CA2083610A1 - Leukotriene antagonists - Google Patents

Abstract

This invention relates to alkanoic acid compounds having phenyl and heteroarylthio substituents which are useful as leukotriene antagonists and pharmaceutical compositions containing such compounds. This invention also relates to methods of treating diseases in which leukotrienes are a factor by administration of an effective amount of the above compounds or compositions.

Description

WO 9~ 889 1 Pcr/us91/0392~
IEUKOTRIENE AN~AGONISTS 2 ~ 8 3 ~ ~ ~
BA~KGRO~p QF T}~ lNVENTIQN
"Slow Reacting Substance. of Anaphylaxis" (SRS-A~ has been shown to be a highly potent bronchoco~stricting substance which is released primarily from mast cells and basophils on antigenic 5 challenge. SRS-A has been proposed as a primary mediator in human asthma. SRS-A, in addition to its pronounced effects on lung tissue, also produces permeability changes in skin and may be involved in acute cutaneous allergic reactions. Fur~her, SRS-A has been shown to effect depression of ventricular contraction and potentiation of the 10 cardiovascular effects of histamine.
The discovery of the naturally occurring leukotrienes and their relationship to SRS-A has reinforced interest in SRS-A and other arachidonate metabolites. SRS-A derived from mouse, rat, guinea pig and man have all been characterized as mi%tures of leukotriene-C4 15 (LTC4), leukotriene-D4 (LTI)4) and leukotriene-E4 (LTE4), the structural formulae of which are represented below.
OH
~C2H
\~CsH ~ ~ SR

~G]u LTC4 R1 1 = Cys-Gly LTD4 Rl l = Cys-Gly LTE4 ~1 l = Cys Leukotrienes are a group of eicosanoids formed from

2 5 arachidonic acid metabolism via the lipoxygenase pathway. These lipid derivatives originate from LTA4 and are of two types: (1) those containing a sulfido- peptide side chain (LTC4, LTD4, and LTE4), and (2) those that are nonpeptidic (LTB4~. Leukotrienes comprise a group of naturally occurring substances that have the potential to O ~ontribute significantly to the pathogenesis of a variety of inflammatory and ischemic disorders. The pathophysiological role of leukotrienes has been the focus of recent intensive studies.
By antagonizing the effects of LTC4, LTD4 and LTE4 or o~her pharmacologically active mediators at the end organ, for examp]e,

3 ~ airway smooth muscle, the compounds and pharmaceutical compositions of the instant invention are valuable in the treatment of .
,, wo 91/1s88~ Q ~ 2 - PCI/US91/03922 diseases in subjects, including human or animals, in which leukotrienes are a factor.
~UMMARY OF ~IN~;~
This invention relates to compounds represented by structural formula (I) (O)qS

A~R~ I

wherein qisO, 1,or2;
R1 is (L)a-(CH2)b-(T)C-B wherein a is O or 1;
bis3 to 14;
c is O or 1;
L and ~r are independently oxygen, sulfur, or CH2 with the proviso that L and T are not sulfur when q is 1 or 2;
B is C14 alkyl, ethynl, trifluoromethyl, isopropenyl, furanyl, thienyl, cyclohexyl, or phenyl optionally monosubstituted with Br, C1, CF3, C14 alkyl, Cl 4 alkoxy, methylthio, or trifluoromethylthio;
2 O R2 and A are independently selected from H, CF3, Cl 4 alkylj C~ 4 alkoxy, F, C1, Br, I, OH, N02 or NH2; or Rl and A a~e H and R2 is (L)a-(CH2)b-(T)C-B wherein a, b, c, I" T, and B are as defined above;
Y is COR3 or ~CHX)n(CH2)p~Z wherein R3 is OH, NH2, aryloxy or Cl-6 alkoxy;
p is O or 1;
x is H, OH, Cl 4 alkyl, Cl 4 alkoxy, or F; and Z is COR3, or te~azolyl;
R is ~~(C) W

m is O to 6, but m is 1 to 6 when w is imidazole;
R4 and Rs are indepe~dently hydrogen or C1 4 alkyl at any position when m is not O;

WO 91/188B9 - 3 - PCI /US91/0392' W is a 5-membered ring heteroaryl group selected 2f~
tetra~olyl, thiazolyl, tnazolyl, thienyl, furyl, oxazolyl, thiadi~zolyl.
pyT~olyl, imida~olyl or pyra~olyl, unsubstituted or substitutecl with one to three of the group s ~(C) j-V;

R4 and Rs a~e as defined above;
j is 0 to 6; and V is hydrogen, Cl 1 alkyl, COR3, S03H, S02H, S02NH2, COCH20H, CHOHCH20H, or tetrazolyl, with R3 as defined above; or a pharmaceutically acceptable sal~ thereof.
This invention further relates to the ester or diester derivatives of the compounds of Formula (~).
This in~ention includes all stereoisomers, racemates, or mixtures thereof. For example, W can be 1,2,3-triazole; 1,3,4-triazole;
1,2,3-thiadiazole, 1,3,4-thiadiazole, and other possible steroisomers.
This invention further relates to pharmaceutical compositions comprising a pharmaceutical carrier or diluent and a nontoxic 2 0 amount of the compound of formula ~I). Such composi~ions are useful fo~ inhibiting the ef~ects of leukotrienes and in treating diseases in which leukotrienes are a factor.
This invention furthe~ relates $o pharmaceutical compositions comprising a pharmaceutical carrier or diluent and non~oxic amounts 25 of a compound of formula (I) and a histamine H1-receptor antagonist.
Such compositions are useful in inhibiting antigen-inducecl respiratory anaphylaxis.
This invention further relates to methods for inhibiting the effects of leuko~ienes. This invention also rela~es to methods for 3 0 inhibiting antigen-induced respiratory anaphylaxis comprising a~ministration of an effective amount of the above-described pharmaceu~ical compositions.

The cs>mpounds of this invention are represen~ed by the 3 5 following general structural formula (1)

- 4 - PCl/US91/0392 , s~ (o)qS R

A JJ~ R 1 wherein q is 0, 1, or 2;
R l is (L)a-(CH2)b-(T)c-B wherein a is 0 or 1;
b is 3 to 14;
c is 0 or 1;
L and T are independently oxygen, sulfur, or CH2 with the0 plOViSO that L and T are not sulfur when q is 1 or 2;
B is C14 alkyl, ethynl, trifluoromethyl, isopropenyl, furanyl, thienyl, cyclohexyl, or phenyl op$ionally monosubstituted with E3r, C 1, CF3, Cl-4 alkyl, Cl-4 alkoxy, methylthio, or trifluoromethylthio;
R2 and A are independently selected from H, CF3, Cl-4 alkyl, Cl-45 alkoxy, F, C1, Br, I, OH, NO2 or NH2; or Rl and A are H and R2 is (L)a-(CH2)b-(T)c-B wherein a, b, c, L, T, and B are as defined above;
Y is COR3 or (CHX)n(CH2)p-Z wherein R3 is OH, NH2, aryloxy or Cl-6 alkoxy;
n is 0 or 1;
p is 0 or 1;
X is H, OH, C-~-4 aLkyl, C1-4 alkoxy, or F; and Z is COR3, or te~azolyl;
. Ris R
~(C~ mW
R~
m is 0 to 6;
R4 and Rs are independenlly hydrogen or C1 4 alkyl at any position when m is not 0;
W is thienyl substituted with one tb three of the group 3~) V;

- 5 - PCI /I,'S91/03922 2~3~1~
j is O to 6; and V is hydrogen, Cl~ alkyl, CO~3, SO3H, S02H, S02NH2, COCH20H, CHOHCH20H, or tetrazolyl, with R3 as defined above; or a 5 pharmaceutically acceptable salt thereof.
This invention includes all stereoisomers, racemates, or mix~ures thereof. For example, W can be 1,2,3-triazole; 1,3,4-triazole;
1,2,3-thiadiazole, 1,3,4-thiadiazole, and other possible steroisomers.
The compounds of this invention further comprise ~he ester 10 and diester derivatives of the compounds of Pormula (I).
The preferred compounds of this invention are ~hos~ where m is 0, 1 or 2; thienyl is substituted with one of the groups --(C) j-V;
'~i where j is O or I and R4 and Rs are hydrogen and V is COR3, SO3H, SO2H, SO2NH2, COCH20H, CHOHCH20H, or tetrazolyl, with R3 as de~lned above; or a pharmaceutically acceptable salt ehereof.
A subgeneric class of these compounds are those represented 20 by structural formula (Ij wherein Y is C02H. Particular members of this subgeneric class are exemplified by the following compounds:
( 1 ) 2 - (2-d odecylphenyl ) -2- [ ( 1 -c arboxymeth yl -5 -tetrazolyl] -thio]acetic acid;
(2) 2-(2-dodecylphenyl)-2-[~1-(3-carboxypropyl)-5-tetrazolyl]-2 S thio]acetic acid;
(3) 2-(2-dodecylphenyl)-2-[(1-sulfomethyl-5-tetrazolyl)-thio]acetic acid;
(4) 2-(2-dodecylphenyl)-2-[~1-methyl-5-tetra~olyl)thio]acetic acid;
3Q (5) 2-[2-~8-phenyloctyl)phenyl]-2-~1-(3-carboxypropyl)-5-~etrazolyl]~hio]acetic acid;

(6) 2-~2-(tetrazol-5-yl)ethylthio]-2-(2-dodecyl-phenyl)acetic acid; and

(7) 2-~2-Dodecylphenyl)-2-(5-carboxy-4-methyl-2-3 ~ thiazolylthio)aceeic acid.
A second subgeneric class of ~hese compounds are ~hose represented by structural formula (I) where Y is CH2COOH.

2~3~
Par~icular members of this subgeneric class are exemplified by tllc following compounds:
(1) 3-(2-dodecylphenyl)-3-[[1-(3-carboxypropyl)-5-tetrazolyl]-propanoic acid;
(2) 3-[2-(8-phenyloctyl)phenyl]-3-[(1-car~oxymethyl-S-tetrazolyl)thio]propanoic acid; and (3) 3-[2-(8-phenyloctyl)phenyl]-3-[[]-(3-carboxypropyl)-5-tetrazolyl]thio]propanoic acid.
A third subgeneric class of these compounds are those 10 represented by structural forrnula (1) where Y is CH(OH)COOH.
Par~icular members of this subgeneric class are exemplified by the following compounds:
(1) 3-[2-(8-phenyloctyl)phenyl]-3-[[1-(3-carboxypropyl)-S-tetrazolyl]thio]-2-hydroxypropanoic acid;
(2) 3-[2-(~-pheslyloctyl)phenyl]-3-[(1-carboxymethyl-5-tetrazolyl)~hio]-2-hydroxypropanoic acid.
(3) 2-hydroxy-3-~2-(8-phenyloctyl)phenyl]-3-[[ 1-(2-carboxypropyl)-S-thienylmethyl]thio]propanoic acid;
(4) 2-hydroxy-3-[2-(8-phenyloctyl)phenyl]-3-[(2-2 0 carboxythien-5-ylmethyl)sulfonyl]propionic acid; and (S ) 2-hydroxy-3 - [2-( 8 -phenyl oc tyl )phenyl~ -3 - [ (2 -carboxythien-S-ylmethyl)thio]propionic acid.
Yet another set of compounds which are preferred are those where Y is CHXCO{)H or derivatives of this acicl such as its esters, amides and salts.where X is C1-C4alkoxy, particularly methoxy.
Exemplary compounds are:
3- [2-(8-phenyloctyl)phenyl] -3 - ~ [ 1-(3 -carboxypropyl)-5 -tetrazolyl]thio]-2-methoxypropanoic acid;
3 0 3-[2-(8-phenyloctyl)phenyl~-3-[(1 -carboxyme~hyl-S-tetrazolyl)-thio]-2-methoxypropanoic acid.
2-rnethoxy-3-[2-(8-phenyloctyl)phenyl]-3-[[1 -(2-carboxy-propyl)-~-thienylmethyl]thio]propanoic acid;
2-methoxy-3-12-(8-phenyloctyl)phenyl] -3 -[(2-carboxythien-3 5 5-ylmethyl)sulfonyl]propionic acid; and 2-methoxy-3-[2-(8-phenyloc~yl)phenyl]-3-[(2-carboxythien-S-ylmethyl)thio]propionic acid.
Some of the compounds of formula ~I) contain one or two asymmetric centers. This leads to the possibility of two or four wo 91/~8889 - 7 - Pcr/US9l/03922 2~36~
stereoisomers for each compound. The present invention includes all such stereoisomers, racemates, or mixtures thereof. The compounds of the present invention, depend;ng upon their structure, are capable of forming salts with known pharmaceutically acceptable bases, 5 according to procedures well known in the art. Such acceptable bases include inorganic and organic bases, such as ammonia, arginine, organic amines, alkaline earth and alkali metal bases. Of particular utility are the dipotassium, magnesium, calcium, diammonium, ~inc, pipeazine~ ethylenediamine l O and disodium salts of the diacid compounds of formula ~1).
The compounds of ~he forsnula (I) wherein Y is CO2H are conveniently prepared from an aldehyde precursor of the following structural formula (II) R2 ~CHC
A R, I I
wherein A, R1 and R2 are described above. A compound of formula (II) is treated with trimethylsilyl cyanide in the presence of ~inc iodide at low ternperatures in an inert solvent to form the 2 0 trimethylsilyl-protected cyanohydrin. Treatment of this with gaseous hydrogen chloride in methanol provides the methyl 2-hydroxyacetate derivative which is converled to the 2-chlcroacetate wi~h thionyl chloride. This valuable intermediate is then reacted with an appropriate thiol selected to give, af~er removal of ester 2 5 protective groups, the desired product of formula (I) The compounds of the formola (I~ wherein Y is CH(X)CO2H, wherein X is H, Cl 4 alkyl, or Cl ~ allcoxy, are prepared by reacting the appropliate aldehyde of the formula (II) and an esterified bromoacetate, conveniently t-butyl bromoacetate, with a mixture of 3 O diethyl aluminum chloride, zinc dust and a catalytic amount of cuprous bromide at low temperatures in an inert solvent to give the esterified 3-hydroxy-propionatc derivative which is reacted directly with a substituted thiol in tlifluoroacetic acid Altematively, a mixture of trimethyl borate and zinc in tetrahydrofuran may be used 3 5 to prepare the 3-hydroxypropionate deriva~ive By ~mploying an esterified 2-bromo-propiona~e in the above reaction with an

- 8 - PCl /US91/~39~_ 2~83~
aldehyde (II), the compounds of the formula (I) wherin Y is CH(CH3)C02H are ob~ained.
Alternatively, the compounds of the formula (I) wherin Y is CH(X)C02H wherein X is H, Cl 4 alkyl, Cl 4 alkoxy, or fluoro are S prepared from a propionate precursor of the following structural formula (IV) R2 ~C2RI I

10 wherin A, R1 and R2 are described above, Rlo is a conventional ester protective group, such as t-butyl or C1 4 alkyl, and R11 is hydrogen, Cl 4 alkyl, Cl 4 alkoxy, or fluoro. A compound of formula (IV) is reacted with a mixture of alkali metal alkoxide, such as sodium methoxide, and an appropriate thiol to give, after removal of the 15 ester protective group, the desired product of formula (I).
The propionate precursors of formula (IV) are prepared from the corresponding aldehydes of formula (Il) by general procedures such as reaction with an alkyl (triphenylphosphoranylidene)acetate or by conversion of the aldehyde to a 3-hydroxypropionate 2 0 derivative, as described above, followed by an elimination reaction to form the double bond. Additiona}ly, the propionate precusor is obtained from a 3-methanesulfonyloxypropionate derivative by treatment with triethylamine.
The compounds of the formula (I) wherein Y is 25 CH(OH)(CH2)pC02H are prepared from an epoxide precursor of the following structural formula (V) R2 ~(CH2)p-C02RI2 A/~Rl (V) 30 wherein A, R1 and R2 are described above, p is 0, 1 or 2 and R12 is a conventional ester protective group, such as t-butyl or Cl 4allcyl such as methyl or ethyl. A compound of formula (V) is reacted in an inert solvent with triethylamine and the appropriate thiol selected to give, after removal of ester protective groups, a desired product of 3 ~ formula ~I).

wo g~ 3889 - 9 - ~ PC~ JS91/0392 The epoxide precursors of formula (Y) where p is 2 are prepared by reaction of the Grignard derivative of a bromobenzene compound of the formula (VI) R2 ~Br S A Rl (VI) with acrolein to give the corresponding enol delivative which is treated with a trialkylorthoaceta~e, followed by epoxidation using m-chloroperbenzoic acid.
The epoxide precursors of formula (V) where p is I can be prepared by Arndt-Eistert homologation of the compound where p is O and R1 2 is H.
The epoxide precursors of formula (V) where p is O are prepared by reaction of an aldehyde of the formula (Il) with a lower l S alkyl chloroaceta~e and an alkali me~al alkoxide, such as sodium methoxide in an appropriate solvent such as diethyl ether or methylene chloride.
The compounds of the formula (I) wherein Y is (CH2)3C02H are prepared from a tetrahydro-4H pyran-2-one preeursor of the 2 0 following structural formula (VII) o 0~
R2 ~f ~,J J

A/~R' (YII) wherein A, Rl and R2 are described above. A compound of formula 2 S (VII) is reacted with a mixture of zinc iodide ancl a substituted thiol in an inert solvent or with a substituted thiol in trifluoroacetic acid to give, after removal of any ester protective group, a product of formula (I).
The teIrahydro-4H-pyran 2-one precursors of formula (VII) 3 0 are prepared by reaction of Ihe Grignard derivative of the bromobenzene compound of formula (VI) with chlorotitanium ~riisopropoxide followed by reaction with 5-oxovalerate alkyl ester.

wo 9~ 889 2 l~ 8 ~ o - Pcr/us9l/039z2 The aldehydes of the formula (Il) are known or readily prepared utilizing the general procedures descTibed as follows.
The aldehyde precursors to the compounds of the formula (I) wherein R1 is, for example, an alkyl radical containing 8 to 13 carbon atoms are prepared from the appropriate 2-alkylphenyl-4-,4-dimethyloxazoline [see Meyers et al. L. rg. ~hem., 43 1372 (1978)].
The aldehyde precursors of the compounds of ~he formula (I) wherein R1 is, for example, an alkoxy radical containing 7 to 12 carbon atoms are prepared by the O-alkyla~ion of the appropriate 2-hydroxybenzaldehyde with the corresponding alkylating agent by standard methods.
The thioalkyl containing aldehyde precursors of the compounds of the formula (I) are prepared by the reaction of the appropriately substituted o-haloalkylthiobenzene (for example, a compound of formula (III):
~X

A SR ( 1 I I ~

wherein X is halo and R is alkyl, with magnesium and 2 0 dimethylformamide by standard methods.
The phenylthioalkyl containing aldehyde precursors of the compounds of the formula (I) are prepared by the reaction of the appropriately substituted haloalkylbenzaldehyde with a thiophenol and ~riethylamine.
2 5 The heteroaryl mercaptan precursors necessary to prepare the compounds of formula (1) are known compounds and are convenient1y prepared employing standard chemical reactions. The mercapto derivatives of these precursors are prepared according to known methods. For example, 5-(2-mercaptoethyl)tetrazole can be 3 0 prepared by adding B-merceptopropionitrile to a mixture of sodium azide and aluminium chloride in tetrahydrofuran. The preparation of tetrazolthiol compounds with various substi~uents is described in U.S.
Patents 4,048,311, 4,220,644; and 4,286,089 of Berges. The prepara~ion of triazolthiols and thiadiazolthiols with various 3 ~ substi~uents is taught in U.S. Patents 3,868,369 and 3,989,694 of Berges. These mereaptans are reacted as described above to yield compounds of formula ~I).

wo gl/188~9 ~ 2~ 8 ~ PCr/US91/0392~
Appropriate modifications of the general processes disclosed, and as further described in the Examples provided hereinbelow, furnish the various compounds defined by formula (1).
The leukotriene antagonist aclivity of the comyounds of this invention is measured by the ability of the compounds to inhibit the leukotriene induced contraction of guinea pig tracheal tissues I n vitro. The following methodology was employed.
ln vi~ro: Guinea pig (adult male albino Hartley strain) tracheal spiral strips of approximate dimensions 2 to 3 mm cross-sectional width an 3.5 cm length were bathed in modified Krebs buffer in jacketed lû ml tissue bath and continuQusly aeraeed with 95% 0~/5 %
C 0 2 . The tissues were connected via silk suture to force displacement transducers for recor~ing isometric ~ension. The tissues were equilibrated for I hr., pretreated for l 5 minutes with mecloferlamic acid ( 1 M) to remove intrinsic prostaglandin responses, and then pretreated for an additional 30 minutes wi~h either the test compound or vehicle control. A cumulative concentration-response curve for LTD4 on triplicate tissues was generated by successive increases in the bath concentration of the LTD4. In order to minimize 2 0 intertissue variability, the cont~actions elicited by LTD4 were standardized as a percentage of the maximum response obtained to a reference agonist, carbaehol 110 M).
Calculations: The avera~es of the triplicate LTD4 concentration-response curves both in the presence and absence of the test 2 5 compound were plotted on log graph paper. The concentration of LTD4 needed to elicit 30% of the con~action elici~ed by carbachol was measured and de~ned as the EC30. The -log KB value for the test compound was determined by the following equations:
EC~o (presence of tes~ com ound) 1. X = dose ratio = EC30 (presence of vehicle cont~ol) 2. KB ~ concentration of test compollnd/(X-1) The compounds of this inve-n~ion possess biosignificant 3 5 antagonist activity against leukotrienes. The aneagonist acti~--ty of representative compounds of this invention is tabulated below in Table I (other data appears in the preparative examples). The -log KB values were calculated from the above test protocol. Where

9 2n~3~ 12- P~/US91/03922 compounds were tested more than once, the -log KB values given herein represent the current average data.
TABLE T
Leukotriene Anta~nist Açtivity 5Compounds_Qf Formula (I)*

-W _ _ _ m -(~ 4~.V Rl~ _Y -Log KB

5-tetrazolyl 0 1-CH2COOH ~12H25 CC)2H 6. 1 5-tetrazolyl 0 1-ClEI2SC)3H C12H25 CO2H . 6.4 5-tetrazolyl 0 1-CH3 C12H25 C~)2H 5.8 5-tetrazolyl 0 1-(CH2)3COOH C12~125 CO2H 7.4 5-tetrazolyl 0 1-(CH2)3COOH (cH2)gph CO2H 6.0 5-tetrazolyl 0 1-(CH2)3~H C12H2s CH~CO2H 6.6 5-tetrazolyl 0 1-CH2COOH (CH2)8Ph CH2CO2H 6-3 5-tetrazolyl 0 1-(CH2)3COOH (CH2)~Ph CH2CO2H 6.0 5-tetrazolyl 2 1-H C12H2~ CO2H 6.5 5-tetrazolyl o 1-(CH2)3CH (CH2)gPhCH(OH)CO2H 7.0 2 0 5-tetrazolyl 0 1-CH2CO~H (CH2)gPhCH(OH)CO2H 7.0 2-thiazolyl 0 4-CH3,5-COOH c~12H25C~ 6.7 (~)qS' A ~R
l4 (l)mW
25 For the compounds listed in the above table R is ~5 wherein R4 and RS are H, W is optionally substituted with -(C)jR4Rs-V; R2 and A
are H; and q is O.
Pharmaceutical compositions of the presen~ invention comprise a pharmaceutical ca~ier or diluent and an amount of a compound of 3 0 the ~rmula ~T) or a pharmaceutically acceptable salt, such as an alkali metal salt thereof, sufficient ~o produce the inhibition of ~he effects of leukotrienes, such as symp~oms of asthma and other hypersellsitivity diseases.

wo 91/18889 2 ~ ~ ~ 6 ~- ~
When the pharmaceutical composition is employed in the forrn of a solution or suspension, examples of appropriate pharmaceutical carriers or di]uents include: for aqueous systems, water; for non-aqueous systems, ethanol, glycerin, propylene glycol, corn oil, 5 cottonseed oil, peanut oil, sesame oil, liquid paraffins and mixtures thereof with water; for solid systems, lactose, kaolin and mannitol;
and for aerosol systems, dichlorodifluoro- methane, chlorotrifluoroethane and compressed carbon dioxide propellants.
Also, in addition to the pharmaceutical ca~ier or diluent, the instant

10 compositions may include other ingredien~s such as stabilizers, antioxidants, preseFvatives, lubricants, suspending agents, viscosity modifiers and the like, provided that Ihe additional ingredients do not have a detrimental effect on the therapeutic action of the instant compositions.
The nature of the composition and the pharmaceutical carrier or diluent will, of course, depend upon the intended route of administration, i.e. parenterally, topically, orally or by inhalation.
In general, particularly for the prophylactic treatment of asthma, the compositions will be in a form suitable for administration 2 0 by inhalation. Thus the compositions will comprise a suspension or solution of the active ingredient in water for administration by means of a conventional nebulizer. Alternatively the compositions will comprise a suspension or solution of the ac~ive ingredien~ in a conventional liquified propellan~ or compressed gas to be 2 5 administered from a pressurized aerosol container. The compositions may also comprise the solid active ingrediene diluted with a solid diluent for administration from a powder inhalation device. In the above compositions, the amount of carrier or diluent will vary but preferably will be the major proportion of a suspension or solution of the active ingredient. When the diluent is a solid it may be present in lesser, equal or greater amounts than the solid active ingredient.
For parenteral administration the ph~rmaceutical composition will be in the form of a sterile injectable liquid such as an ampule or an aqueous or nonaqueous liquid suspension, 3 5 For ~opical administration the pharmaceu~ical composi~ion will be in the form of a cream or ointment.
Usually a compound of formula I is administered to an animal subject, including humans, in a composition comprising a nontoxic amount sufficient to produce an inhibiiion of the syrnptoms of an WO gl/18889 2 ~ 4 - PCI/US91/0392 allergic Tesponse. When employed in this manner, the dosage of ,.le composition is easily determined by those skilled in the art and are generally selected from the range of from 350 mg. to 700 mg of active ingredient for each adminis~ration. For convenience, equal 5 doses will be administered 1 to 4 times daily with the daily dosage regimen being selected from about 350 mg. to about 2800 mg.
The pharmaceutical preparations thus described are made following the conventional techniques of the pharmaceutical chemist as appropriate to the desired end product.
Included within the scope of this disclosure is the me~hod of inhibiting the symptoms of an allergic response which comprises administering to an animal subject a therapeutically effective amount for producing said inhibition of a compound of formula 1, preferably in the form of a pharmaceutical composition. The administration 15 may be carried out in dosage units at suitable intervals or in single doses as needed. Usually this method will be practiced when relief of allergic symptoms is specifically required. However, the method is also usefully carried out as continuous or prophylactic treatmen~. It is within the skill of the art to determine by routine experimentation 2 0 the effective dosage to be administered from the dose range set forth above, taking into consideration such factors as the degree of severity of the allergic condi~ion being treated, and so forth.
Compounds of this invention, alone and in combination with a histamine H1-receptor antagonist, inhibit antigen inàuced contraction 2 5 of isolated, sensitized guinea pig trachea (a model of respiratory anaphylaxis) .
Pharmaceutical compositions, as described herein-abo~e, of the present invention also comprise a pharmaceutical carrier or diluent and a combination of a compound of the formula (I) or a 3 0 pharmaceutically accep~able salt thereof, and a his~amine Hl -receptor antagonist in amounts sufficient to inhibit antigen-induced respiratory anaphylaxis. Examples of histamine H1-receptor antagonists include mepyrarnine, 2-~4-(5-bromo-3-methyl-pyrid-2-yl~utylamino]-5-[(6-methyl-pyrid-3-yl) methyl]-4-pyTimidone and 3 S other known H1-receptor antagonists. Tlle above-defined dosage of a compound of formula I is convenien~ly employed for ~his purpose with the known effectiYe dosage for the histamine Hl-receptor antagonist. The methods of administra~ion described above for the Wo 9l/188~9 - 1 5 - ~ ~ ~ P~ ssl/o3922 single active ingredient can similarly be employed for the combination with a histamine Hl-receptor antagonist.
The following examples illustrate the preparation of the compounds of this invention and their incorporation into pharmaceutical compositions and as such are not to be considered as limiting the invention set forth in the claims appended hereto.
EXAMPlo~E 1 mçthyl-S-tetE~zQlYl)thiQla~e~iç acid 1 Q L~) 2-L2-DQd~cyJ~h~nYl~-4~-~lim~hYlQxazQ~n~
To freshly prepared dodecylmagnesium bromide (frQm 30.l3 mmol of dodecyl bromide and 26.20 mmol of m~gnesium) in distilled tetrahydrofuran (50 ml) was added 2-(2-methoxyphenyl)-4,4-dimethyloxazoline [A.I. Meyers et al., L~e~h~. 43. 1372 (1978~]
l S (17.88 mmol) in ~etrahydrofuran (30 ml). The resultant yellow solution was stirred under argon at ambient temperature for 20 hours. The solution was cooled in an ice water bath and quenched with aqueous ammonium chloride ( l 00 ml) . The reaction product was extracted into diethyl ether (lO0 ml) and the organic phase was 2 0 washed with saturated sodium chloride solution (50 ml) and then dried over anhydrous magnesium sulfate. Evaporation of the organic phase afforded a colorless oil which was purified by flash chromatography over silica gel with ~ percent ethyl acetate in hexane as eluant to afford the desired product as a pale yellow oil.
Analysis for C23H37NO: Calculated: C, 80.41; H, 10.85; N, 4.08.
~ound: C, 80.22; H, lO.SS; N9 3.87.
(b) 2-(2-Dodecylphen~1-3~4~-trimethyloxazolinium iodide A solution of the compound of ~xample l(a) (17.2 mmol) in me~hyl iodide (20 ml) was refluxed under argon for 18 hours. The 3 0 volatiles were removed under vacuum and the solid residue tnturated with ethyl acetate (25 ml) to afford the desired product as white crystals (mp 78-84C).
2-Do~lb~nz~ldehvdç
To an ice cold solution of the compound of Example l(b) (10.0 35 mmol) in methanol (~0 ml) over a period of 1~ minutes was added in small portions sodium borohydride (lO.0 mmol). The reactis,a mixture was allowed to stir for 30 minutes and was then quenched - with ~ percent sodium hydroxide (S0 ml). The reaction mixture w~s extracted with diethyl ether (2 x 50 ml) and the extract was washed wo 91/18889 2 ~ ~ 3 6 ~ ~ - I 6 - pcr/uss1lo392 with brine (50 ml) and dried over anhydrous magnesium sulfate.
Evaporation of the extract afforded an oil which was dissolved in acetone (50 ml) and 3N hydrochloric acid ( 10 ml) was added. The mixture was flushed with argon and stirred for 16 hours at ambient 5 temperature. The volatiles were removed under vacuum and the residue partitioned between diethyl ether (50 ml) and water (50 ml).
The aqueous phase was extracted with more diethyl ether (50 ml).
The combined organic phase was washed with brine (50 ml) and dried over anhydrous magnesium sulfate. Evaporation of ~he organic 10 phase yielded an oil which was purified by flash chromatography over silica gel with 2 percent ethyl acetate in hexane as eluant to afford the desired product as a colorless oil.
Analysis for ClgH30O: Calculated: C, ~3.15; H, 11.02. Found C, 82.59; Hl 10.65.
15 (d? Methvl 2-(2-dodecvlphen~ll 2 llv~
The compound of Example l(c) (17.2 mmol) was dissolved in me~hylene chloride (20 ml) and s~irred at 0C under argon. Zinc iodide ( 1.87 mmol) was added, followed by ~he dropwise addition of trimethylsilyl cyanide (2.45 ml, 18.3 mmoles) dissolved in methylene 20 chloride (30 ml). After 1 hour at 0C the ice bath was removed and the mixture stirred for 1 hour at room temperature. The solvent was stripped and methanol ( 100 ml) was added after the residue was cooled in an ice bath. Excess hydrogen chloride was bubbled into the solution wh;le the mixture was stilTed at ice bath temperature. The 2 5 ice bath was ~hen removed and the mixture stirred a~ room temperature for 18 hours. Water (20 ml) was added and the mixture stirred for 2 hours. The solvent was evaporated and the aqueous residue exeracted with ethyl acetate. The combined organic phases were dried over anhydrous sodium sulfate, fil~ered and evaporated.
3 0 The crude product was flash chroma~ographed on silica gel, eluted with 20% ethyl acetate/hexane, to give the product as a clear colorless liquid.
(e~ Me~hvl 2-Çhloro-2-(2-dodecvlphen~ cetate The compound of Example l(d) (12 mmol) was stirred under 35 argon in an ice bath and thionyl chloride (20 ml) was added in a single portion. The ice bath was removed and the mixture was stirred under argon for 18 hours. The solvent was stripped and the residue was flash chromatograpbed on 200 grams of silica gel with wo 91/18889 -17 - ~ 3 ~ ~ ~ Pcl/us9l/o3922 20% methylene chloride/carbon tetrachloride as eluant to give the product as a clear colorless liquid.

thi ol acetate The compound of Example I (e) was mixed in an amount of 0.7 g (0.002 mol) with 0.37 g (0.0027 mol) of 5-mercapto-1-methyltetrazole sodium salt, I ml of triethylamine and 20 ml of methylene chloride. The mixture was stirred at ambient temperature under argon for 3 days. The re action mixture was washed with water, dried over anhydrous magnesium sulfate and filtered. The filtrate was concentrated under vaccum to yield 0.7 g of product.
(g~ 2-C2-DQdecvlphenvl~-2-~(l-m~thyl-s-tetrazolvl)thiolacetic acid The compound of Example l(f) in an amount of 0.7 g (0.0016 mol) was dissolved in 5 ml of methanol, mixed with 0.4 g (0.01 mol) of sodium hydroxide, and stirred foT 2 hours. The reaction mixture was concentrated under vacuum and the residual oil was redissolved in 5 ml. of water. The pH of the solution was adjusted to 4.8 or until a precipitate formed. The precipitate was filtered, redissolved in 2 0 methylene chloride, washed with water, dried over anhydrous magnesium sulfate, filtered, and concentrated under vacuum. The resulting 0.3 8 of oil was flash chromatographed on silica gel, with methylene chloride, 0.05% me~hanol, and û.001% formic acid as eluent to yield O.lB g of oil which solidi~ied to a low melting wax.
2 5 Similarly, the following compounds are prepared according to the general method of Example 1 from the 2-(2-meehoxyphenyl)-4,4-dimethyloxazoline and the appropriate alkyl halide:
2-(2-Tetradecylphenyl)-2-~(1 -methyl-5-~etrazolyl)-~hio]acetic acid; and 3 0 2-(2-Oc~ylphenyl)-2-[(1 -methyl-5-tetrazolyl)thio]acecic acid.
EXAMPI,E 2 Preparation_of 2-(2-DodecYlphenvl)-2-~(1-carboxymethyl-5-letrazQlvl~thiolacetic acid.
(a) MQthyl_2-(2-Dodecy 3 5 te~ra~olvl)lthiolac~tate The compound of Example l(e) (3S2 mg, 1 mmol), triethylamine (0.21 ml, 1.5 mmol) and 5-mercapto-1-carbethoxy-methyltetrazole (250 mg, 1.33 rnmol) were combined in 25 ml of methylene chloride. The mixture was stilTed under argon at ambient wo gl,l8889 2 ~ ~ 3 6 ~ ~ - 1 8 - ~r/ussl /039~2 tempera~ure for 2 days. The solvent was stripped and the residu~
flash chromatographed on 50 grams of silica gel eluted with 15%
ethyl acetate/hexane to give the product (500 mg, 99%) as a clear colorless liquid.
s (~ 2-l2-Po~çnvl~-2-~2-~ç~rboxym~tra açetic ~cid The compound of Example 2(a) (260 mg, 0.52 mmol) was dissolved in 4.2 ml of methanol and st;rred under argon in an ice bath. A lN solution of sodium hydroxide (2.1 ml, 2.1 mmol) was added. The ice bath was removed, and the mixture stinred for 1 hour a~ ambient temperature during which time a white precip;tate formed. The methanol was evaporated and an additional 4 ml of water added to give a slightly lturbid mixture which was stirred overnight a~ ambient ~emperature. The mixture was acidified with dilute hydrochloric acid, extracted with ethyl acetate, dried over anhydrous sodium sulfate, filtered and evaporated. The crude product was recrystallized from ethyl acetate/hexane to yield the desired product (204 mg, 86%) as a white crystalline solid with m.p.
1a,7 148C.
Analysis for C23H34N4O4S- Calculated: C,59.72; H,7.41;
N,12.11. Found: C,59.61; H,7.27; N,12.15.

~~razolyllthiol~çetiç_~c.id 2 5 (a) Methyl 2-(2-dodecYlphenYI)-2~ -carbQxyFro~vl~-5 tç~r~azolyllthiol~ce~ate The compound of Example l(e) (325 mg, 1 mmol), triethylamine (0.42 ml, 3 mmol), ~-mercapto- 1 -(3-carboxy-propyl)tetrazole (2S0 mg, 1.33 mmol) and 25 ml of methylene 3 0 chloride were combined and stirred under argon at ambient temperature overnight. The solvent was s~ripped and the residue flash chromatographed on 50 grams of silica gel, eluted with 70:30-1 (hexane: ethyl acetate: formic acid) ~o yield the desired product (411 mg, 82%).
3 ~
thiolacetic acid The compound of Example 3(a) (411 mg, 0.82 mmol) was dissolved in 10 ml of methanol and stirred under argon in an ice bath. A 1 N solution of sodium hydroxide (3.2 ml, 3.2 mmol) was WOgl/188~39 -19- ~83~a l~cr/US9l/03922 added dropwise, the ice bath removed and the mixture stilTed overnight at ambient temperature. The solvent was stripped and the residue acidified with dilute hydrochloric acicl at ice bath temperatuTe. The crude product was extracted with ethyl acetate, 5 dried over anhydrous sodiu m sulfate, filtered and evaporated. The Tesidue was flash chromatographed on 50 grams of silica gel eluted wth 30:70:1 (ethyl acetate: hexane:formic acid) followed by 50:50:1 (ethyl acetate:hexane:formic acid). Recrystallization from ethyl ether/hexane gave the desired product (285 rng, 71 %) as a white 10 crys~alline solid with m.p. 86-~38DC.
Analysis for C2sH3gN4O4S - Calculated:
C,61.20; H,7.81; N,11.42; S,6.53. Found: C761.43; H,7.83; N,11.59;
S,6.60.

PTeparation o_2-(2-D~g~E~2-[~-sulfometh~1-5-tetrazol~ hiolaceti~ acid ammQni~m salt hYdrate The compound of Example l(e) (200 mg, 0.57 mmol) and 5-rnercapto-1-slllfomethyltetrazole disodium salt (136 mg, 0.57 mmol) were dissolved in 4 ml of dimethyl- formamide and stirred at 2 0 ambient temperature overnight. The solvents were pumped off and the residue dissolved in S ml of water. A lN solution of sodium hydroxide (2 ml, 2 mmol) was added at O and the mixture stirred overnight at ambient temperature. The solvents were pumped off and the residue flash chromatographed on 50 grams of silica gel 2 5 eluted with 6:3:1 (methylene chloride: ethanol: ammonium hydroxide) The solvents were stripped and the residue taken up in water and lyophilized to give the desired compound ~180 mg 58%) as an amorphous white solid.
Analysis for C22H34N4OsS2 2NH3 312 H2O -Calculated: C,47.21; H,7.74; N,15.01.
Pound: C,47.57; H,7.39; N 14.70.

EXAMPLE S

(a~2-(8-PhenvlQct,yl~benzaldeh,yde 8-Phenyloctyl bromide was prepared from 8-phenyloctanol, carbon tetrabromide and triphenylphosphine in methylene chloride.
A solution of 8-phenyloctanoic acid (19.8 mmol) in sieve dried w{~ 91/18889 2 ~ 20- Pcr/US91/0~2 tetrahydrofuran (5 ml) was reduced with diborane in tetrahydrofuran (30 ml, 29.1 mmol) al 20C for 4 hours to give ~-phenyloctanol. To an ice cold solution of ~he oclanol (ca. 19.8 mmol) and carbon tetrabromide (21.98 mmol) in rnethylel1e chloride (50 ml) 5 was added ~riphenylphosphine (22.30 mmol) in methy~ene chloride (50 ml) and the resulting solution was stirred for 2.5 hours. The volatiles were evaporated and the residue was taken up in ether (100 ml), cooled in ice, and filtered. The filtrate was evaporated and di~tilled to afford 8-phenyloc~yl bromide as an oil. To 8-phenyloctyl î O bromide ~nd 21.27 mrnol of magnesium) in distilled tetrahydrofuran (40 ml) was added 2-(2-methoxyphenyl)-4,4-dimethyloxazoline (17.10 mmol) in tetrahydrofuran (20 ml). After stirring for 24 hours, the reaceion mixture was similarly worked up to yield 2-[2-(8-phenyloctyl)phenyl]-4,4-dimethyloxazoline as an oil. A solutis)n of the oxazoline (1 1 .58 mmol) in methyl iodide (20 ml) was refluxed under argon for 18 hours. Removal of the volatiles afforded the corresponding 3,4,4-trimethyloxazolinium iodide as a white solid (mp 76.5-78C). To an ice cold solution of the iodide (9.46 mmol) in methallol (35 ml) was added in portions sodium borohydride ~9.20 mmol). Treatment of the reaction mixture as in Example l(c) results in the isolation of the desired product as an oil.
Analysis for C21H26O: Calculated: C, 85.67;
H, 8.90. Found: C, 85.12, ~5.22; H, 8.941 8.96.
~rnative prç~ar~tion of 2~ henvloctvlLbenzald~h~de 2 5 A solution of ~-hexynyl alcohol (102 mmol) in pyridine (150 ml), under argon, was cooled to 0C and p-toluenesulfonyl chloride (204 mmol) was added. The reaction mixture was Icept at about 4C
for 18 hours, poured into ice-water and then taken up in ether. The ether extract was washed with cold 10% hydrochloric acid, water and briIle. The organic layer was dried and concen- trated in vacuo to give 5-hexynyl p-toluenesulfonate. A solution of phenylacetylene (97 mmol) in tetrahydrofuran (200 ml) containing a trace of triphenylrnethane was cooled to O~C and then n-butyl lithium (37.3 ml of 2.6 mol in hexane) was added dropwise. The resulting solution 3 5 was stirred at 0C for 10 minutes and hexamethylphosphoramide (21 ml) was added dropwise. Af~er stirring for 10 minutes a solution of 5-hexynyl p-toluenesulfonate (97.1 mmol) in tetrahydrofuran (200 ml) was added. The reaction mixture was stirred at room tempera~ure for 18 'nours, diluted with ether and ~he organic layer wo 91/18889 - 21 ~ 2 ~ 8 3 6 ~ ~ Pcrl~s9llo3922 was washed with water and brine. The dried organic solution was concentrated and the produc~ was purified by flash chromatography to give 1-phenylocla-1,7-diyne. A mixture of this compound (43 mmol), 2-bromoben~aldehyde (35.8 mmol), cuprous iodide (0.5 mmol) and bis(triphenylphosphine) palladium (Il) chloride (0.7 mmol) in triethylamine (100 ml) was heated in an oil bath (95C) for one hour. The reaction mixture was cooled to 0C, filtered and the filtrate was concen- trated. The residl~e was dissolved in ether, washed with 10% hydrochloric acid, water and brine. The organic layer was dried and concentrated to give a product which was purified by flash chrvmatography to yield 2-(8-phenyl- 1,7 -octadiynyl)benzaldehyde. A solution of this compollnd (24.1 mmol) in ethyl acetate (100 ml) and 10% palladium on charcoal (I g) was hydrogenated (40 psi of hydrogen) at room temperature for 15 minutes. The catalyst was filtered off and the filtrate concentrated to give the 2-(8-phenylocsyl)benzaldehyde.
(~ Methvl 2-[2-(~-phenvloc~y~henyl1-2-hvdrQxY~çetate The compound of Example 5(a) or S(b) (10 mmol) was dissolved in methylene chloride (10 ml) and stirred at 0C under argon. Zinc iodide (1.1 mmol) was added followed by the dropwise addition of trimethylsilyl cyanide ( 1.47 ml, 11 mmol) dissolved in methylene chloride (20 ml). After 1 hour at 0C the ice baeh was removed and the mixture stirred for 1 hour at room ~emperature.
The solvent was stripped and me~hanol (60 ml~ was added at ice 2 5 bath temperature. Fxcess hydrogen chloride was bubbled into the solution while stirring. The ice bath was removed and the mixture sti~Ted at room ~emperature for 18 hours. Water ( 12 ml) was ~dded and the mixture stirre~ for 2 hours. The solvent was evaporated and the residue extracted with ethyl acetate, dried over anhydrous 3 0 sodium sulfate, filtered and e~aporated. The crude product was flash chromatographed on 200 grams of silica gel with 20% ethyl acetate/hexane as eluant to give the product as a clear colorless liquid .
(d~ _Me~hyl 2-chlor~2-l2-L~henyloct~ ?henylla~tate 3 5 The compoul~d of Example 5(c) (6.8 mmol) was stirred under argon in an ice bath and thionyl chloride (15 ml) was added in a single porlion. The ice bath was removed and the reaction mixture was stirred for 18 hours. The solvent was stripped and the residue flash chromatographed on 100 grams of silica gel with 20%

wo 91/188~9 2 ~ ~ ~ 6 ~ ~ - 2 2 - PCr/~91/03922 methylene chloride/carbon tetrachloride as eluans to give the product as a clear colorless liquid.
Le) Methyl, 2-~2~ henYloçtyl~phenyll-2-[~1-(3-carboxylpropvl)-~etrazQlvllthiolaceate S The compound of Example 5(d) (744 mg~ 2 mmol) was dissolved in methylene chloride (25 ml) and stirred under argon at room tempeTature. 5-mercapto-1-(3-carboxy-propyl)tetrazole (376 mg., 2 mmol) and triethylamine (~.84 ml, 6 mmol) were dissolved in methylene chloride (25 ml) and added to the solution of the compound of Example 5(d). The mixture was stirred under argon for 24 hours. The solvent was stripped and the residue was flash chromatographed on 100 grams of silica gel with 70:30:1 hexane:
ethyl acetate: formic acid as eluant to give the desired product (830 mg9 79%)-(f!_ 2-~2-(8-phenvlo~ ,phenyll-2-~L1-(3-çarboxvprop~
tetrazo!Yllthiola~ic acid.
The compound of Example 5(e) (524 mg, 1 mmol) was dissolved in methanol (12 ml) and stirred under argon in an ice bath, A lN
solution of sodium hydroxide (4 ml, 4 mmol) was added dropwise, 2 0 the ice bath removed, and the mixture stirred overnight at room temperature. The solvent was stripped and the residue was cooled in an ice bath and acidified with dilute hydrochloric acid. The crude product was extracted with ethyl acetate, dried over anhydrous sodium sulfate, fi1ltered, and evaporated. The crude product was 2 5 flash chromatographed on 70 grams of silica gel eulted with 30:70:1 ethyl acetate: hexane: formic acid followed by SO:S0:1 ethyl acetate:hexane:formic acid to eluent to give the desired product (495 ~g; 97%)-(~,) 2~ Phenvlo~tYl)~henvll-2-~1-(3-c~rboxy.pr~pvl~-5-3 0 ~etrazolYllthiolacetic acld. dipo~assium sa~hvdrate.
The compound of Example 5(f) (495 mg, 0.97 mmol) was treated with a solution of potassium carbonate (415 mg, 3 mmol) in 10 ml of water under argon at ice-bath temperature. The ice ba~h was removed and the mixture allowed to stir for lS minutes at room 3 5 tempera~ure. The so~ution was then chromatographed on a Cl 8 eolumn; elution was with water to remove the excess base and then with 1:1 (acetonitrile:water~. Lyophilization gave the desired compcund (~24 mg, 92%) as a white hygroscopic solid.

wo gl/18889 2 3 ~ 2 ~ ~ 3 ~ ~ ~ PCr/U~91/0392~
Analysis for C27H32N404S 2K H20 - Calculated: C,53,62; H,5.67;
N,9.26. Found: C,53.81; H,5.51; N,9.36.
Similarly the following compounds are prepared according to the general method of Example 5 from 2-(2- methoxyphenyl)-4,4-5 dimethyloxazoline and the appropriate alkyl halide:
2-[2-(4-phenylbutyl)phenyl] -2-[[1 -(3-carboxypropyl)-5-tetrazolyl]thio]acetic acid, dipotassium salt, hydrate; and 2-[2-(1 0-phenyldecyl)phenyl]-2-[[ 1 -(3-carboxypropyl)-5-tetrazolyl~thio]acetic acid, dipotassium salt, hydrate.

wo gl/18889 - 2 4 - Pcr/ussl/o3922 2~3~ ~ EXAMPLE 6 Prepara~ion o~-!2-Dodeçy!phenv~ -(3-carboxvpr 5-te~razol llthioll?ro~anoic acid Butvl ~-hvdroxy-~-(2-dQ~çcvl~pbenyl)propionate A solution of diethylaluminum chloride (54.7 mmol) in hexane was added to a slurry of zinc dust (74.5 mmol) and a catalytic amount of copper (I) bromide (2.5 mmol) in anhydrous tetrahydrofuran (300 ml) while stirring under argon at 20C. The resulting mixture was then cooled to 0C in an ice-methanol bath. A
10 solution of t-butyl bromoacetate (49.8 mmol) and 2-dodceylbenzaldehyde of Example 1 (c) (54.7 mmol); in anhydrous tetrahydrofuran was added slowly over 60 minutes. The reaction was stirred for about 24 hours and was permitted to warm slowly to room temperature. The mixture was filtered to remove zinc, 15 concentrated, acidified with 3N hydrochloric acid and extracted with ether. Organic extracts were dried over magnesium sulfate, filtered, and evaporated to afford crude product. This material was then ~lash chromatographed on silica using 8% ethyl acetate in hexane to give the desired product in 79% yield.
2 0 (b) _ 3-~2-dQ~eçY.lDhçnvl)-~ (3 -carbQxxpro~v!l-5 -tetrazolyllthiQl~rop~noiç acid.
A 3 neck 100 ml round ~ottom flask e~uipped with a thermometer and stiTring bar was cooled to -30C using a dry ice-acetone bath followed by an ice-methanol bath. The flask was 2 5 charged with trifluoroacetic acid (20 ml) followed by 5-mercapto-1-(3-carboxypropyl~tetrazole (0.00128 mol, 0.1923 8)- The mixture was permitted to cool under argon for 10 minutes~ To this was added the compound of Example 6(a) (0.001282 mol, 0.~ g), in me~hylene chloride (5 ml). The reaction was stirred for 2 hours al 1~C and was 3 0 pérmitted to warm to ambient temperature. The trifluoroacetic acid was evaporated and the resulting oil was chromatographed on C1 8 packing using 20% water in methanol wi~h 1.0% forrnic acid to yield the desired product.
Analysis for C26H40N4O4S - Calculated: (with 0.5 mole H2O) 3~ C:60.81; H:7.79; N: lO.g2. ~ound: C: 60.24; H: 7.79; N: 10.62.
Similarly, the following compounds are prepared according to the general method of Example 6.
3-(2-tetsadecylphenyl)-3-[[ 1 -(3-carboxypropyl)-5-tetrazolyl]thio]propanoic acid; and , .~

wo ~ 889 - 2 5 - 2 ~ 8 3 ~ 1 ~ PCI /US91 /03922 3-(2-octylphenyl)-3 - [ [ 1-(3 -carboxypropyl)-5 -tetrazolyl]thio~psopanoic acid.
E~AMPLE 7 Prep~rat on Qf~-[~-Phenvlo~tv~ç~ll-3-LLL-ç~rboxvmethYI-~-S
~a) ~B~l ~ -hydrox~2~
A solution of diethylaluminum chloride (0.0082 moles, 8.2 ml) in hexane was added to a slurry of zinc dust (0.0111 mol; 0.7248 g) and a catalytic amount of copper (I) bromide (0.0004 mol; 0.0585 g) 10 in anhydrous tetrahydrofuran (40 ml) while stirring under argon at 20C. The resulting mixture was then cooled to -20C in an ice-methanol bath. A solution of t-butyl bromoacetate (0.0082 mol; 1.32 ml~ and 2-(8-phenyloctyl)benzyladehyde of Example S(a) or (b) (0.0082 moles; 2.3969 g) in anhydrous te~ra- hydrofuran was added 15 slowly over 60 min. The reaction was stirred overnight and was permit~ed to warm up slowly to room temperature. The mixture was fillered to remove zinc, concentrated, and azeotroped with methylene chloride. The resulting oil was then chromatographed on silica using 5% ethyl acetate in hexane with 0.5% formic acid to yield the desired 2 0 product.
(b~ Pre~a~iQn of 3-~2-(~-12hçny~ phçnyl1-3~

A 3-neck 1 ûO ml round bottom flask equipped with a thermome~er and s~irring bar was cooled to -30C using a dry ice-25 acetone bath followed by an ice melhanol bath. The flask wascharged wi~h trifluoroacetic acid ( 10 ml) followed by 5-mercap~o- 1-carboxymethyltetrazole (0.00124 mol, 0.2 g). The mixture was permitted to cool under argon for 10 minutes. To this mixture was added the compound of Example 7(a) (0.00113 mol, 0.4634g, in 30 methylene chloride (3 ml). The reaction was stirred for 2 hours at -15C and was then permittecl to warm to room temperature. The trifluoroacetic acid was evarporated and azeotroped with methylene chloride. The resulting oil was chromatographed using 30% ethyl acetate in hexane with 0.5% iFonnic acid to provide a 26% yield of 3 5 desired product.
Analysis for C26H32~4O4S 3/4H20~
Calculated: C:61.21; H:6.32; N:10.98; S:6.29;
Found: C:61.22; H:6.47; N:11.87; S:6.38 W~ 91/~8B89 - 2 6 - P~/U!~91/03922 2~ 6~ ~
Similarly, the following compounds are prepared according .~, the general method of Example 7:
3-[~-(4-phenylbutyl)phenyl]-2-[(1 carboxypropyl-5-tetrazolyl)thio]propanoic acid; and 3-[~-(10-phenyldecyl)phenyl]-2-[(1 -carboxypropyl-5-tetrazolyl)thio]propanoic acid.

prepara~ of ~-L2-(~Ph~nyloctyl~phenyll-3-L~ rboxypropYl)-A 3 -neck 1 00 ml round bottom flask equipped with a thermometer and s~irring bar was cooled to -30C using a dry ice-acetone bath followed by an ice-methanol bath. The flask was charged with trifluoroacetic acid (20 ml) followed by 5-mercapto- 1-~3-carboxypropyl)tetrazole (0.0024 mol, 0.4S74 g). The mixture was 15 pennitted to cool under argon for 10 minutes. To this was added the compound of Example 7(a) ~0.0024 mol, 0.9069g) in methylene chloride (5 ml). The reaction was stirred for 2 hours at -15C and then was permitted to warm to room temperature. The trifluoroacetic acid was evaporated and the resul~ing oil was 20 chromatographed on reverse phase C18 packing using 20% water in methanol with 0.5% formic acid to give the desired product in 26%
yield.
Analysis for C2~H36N404S . 1/2 H2O -Calculated: C:63.02; H:6.80; N:10.5.
Found: C:63.08; H:6.74; N:10.4 Prçpara~iQn_ of 2-(2-unde~l oxYphenyl~ -2-~ ( I -c arboxvm eth Yl -~ -tetrazolv~ ce~iç acid (a) _ 2-~ndeçyloxy~enzaldeh~
3 0 To a stirred suspension of sodium hydride (lO.0 mmol~, which was prewashed with petroleum ether, in sieve dried dimethylfonnamide (10 ml) was added dropwise a solution of salicylaldehyde (10.1 mmol3 in dirnethyl- formamide (1 ml). To the reaction mixture was then added undecyl bromide ( 10.0 mmol) and 3 5 the mixture stirred for 16 hours at ambienl temperature under nitrogen. The reaction mixture was ~aken up in hexane (50 ml) and washed with 10 percent sodium hydroxide (2 x 50 rnl) and saturated sodium chloride (50 ml). The organic phase was dried over anhydrous magnesium sulfate and charcoal. Evaporation of the wv gl/l88~9 ~ ~ ~ 3 ~ 1 ~ Pc~ 's9l/o392~
volatiles yielded a colorless liquid which was purified by flash chromatography over silica gel with 2 percent ethyl acetate in hexane as eluant to afford the desired product as an oil.
Analysis for ClgH2gO2: Calculated: C, 78.21; H, 10.21. Found:
C, 77.92; H, 9.9S.
~b) 2-~-~Jndçç~loxv henvl)-2-~(1-car~Qxvmeth~/1-5-tetrazolyl)-Employing the general methods of Example l (d)- l (g), the compound of Example 9(a) is converled to the desired product.
The following compounds are prepared according to the general methods described above ~rom the appropriately substituted hydroxybenzaldehyde and the appropriate alkyl halide:
2-(2-Heptyloxyphenyl)-2-[(1 -carboxymethyl-5-tetrazolyl)thio]acetic acid;
1 5 2-(2-Dodecyloxyphenyl)-2- [( 1 -carboxymethyl -5 -tetrazolyl)thio~acetic acid;
2-(S-Methoxy-2-dodecyloxyphenyl)-2-[( 1 -carboxy-methyl-~-tetrazolyl)thio]acetic acid;
2-(S Methyl-2-dodecyloxyphenyl)-2- [( I -carboxylmethyl -5 -Z 0 tetrazolyl)thio]acetic acid;
2-(5-Fluoro-2-dodecyloxphenyl~-2-[(1 -carboxymethyl-5-tetrazolyl)thio]acetic acid;
2-(S -Chloro-2-dodecyloxyphenyl)-2- [( 1 -carboxymethyl-5 -tetrazolyl)thio]acetic acid;
2-(5-Iodo-2-dodecyloxyphenyl)-2-~(1-carboxymethyl~5-tetrazolyl)thio~acetic acid;
2-(5 -Bromo-2-dodecyloxyphenyl)-2- [( 1 -carboxymethyl -S -tetrazolyl)~hi o] acetic acid;
2-(5-Hydroxy-2-dodecyloxyphenyl)-2-[( 1 -carboxy-methyl-5-3 0 tetrazolyl)thio]acetic acid;
2-(5 -Nitro-2-dodecyloxyphenyl)-2- [( I -carboxymethyl -5 -tetrazolyl)thio~acetic acid;
2-(5 -Amino-2-dodecyloxyphenyl)-2 - [ ( 1 -carboxyme~hyl -5 -~etrazolyl)thio]acetic acid; and 3 S 2-(5-Trifluorometnyl-2-dodecyloxyphenyl)-2-~(1-carboxymethyl-S -tetrazolyl)thio] acetic acid .
2-(2-Dodecylthiophenyl)-2-[( 1 -carboxymethyl-5 -tetrazolyl)thio]acetic acid is prepared from 2-(dodecyl-thio~benzaldehyde .

wo 91ilB889 ~ ~ 8 3 ~ ~ - 2 8 - Pcr/uss1/o392~
2-(2-Heptylthiophenyl)-2-[( 1 -carboxymethyl-5 -tetrazolyl)thio]acetie acid is prepared from 2-(heptylthio)benzaldehyde.

S ~}~r~2n _Q~2-~2-6-l~henylhçxvlox~ph~vll-2-~l-(3 ca,Iboxvpro~yl-~-tetrazolyl~hiola~j5 (a! 2-L~-phenvlhexylQ~)be~nzal~hy~e A solution of 6-phenylhexanoic acid (19.8 mmol) in sieve dned tetrahydrofuran (5 rnl) was reduced with diborane in te~ahydrofuran (30 ml, 29.1 mmol) at 0C for 4 hours to give 6-- phenylhexanol. To an ice cold solution of the hexanol (ca. 19.8 mmol) and carbon tetrabromide (21.98 mmol) in methylene chloride (50 ml) was added tripherlylphosphine (22.30 mmol) in methylene chloride (50 ml) and the resulting solution was stirred for 2.5 hours. The volatiles were evzporated and the residue was taken up in ether ( 100 ml), cooled in ice, and filtered. The filtrate was evaporated and distilled to afford 6-phenylhexyl bromide as an oil. A mixture of the bromide (8.00 mmol), salicylaldehyde (8.19 mmol) and potassium carbonate (9.33 mmol) in dimethylformamide (la ml) was heated to 2 0 100C and maintained at that temperature for one hour. The cooled reaction mixture was taken up in hexane (50 ml) and washed with 5% sodium hydroxide (~0 ml) and saturated sodium chloride (50 ml).
The organic phase was dried over anhydrous magnesium sulfate and charcoal. Evaporation yielded a colorless oil which was puri~ied by 2 5 flash chromatography over silica gel with 5% ethyl acetate in hexane as eluant to afford the desired product as an oil.
Analysis for Cl~H22O~: Calculated: C, 80.82; H, 7.85. Found: C, 80.62; H, 7.72.
(b) 2-~2-(6-PIlQny~h~oxy)~henvll-2-~ -carboxy~ropyl) 3 0 tçtrazQ!~llthiclacetic acid Employing the general methods of Example 5(c) through 5(g) the compound of Example 1 U(a) is converted to the desired product.
The following compounds are prepared according to the general methods described above from the appropriately substituted 3 5 phenyla1kyloxy benzaldehyde.
2-[2-(3^Phenylpropyloxy)phenyl]-2-1 [ 1-(3 -car~oxypropyl)-5 -tetrazolyl)thiolacetic acid; and 2-[2-(9-Phenylnonyloxy)phenyl]-2-[[1 -(3-carboxypropyl)-5-tetrazolyl)thio]aeetic acid.

WO 91 /18889 - 2 9 - PCr/US91 /()392' EXAMPLE I 1 2 ~ ~ 3 ~ ~ ~
Prçparation of ~-methvl-~2-~ 3-~(1-carboxymethvl-~-tetrazo]yl~thiQlpropanoic acid (a) Me~hyl 2-me~hyl-3-h~drD ~ _ To a suspension of zinc dust (15 mmol) and copper (I) bromide (5 mmol) in distilled tetrahydrofuran (10 ml) at 25C was added diethylaluminum chloride ~10 mmol). The mixture was stirred for 5 minutes, then cooled to 0C in an ice-methanol bath. A solution of the compound of Example 1 (c) ( 10 mmol) and me~hyl dl-2-10 brs>mopropionate (10 mmol) in tetrahydrofuran (10 ml~ was added dropwise to the co1d suspension. The resulting mixture was stirred for 3 hours at 25C. The reaction mixture was filaered and the filtrate was washed with wa~er, dried over magnesium sulfate, and evaporated to give the product.
Methvl 3-(2-dQdeçylphenv!~-3-l~1-carbethQxymethy~-S-tetr~zQlvl~hiolpropano~te To a solution of trifluoroacetic acid ( 15 ml) and 5-mercapto- 1-carbethoxymethyltetrazole (2.4 ml) at 0C is added the compound of Example 1 1 (a). The reaction mixture stirred for 3 hows and 2 0 evaporated. The resulting residue is flash chromatographed on silica, and ~luted with 20% ethyl acetate in hexane, to give the product.

t~r~hi~Q~noic açid To a solution of 10% sodium hydroxide (50 ml)9 methanol (12 2 5 ml~ and ethylene glycol dimethyl ether is added the compound of Example 12(b) (93.9 mmol~. The mixture is stirred for 24 hours at 25C. The reaction mixture is then cooled in an ice-methanol bath to 0C and is acidified with hydrochloric acid to pH 3.5, is extracted with diethyl ether, is dlied over magnesium sulfa~e, filtered and 3 0 evaporated. The resulting mixture of isomers is flash chromatographed on silica, and eluted with 30% ethyl acetate in hexane, ~o give the product.

Preparation of 2-L2-dodecYlphen~!~ [~Ltetrazol-S-vl)-3 5 ethvlthiol~cetic ~çid (a~ ~-t2-MeTca~Qe~hvUt~t~zQle To 100 ml of te~rahydrofuran at 0 was added aluminum - chl~ide ~6.65 g, 0.05 mol) and sodium azide (9.75 g, 0.15 mol). This mixture was allowed to come to 22, and s~irred 30 minutes.

2~ PCI/lJS91/0392 ~-Mercaptopropionitrile (4.35 g, 0.05 mol) was added, and the mixture heated to reflux for 24 hr. The mixture was cooled, acidified carefully with excess 15% aqueous hydrochloric acid, and the solvents removed under reduced pressure. The residue was 5 extracted with ethyl acetate, the extracts washed with water, dried, and the solvent evaporated. The residue was recrystallized from 1,2-dichloroethane9 and gave 3.7 g (57%) of 5-(2-mercaptoethyl)tetrazole.
fb! Methvl 2-(2-dQdeçylphenYl)-2-l2-LtetrazQI-~-vl)-ethylthiQlaçetat3~
A mixture of methyl 2-chloro-2-(2 doclecylphenyl)ace~ate (0.615 g, 1.75 mmol), 5-(2-mercaptoethyl)tetrazole (0.227 g, 1.75 mmol), and triethylamine (0.48 ml, 3.5 mmol) in 20 ml of methylene chloride was stirred 18 hours at 22 and the solvent was evaporated.
The residue taken up in diethyl ether, washed with lN hydrochloric 15 acid, dried and the solvent removed. The residue was chromatographed over a silica gel column. Impurities were eluted with ethyl acetate/hexane (1:3), and the product was eluted with methanol/ethyl acetate (1:19). Evaporation of the solvents from these fractions gave the desired product, 0.570 g (73%).
2 0 (ç) 2-(2-Dodeçvlphçnyll-2-~2-(tetrazol-~-vl)-ethylthiol;~tiç açid A stirred suspension of methyl 2-~2-dodecylphenyl)-2-[2-(tetrazol 5-yl)ethylthio]acetate (0.57 g, 1.28 mmol) in 10 ml of methanol and 8 ml of water at 70 was trea~ed with 3 ml of 2.5N
sodium hydroxide. After 30 minutes at 70, the mixtul e was cooled, 2 5 diluted with 10 ml of water and filtered. The filtrate was acidified, extracted wi~h ethyl acetate, and the extracts were dried and the solvent ev~porated. The residue was recrystallized from methanol and gave the desired product 0.415 g (74%). NMR (CDCl3/Me2CO):
10.23-11.70 (broad,2H), 7.52-7.62 (m,lH), 7.27 (s,3H), 5.04 (s,lH), 3 0 3~0~3r44 (m~4H)~ 2.S3-2.92 (t,2H), 1.12-1.84 (m,2ûH), 0.62-1.02 (t,3H)-~ !
35 __~
2-(8-phenyloctyl)benzaldehyde was prepared as described in Example S(a) or (b).

WOgl/18889 -31- 2383~dUS9~'392' (b~ M~thyl ~12~ Phenyl.~ç¢~lQhenvll-2~-~Q~
The compound of Example 7(a) (2.94 g, 10 mmol) was dissolved in diethyl ether (25 ml) and the solution was stirred under argon at 0C. Methyl chloroacetate (1.32 ml, 15 mmol) was added, followed by the addition of sodium methoxide (810 mg, 15 mmol). The mixture was stirred for 2.5 hours at ice bath temperature. A small quantity of water was added, the ether phase separated, dried over anhydrous sodium sulfate, fil~ered and evaporated. The residue was flash chromatographed on 80 grams of silica gel elu~ed wi~h ~-30%
ethyl acetate/hexane tO give the product.
~ `~
tetrazo!vllthiol-~-hy~roxy,~rQpion~te The compound of E~xample 13(c) (549 mg, 1.5 mmol) was dissolved in methanol (6 ml) containing 2% ~riethylamine and the solution was stirred under argon at room ~emperature. The 5-mercapto-1-(3-carboxypropyl)tetrazole (282 mg, 1.5 mmol) and trie~hylamine (0.84 ml, 6 mmol) were dissolved in methanol (9 ml) - and added dropwise to the reaction mixture which was then stirred for 5 days at room temperature. The solvent was stripped and the 2 0 residue was flash chromatographed on 50 grams of silica gel eluted with 70:30:1 (hexane:ethyl acetate:formic acid) to give the desired product.

The compound of Example 13(d) (554 mg, 1 mmol) was dissolved in methanol ( 15 ml) and stirred under argon at ice bath temperature. A lN solution of sodium hydroxide (4 ml, 4 mmol) was added dropwise, the ice ba~h removed and the mixture stirred at room temperature overnight. The methanol was stripped and the 3 0 residue acidified with dilute hydrochloric acid. Extraction with ethyl acetate followed by drying over anhydrous sodium su1fate, filtration and evaporation gave the crude product which was flash chromatographed on 25 grams of silica gel eluted with 50:50:1 (ethyl acetate:hexane:formic acid) to give the desired product.

WO 91/lB889 - 3 2 - PCI IUS91/039"
2 ~ EXAMPLE 14 Pr~r~Li-on of ~-L2-(~hen~octyl~phenvll-3-[(1-~arb~xYmeth~l~-tetr_o~h~ 2~h~roxYpr~noic_ acid (a) Methvl ~-e~2-~8-~henvloçtYl~phen~lpropanQate 2-(8-phenyloctyl)benzaldehyde prepared as in Example ~(a) or (b) (15 g) was dissolved in methylene chloride (58 ml) under an argon atmosphere a~ 25C. Methyl chloroacetate (6.2 ml) was added in one portion. The resulting solution was cooled ~o -28C using a dry ice/isopropanol bath. Sodium methoxide ( 13 5 ml) was ~hen added slowly, maintaining temperature between -28C to -15C. The reaction mixture was then stirred for 45 minutes, and permitled to warm slowly to 0C. The reaction was then stirred for 1.5 hours at 0C. The reaction mixture was quenched with a solution (117 ml) of aqueous buffer (pH 7) and 117 ml of hexane. The layers were separated, the aqueous portion was filtered and washed with hexane.
The combined organic extracts were dried over sodium sulfate, filtered and concentrated. The residue was flash chromatographed on silica and eluted with 3 :97 ethyl acetate/hexane to give the desired product.
2 0 (~) ~IÇtk~-h~Q~ ~arbQ~ymethyl-5-tetrazolYl)ehiol-pro~anQa~e The methyl 2,3-epoxy-3-[2-(8-phenyloc~yl)-phenyl~propanoate (0.4g) of Example 14(a) was dissolved in 5 ml of methanol containing 2% triethylamine under an argon atmosphere. The reaction mixture was cooled using an ice/methanol bath. A solution of 0.205 g of 5-mercapto-1-carbethoxymethyltetrazole and 0.61 ml of triethylamine in 5 ml of methanol containing 2% triethylamine was added. The bath was removed and the reaction was permitted to warm to room temperature. The reaction was concentrated and flash 3 0 chromatographed on silica with 20% ethyl acetate/hexane as eluant to give the desired product.
(c) 3-12~ ~n~oct~henyll-3-~(1-carboxvmethYI-S-The methyl 2-hydroxy-3-[(1-carboxymethyl-5-tetrazolyl)thio]-3 5 propanoate (0.45 g) of Example 14~b) was dissolved in methanol (6.6 ml) and cooled with an ice and methanol bath to which was added 3.3 ml of a 4% solution of sodiun hydroxide. The bath was removed and reaction permitted to warm to room temperature overnight. The methanol was then stripped, and the residue was diluted with dilute WO gl/18889 ~ 3 3 ~ PCr/ussl/03~2z 6 ~ ~
hydrochlonc acid. ExtTaction with ethyl aceta~e was followed by drying over magnesium sulfate, filtration, and evaporation. The crude product was flash chromatographed on silica with 30-50%
ethyl aceta~e/hexane. The column was washed with 100~ methanol.
5 40 mg of desired product was recovered.
EXAMPLE 1~.
Prç~tiQn of 2-(2-Dodeçvlphenvl~-2-(5-çar~x~ 4-meIhyl-2-thi~zQlvlthiQ~L~
(aL~ thYI 2-(2-DQdç~ylph-ç~2--(~-çalr~c)me~hoxv-4-methyl-2 10 thia~ Llthio`Lacelate A mixture of 0.53g (0.0015 mol) of the compourld of.Example 1 (e), 0.39 g (0.00195 mol) of methyl 4-methyl-2-mercaptothiazole-5-carboxylate, 0.23 g (0.0023 moles) of triethylamine and 10 ml of methylene chloride were stirred undeT argon a~ room temperature 15 for 48 hours. The reaction mixture was washed 3 times with 5 ml of 5% sodium carbonate solution followed by water, dried over magnesium sulfate, filtered and concentration under vacuum. The oily residue was flash chromatographed using hexane - 5% ethyl acetate yielding 0.58 g of oil.
20 (b~ 2-l2-Dodecvll~henyl~-2~ oxy-4-methvl-2-thiazolYlthio)-~etic acid A mixture of 0.24 g (0.0046 mol) of ~he compound of Example 1 5(a), 0.11 g (0.0028 mol) of sodium hydroxide, 10 ml of methanol and 1 ml of water were stirred at room temperature and under argon 2 5 for 2 days. The reaction mixture was concentrated under vacuum and the residual oil was redissolved in 5 ml of water. The aqueous solueion was adjusted ~o pH 3.83 with dilute phosphoric acid or until a cloudy solution was obtained; and then was extracted 3 times with 10 ml of ethy1 acetate. The extracts were washed with S nnl of water, 3 0 dried over magnesium sulfate, filtered and concentrated. The oily residue obtained was triturated with petroleum ether. The desired product ~0.054 g) was ob~ained as a powder haYing a m.p. of 88-91C.
Analysis for C2sH3~NO4S~,:
Calculated: C:62.86; H:7.38; N:2.93;
Found: C:62.~; H:7.47; N:2.88.

WO 91/18889 ~ 3 4 ~ pcr/u59l/û3922 2 ~ EXAMPI~E ~
PreF~ratiQn of 2-l2-(8-phenYlQct~l~hçn~ -2-l ~2-carboxv-~-(l~3~4-thiadiazQ~ thiQl~cetic acid (a~ Mçthvl 2 L2~(8-phen~lo~ ~2-LL2-~arbe-thoxv-s-(l~3~4 thi~dl~zQlvl)lthiolacetate The compound of Example 5(d) (744 mg, 2 mmol) is dissolved in 25 ml of methylene chloride and stirred under argon at room temperature. Ethyl S-mercap~o-1,3,4-thiadia;zole-2-carboxylate (380 mg. 2 mmol) and triethylamine (0.84 ml, 6 mnnol) is dissolved in 25 ml of methylene chloride and added to the solution of the compound of Example S(d~. The mixture is stirred under argon for 48 hours.
The solvent is evaporated, and the residue is llash chromatographed on silica gel eluted with ethyl acetate/hexane to give the product.

thi~diazQl-~l~hiQl~çetic ~cid The compound of Example 1 6(a) (526 mg, 1 mmol) is dissolved in 10 ml of methanol and stirred under argon in an ice bath. A lN
solution of sodiurn hydroxide ~4 ml, 4 mmol) is added. The ice bath is removed and the mixture is stirred for 24 hours at room 2 0 temperature. The solvent is evaporated, the residue cooled in an ice ba~h, acidified with dilute hydrochloric acid, extracted with ethyl acetate, dried over anhydrous sodium sulfate, filtered and evaporated to give the product.

2 5 Preparation of 2-[2-(8-phenyloctyl)phenyl]-2-[(2-carboxy-~-f~ryl~hiQlacçtiç ~cid (~ Methvl 2-~2-L~ henyloçtYl)phenyll-2-l(2-çarb furf~ryl)th~Q~te The compourld of Example 5(d) ~744 mg, 2 mmol~ is dissolved 3 0 in 25 ml of methylene chloride and stirred under argon at room temperature. 5-mercaptomethylfuroic acid (316 mg, 2 mmol) and triethylamine (0.84 ml, 6 mmol) is dissolved in 25 ml of methylene chloride and added to the solution of the compound of Example 5(d).
The mixture is stirred under argon for 4B hours. The solvent is 3 5 evaporated, and the residue is flash chromatographed on silica gei elu~ed with ethyl ace~ateihexane/formic acid IO give the product.

W(~ 91 /1 ~889 - ~ 5 - PCl /lJS9~ tO3922 2~8~
(b) 2-l2-(~-phenyloc~phenv]l~ (2-carboxY-S-furfurYl)thiol-acetic ~çid The compound of Example 1 7(a) (494 rrlg, 1 mmol) is dissolved in 10 ml of me~hanol and stirred under argon in an ice bath. A lN
S solution of sodium hydroxide t4 ml, 4 mmol) is added. The ice bath is removed and ~he mixture is s;tirred for 24 hours at room temperature. The solvent is evaporated, the residue cooled in an ice bath, acidified with dilute hydrochloric acid, ex~racted with ethyl acetate, dried over anhydrous sodium sulfate, filtered and 10 evaporated to give the product.
~,~I,E 1B
Preparation Qf 3-~2-(6-thiophenoxyhexvlthio)Dhenvll-3-~1-(3-carboxyprop~ ~olyl~thiol-2-hvdroxypr~panoic açid 15 La~ Preparation of 2-(6-thiophQnoxYhexYlthio~ben-zoic acid Thiosalicylic acid (1.2 g, 0.008 mole) and 6-thio-phenoxyhexylbromide (2.5 g, 0.009 mole) are dissolved in dimethylformamide (50 ml) and the solution is stirred under argon.
Potassium carbonate (1.5 g, 0.011 mole) is added carefully to the 20 reaction. After the addition is complete the mixture is slowly warmed to 100C. The solvents are evaporated, and the residue is dissolved in water, aci~ified with dilute hydrochloric acid, extracted with e~hyl acetate, dried s)ver anhydrous sodium sulfate, filtered, and evaporated. The residue is flash chromatographed on silica gel to 2 5 give the desired product.

(b) Preparation Qf 2-(6-thiophenoxyh~xvlt~hio~benzyl_ alcobol.
To a suspensionof lithium aluminum hydride (.292 g, 0.007 mole) in tetrahydrofuran (30 ml) is added a solution of 2-(6-3 0 thiophenoxyhexylthio)benzoic acid ~2.42 g, û.007 mole) intetrahydrofuran ~30 ml). The reaction is conducted as in Example lb).
(~ PreparatiQn 2-(6-lhio~henoxy~b~b~l9~
To a suspension of manganese dioxide (11.78 g, û.135 mole~ in 3 5 ethyl aceta~e (30 ml) is added a solution of 2-(6-thiophenoxyhexylthio)benzyl alcohol (1.23 g; 0.0037 mole) in ethyl acetate (20 ml). The reac~ion is condueted as in Example lc).

wo sl/l88Bs 2 ~ 3 6 - Pcr/us9l/0392 çpoxv~rQpiQnate The compound of Example 18(c) (10 mmol) is dissolved in diethyl ether (25 ml) and the solution is stirred under argon at 0C.
Methyl chloroacetate (15 mmol) is added followed by the addition of sodium methoxide ( 15 mmol). The mixture is stirred for 2.5 hours at ice bath ~emperature. A small quantity of w,ater is added, the ether phase is separated, dried over anhydrous sodium sulfate, filtered, and evaporated. The residue is flash chromatographed on 80 grams 1 O of silica gel eluted with 5-30~b ethylacetate/hexane to give the produce.

çarbQxvprQpvl - ~-tetra~zQlylLthi ol - 2-h ydrQx YI?rop_on ate The compound of Example 18(d) ~1.5 mmol) is dissolved in 15 methanol (6 ml) containing 2% trie~hylamine and the solution is stirred under argoll at room temperature. 5-Mercapto 1-(3-carboxypropyl)tetra~ole (1.5 mmol) and triethylamine (6 mmol) are dissolved in methanol (9 ml) and are added dropwise to the reaction mixture which is then stirred for S days at room temperature. The 2 0 solvent is stripped and the residue is flash chromatographed on 50 grams of silica gel eluted with 70:30:1 (hexane: ethylacetate: fo:rmic acid) to give the desired product.

The compound of Example lB(e3 (lmmol) is dissolved in methanol (15 ml) and is stirred under argon a~ ice bath temperature.
A lN solution of sodium hydroxide 94 mmol) is added dropwise, the ice bath is removed and the mixture is stirred at room temperature overnight. The methanol is stripped and the residue is acidified with 3 0 dilute hydrochloric acid. Extraction with ethyl acetate followed by drying over anyhydrous sodium sulfate, filtration, and evaporation gives the crude product which is flash chromatographed on 25 grams of silica gel eluted with 50:50:1 (ethyl acetate: hexane: formic acid) to give the desired product.

Wo 91/18889- 3 7 ~ 2 a 8 3 ~ TtUS91/0392 _r~ion of Mç~hy! 2-h~droxy-3-mercapto-3-J2-L8-~henYloctvl~phenvllpropionate (a~ Methvl 2-hv~r~xv-3-(4-mçthQ~henylmethyl~hio~-3-~2-(8-5 ~h~nylo~tyl~*henyll~ro~ionate A solution OI 4.~4 gm (12.4 mmol) of the epoxyester of Example 13(b) and 1.91 gm (12.4 mmol) of p-methoxybenzyl mercapton in a mixture of 1.7 ml of triethylamine and 20 ml of methanol were stirred for 42 hours, and the solvents evaporated. The residue was 10 dissolved in diethyl ether and washed with O.lN HCl. Thç organic layer was dried, and the solvent evaporated. The residue was chromalographed over an alumina column. Impurities and the undesired regioisomer were eluted with hexane/ethyl acetate/methanol (60:40:1). The desired product was eluted with 1~ hexane/ethyl acetate/methanol (40:60:2), 2.6 gm (40%). nmr (CDC13 ) :
7.68(m, lH), 7.20(m, lOH), 6.82(d, 2H), 4.58(t,1H), 4.45 (d~ lH), 3.82 (s, 3H), 3.72 (s, 2H), 2.66(s, 3H), 3.14(d, lH), 2.24-2.72 (m, 4H), 1.10-1.72 (m, 1 2,H).
~'~) Methyl 2-hy~roxy~-merça~2to-3-l2-~8-phenYloctvl)phenvll-2 0 propiQnatç
A solution of 1.1 gm (2.12 nlmol) of the compound in Example19(a) in 25 ml of methanol was ~ea~ed with a solution of 2.02 gm (6.35 mmol) of mercuric acetate in 100 ml of methanol. After stirring 16 hours, the white precipitate was filtered and washed with 25 diethyl ether. This mercuric salt was dissolved in 2~ ml of hot dimethylformamide, 50 ml of me~hanol was added, and H2S was bubbled into the solution for 30 minutes. The black precipitate was filtered, the filtrate was concentrated, diluted with water, and extracted with diethyl ether. The diethyl ether layer was washed 30 well with water, dried, and the solvent evaporated. The residue was chromatographed over a silica gel column, and the product was eluted with a mixture of ethyl acetate /hexane (40:60), 370 mg (44%). nmr (CDC13/D2O): 7.02-7.78 (m, 9H), 4.62 (s, 2H), 3.70 ~s, 3H), 2.50-2.88 (m, 4H), 1.20~ 2 (m, 12H).

wo9l~l8889 2~ 38- Pcr/ussl/o3922 I~IE~

ar~Qx~hien-5-vl~ hvl)sulfonYl~prQ~Qiç ~çid 2Q(al ~thyl 2L~-~nçthQ~y-~R~-14 S methQxvphenyl~ethvllthiG-~ -(8-~henylQç~l~
A cold (0) solution of 5.58gm (O.Ollmole) of methyl 2(S)-hydroxy-3(R)-(4-methoxyphenylmethyl)thio-3-[2-(8-phenyloctyl)phenyl]propionate (prepared as described in EPO
published application 88305188.0) and 0.67 ml of iodomethane in 10 1 50 ml of tetrahydrofuran and ~0 ml of dimethylformamide was treated with 430 mg of a SO% NaH dispersion. The ice bath was removed, the reaction stirred at 23 for 2 hours, poured into cold aqueous 05N HCI, and extracted with diethyl ether. The extracts were washed with wa~er, dried and evaporated. The Tesidue was 15 chromatographed over a silica gel comurnn, and the pToduct eluted with 20% ethyl acetate in hexane. NMR (CDC13): ~ 7.09-7.54(m, llH), 6.80(d, 2H), 4.37(d, lH), 4.11(d, lH), 3.75(s, 3H), 3.73(s, 3H), 3.63(d, lH), 3.52(d, lH), 3.28(s, 3H), 2.60(t, 2H), 2.42(t, 2H), 1.23-1.68(m, 1 2H), 2 0 20(b~ _ Meth~l LR-(R*,~:mçrç~ o-2-methQx~-2-(3-Rhenyloctyl~ohçn~l ~ropanoat~
A solution of 3.42 of methyl 2(S)-methoxy-3(R)-(4-methoxy phenylmethyl)thio-3-[2-(8-phenyloctyl~phenyl] propionate in 1 OOml of methanol was treated with 5.3gm of Elg(VAc)2 and stirred 2 5 overnight. The solveslts were removed and the residue stirred with a mixture of 25% diethyl ether in hexane. The solid remaining after titura~ion was dissolved in lOOml of rnethanol, and H2S was bubbled through for 1 hour. HgS was filtered out and the filtrate was evaporated. The residue was chromatographed over silica gel, and 30 the product eluted with a mixture of 30% ethyl acetate in hexane.
NMR (CDC13): ~ 7.12-7.46(m, 9H), 4.52(t, lH), 4.16(d, lH), 3.74(s, 3H), 3.31 (s, 3H), 2.55-2.78(m, 4H), 2.10 (d, lH), 1.28-1.69(m, 12H).
20~ç) h~hYL~*~ -2-methoxv-3-L2-L~-phen~lo~yl) 3 5 A solution of 1.42gm (3.43 mmole) methyl [R-(R*,S~)]-3-mercapto-2-methoxy-2-(8-phenyloctyl)phenyl propanoate in a mixture of 60ml tetrahydrofuran and 30ml dimethylformamide was treated with 137mg of a 60% dispersion of NaH in mineral oil at 0.
When salt formation was complete, 806mg of methyl wo gl/18889 ~ 3 9 ~ pcr/us9lto392~
2~33~
5-bromomethylthienyl-2-carboxylate was added. S~irring was continued for 2 hours at û and then 1 hour at 23. The reaction was poured into H20 and extracted with diethyl ether. The extracts were washed with H207 dried and the solvent removed. Partial purification of the intermediate product was achieved by chromatography over a silica gel column and eluting with 20% ethyl acetate in hexane. The crude methyl [R-(lR*,S*)]-~-[(5-carboxymethyl-2-thienylmethyl)thio]--methoxy-2-(8-phenyloctyl)benzene propanoate, 1.1 gm, was dissolved in lOOml of CH2C12 and ~eated at 23 with .833gm 85%
1 0 m-chloroperbenzoic acid. Af~er 11/2 hours, the reaction was washed repeatedly with aqueous Na2C03. The organic layer was dned and the solvent evaporated. The residue was chromatographed over a silica gel column and the product was eluted 30% ethyl acetate in hexane and recovered from the eluant.
NMR (CDC13): ~ 7.98(m,1H), 7.62(d, lH), 7.40-7.06(m, 8H), 6.90(d, lH), .lO(d, lH), 4.82(d, lH), 4.62(d, lH), 4.27(d, lH), 3.86(s, 3H), 3.64(s, 3H), 3.57(s, 3H), 2.92-2.40(m, 4H), 1.82-1.02(m, 12H).
20(d~ [R-(R*~S*)l-2-methoxv-3-l2-(8-phenyloctvl~phenyll-3-(2 c~xvthiçr~ ylmethY~sulfonyl)pro~n~ic acid 2 0 A solution of the dimethylester from 1 9(a), 0.98gm, in 20ml of glacial acetic acid and 8ml of concentrated HCI was refluxed for 6 hours. The reaction was cooled, poured into water, and extracted with CHC13. l`he CHC13 was washed repeatedly with H20, dried, and the solvent removed, giving 0.82gm of the product. NMR (CnC13-Me2CO-D6) 8 8.03(m, lH)> 7.63(d, lH), 7.40-7.10(m, 8H), 7.00~d, lH), 6.63(broad s, 2H), 5.20(d, lH), 4.82td, lH), 4.72~d, lH), 4.33(d, lH), 3.67(s, 3H), 2.46-2.96(m, 4H), 1.10-1.76(m, 12H).

LR-(R*~ -2-h~lL xv-~:L2-(~-nh~nvlQ~yl~he,nvll-3-(2-3 0 car~ox~hien~-vlrD~hyl~thio~ropanoi a.cid 21 La) Me~hYl ~-(~ ~-LdrQxy-.~ -phenvl~ct~)-ph_nvll-~-(2-çarbQ~hQxvthien-S-ylmelh~)-thiQ)3~rQ~pionate .A solution of 1.26gm(3.17mmole) me~hyl 2(S)-hydroxy-3~R)-mercapto-3-~2-(8-phenyloctyl)phenyl]propionate mercapto in a 3 5 mixture of 30ml tetrahydrofuran and 20ml dimethylformamide was treated with 3.17mmole of NaH. When the anion had completely formed, 3.52mmole of methyl 5-bromomethylthienyl-2-carboxyla~e was added, the ice bath removed, and ~he reaction stirred for 2 hours. The mix~ure was poured into H20, extracted with diethyl WO 91 /~X889 - 4 - PCl /US~I /03922 e~her, the extracts washed with H2O, dried and the solvent removeu.
The residue was chromatographed over a silica gel column, and the product eluted with a mixture of 30% ethyl acetate in hexane to give, after removing the solvent, ~he title compound.
NMR (CDC13): ~ 7.65(d, lH, 7.32-7.10(m, 9H), 6.91(d, lH), 4.6-4.55(m,2H), 3.97(d, lH), 3.88(s, 3H), 3.81(d, IH), 3.66(s, 3H), 3.00(d, lH), 2.62(t, 2H), 2.48(m, 2H), 1.62(m, 2H), l.S0-1.26(m, 10H).
21(b) IR-fR*~S*~l-2-hY~rQxv-~-~2-~-phenylQctvl~henvll-3-(~-A solution of 145mg of the preceding diester in a mixture of 3ml of tetrahydrofuran and 2ml of methanol was treated with excess LiOH in H2O. After 3 hours. at 23 the solution was diluted with water and filtered. The filtrate was acidified and extracted with ethyl acetate. The extlacts were washed with H20, dried and the solvent evaporated to give the captioned compound. NMR ~CDCI3) 7.58(d, lH), 6.88(d, lH), 4.57-4.42(m, 2H), 3.94(d, lH), 3.78(d, lH).
EX~MPLE 22 As a specific embodiment of a composition of this invention, an active ingredient, such as the compound of Example 4, 13, or 14 is dissolved in isotonic saline a~ a concentration of 1 to 10 mgJml and aerosolized from a nebulizer operating at an air flow adjusted ~o deliver the desired aerosolized weight of drug.
I~XAMPLE ~
As an additional embodiment of a composition of this inven$ion 100 to 1000 mg of an active-ingredient, such as the compound of Example 4, 13, or 14 is combined with 4 mg of chlorpheniramine maleate with a suitable carrier or excipient.

Claims (11)

What is claimed is:

1. A compound of formula I

I
wherein q is 0, 1, or 2 R1 is (L)a-CH2)b-(T)c-B wherein a is 0 or 1, b is 3 to 14, c is 0 or 1;
L and T are independently oxygen, sulfur or CH2 with the proviso that L and T are not sulfur when q is 1 or 2; and B is C1-4 alkyl, ethynyl, trifluoromethyl, isopropenyl, furanyl, thienyl, cyclohexyl, or phenyl optionally monosubstituted with Br, Cl, CF3, C1-4 alkyl, C1-4 alkoxy, methylthio, or thrifluoromethylthio;
R2 and A are independently selected from H, CF3, C1-4 alkyl, C1-4 alkoxy, F, Cl, Br, I, OH, NO2 or NH2; or R1 and A are H and R2 is (L)a-CH2)b-(T)c-B wherein a, b, c, L, T, and B are defined above;
Y is COR3 or (CHX)n(CH2)p-Z wherein R3 is OH, NH2, aryloxy or C-6 alkoxy;
n is 0 or 1;
p is 0, 1;
X is H, OH, C1-4 alkyl, C1-4 alkoxy, or F; and Z is COR3 or tetrazolyl;
R is m is 0 to 6;
R4 and R5 are independently hydrogen or C1-4 alkyl at any position when m is not 0;
W is thienyl substituted with one to three of the group ;

j is 0 to 6; and V is hydrogen, C14 alkyl, COR3, SO3H, SO2H, SO2NH2, COCH2OH, CHOHCH2OH, or tetrazolyl with R3 as defined above;
a pharmaceutically acceptable salt thereof.

2. A compound of claim 1 where q is 0, j is 0 or 1, m is 1; V
is COR3 and Y is COR3.

3. A compound of claim 2 where R2 is hydrogen, A is hydrogen and R1 is docecyl or 8-phenyloctyl .

4. A compound of claim I where q is 0, j is 0 or 1, V is COR3 and Y is -CH2COR3.

5. A compound of claim 4 where R2 is hydrogen, A is hydrogen and R1 is docecyl or 8-phenyloctyl.

6. A compound of claim 1 where q is 0, j is 0 or 1, V is COR3 and Y is -CH(OH)COR3.

7. A compound of claim 6 where R2 is hydrogen, A is hydrogen and R1 is docecyl or 8-phenyloctyl.

8. A compound of claim 7 which is 2-hydroxy-3-[2-(8-phenyloctyl)phenyl]-3-[(2-carboxythien-5-ylmethyl)thio]propionic acid, or a pharmaceutically acceptable salt thereof.

9. A compound of claim 1 where q is 0 or 2, j is 0 or 1, V is COR3 and Y is -CH(OCH3)COR3.

10. A compound of claim 9 where R2 is hydrogen, A is hydrogen and R1 is docecyl or 8-phenyloctyl.

11. A compound of claim 10 where q is 2, the compound 2-methoxy-3-[2-(8-phenyloctyl)phenyl]-3-[(2-carboxythien-5-ylmethyl)sulfonyl]propionic acid, or a pharmaceutically acceptable salt thereof.