~I-LC, no star~ing material present), cooled (ice bath) and slowly treated with cold water (130 mL) followed by dilute H2SO4 (1:1, v/v, 25 mL). The decom-posidon of the complex is completed by refluxing the mixture for 4 hours (followed by TLC, 8:2 ether-ethyl acetate). Following sdrring overnight at room temperature, the layers are 15 separated and extracted with ether (3x). The extracts are washed with 5% NaHCO3, dried (MgS04) and evaporated to dryness. The crude material (arnber oil, 13.93 g) is purified by flash chromatography (on silica Merck-60, eluted with 8:2 petrolether-ethyl acetate) to pr~
vide the title compound as a light yellow oil (12.5 g, 73.5%).

NMR (CDCI3, 400 MHz): o 2.4 (s, 3H, CH3), 3.9 (s, 3H, OCH3), 6.96 (d, J 8.8 Hz, 2H, ArH), 7.38 (m, 2H, ArH), 7.53 (d, J 6.9 Hz,lH, ArH), 7.57 (s, 1~1, ArH), 7.82 (d, J 8.7 Hz,2H, ArH).
MS (EI, m/z): 226 (M)~, 135 (b.p.), 91.

. . .
.:'' ; : ' ' '. ' : . : ~ . , -.: .. . . . : . -Wo 9l/06538 2 ~ ~ 7 1 3 8 Pcr/US90/06252 B. 3-Bromomethvl-~4'-methoxvl-benzophenone A solution of the benzophenone (17.5 g, 77.4 mmole) of Step A in ethylene bromide (26.5 mL) (containing a small amount of benzoyl peroxide) is heated at reflux. A
solution of bromine (12.7 g, 79.6 mmole) in ethylene bromide (15 mL) is added dropwise ~ver 30 minutes while the mixture is irradiated with a Photolamp (300 W). Reflux is con-tinued for 17 hours (TLC, 9:1 petrolether-ethylacetate, traces of starting material still present).
The solvent is removed in vacuo and the residue (brown oil, 39.22 g) is purified by flash chromatography (on silica Merck-60, preabsorbed in dichloromethane, eluted with 9: 1 petrol-ether-ethyl acetate) to give unreacted starting material (2.59 g, ca 15%) along with the desired product (14.36 g, 61% or 71.5% based on recovered unreacted starting mate}ial) and some mixed fractions (ca 5.20 g). The light yellow so~id melts at 58-61C and it is used as such in the next step.

NMR (CDCI3, 400 MHz): o 3.88 (s, 3H, OCH3), 4.52 (s, 2H, CH2Br), 6.96 (d, J8.8Hz,2H,ArH)7.44(t,J7.6Hz,lH,ArH),7.58(d,J7.8Hz,lH,ArH), 7.66 (d, J 7.6 Hz, lH, ArH), 7.76 (s, lH, ArH), 7.81 (d, J 8.8 Hz, lH, ArH).
MS (EI, m/z): 306/304 (1 bromine, M)+, 225, 135 (b.p.). Trace of dibromo at 386/3841382 ibl CH3O~

C, 3-CyanomethYl-r4'-methoxvl-benzophenone The bromo compound (14 g, 45.9 mmole) of Step B, is dissolved in dioxane (30 mL) and a solution of NaCN (7 g) in water (28.5 mL) is added. The rnixture is refluxed for 6 hours (TLC, petrolether-ethyl acetate 8:2), charcoalized if needed and extracted with ether (3x). Tne extracts are dried (MgSO4) and evaporated to dryness to yield a brown oil (13.44 g). The crude product is purified by flash chromatography (on silica Merck-60, pre-absorbed in dichloromethane, eluted with 6:4 hexane-ethyl acetate) to provide the pure product (10.69 g, 92%) as a light yellow oil that sets up upon standing. The nearly colorless solid melts at 7~71C.

NMR (CDC13, 400 MHz): ~ 3.80 (s, 2H, CH2CN), 3.87 (s, 3H, OCH3), 6.95 (d, J 8.6 Hz, 2H, ArH), 7.48 (t, J 7.7 Hz, lH, ArH), 7.54 (d, J 7.6 Hz, IH, 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)+, 135 (b.p.).

:~ -.~ .: - : ~ . - , . .

O 91/06~38 2 ~ 6 7 1 3 8 PCr/US9~/062~2 D . 3-~4-MethoxvbenzovlI-phenvlaceic acid The nitrile (4 g, 15.9 mmole) of Step C, is dissolved in 40% NaOH (40 mL) ~md the solution is heated at reflux under nitrogen for 7 hours (TLC, toluene-methanol 9: 1).
Water is added while cooling in an ice bath. The solution is washed with ethyl acetate and then acidified in the cold with concentrated HCI (to pH 2). The acid is extracted with ethyl acetate (3x) and the extracts are dried (MgSO4) and evaporated tO dryness IO yield the crude product (yellow solid, 3.56 g, 82%), m.p. 138-140C.

NMR (CDCI3, 400 MHz): o 3.72 (s, 2H, CH2COO), 3.88 (s, 3H, OCH3), 6.95 (d, J 8.8 Hz, 2H, ArH), 7.43 (t, lH, ArH), 7.48 (d, lH, ArH), 7.65 (d. lH, ArH), 7.68 (s, lH, ArH), 7.81 (d, J 8.6 Hz, 2H, ArH).
MS (El, m/z): 270 (M)+, 211 (M-CH2COOH)+, 135 (b.p.), 107.

E. 3-r4-Hydroxvbenzovll-phenvlacetic acid An intimate mixture of the acid (8.1 g, 0.030 mole) of Step D, and pyridine hydrochloride (13.87 g, 0.120 mole) is stirred under nitrogen in an oil bath heated at 200-1 ;, 210C for 7 hours (TLC, toluene-methanol 9:1, dichloromethane-rnethanol 9:1). After cooling, the mLxture is dissolved in dichloromethane. The solution is extracted with IN-NaOH, the extract æidifled in the cold with concentrated HCI and extracted with ethyl acetate (3x), After drying (MgSO4) the solvent is removed to provide the crude title compound as a tan solid (7.61 g, quantitative yield), m. p. 147-149C.

NMR (DMSO-d6, 400 MHz): ~ 3.67 (s, 2H, CH2COO), 6.88 (d, J 8.84 Hz, 2H, ArH), ca. 7.5 (m, 4H, ArH), 7.65 (d, J 8.8 Hz, 2H, ArH), 10.4 (s, lH, OH), ca. 12.3 ~s, lH, COOH).
MS (m/z): 257 (M+H)+, 217, 131, 91 (b.p.).

-, ~ .

' ' ~ ' ' ' wo 91/06538 ~ ~ 6 7 1 3 8 PCI/US~0/06252 F. 3-r4-Hvdroxvbenzovll-phenvlacetic acid methvlester A mixture of the acid (8.56 g, 33.4 mmole) of Step E and p-toluenesulfonic acid monohydrate (1.05 g, 5.6 mmole) in methanol (70 mL) is refluxed for 2.5 hours (TLC, rnethanol-toluene 1 :9). The methanol is evaporated and the residue is dissolved in ethyl S acetate and washed with brine. After drying (MgSO4) the solvent is removed to yield a tan solid (8.68 g, 96.2%, m.p. 111-113C). The crude product is used as such in the next step.

NMR (CDCI3, 400 MHz): o 3.69 (s, 2H, CH2COO), 3.69 (s, 3H, COOCH3), 6.86 (d, J 8.4 Hz, 2H, ArH), 7.41 (t, lH, 7.58 Hz, lH, ArH), 7.47 (d, J 7.56 Hz, lH, ArH), 7.62 (d, J 7.4 Hz, lH, ArH), 7.64 (s, lH, ArH), 7.74 (d, 2H, J8.4Hz,ArH).
MS (m/z): 271(M+H)+, 217, 131, ~1 (b.p.).

G. 3-r4-(2-Ouinolinvlmethoxv!benzoyllbenzene acetic acid methvlester A mixture of the phenol (4 g,l4.8 mmole) of Step F, powdered anhydrous K2CO3 (2.05 g, 14.8 rnrnole) and 18-crown-6 (0.4 g, 1.48 mmole) in acetonitrile (35 mL) 15 is stirred at room témperature under nitrogen for 15 minutes. 2-Chloromethylquinoline (2.9 g, 16,28 mmole, freshly prepared from the hydrochloride salt) is added in one portion and the mixture is heated in an oil bath kept at 65-70C for 8 hours (ll C, toluene-methanol 9:1). A 10 % excess of K2CO3, crown ether and chloromethylquinoline is added and the heating is continued for another 8 hours. The acetonitrile is evaporated and the residue is 20 partitioned between water and ethyl acetate. The organic layer is dried (MgS04) and evaporated to yield a tan solid (6.57 g). The crude product is purified by flash chromatography (on silica Merck-60, preabsorbed in dichloromethane, eluted with petrolether-ethyl acetate 7:3) to give the title compound as a light yellow solid (5.03 g, 82.7%) m.p. 93-95C.

NMR (CDC13, 400 MHz): o 3.67 (s,SH, CH2COO + OCH3), 5.45 (s, 2H, ArCH20), 7.08(d,J8.8Hz,2H,ArH),7.40(t,J7.8Hz,lH,ArH),7.46(d,J7.7Hz, lH, ArH), 7.55 (t, J 7.3 Hz, lH, ArH), 7.6-7.66 (m, 3H, ArH), 7.7-7.82 (m, 4H, ArH), 8.07 (d, lH, ArH), 8.20 (d, J 8.4 Hz, lH, ArH).
MS (EI, m/z): 411 (M)+, 142, 121 (b.p.).

:
.

:' ' . ~ : ' : .

. ) 91/~6538 2 ~ 6 7 1 3 ~ PCT/US90/06252 H. 3-~4-(2-OuinolinYlmethoxv)benzoyll~enzene acetic acid To a solution of the ester (5 g, 12.16 mmole) of Step G, in dry tetrahydro-furan (66 mL) is added IN-LiOH (37 mL, 37 mmole) and the mixture is stirred under nitrogen at room temperature for 2.5 hours ~TLC, toluene-MeOH 9:1). The tetrahydrofuran 5 is evaporated and the residue is diluted with water, acidified (~o pH 6.5) with 10% acetic acid and extracted with ethyl acetate (3x). The extracts are washed with brine, dried (MgSO4) and evaporated to dryness. The crude product (4.94 g, pale yellow solid) is reclystailized from ethyl acetate to provide 3.65 g (75%) of the pure title compound (white solid, m.p.
146- 147C).

NMR (DMSO-d6, 400 MHz): o 3.68 (s, 2H, CH2COO), 5.48 (s, 2H, ArCH2O), 7.22 (d, 2H, J 8.8 Hz, ArH), 7.47 (m, lH, ArH), 7.53 (m, 2H, ArH), 7~62 (m, 2H, ArH), 7.69 (d, J 8.4 Hz, lH, ArH) 7.74-7.82 (m, 3H, ArH), 8.2 (m, 2H, ArH), 8.43 (d, J 8.5 Hz, lH, ArH), 12.39 ( lH, COOH).
MS (EI, m/z): 397 (b.p., M)+, 380 (M-OH)f, 142 .
15 Analvsis for: C25HIgNO4 Calculated: C, 75.57; H, 4.78; N, 3.53.
Found: C, 75.22; H, 4.76; N, 3.39.

~xample 4 ~L4 (2~Naphthalenylmethoxv)benzovllbenzene acetic acid , A. 3-~4-(2-NaphthalenYlmethoxv)benzovllbenæne acetic acid methvlester A rnixture of the phenol (1 g, 3.7 mmole) of Example 3F, powdered anhhy-drous K2CO3 (0.48 g, 3.7 mrnole), 18-crown-6 (0.098 g, 0.37 mmole) and acetonitrile (10 mL) is st*ed under nitrogen for 15 minutes. 2-Bromomethylnaphthalene (0.496 g, 4.07 mmole) is added and the mixture is placed in an oil bath heated at 65-70C for 10 hours (TLC, dichloromethane-ethyl acetate 8:2). A 10% excess of K2CO3, crown ether and bromo-methylnaphthalene is added and the heating is continued for another 4 hours. The acetonitrile is evaporated and the residue dissolved in water and extracted with ethyl acetate (3x). The extracts are washed with lN-NaOH and brine, dried (MgSO4) and evaporated to dryness.
The crude product (1.49 g, waxy solid) is used as such in the next step.

NMR (CDC13, 400 MHz): o 3.7 (s, SH, OCH3+ CH2COO), 5.32 (s, 2H, ArCH20), 7.08 (d, J 8.7 Hz, 2H, ArH), 7.4-7.56 (m, SH, ArH), 7.63-7.68 (m, 2H, ArH), 7.82-7.92 (m, 6H, ArH).
MS 'rn/z): 410 (M)+, 141 (b.p.).

WO 9 l /06538 2 ~ ~ 7 1 3 ~ pcr/ us 90/ 06252 ~ ~

B. 3-f4-(2-NaphthalenvlmethoxY)benzoYllbenæne acetic acid A solution of the ester (1.29 g, 3.15 mmole) of Step A, is treated dropwise with lN-LiOH and the mLxture is stirred under nitrogen overnight. The solvent is evaporated and the residue is dissolved in water, acidified in the cold with 10% aceic acid (to pH 3) and extracted with ethyl acetate (3x). The extracts are dried (MgS04) and evaporated to dryness.
The residue (1.24 g, quantitative yield) is recrystallized by dissolving it in a relatively large volume of warm ethyl acetate-dichloromethane followed by concentrating to half volume.
The precipitate is collected and dried at 45C in vacuo (0.610 g, 48.8%), m.p. 150-152C.

NMR (DMSO-d6, 400 MHz): o 3.70 (s, 2H, CH2COO), 5.40 (s, 2H, ArCH2), 7.20 (d, 2H, ArH), 7.45-7.60 (m, 7H, ArH), 7.75 (d, 2H, ArH), 7.95 (m, 3H, ArH), 8.02 (s, lH, ArH), 12.47 (broad s, lH, COOH).
MS (+FAB, m/z): 397 (M+H)+, 217, 141.
Analvsis for: C26H204 Calculated: C, 78.78; H, 5.09.
1;~ EQ~: C, 78.12; H, 5.13.

Ex~n1p1e 5 5-PhenYI-4-r4-(Z-()uinoUnvl~nethoxv)-DhenYIl-2-oxa7ole proDanoic acid A. 4-Methoxvbenzoin To a solution of KCN (5 g) in water (35 mL) is added 4-methoxybenzalde-hyde (27.2 g, 0.2 mole), benzaldehyde (21.2 g, 0.2 mole) and 95% ethanol (70 mL). The mixture is refluxed under nitrogen for 4.5 hours and the ethanol removed in vacuo. Water (200 mL) is added to the residue and then distilled off at reduced pressure ~to remove remaining unreacted bezaldehyde). The procedure is repeated twice and the residual water azeotroped with ethanol. The crude product (56.3 g, orange semi-solid) is purified by flash-2s ch~omatography (on silica Merck-60, preabsorbed in dichloromethane-ethyl acetate and eluted with hexane-ethyl acetate 8:2) to yield a light yellow solid (20.1 g, 41.5%), m.p. 99-101C.

NMR (CDCI3, 400 MHz): o 3.82 (s, 3H, OCH3), 4.62 (broad s, lH, OH), 5.88 (s, lH, C_OH), 6.86 (d, 2H, J 8.94Hz, ArH), 7.22-7.38 (m, 5H, ArH), 7.91 (d, 2H, J 8.94Hz, ArH).
MS ( C I, m/z): 243 (b.p., M~H)+, 225, 197, 137 (M-PhCO)+

- . ,: . .
. . , . , , 91/06538 2 ~ ~ 7 1 3 ~ Pcr~usso/06252 B ~MethoxYbçnzoin hemisuccinate A mixture of 4-methoxybenzoin (20 g, 0.083 mole) and succinic anhydride (9. l g, 0.091 mole) in toluene (6 mL) is heated for 7 hours under nitrogen at 135C (intemal temp.). The solution is poured into 0.5N-NaHCO3, the organic layer was separated and re-5 extracted with 0.5N-NaHCO3. The combined extracts are washed with ether and then acidified in the cold with concentrated HCI. The liberated oil is extracted with ethyl acetate (3x), the extracts washed with water and dried (MgS04). Removal of the solvent yields a yellow solid (20.89 g, 73.8%), m.p. 104-108C. It is used in the next step without further purification.

NMR (CDCI3, 400 MHz): ~ 2.72-2.82 (mm, 4H, CH2CH2COO), 3.82 (s, 3H, OCH3), 6.86 (d, 2H, J 9.1Hz, ArH), 7.34-7.46 (m, SH, ArH), 7.92 (d, 2H, J 9.~Hz, ArH).
MS (EI. m/z): 342 (M)-+, 135 (b.p.).

C. 4-(4-Methoxyphenvl)-5-phenyl-2-oxazole-propanoic acid 1 ~ A mixture of the crude 4-methoxybenzoin hemisuccinate (20.8 g, 0.061 mole) of Step B, urea (8.7 g, 0.146 mole) and acetic acid (60 rnL) is heated at reflux under nitrogen for 5.5 hours. The rnixture is cooled and poured into ice water. The liberated oil is extracted with ethyl acetate (3x). The extracts are washed with water until neutral and then extracted with saturated sodium carbonate. The combined aqueous extracts are carefully acidified in the cold with concenllated HCI and extracted with ethyl acetate. The organic extract is dried (MgSO4) and evaporated to dryness to provide a waxy yellow oil ~19.6 g).
Purification of the residue by flash chromatography (on silica Merck-60, eluant: dichloro-methane-ethyl acetate 8:2) yields a pale yellow solid (14.3 g,72.7%), m.p. 100-101C.

NMR (CDCI3, 400 MHz): ~ 2.96 (t, 2H, CH2C), 3.20 (t, 2H, CH2COO), 3.83 (s, 3H, OCH3), 6.90 (d, 2H, ArH), 7.28-7.38 (m, 3H, ArH), 7.54-7.62 (m, 4H, ArH).
MS (EI, m/z): 323 (M)+, 278 (b.p., M-COOH)+, 152, 77.

.

wo gl/06~38 2 0 ~ 7 1 3 8 Pcr/US90/06252 I). 4-(4-Hvdroxv*henvl)-5-phenvl-2-oxazole-~ropanoic acid To a solution of the methoxyacid (5.6 g, 17.3 mmole) of Slep C, in acetic acid (55 mL) is added 48% HBr (84 mL) and the mixture is heated at reflux under nitrogen for 8 hours (TLC, 1: 1 hexane-ethyl acetate). After cooling, water is added and the solution extracted with ethyl acetate (3x). The extract is dried (MgS04) and evaporated to dlyness. The residue (brown waxy oil, 5.25 g, g9%) is used in the next step without further purification. For analytical characterization a small sample is flash-chromatographed (on silica Merck-60, eluant: dichloromethane -methanol 98:2 and 95:5).

NMR (DMSO-d6, 400 MHz): o 2.75 (t, 2H, J 7.14Hz, CH2C), 3.02 (t, 2H, J 7.1Hz, CH2COO), 6.77 (d, 2H, J 8.7Hz, ArH), 7.35 (d, 2H, J 8.55Hz, ArH), 7.41 (t, 3H, J 7.12Hz, ArH), 7.50 (d, 2H, J 7Hz, ArH), 9.64 (broad s, exchangeable).
MS (EI, m/z): 309 (M)~, 264 (M-COOH~+, 121, 105, 77.

E. 5-Phenvl-4-(~hYd,roxvpheny!)-2-oxazole propanoic acid methylester A solution of the crude acid (5 g, 16.18 mmole) of Step D, in methanol (40 mL), containing a small amount of p-toluenesulfonic acid H2O (0.58 g) is refluxed for 2.5 hours. The methanol is evaporated and the residue is partitioned between ethyl acetate and 20~o NaCI. The extracts are washed, dried (MgSO4) and evaporated to yield a thick oil (ca.

4.8 g). The residue is flash-chromatographed (on silica Merck-60, preabsorbed in dichloro-methane, eluted with a dichloromethane-ethyl acetate gradient from 90:10 to 75:25) to yield a white solid (3.56 g, 68%), m.p. 115-116C.

NMR (CDC13. 400 MHz): o 2.92 (t, 2H, J 7.4 Hz, CH2C), 3.20 (t, 2H, J 7.4Hz, CH2COO), 3.71 (s, 3H, OCH3), 6.74 (d, 2H, J 8.59 Hz, ArH), 7.26-7.36 (m, 3H, ArH), 7.40 ~d, 2H, J 8.7Hz, ArH), 7.54 (d, 2H, J 7.56Hz, ArH).
MS (EI, m/z): 323 (M)+, 264 (M-COOCH3)+, 105, 77 (b.p.).

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;' ,~ ' . : ' ~ ) 91/06538 2 0 ~ 713 8 PCr/US90/062~2 F. S-Phenvl-4-~4-(2-quinolinylmethoxv)phenvll-2-oxazole propanoic acid methvlester A mixture of the ester (2.46 g,7.61 mmole) of Step E, powdered anhydrous K2CO3 (1.05 g, 7.60 mmole), 18-crown-6 (0.223 g, 0.843 mmole) and acetonitrile (33 mL, ex-sieves) is stirred at room temperature under nitrogen for lS minutes. 2-Chloromethyl-quinoline (free base, freshly prepared from the hydrochloride salt, 1.35 g, 7.60 mmole) is added and the mixture is placed in an oil ba~h heated at 65C for 10 hours (N.B. A 10%
excess of the chloromethylquinoline, 18-crown-6 and K2CO3 is added after 6 hours). The solvent is removed and the residue is partitioned between ethyl acetate and water. The extracts are washed (brine), dried (MgS04) and evaporated to yield a yellow solid. The crude 10 product is flash chromatographed (on silica Merck-60, eluant: toluene and then toluene-methanol 97.5:2.5) to provide the title compound (3.5 g, quantitative yield).

NMR (CDC13, 400 MHz): ~ 2.90 (t, 2H, J ca. 7.2Hz, CH2C), 3.16 (t, 2H, J 7.2 Hz, CH2COO), 3.72 (s, 3H, OCH3), 5.41 (s, 2H, ArCH2O), 7.02 (d, 2H, J 8.8Hz, ArH), 7.29-7.36 (m, ca. 4H, ArH), 7.54-7.84 (m, ca. 6H, ArH), 7.83 (d, lH, lS J 8.1Hz, ArH), 8.08 (t, lH, J 8.5Hz, ArH), 8.19 (d, lH, J 8.5Hz, ArH).
MS (+FAB, rn/z): 487 (M+Na)+, 465 (M+H)+

-. - .
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.

- ' wO 91/0653~ ~ ~3 6 7 1 3 8 Pcr/usso/o62;2 G. S-Phenvl-4-r4-(2-quinolinvlmethoxv)phenvll-2-oxazole propanoic acid A solution of the ester (3.4 g,7.32 mmole) of Step F, in dry tetrahydrofuran (37 mL) is treated dropwise under nit~ogen with IN-LiOH (21.98 mL, 3 equiv.) and stirred at room temperature for 3 hours (TLC, dichloromethane-methanol 97:3 or toluene-methanol 5 9S:S). The solvent is evaporated, the residue is dissolved in water, neutralized in the cold with 10% acetic acid (to pH 5.5-6) and extracted with ethyl acetate. The extracts are washed with brine, dried (MgSO4) and evaporated to yield a pale yellow solid (3.18 g, quantitative yield). The crude product is recrystallized f~m warm ethyl acetate (containing enough dichloromethane to obtain a clear solution) to yield a first crop of crystals (2.63 g, m.p.
dec. 192-194C). A second crop is obtained by concentrating the mother liquors (0.327 g, m.p. dec. 192-193C). Tlie combined yield is 8S.8%.

IR (KBr, cm~l): 1720 (CO).
NMR (DMSO-d6, 400 MHz): ~ 2.76 (t, 2H, J 7Hz, CH2C), 3.03 (t, 2H, J 7Hz, CH2COO), 5.38 (s, 2H, ArCH2O), 7.11 (d, 2H, J 8.8Hz, ArH), 7.36-7.56 (m, 7H, ArH), 7.61 (t, IH, ArH), 7.69 (d, IH, J 8.5Hz, ArH), 7.78 (t, IH, ArH), 8.00 (t, J 7.9Hz, 2H, ArH), 8.42 (d, IH, J 8.5Hz, ArH).
MS (EI or C I, m/z): 451 (M+H)+, 310 (b.p.).

Analvsis for: C28H22N2O4 l~l~g: C, 74.65; H, 4.92; N, 6.22.
e~ C, 74.20; H, 4.86; Il, 6.00.

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' : , - . -.: '"'"~' ' .lO 91/û6538 ~ i 7 1 3 8 PCI/US90/06252 ~ 4-~2-~aphthalenYlmethoxvlrhenvl1-~-vhenvl-2.oxa7ole Dropanoic acid A. 4-~4-r2-Naphthalenvlmethoxvlphenvll-5-phenvl-2-oxazole ~ropanoic acid methvlester A mixture of the hydroxyester (1.5 g, 4.6 mmole) of Example 5E, powdered anhydrous K2CO3 (0.636 g,4.6 mmole), 18-crown-6 (0.123 g, 0.46 mmole) and acetonitrile (18 mL) is stirred at room temperature under nitrogen for 15 minutes. 2-Bromo-methylnaphthalene (1.13 g, 5.1 mmole) is added and the mixture is placed in an oil bath heated at 70C for 8-9 hours ~TLC, hexane-ethyl acetate 9: 1 or dichloromethane-methanol 9: 1). The solvent is evaporated and the residue dissolved in water and extracted with ethyl acetate. The extracts are washed and dried (MgSO4). Removal of the solvent yields a tan solid (2.17 g, quantitative yield). A sample is recrystallized from methanol (containing enough dichloromethane to obtain a clear solution) by concentrating to small volume and cooling in an ice bath. The white solid is collected and dried overnight in vacuo, m.p.134-1 ~ 135C.

IR (KBr, cm ~): 1740 (CO).
NMR (CDCI3-400 MHz): o 2.89 (t, 2H, J 7.5Hz, CH2C), 3.16 (t, 2H, J 7.5Hz, CH2COO), 3.71 (s, 3H, OCH3), 5.2 (s, 2H, ArCH20), 7.00 (d, 2H, J 8.6Hz, ArH), 7.25-7.35 (m, 3H, ArH), 7.46-7.58 (m, 7H, ArH), 7.8-7.9 (m, 4H, ArH).
MS (CI, rn/z): 464 (M+H)+, 324.

Analvsis for: C30H25NO4 Calculated: C, 77.73; H, 5.44; N, 3.02.
Found: C, 77.44; H, 5.36; N, 3.03.

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wo 9l/0653X ,~ 7 i 3 8 Pcr/US9()/062~2 ~, B. 4-~4-(2-Naphthalenvlmethoxv)phenyll-5-phenvl-2-oxazole ~rODanOiC acid A solu~on of the ester (1.49 g, 3.21 mrnole) of Step A, in dry tetrahydrofuran (18mL) containing I N-LiOH (9.6 mL) is stilTed under nitrogen overnight at room temperature (I~C,75:25 hexane-ethyl acetate). The solvent is evaporated, the residue dissolved in water 5 and acidified (to pH 5) with dilute HCI. The mixture is extracted with ethyl acetate, the extracts are dried (MgS04) and evaporated to yield the crude product (1.39 g, m.p. 145-150C). For purification, it is dissolved in hot ethyl acetate (containing enough dichloro-methane to obtain a clear solution), concentrated to half volume and precipitated with ether.
The white solid melts at 151-152C (1.07 g, 58%).

10 IR (KBr, cm~l): 1720 (CO).
NMR (DMSO-d6, 400 M~z): ~ 2.78 (t, 2H, CH2C), 3.03 (t, 2H, J 7Hz, CH2COO), 5.29 (s, 2H, AICH20), 7.10 (d, 2H, J 8.9Hz, ArH), 7.34-7.60 (m, 10H, ArH), 7.90-8.00 (m, 4H, ArH), 12.28 (s, lH, COOH).
MS (EI, m/z): 450 (M+H)+, 310.

1 ~ Analvsis for: C29H23NO4 Calculated: C, 77.48; H, 5,15; N, 3.11.
Fol~nd: C, 76.40; H, 5.16; N, 3.04.

.';091/06~38 2a67~38 PCr/US90/06252 4-r4~ Methvl-l H ben7imidazol-2-vl)methoxvl-Rhenvll-s-nh~nvl-2-oxazole eroDanoic acid A. 4-~4-~(]-Methvl-1 H-benzimidazol-2-yl~methoxYlphenYll-5-phenYl-2-oxazole propanoic acid methvlester A mixture of the ester (0.5 g, 1;55 mmole) of Example 5E, powdered anhy-drous K2CO3 (0.214 g, 1.55 mmole), 18-crown-6 (0.0416 g, 0.155 mmole) and acetonitrile (6 mL) is stirred under nitrogen at room temperature for 15 minutes. 2-Chloromethyl-1-methylbenzimidazole (0.307 g, 1.7 mmole) is added and the mixture is placed in an oil bath l o heated at 65-70C for 4 hours (TLC, dichloromethane-ethyl acetate 9: 1, iodine visualization).
A 10% excess of K2CO3, 2-chloromethyl- l -methyl-benzimidazole and 18-crown-6 is added at this point and the heating continued for another 10 hours. The solvent is evaporated, the residue dissolved in water and extracted with ethyl acetate. The extracts are washed, dried (MgSO4) and evaporated to dryness. The residue (1.64 g) is purified by flash-chromato-graphy (on silica Merck-60, preabsorbed in dichloromethane containing a small amount of methanol, eluted with dichloromethane-ethyl acetate 8:2) to yield 1.03 g (71.2%) of a light yellow solid, m.p. 142-144C (dec).

NMR (CDCI3, 400 MHz): o 2.87 (t, 2H, J 7.1Hz, CH2C), 3.16 (t, 2H, J 7.8Hz, CH2COO), 3.72 (s, 3H, COOCH3), 3.90 (s, 3H, NCH3), 5.41 (s, 2H, ArCH2O)~ 7.07 (d, 2H, J 8.8Hz, ArH), 7.25-7.40 (m, 7H, ArH), 7.5-7.6 (m, 3H, ArH), 7.78 (d, IH, ArH).
; MS (+C I, m/z): 468 (M+H)+, 324, 293, 147.

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wo 91/06538 2 0 5 7 1 3 8 P~r/US90/06252 B . 4-~4-r( l -Methvl- I H-benzimidazol-2-vl)methoxvl-5-phe_vl-2-oxazole propanoic acid A solution of the esler (I g, 2.14 mrnole) of Step A, in tebahydrofuran (13 mL) containing lN-LiOH (6.42 mL) is stirred under nitrogen at room temperature for 1 hour (~ll C, dichloromethane-ethanol 9: 1). The solvent is evaporated, water added and the pH
s adjusted to 6.5 with 10% acetic acid. The light ycllow precipitate is collected, washed with water and dried vacuo. It is redissolved in hot ethyl acetate (~ontaining enough methanol to obtain a clear solution), concentrated to a smaller volume and cooled in an ice bath. The crystals are collected and dried (0.642 g, 66.2%, m.p. 222-224C).

NMR (DMSO-d6, 400 MHz): ~ 2.76 (t, 2H, J 7Hz, CH2C), 3.03 (t, 2H, J 7Hz, CH2COO), 3.86 (s, 3H, NCH3), 5.43 (s, 2H, ArCH20), 7.14-7.66 (m, 13H, ArH).
MS (CI, mJz): 454 (M+H)+, 147 (b.p.).

Analvsis for: C27H23N3O4 Calculated: C, 71.51; H, 5.11; N, 9.27.
15 ~Q~: C, 71.62; H, ~ 17; N, 9.40.

E~m~
The compounds 5- and 12-hydroxyeicosatetraenoic acid (5-HETE and 12-HETE) and LTB4 are early arachidonic acid oxidation products in the lipoxygenase cascade, which have been shown to mediate several aspects of inflammatory and allergic response. This is especially true with respect to 5,12-diHETE, which is also denoted as LTB4 [see Ford-Hitchinson, J. Roy. Soc. Med., 74, 831 (1981)]. Compounds which inhibit the PLA2-mediated release of arachidonic acid thereby effectively prevent the oxidation of arachidonic acid to the various leukotriene products via the lipoxygenase cascade. Accordingly, the specificity of action of PLA2 inhibitors can be determined by the activity of ~est compounds in 2s this assay, which measures the ability of compounds to inhibit the synthesis of LTB4 by rat glycogen-elicited polymorphonuclear leukocytes (PMN) in the presence of exogenous substrate.
The assay is carried out as follows:
Rat polymorphonuclear leukocytes (PMNs) are obtained from female Wistarrats (150-200 g) which receive an injection of 6% glycogen (10 ml i.p.). Rats are sacrificed 18-24 hours post injection by CO2 asphyxiation and the elicited cells are harvested by peritoneal lavage using physiological saline (0.9% NaCI). The exudate is centrifuged at 400 xg for 10 ., , ~ - ,.

, . o 91/06538 2 ~ ~ 71 ~ ~ PCr/US90/062;2 minu~es. The supematant f}uid is discarded and ~he cell pellet is resuspended to a concentration of 2.0 x 107 cells/mL in HBSS containing Ca~ and Mg++ and 10 ~,lM L-cysteine.
To 1 mL aliquots of cell suspension, test drugs or vehicle are added, then preincubated at 37'C for 10 minutes. A23187 (1 IlM), [3H]-AA (3.0 llCi/mL) and unlabeled AA (1 ~lM) S are then added and the samples are further incubated for 10 minutes. The reaction is terminated by centrifugation and pelleting cells. Supernatants are then analyzed by HPLC analysis on a 15 cm x 4.6 mm ID supelcosil LC-18 (Supelco)(3M) column, using a two solvent system at a flow rate of 1.4 rnL total flow as follows:
Solvent A: 70:30 17,4 mM H3PO4:CH3CN
10Solvent B. CH3CN
Gradient: (system is equilibrated with Solvent A) Time Percent A Percent B -1515.0 100 0 20.0 65 35 40.0 65 35 42.0 10 90 50.0 10 90 2050.1 100 0 Percent solvent changes are accomplished in a linear fashion.

Injections: 140 llL of each supernatant is injected directly onto column and 3H arachidonic acid metabolites are monitored using an on-line radioactivity detector (Ramona, IN/US, Fairfield, NJ).
Standards: 104 - 2.0 x 104 dpm of eicosanoids of interest are injected in 90 ~L EtOH
cocktail.

Co-chromatography with standard 13H] leukotriene B4 (LTB4) in medium of stimulated 30 PMN exposed to drug is compared to that found in medium of stimulated cells exposed to no drug, generating percent inhibition.

Results are expressed as percent inhibition at a given compound dose or as an ICso value.
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Wo 9l/06538 2 ~ 6 7 1 3 ~ PCI/US90/062~ ~

Testing compounds of the invention m this assay gave the following results:

Compound of Example No. % Inhibition ketoprofen -50* (at IO~lM) 95 (at 0.5~LM) 2 91 (atO.5~,1M) 3 87 (at IO~M) 38 (at 0.511M) 4 8 (at I O~LM) 96 (at lOIlM) 6 95 (at lO~,lM) 81 (atO.511M) 6A 94 (at lOIlM) 63 (at 0.511M) 7 85 (at lO~M) 20 * a negative value denotes potentiation of cyclooxygenase (PGE2 synthesis) 13~ample 9 The procedure of Example 8 is also employed for the determination of the extent to which compounds of the invention inhibit the synthesis of the arachidonic acid 25 cyclooxygenase oxidation product PGE2.
In this assay, the procedure of Example 8 is carried out as described.
However, in order to determine cyclooxygenase activity, the samples are co-chromatographed with authentic reference [3H]-PGE2-` The results are calculated as in Example 8 and presented below:
.~ 30 Table II
C~ompound of ~, Example No. % Inhibinon ' 35ketoprofen 87 (at lOIlM) -13* (atO.511M) 2 -22* (at 0.511M) 3 - 8 (at IOIlM) . , .

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- 0 91/06~f38 2 ~ ~ ~ 1 3 g PCT~US90/06252 Table II fcont.) Compound of Example No. % Inhibition -8* (at O.511M) 4 -31~ (a~ lO~LM) -275* (al lO,~M) 6 -l91* (at lOIlM) ' -12* (atO.5~1M) 6A -79* (at lO~lM) -29* (at O.5~M) 7 -268* (at lO~,lM) * Negative values denote a potentiation of cyclooxygenase (PGE2 synthesis).

,ExamDle 10 The compounds of the invention are tested in an in vitro isolated phospholipase A2 assay to deterrnine the ability of the test compounds to inhibit the release of arachidonic acid from an arachidonic acid-containing substrate by the action of phospholipase A2 enzyme from human and non-human sources.
This assay is carried out as follows:
Into a 15 rnL polypropylene tube are added the following:

~ Volume. ~lL Final Conc.
3H-AA E. coli substrate l 25 5 nrnoles PL
CaCl2(0.lM) 2 5 SmM
Tris-HCl (O.SM) pH 7.5 3 20 100 mM
Water 4 25 Drug/vehicle 5 1 50,uM
PLA2 25 Volumeyielding 12%
hydrolysisin lOmin.

* pre-incubate at room temperature 30 min prior to substrate addition.
;' Prepared by adding 2 mL deionized and distilled water to 2 rnL 3H-arachidonate 35 labeled E. coli (lower count), to which is added l mL of 3H-arachidonate labeled E. coli (higher count) to yield a total of S m substrate (containing lOOO nmoles phospholipid).
2 Stock 0.1 m CaCl2, required for enzyme activity.
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W o 91/06538 PC~r/US90/062~2 20~7138 3 Slock 0.5 m Trisma-Base.
S~ock 0.5 M Trisma-HCl. Adjust pH to 7.5 (optimum for enzyme).
4 Deionized and distilled water.
S Stock 10 mM prepared in dimethyl sulfoxide. Make 1:2 dilution with dimethyl S sulfoxide and add 1 IlL to 100 ',IL assay tube.
6 Two human PLA2 enzymes are used:
a) Semi-purified human platelet acid extract PLA2 (in 10 mM sodium acetate buffer, pH 4.5). Remove protein precipitate by centrifugation at about 2200 rpm for 10 rninutes.
b) Purified human synovial fluid.

Incubate the 100,uL reaction mixture for 10 minutes at 37 C in a shaking water ba~h.
The reaction is terminated by the addition of 2 mL tetrahydrofuran, followed by vortexing.
NH2 columns (100 llg/mL - Analytichem International) are conditioned with 0.5 mLtetrahydrofuran followed by 0:5 mL tetrahydrofuran/water (2 mL:0. 1 mL, vh).
The sample is loaded onto the columns and slowly drawn through them. The hydrolyzed arachidonic acid retained in the columns is eluted therefrom with l mL
tetrahydrofuran/glacial acetic acid (2%), The arachidonic acid is transferred to scintillation vials and quantitated by ~-counting analysis. A "total counts" sample is prepared by pipetting 20 25 ~LL 3H-arachidonate E. coli directly into a scintillation vial to which is added 1 mL
tetrahydrofuran. 10 mL aquasol (scintillation cocktail) is added to all samples. Calculations:
:
[3H]AA dpm (sample) - [3H]AA dpm (nonspecific hydrolysis) % hydrolysis= x 100 total counts dpm vehicle dpm - drug dpm % change = x 100 vehicle dpm Activitv of Standard Drugs:
IC~o (~ M) Human Platelet Hurnan Synovial Dru~ P L A ~ _ P L ~
Arachidonic Acid 8.6 3.2 Monoalide 25.2 0.14 ;, .
,'' : ' . :
, - o 91/06538 2 ~ 6 7 ~ 3 8 PCT/US9o/06252 When ~ested in this assay, the compounds of the invention gave the following results:
Table m % Inhibition atlO ~IM
Compound of Example l~o. HSF*
ketoprofen -16.9* *
3 25.5 4 l5.l 10 * human synovial fluid ** negative values denote a potentiation of HSF

l~am~le 1 1 The compounds of the invention are evaluated for their ability to inhibit the l 5 lipoxygenase andlor cyclooxygenase pathways of arachidonic acid metabolism in the in vivo murine zymosan peritonitis assay.
This assay is carried out as follows:
Male CD-1 mice (8 weeks old) are placed in plastic boxes in groups of six. Animals are injected with l mL i.p. of either 1% zymosan in pyrogen free 0.9% saline or saline 20 (unstimulated control). Compounds are dosed orally l hour prior to zymosan injection.
Twenty minutes after zymosan injection, the mice are asphyxiated by C02 ir~halation and the peritoneal cavity is lavaged with 2 mL ice cold Hanks Balanced Salt Soluion (HBSS) without CaC12, MgSO4 7H2O and MgCl2- 6H2O. Peritoneal lavage fluid from each mouse is removed by syringe and placed in S mL plastic test tubes put on ice and volume is noted.
25 Preparation of samples for evaluation by ELISA is as follows: Samples are centrifuged at 800 xg for 15 minutes, l mL of the supernatant is added to 8 mL ice cold methanol and kept at -70 C overnight to precipitate protein; and samples are then centrifuged at 800 xg for lS
minutes, followed by a drying procedure in a Savant speed vac concentrator. The samples are reconstituted with l mL ice cold ELISA buffer and stored at -70 C until assayed. The assay 30 for eicosanoids (LTC4 and 6-keto-PGFIa) is performed according to conventional ELISA
procedures.
Compounds to be tested orally are suspended in 0.5% Tween 80. Compounds to be tested intraperitoneally are suspended in 0.5% methylcellulose in 0.9% saline.
The total metabolite level in lavage fluid/mouse is calculated and the significance 35 is determined by a one-way analysis of variance with LSD comparisons to control (p 5 0.05).
Drug effects are expressed as a percent change from control values.

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wo 9l/06538 ;~ O li 7 1 3 8 PCr/USso/062s~

The activity of standard drugs in this assay is as follows: -Compound EDsomg/k~ p.o.
LTC4 6-keto-PGFl,./TxB~
BW755C ~10 22.0 Phenidone 24.0 ~30.0 IndomethacinNot Active 0.126 IbuprofenNot Active 7.0 When tested in this assay a compound of the invention and the anti-inflammatory compound etodolac gave the following results:

Table Vl Compound of Dose % In hi bition Examp]e No. ~ LTC4 6-keto-PGF

5 10 (i.p.) * 86 -27 **

20 * ~ntraperitoneally administered ** negative values denote potentiation The results show that the compound of the invention exerts a potent inhibitory effect on the lipoxygenase pathway but not on the cyclooxygenase pathway.

,, .

-, : -.
, . . .. . .... .

Claims (11)

?O91/06538 PCT/US90/06252 WHAT IS CLAIMED IS:

1. A compound having the formula A(CH2)nO-B
wherein A is phenoxyethyl, phenoxyphenyl or a group having the formula wherein X is -N- or ;

Z is , , , , -S- or -O-;
R1 is hydrogen, lower alkyl or phenyl;
R2 is hydrogen or lower alkyl; or R1 and R2 taken together form a benzene ring;
R3 is hydrogen or lower alkyl;
n is 1 -2;
B is , or wherein R4 and R5 are each, independently, hydrogen or lower alkyl;
R6 is halo or nitro;

R7 is -?-R8 or ;
R8 is lower alkyl;
m is 0 - 3 ;
and the pharmacologically acceptable salts thereof.

2. The compound of Claim 1, having the name 1-[2-nitro-4'-(2-quino-linylmethoxy)- [ l,l '-biphenyl]4-yl]ethanone.

3. The compound of Claim 1, having the name 2-fluoro-4'-(2-quinolinyl-methoxy-[1,1'-biphenyl]-4-acetic acid.

4. The compound of Claim 1, having the name 3-[4-(2-quinolinyl-methoxy)benzoyl]benzene acetic acid.

5. The compound of Claim 1, having the name 3-[4-(2-naphthalenyl-methoxy)benzoyl]benzene acetic acid.

6. The compound of Claim 1, having the name 5-phenyl-4-[4-(2-quino-linylmethoxy)-phenyl]-2-oxazole propanoic acid.

7. The compound of Claim 1, having the name 4-[4-[2-naphthalenyl-methoxyjphenyl]-5-phenyl-2-oxazole propanoic acid.

8. The compound of Claim 1, having the name 4-[4-[2-naphthalenyl-methoxy]phenyl]-5-phenyl-2-oxazole propanoic acid methylester.

9. The compound of Claim 1, having the name 4-[4-[(1-methyl-1 H-benz-imidazol-2-yl)methoxy]phenyl]-5-phenyl-2-oxazole propanoic acid.

10. A process for preparing a compound of Claim 1 comprising a) etherifying a compound of formula , or wherein R4, R6 and m are as defined above; R7 is -?-R8 or ; and -COOR5 is an ester function;
with a compound of formula A(CH2)n-X
where A and n are as defined above and X is a leaving group, and, if necessary, hydrolyzing the product, to give a product wherein R5 is hydrogen or lower alkyl; or b) hydrolyzing a compound of formula I, II, III or IV which contains the -COOR5 moiety, wherein R5 is lower alkoxy, to give a compound wherein R5 is hydrogen or a pharmacologically acceptable salt thereof;
c) converting a compound of formula I, II, III or IV to a pharmacologically acceptable salt.

11. A process as claimed in Claim 10 substantially as hereinbefore described and illustrated in any one of Examples 1C, 2F, 2G, 3G, 3H, 4A, 4B, 5F, 5G, 6A, 6B, 7A and 7B.