CA1232917A - Leukotriene antagonists - Google Patents

Abstract

LEUKOTRIENE ANTAGONISTS

ABSTRACT OF THE INVENTION

The compounds represented by the formula (I) (I) wherein n is 1 or 2; m is 0, 1 or 2; p is 9, 10, 11, 12 or 13; X is hydrogen or hydroxyl; R is hydroxyl or amino;

R1 is hydrogen, amino or ; and R2 is hydroxyl, amino, -NHCH2CO2H, ,

Description

3291~

-- 1 ~

LEUKOTRIENE ANTAGONISTS

BACKGROUND OF THE INVENTION
"Slow Reacting Substance of Anaphylàxis" (SRS-A) has been shown to be a highly potent broncho-constricting subs~ance which is released primarily from mast cells and basophils on antigenic 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 inYolved in acute cutaneous allergic reactions. Further, SRS-A has been shown to efect depression of ventricular contraction and ; potentiation o~ cardiovascular effects of histamine.
; ~ The discovery of the naturally occurring leukotrienes and ~heir relationship to SRS-A has rein~orced interest in SRS-A and other arachidonate metabolites. SRS-A derived from mouse, rat, guinea pig and man have all been characterized as mixtures of leukotriene-C4 (LTC4), leukotriene-D4, (LTD~) and leukotriene-E4, ~LTE4); the structural ~ormulae of which are represented below.

OH ~-Glu CO2H LTC4 R'7 = Cys-Gly 35 ~ ~ C H ¦ ~LTD4 R" = GYs-GlY
5 11 ~ SR" LTE4 R~ = Cy~s ,,, : : :

~ ~ ' ' ~ '. ''' ~ ~Z.~Z9~7 By antagonizing the effects of LTC4, LTD4 and LTE4 or other pharmacologically active mediators at the end organ, airway smooth muscle, the compounds and pharmaceutical compositions of the instant invention are valuable in the treatment of diseases in which leukotrienes are a factor, such as asthma.
: SUMMARY OF THE rNVENTION
The invention provides compounds of formula (I) Il 1( 2)m C 2 CH3(CH2)p /CHCIH(CH2)nC (I) /C=C\ X
H H

wherein n is 1 or 2; m is 0, 1 or 2; p is 9, 10, 11, 12 or 13;
: X is hydrogen or hydroxyl; R is hydroxyl or amino;
O
R1 is hydrogen, amino or -NHCCH3; and R2 is hydroxyl, amino, -NHCH2CO2H, -NCIH2CO2H, -NHCHCO2H, -NHICHCO2H or -NHCH2CONH2 CH3 CH3 CH(CH3)2 ~ with the proviso that when m is 0, R1 is hydrogen or a :, pharmacQutically acceptable salt thereof and a method of ~, making same. The method comprises:
(A) when X is hydroxyl and n is 2, reacting : ~O\
CH3~CH2)p / CH-CHCH2CH2COR3, wherein COR3 is COR or ~ C C ~
H H
: a radical such as CO2-alkyl easily convertible into an acid 30~ Rl' ~:~":: ~ moiety with HS(CH2)nCHCOR2', wherein Rl' and R2l are respectively Rl and R2 or a radical such as an alkyl ester of trifluoromethylacetamide easily convertible into R
or R2;
(B) when X is hydroxyl and n is 1, reacting :
~, I

~32g~7 - 2a -S ~CH2 ) mCHCR2 ' CH3(CH2)p /CHCHO with C~3COR3; or ~C=C\
H H
(C) when X is hydrogen, reacting R~ l ' 7 (CH2, mCHCOR2 HCCHCH2(CH2)nCOR3 with CH3(CH2)pCH P03;

and then optionally converting Rl', R2' and R3 into Rl, R2 and R and optionally forming the pharmaceutically acceptable salt of the compounds.
DETAILED DESCRIPTION OF THE INVENTION
The compounds of this invention are represented by the following general structural formula (I) Rl (CH2)mCHCR2 (CH2) ~CHCH(CH2)nCOR (I) l ~ 20 / C~C \ X
:~ H H
I wherein n is 1 or 2; m is 0~ 1 or 2; p is 9, 10, 11, 12 or 13; X is hydrogen or hydroxyl; R is hydroxyl or amino;
,1 : O
Rl is hydrogen/ amino or -NHCCH3; and R2 is hydroxyI, amino, NHCH2CO2H, -NfH2CO2H; -NHCHCO2H, -NHCHCO2H, or -NHCH2CONH2 CH3 CH3 CH(CH3)2 : with the proviso that when m is 0, Rl is hydrogen or a pharmaceutically acceptable salt thereof.
30 ~ The stereochemistry of the compounds of:formula~
: is such that the double bond adjacent to the alkyl moiety is in the cis:configuration. The compounds~of formula (I) are the result of a Wittig reaction between appropriate alkyltriphenyl-phosphonium ylid with an appropria~e inter-3~5 ~;mediate compound~. The compounds of formula (~) wherein n is 2and p is:~ have~a Clg carbon skeleton and are designated ~L23;~17 - 2b -6(Z)-nonadecenoic acid derivatives. Similarly, when n is 1 and p is 11, the compounds of formula (I) have a C18 carbon skeleton and : 15 :
~, ' " .

: ~: :.: : : ~: :
~ 35 ~ :

` ~2329~7 1 are designated 5(Z)~octadecenoic acid derivatives. When n is 2 and p is 9 the compounds of formula (I) have a C17 carbon skeleton and are designated 6(Z)-heptadecenoic acid derivatives. Also, when n is 2 and p is 13 the compounds of formula (I) have a C21 carbon skeleton and are designated 6(Z)-heneicosenoic acid derivatives.
The 6(Z)-nonadecenoic acid derivatives of the compounds of formula (I) wherein R is hydroxyl and X is hydroxyl are represented by the following general structural formula (II) ~ 1 7 'CH2 ) mCHCOR2 C12H25~ /CHlHcH2cH2cO2H
~C=C\ OH (II) H H

wherein m, Rl and R2 are described above. The compounds of the formula (II) contain two asymmetric centers, one of which is at carbon atom 4 (i.e. the hydroxyl substituted carbon atom "C4") and one of which is at carbon atom 5 (i.e. the thiol substituted carbon atom-"C5"). This leads to the possibility of four stereoisomers for each compound. In practice, the compounds of this invention of formula ~II) have been , prepared in a mixture of two stereoisomers, that is tha 4R, SS isomer and the 4S, SR isomer. The individual pure stereoisomers are obtainable by preparative high pressure liquid chromatography (~PLC) separation of the appropriate intermediate compounds if those compounds possess a third asymmetric cente~r. In the cases where a third asymmetric center is~not available in the synthetic pathway of the desired compounds, one may be introduced by employing an 35~ asymmetric protecting group, such as an N-trifluoroethoxy-arbonyl-i-prolyl ester. After separation of the : ~ .
:~:: :

1232~7 1 individual stereoisomer, the protecting group is removed by standard procedures.
The 6(Z)-nonadecenoic acid derivatives of formula (II) are exemplified by the following compounds as the 4R, SS isomer, the 4S, 5R isomer or mixture of the two isomers:
4-hydroxy-5-[(2-carboxyethyl)thio]-6(Z)-nonadecenoic acid, wherein m is l, Rl is hydro~en and R2 is hydroxyl;
5-l(3-carboxymethylamino-3-oxopropyl)-thio]-4-hydroxy-6(Z)-nonadecenoic acid, wherein m is l, Rl is hydrogen and R2 is -NHC~2CO2H;
5-[(2-amino-3-carboxymethylamino-3-oxopropyl)thio]-4-hydroxy-6(Z)-nonadecenoic acid, wherein m is l, Rl is amino and R2 is -N~CH2CO2H;
5-[(3-car~oxymethyl-N-methylamino-3-oxopropyl thio]-4-hydroxy-6(Z)-nonadecenoic acid, wherein m is l, Rl is hydrogen and R2 is -ICH2CO2H;

5-~(2-amino-3-carboxamidomethylamino-3-oxo-20 propyl)thio]-4-hydroxy-6(Z)-nonadecenoic acid, wherein m is 1, Rl~is amino and R2 is -NHC~2CONH2;
4-hydroxy-5-[(carboxymethyl)thio]-6(Z~- :
: ~ nonadecenoic acid, wherein m is 0, Rl is hydrogen and R2 is hydroxyl;
5-[[2-(aminocarbonyl)ethyl]thio]-4-hydroxy-6(Z)-nonadecenoic~acid, wherein m is 1, Rl is hydrogen and ~: R is amino;
~ 2 :~ ~ 4-hydroxy-5-[(2-amino-2-carboxyethy~)thi~]-6(Z)-nonadecenoic acid, wherein m is 1, Rl is amino and R2 : 30 is hydroxyl; and 4-hydroxy-5-[(3-carboxypropyl)thio]-6(~)-nonadecenoic acid, wherein m is 2, Rl is hydrogen and R2 is hydroxyl.
The 5(Z)-octadecenoic acid derivatives of the 35:compounds of formula (I) wherein R is hydroxyl are represented by the following general structural formula .; ~
: ~ :
. .

1~23~9~7 ~1 t (CH2)mCHCR2 C12H25~ H7HCH2C02H
C=C X (III) H \H

wherein m, X, Rl and R2 are described above. Like the 6(Z)-nonadecenoic acid derivatives of formula (II), the 5(Z)-octadecenoic acid derivatives contain two asymmetric centers at carbon atoms C3 and C4 and thus the possibility of four stereoisomers for each compound exists.
Specific compounds of the formula (III) are those 5(Z)-octadecenoic acid derivatives exemplified by the following compounds as a mixture of the four isomers:
3-hydroxy-4-[(2-carboxyethyl)thio]-5(Z)-octadecenoic acid, wherein m is 1, X is hydroxyl, ~1 is hydrogen and R2 is hydroxyl.
~ lso representative of the compounds of formula

2~ (III) is 4-~(2-carboxyethyl)thio]-5(Z)-octadecenoic acid, wherein m is 1, X is hydrogen, Rl is hydrogen and R~
is hydroxy.
The compounds of the formula (I) wherein X is - hydrogen are represented by the general structural formula , .
(IV):
R
7 (CH2)mCHCR2 (GH2)~ ~CHCH2(CH2)ncoR
C=C (IV) ; H ~H
~ ~ , : : :
wherein n,~ m, p, R, Rl and R2 are described above.~ ~
Speclfic compounds of formula (IV) are those 6(Z)-non-adecenoic acid derivatives wherein n is ~ p is ll~and R is ~hydroxyl which are exemplified by the following compound:
5-~(2-carboxyethyl)thio]-6(Z)-nonadecenoic acid, wherein m~is 1, Rl is hydrogen and R2~is hydroxyl.

. : :

~L~3Z9~7 1 Specific compounds of the formula (I) are those 6(Z)-nonadecenoic acid derivatives wherein R is amino and X is hydroxyl which are represented by the general formula (V) as follows:
~1 1( 2)mCHCOR2 12H25~ ~caC~HCH2CH2CONH2 ~C=C H (V) wherein m, Rl and R2 are described above. The 6(Z)-nonadscenoic acid derivatives o~ ~ormula (V) are exempliied by the following compounds as a mixture of isomers:
5-[~3-carboxymethylamino-3-oxopropyl)thio]-4-hydroxy-6(Z)-nonadecenamide, wherein m is 1, Rl is amino and R2 is -NHCH2CO2H; and 4-hydroxy-5-[(2-carboxyethyl)thio]-6(Z~-non-adecenamide, wherein m is 1; Rl is hydrogen and R2 ishydroxyl.
Additional compounds of the formula (I) are those 6(Z)-heptadecenoic acid derivatives wherein R is hydroxyl -~ and X is hydroxyl which are represented by the general structural formula (VI) as follows:

(CH2)mcHcoR2 CloH21~ ~CH~HcH2cH2cO2H
30~C=C\ OH (VI) H H

wherein m, Rl and R2 are described above. The 61Z)-heptadecenoic acid derivatives of formula (VI) are exemplified as a mixture of isomers:

lZ3~7 1 4-hydroxy-5-[(2-carboxyethyl)thio~-6(Z)-heptadecen-oic acid, wherein m is 1, Ri is hydrogen and R2 is hydroxyl.
Further compounds of the formula (I) are those 6(Z)-heneicosenoic acid derivatives wherein R is hydroxyl and X is hydroxyl which are represented by ~he general structural formula (VII) as follows:

Rl 1 (CH2)mCHCR2 C14~21~ ~CHcHcH2cH2c02H
~C=C 1H (VII) H H

wherein m, Rl and R2 are described above. The 6(Z)-heneicosenoic acid derivatives of the formula (VII) are exemplified as a mixture of isomers:
4-hydroxy-5-[(2-carboxyethyl)thio]-6(Z)-heneicosenoic acid, wherein m is 1, Rl is hydrogen and R2 is hydroxyl.
The compounds of the form~la (II) are readily : prepared by reacting the appropriate thiol containing compound of the formula ~A) 1~ 1 HS(cH2)mcHcoR2l (A~

: wherein m is described above and Rl' and R~' are : respectively Rl and R2 or a pro~ecting radical easily ~: ~ 30 convertible to the desired substituent, such as an alkyl ester or trifluoromethylacetamide, with a 4,5-epoxy-6(Z)-nonadecenoic acid derivative of the formula (B) ~ C12H25~~CH~ CH~H2CH2COR3 (B) : :: : : :

:

:
~ :: :

1~3~917 1 wherein -COR3 is a radical which is easily convertible into the desired acid moieties, such as CO2alk wherein alk is a lower alkyl group. The thiol containing compounds of formula (A) are known or easily prepared from known compound utilizing standard chemical transforma-tions. The 4,5-epoxy-6~Z)-nonadecenoic acid derivatives (B) are readily prepared from monoalkyl succinate (1) via the following synthetic pathway:
lo R
H~CCH2CH2C02alk ~ ClCCH2CH2C2 (1) (2) OHCCH2CH2C02alk >OHCCH=CHCE~2CH2C02alk -15(3) (4) /o\
/ \ C12H25\ ~CH CHCH2CH2CO2alk OHCCH -C~CH2CH2CO2alk ~ ~C=C\
(5) H H (6) Mono-methyl succinate tl), wherein alk is methyl, was chlorinated with oxalyl chloride in dimethyl formamide ~ and methylene chloride to afford 3-carbomethoxypropionyl : 25 chloride (2). Compound (2) was catalytically hydrogenated over palladium-charcoal catalyst in the presence of ~- :2,6-lu~idine to yield 3-carbomethoxypropionaldehyde (3) which was reacted with formylmethylene triphenyl-~ phosphorane under Wittig reaction conditions to obtain :~ 30 S-carbomethoxy-2-pentenal (4). Compound 4 was epoxidized :~ ~ : :with aqueous hydragen peroxide in the presence of lN
~ : sodium bicarbonate to afford methyl-4,5-epoxy-6-oxo-: : : :
hexanoate (S). Compound (5) was reacted with tridecyl-: tripheny}phosphonium ylid to give methyl-4,5-epoxy-6(Z)-35~ nonadecenoate (6).

:~ , ., : : ~

, : ~ :' : , , `:

~3;Z~7 g 1 The 5(Z) octadecenoic acid deriva~ives of the formula tIII) wherein X is hydroxy are prepared via the synthetic pathway starting from 2(E),4(Z)-heptadecadienol (7) as follows:
H\ ~CH2O~

/ =C H . ~ C--C ~CH2~
H H H H
(~) (8) I(CH2)mCHCOR2' l(CH~)mCHCOR2' C12H25~ ~CH~HCH2OH 12 25~
15 H H H \H
(9) (10) ~ ll : ~ 7 ~C~2 ) mCHCOR2 : C12H25~ ~CHcHcH2Co2alk . ~C=C~ 1H ~ (III) (11) Compound (7) was reacted with m-chloro-: perbenzoic acid (MCPBA) to afford 2,3-trans-epoxy-4(2)-: : heptadecenol (8). Compound 8 was reacted with methyl 3-mercap~opropionate (m c 1, Rl' = H and R2' = -OCH3) to yield 3-[(2-carbomethoxyethyl)thio]-1,2-dihydroxy-4(Z)-heptadecene (9). The 1,2-diol moiety of Compound (9) was cleaved by periodate to give 2-[(2-carbomethoxyethyl)thio]-

3(Z)-hexadecenal (10). Compound (10) was reacted with 35 methyl acetate (alk = -OCH3) in the presence of lithium diisopropylamide to give methyl-3-hydroxy~-4-~(2-carbo-. .

- .
: .: .

~ - . .
: -~,~ : . ~ -: -~Z3;~9~L7 1 methoxyethyl)thio]-5(Z)-octadecenoate tll) which was saponified to afford compound of the formula (III) wherein m is 1, Rl is hydroxyl, Rl is hydrogen and R2 is hydroxyl.
The 6(Z)-nonadecenoic acid derivatives of formula (IV) are prepared via the pathway starting from 5-carboxy-alkyl pentanoic acid (12) as follows:
Ir H02CCH2CH2CH2CH2C02alk ~ Ho2ccHcH2cH2cH2co2alk -- >
(12) (13) .
R ' R ' I(CH2)mC CO 2 ( 2)m C R2 lS HO2CCHCH2CH2CH2CO2alk ~HI~HCH2CH2CH2C2alk (14) (1;) ~ 1 l(c~2)mcHcoR2l 12H25~ 5HCH2C}I2C~I2C02alk ~C=C ~ (IV) H ~ .
(16) --5-Carbomethoxypentanoic acid (12), wherein alk is ~: ~ methyl, was reacted with ~hionyl chloride and then N-bromosuccinimide (NBS) followed by base hydrolysis to give 2-bromo-5-carbomethoxypentanoic acid (13). Compound ~: (13) was reacted with methyl-3^mercaptopropionate (m = 1, 3~ Rl' = H and R2' = OCH3) to yield 5-carbomethoxy-2-~[~(2^carbomethoxyethyl)thio]^pentanoic acid (14~. Compound ~ ~ (14) reac:ted first with diborane and then with dimethyl-:; : sulfoxide and trifluoroacetic anhydride to afford 5-carbomethoxy-2-~(2-carbomethoxyethyl)thio]-pentanal : : 35 (15). Compound (15) was reacted under Wittig conditions : with tridecyltriphenylphosphonium ylid to yield methyl-; 5-[(2-carbomethoxyethyl)thio]-6(Z)-nonadecenoate ~16) : whlch was saponified to afford a compound of ~he formula :
:, :, :: - :, -, ,.. ~ - .~

(IV) wherein m is 1, R1 is hydrogen, and R2 is hydroxyl.
The 5(Z)-octadecenoic acid derivatives of the formula (III) wherein X is hydrogen can be prepared via the general synthetic pathway utilized for the preparation of the 6(Z)-nonadecenoic acid derivatives of formula (IV) but starting from 4-carboxyalkyl butanoic acid.
The compounds of the formula (V) can be prepared by initially forming the .gamma.-lactone of the appropriate compound of formula (II) with trifluoroacetic anhydride and then reacting the .gamma.-lactone with ammonia to afford the desired amide.
The compounds of formulae (VI) and (VII) are conveniently prepared employing procedures analogous to those utilized in the preparation of compounds of the formula (II) by substituting the appropriate C11 and C15 alkyl triphenyl phosphonium ylid for the tridecyltriphenyl phosphonium ylid in the reacdtion with the epoxide aldehyde, compound (5) to obtain intermediate compounds of the structural formula (C) as follows:

(C) wherein n and p are descreibed above and R' is R or a protecting radical easily convertible to R such as, -Oalk wherein alk is an alkyl radical containing one to six carbon atoms.
The instant invention also includes the compounds of formula (C) above which possess the double bond in the cis conformation. The structural similarities between the compounds of formula (C) and the compounds of formula (I) contribute the leukotriene antagonist properties of the compounds of formula (I).

~L o 1 The leukotriene antagonist activity of the compounds o this invention is measured by the ability of the compound to inhibit leukotriene induced contraction of guinea pig tracheal ~issues in vitro and to inhibit leukotriene induced bronchoconstriction in guinea pigs in vivo. The following me~hodologies were employed:
In vitro: Guinea pig (adult male albino Hartley strain) tracheal spiral strips of approximate dimensions 2 to 3 mm cross-sectional width and 3.5 cm length were bathed in modified Krebs buffer in jacketed lO ml tissue bath and continously aerated with 95% 2/5~ CO2. The tissues were connected via silk suture to force displacement transducers for recording isometric tension. The tissues were equilibrated for l hr., pretreated for lS minutes with meclofenamic acid (1 ~) to remove intrinsic prostaglandin responses, and then pretreated for an additional 30 minutes with 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 conce~tration of the ~TD4. In order to minimize intertissue variabili~y, the contractions elicited by LTD4 were standardized as a percentage of the maximum response obtained to a reference agonist, carbachol (10 ~M).
Calculations: The averages of the triplicate LTD4 concentration-response curves both in ~he presence and ab~ence of the test compound were plotted on log graph paper. ~he concentration of LTD4 needed to elicit 30% of the contraction elicited by carbachol was measured and de~ined as the EC30. The PA2 value ; ~ for the test compound was determined by the following ~; equations:
1 EC30 (presence of test compound)_ = dose ratio = X
EC30 (presence of vehicle control) 2. KB = concentration of test compound/(X-l) 3. pA2- - log KB

,, , .,., :. . .

~;~32~17 1 In vivo: Anesthetized, spontaneously breathing guinea pigs (Adult male albino Hartley strain) were monitored on a Buxco pulmonary mechanics computer. Changes in airway resistance (RL) were calculated by the computer on a S breath-by-breath basis at isovolumic points from signals measuring airflow and transpulmonary pressure using differential pressure transducers. Animals received either test compound or vehicle control intravenously via the jugular vein. LTD4 was then injected into the jugular vein. The bronchoconstriction produced was reflected by ~ changes in airways resistance relative to the baseline values obtained prior to injection of the test compound or vehicle control. Each guinea pig received either vehicle control or test compound.
Calculations: The average of 3 - 6 animals per treatment was calculated using the % changes in the pulmonary parameters for control and test compound-treated animals. The average % inhibition by ~; the test compound was calculated from the following 2G equation:
RL RL
(vehicle control) - (test comPOund) _ x 100 RL
(vehicle control) The compounds of this invention possess biosignificant antagonist activity against leukotrienes, primarily leukotriene D4. Representative of the antagonist activity o the compounds of this invention, tabulated below are a number of claimed compounds and the P~2 values and the RL values calculated from the above test protocols.

~ 35 ;~: :
~: :

;: ` :
.
:~ : . --. :

~232~3~7 1 Compounds of the Formula_(II) In Vitro In Vivo PA2 Concentration RL
m R1 R2 1 ~ -OH 6.2 5 mg/kg 91 5 1 H -NHCH2CO2H 6.2 5 mg~kg 36~
(a)l -NH2 -NHCH2c2H 5.2 5 mgjkg 96%
(b)l -NH2 2 2 5 O(c) _ _ 1 ~ -N(CH3)CH2CO2H 6.1 5 mg/kg 90%
1 -NH2 -NHCH~CONH2 5.5 5 mg/kg 98%
100 H -OH 5.9 1 H -N~2 5.1 5 mg/kg 38%
(a,d)l -NH2 -OH 5.1 - -(b,e)l -NH2 -OH 5.1 2 H -OH 6.2 S mg/kg 88 ( ) 4(R),5(S) isomer (b) 4(S),5(R) isomer (c? partial agonist (d) 1.0 Na salt (e) 1.5 Na salt Compounds of_the Formula (III) In VitroIn:Vivo A2 Concentration RL

m X R1 R2 , 1 OH H -OH 6.1 10 mg/kg 88%
1 H H -OH 6.6~0.4 - -.

: Compounds of the Formula (IV):

n m p R R1 R~
2 1 ~ 11 OH X OH 5.5 10 mg/kg 98 , j :

~ 35~:

: ::: : , ~ ~ ' ::: : , : , ~3~ 7 1 Compounds of the Formula (V) In Vitro In Yivo PA2 Concentration RL

m Rl R
(f)l -NH2 -NHC~2CO2H 4 9 - =

(f) Na Salt (g) 0.3M NH3 Compounds of the Formula (VI) m Rl R2 1 H -OH 5.3 - - -~?Unds of the Formula (VII) m R1 R2 : l H -OH 5.1 ~ 20 ~ '~
The specificity of the antagonist activity of a ~ number of the compounds of this invention is demonstrated : ~ by relatively low levels of antagonism toward agonis~s such as potassium chloride, carbachol, histamine and PGF2a.
Pharmaceutical compositions of the present inven-tion comprise a pharmaceutical carrier or diluent and an : amount of a compound of the formula (I~ or a pharma-::30 ceutically acceptab~e salt, such as an alkali metal salt thereof sufficient to produce the inhibition of the effects of leukotrienes, such as symptoms of asthma and : other allergic diseases.
When the pharmaceutical composition is employed in the fo~m of a solution or suspension, examples of appropriate pharmaceutical carriers or diluents include:
: ~ :
`~ : :

., , ~ ~ . '.
`: ~ , :' `

~;~3~91~

1 for aqueous systems, water; for non-aqueous systems, ethanol, glycerin, propylene glycol, corn oil, cottonseed oil, peanut oil, sesame oil, liquid parafins and mixtures thereof with water; for solid systems, lactoser kaolin and S mannitol; and for aerosol systems, dichlorodifluoro-methane, chlorotrifluoroethane and compressed carbon dioxide. Also, in addition to the pharmaceutical carrier or diluentf the instant compositions may include other ingredients such as stabilizers, antioxidants, preservatives, lubricants, suspending agents, viscosity modifiers and the like, provided that the-additional ingredients do not have a detrimental effect on the ~herapeutic 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 or by inhalation.
In general, par~icularly for the prophylactic treatment of asthma, the compositions will be in a form 2Q suitable for administration by inhalation. Thus the com-positions 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 active ingredient in a conventional liquified propellant or ~ compressed gas to be administered from a pressurized ; ~ aerosol container. The compositions may also comprise the solid active ingredient 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 less, equal or greater amounts than the solid active ingredient.
For parenteral administration the pharmaceutical composition will be in the form of a sterile injectable ~ ; ~

..
: ~
: ~ - :

` ' -- ~Z3~917 1 liquid such as an ampul or an aqueous or nonaqueous liquid suspension.
Usually a compound of formula I is administered to an animal subject in a composition comprising a nontoxic amount sufficient to produce an inhibition of the symptoms of an allergic response. When employed in this manner, the dosage of the composition is selected from the range of from 350 mg. to 700 mg. of active ingredient for each administration. For convenience, equal 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 method of inhibiting the symptoms of an allergic response resulting from a mediator release which comprises administering to an animal subject a therapeutically eff~ctive amount for producing said inhibition of a compound of formula I, preferably in the orm of a pharmaceuticaL composition. The administration may be carried out in dosage units at suitable intervals or in singLe doses as needed. Usually this method will be practiced when relief of allergic symptoms is ~pecifically required, however, the method is also usefully carried out as con~inuous or prophylactic treatment. It is within the skil} of the art to determine by routine experimentation the effective dosage to be administered from the dose ; 30~ range set forth above, taking into consideration such factors as the degree of severity of the allergic condition being treated, and so forth.
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.
~ J

:~ :

~23~9~

1 EXAMPLE_l Preparation of 5-[(2-amino-3-carboxymeth~lamino-3-oxopropyl)thio]-4 hydroxy-6(Z)-nonadecenoic acid [Formula (II) wherein m is 1, Rl is amino and R2 is S -NHCH2CO2~] as the 4(R),S(S) isomer, 4(S),5(R) isomer and the mixture of the two isomers.
(a) 3-CarbomethoxYpropionyl chloride l(a) To an ice~cold solution of mono-methyl succinate, (150 g, 1.135 mol) in 900 ml of methylene chloride and 3 10 ml of N,N-dimethylformamide was added 119 ml (1~26 mol) of oxalyl chloride, while keeping the pot temperature below 5C. After the addition was complete, the reaction mixture was stirred at 0C for 1 hour and then concentrated in vacuo to give a crude yellow liquid, which 15 was used without purification in the Rosenmund reduction.
(b) 3-CarbomethoxYpropionaldehyde l(b) To the acid chloride, l(a) (8.9 9, 0.059 mol) in 200 ml of sieve-dried tetrahydrofuran was added 6.9 ml (0.059 mol) of 2,6-lutidine. After standing at room temperature for 30 minutes, the mixture was filtered and ; 0.7 9 of 10% palladium on carbon was added to the filtrate. The mixture was hydrogenated at 50 psi for 2 --hours, the solids were filtered off and the filtrate concentrated. The residue was dissolved in methylene chloride, washed twice with 10~ hydrochloric acid solution and then twice with 5% sodium bicarbonate solution. The organic extract was dried with anhydrous sodium sulfate and concentrated in vacuo. Kugelrohr distillatisn (40-55C/~05 mm) yielded a colorless liquid.
30~ (c) 5-Carbomethoxy-2-pentenal l(c) ~; To a mechanically-stirred solution of l(b) (72.1 g, 0.620 mol) in 750 ml of toluene under argon was added 235.g g (0.776 mol) of formylmethylene triphenyl-phosphorane. The reaction mixture was refluxed for 1.5 hou~s, cooled to room temperature and then concentrated in vacuo. The residue was left standing under die~hyl ether ~, ~
: -~: ~ : ::

::::

.

~L~3~917 l at 0C overnight. The mixture was filtered and the solidwas washed with cold ether. The filtrate was concentrated and the resulting maroon oil was subjected to Kugelrohr distillation. The product was collected at 65-80C/.05 mm.
(d) MethYl 4,5-Epoxy-6-oxohexa-oate l(d) To a solution of 36.4 ml of a 30% hydrogen peroxide solution and 58.2 ml of lN sodium bicarbonate in 800 ml of ~ethanol and 400 ml of water under argon was added dropwise over 45 minutes 41.2 g (0.29 mol) of l(c) in 400 ml of methanol. After the addition was complete, the reaction was stirred for 2.5 hours at room temperature, while maintaining the pH between 9 and 9.S by the addition of sodium bicarbonate solution. The reaction mixture was then poured into one liter of saturated ammonium sulfate solution, the methanol was removed in vacuo, the solids were filtered off and the produc~ was extracted into methylene chloride. The aqueous layer was - back extracted ~wice, the combined extracts were dried with anhydrous sodium sulfate and then concentrated to give a crud golden oil. The product was Kugelrohr distilled at 82-9C/0.1 mm.
te) Meth~l 4,5-epoxY-6(Z)-nonadecenoate l(e) To an ice-cold solution of tridecyl triphenyl-phosponuim bromide (preparation described below) (97.2 g, 0.185 mol) in 600 ml of sieve-dried tetrahydrofuran under argon was added dropwise 17.1 ml of a 2.4 M solution of ; ~ n-butyl }ithium in hexane; the temperature was maintained ;~ at O~C during the 30 minute addition. The reaction mixture was stirred at this ~emperature for an additional 15 minutes, cooled to -i80C (dry ice/isopropanol) and then 2605 9 (0.168 mol) of l(d) in 150 ml of dried tetrahydro-furan was added dropwise over 30 minutes. After the addition was complete, the reaction was stirred for l hour at -78C and then concentrated in vacuo at 24~C. The residue was triturated with hexane and then left standing ., ~ at 0C overnight. The hexane was decanted and then the `:
; ~ ~, f ;

:

' ~23~7 1 residue was sonicated four times with hexane. The combined extracts were concentrated in vacuo and the crude - product purified~by preparative HPLC (2 Waters Prepak silica columns, elutiny with 8% ethyl acetate in hexane), S giving lte) as an oil.
(f) Tridecyl triphenyl phosphonium bromide A solution of l-bromotridecane (100 g, 0.4 mol) and triphenyl phosphine (10~ g, 0.4 mol) in 500 ml of xylenes was refluxed overnight. The reaction mixture was then cooled to room temperature and poured into diethyl ether. The resulting oil was washed ~hree times with ether, dissolved in methylene chloride and then concentrated in vacuo. The oil was left standing in diethyl ether at 0C overnight and then concentrated in vacuo to give a white solid.
(g) 4-Hydroxy-5-[(2-trifluoro_cetamido-3-carbomethoxy-methylamino-3-oxopropyl)thio]-6(2?-nonadecenoic acid, y-lactone and methvl ester To the epoxide, l(e) (12.9 g, 0~04 mol) under argon at room temperature was ~dded dropwise over 1 hour a solution of 14.5 g (0.05 mol) of 2-trifluoroacetamido-3-carbomethoxymethylamino-3-oxopropylmercaptan o F3C~NH
(HSCH~CH ~ HCH2CO2CH3) in triethylamine (6.7 ml)/methanol (150 ml). The reaction mixture was stirred at room temperature overnight and then concentrated in vacuo. The re idue was dissolved in a minimum volume of methylene chloride, 100 ml. of hexane was added and then this solution was stored at -15C for 1 hour. The resulting solid was filtered off and then the filtrated was concentrated in vacuo, to give a crude oil, consisting of a mixture of the desired products.

. :

-~ ;

. ~ .

1 (h) 4(R)_-Hydroxy-5(S)-[(2-trifluoroacetamido-3-carbo-methoxymethy~amino-3-oxopropyl)thio]-6(Z)-nonadecenoic acid y-lactone and 4(S)-Hydroxy-5(R)-[(2-trifluoro-acetamido-3-carbomethoxymethylamino-3-oxopropyl)-thio]-6(Z)-nonadecenoic acid, ~-lactone ~he crude mixture from Example l(g) (20.4 g.) in 200 ml of toluene was heated to 80C in the presence of 75 mg of p-toluenesulfonic acid for 15 minutes. The reaction mixture was then concentrated in vacuo and the resulting crude oil was purified by preparative HPLC (2 Waters Prepak silica columns, eluting with 45~ ethyl acetate in hexane) to give the desired products as the pure 4(R), 5(S) isomer and the pure 4(S), 5(R) isomer.
(i) 4(R)-Hydroxy-5(S) 1(2-amino-3-carboxymethvlamino-3-oxo-~opyl~thio]-6(Z)-nonadecenoic acid, hYdrate The lactone of Example l(h) in the 4(R), 5(S) isomer form, (1.9 9, 0.00327 mol) in aqueous sodium hydroxide (0.72 g, 0.018 mol in 50 ml water) was stirred at room temperature overnight. The pH of the reaction mixture was then adjusted to 3.S with concentrated hydrochloric acid and the resulting solid was collected and dried: mp 144-146C; Anal. Calcd. for C24H44N2O6S 1 1/4 H2O: C, 56.38; H, 9.17; N~
5048. Found: C, 56.16; H, 8.83; N, 5.54.
; 25 Similarly, the lactone of Example l(h) in the

4(S), 5(R) isomer form was converted to the desired 4(S),

5(R) isomer of the above noted compound: mp 141-143C;
AnalO Calcd- for C24H44N26S 1 1/4 ~2 C~
56.39~; H, 9.17; N, 5.48; Found: C, 56.20; H, 8.88; N, 3~ 5.18.
The isomeric mixture of the desired product was obtained treating ~he mixture of products prepared according to Example 1(9) without separation as described above: mp 140-143C; Anal. Calcd for C24H44N2O6S-1 1/4 H2O: C, 56.39; H, 9.17; N, 5.48; Found: C, ; 56.44; H, 8.75; N, 5nl6~

, 1: :
`: : :

.

:

~L23~917 Preparation of 4-hydroxy-5-~(2-carboxyethyl)thio]-

6(Z)-nonadecenoic acid (Formula II) wherein m is l, R
is hydrogen and R2 is hydroxyl) as a mixture of the 5 4(R), 5(S) and the 4(S), 5(R) isomers.
A solution of 1~32 g (4.1 mmoL) of 4,5-epoxy-6(Z)-nonadecenoic acid methyl ester and 1.46 g (12.2 mmol~ of methyl-3-mercaptopropionate in 20 ml of methanol was treated with 1.62 g (16.1 mmol) of triethylamine in 10 ml of methanol under an argon atmosphere overnight The reaction mixture was concentrated in vacuo and purification was carried out by chromatography on silica gel with hexane/ether (60/40) to give diester which contained some mono ester/lactone as evidenced by IR and tlc (CH2C12/hexane/ace~one, 47/47/5).
A solution of this diester/lactone mixtur~ (1.6 g, 3.6 mmol) in 30 ml o~ methanol was txeated overnight with 1.43 g (36 mmol) of sodium hydroxide in 5 ml of water at room temperature, under an argon atmosphere. The reaction mixture was partly concentrated in vacuo, redissolved in 20 ml of water, and acidified with dilute phosphoric acid. The aqueous solution was extracted 3 times with 50 ml of ether, and the extracts were dried over magnesium sulfate, filtered and concentrated in vacuo to give white crystalline solid; m.p. 78-80C,. Anal.
calcd. for C22H40O5S: C, 63.43, H, 9-68- Found:
C, 63.11, H, g.69.

Preparation of 5-[(3~carboxymethylamino-3-oxo-propyl)thio]-4-hydroxy-6(Z)-nonadecenoic acid, ~Formula (II) wherein m is l, Rl is hydrogen and R2 is -NHC~2C02H]
The above named compound was prepared by the general method of Example 2 using 3-carboxymethylamino-3-oxopropyl mercaptan, which in turn was prepared by the coupling reaction between 3,3'-dithio-dipropionic acid and : ~' : ::

~, :

, .

- :1232~7 1 glycine methyl ester using DCC, followed by the reduction of the disulfide wi~h tri-n-octyl phosphine in acetone/water 1:1 mixture. Product was obtained as an oil. Anal. calcd~ for C24H43NO~S: C, 60.85;
H, 9.15~ N, 2.95. Found: C, 60,65; H, 9.07; N, 2.89.

Prepara~ion of 5-[(3-carboxymethyl-N-methylamino-3-oxopropyl)thio]-4-hydroxy-6(Z)-nonadecenoic acid, ~Formula (II) where m is 1, Rl is hydrogen and R2 i5 ~H3 The above named compound was prepared by the general method of Example 2 using 3-carboxymethyl-N-methylamino-3-oxopropyl mercaptan. Mercaptan was prepared by coupling reaction between p-methoxy benzylthiopropionic acid and N-methyl glycine methyl ester in the presence of DCC, followed by treatment with HF at -78C. Product obtained had a melting point of 80-82C.
Anal. calcd.: C, 6L.57; H, 9.30; N, 2.87. Found: C, 20 6L.55, H, 9.65; N, 3.02.
EXA~PLE 5 Preparation of 5-[(carboxymethyl~thio]-4-hydroxy- -6(Z)-nonadecenoic acid [Formula (II) wherein m is 0; R
is hydrogen and R2 is hydroxyl]
The above named compound was prepared by the general method of Example 2 using commercial methyl thioglycolate: mp 50-52C. Anal. calcd. for C21~38Q5S: C, 62.65; H, 9.51. Found: C, 62.74; H, 9.57.

Preparation of 5-[(3-carboxypropyl)thio]-4-hydroxy~6(Z)-nonadecenoic acid, [Formula (II) wherein m is 2; Rl is hydrogen and R2 is hydroxyl]
The above named compound was prepared by the general method of Example 2 using commercial 4~mercapto-' :
1~

.' .

.
, .. ` iL~32~

1 butyric acid, mp 56-58C. Anal. calcd~ for C23H42O5S- 1/4 H2O: C, 63~48; H, 9.96. Found:
C, 63.29; H, 9.75.

S Preparation of 5-[[2-(aminocarbonyl)e~hyl]thio]-4-hydroxy-6(Z)-nonadecenoic acid, [Formula (II) where m is l; Rl is hydrogen and R2 is amino]
The above named compound was prepared by the general method of Example 2 using 2-(aminocarbonyl~ethyl mercaptan. Mercaptan was prepared by treatment of 3,3'-dithiodipropionyl iodide with concentrated ammonium hydroxide followed by the reduction of disulfide with tri-n-octylphosphine and acetone/water 1:1 mixture.
Product obtained had a melting point of 84-86C. Anal. `-15 caLcd. for C22H41NO4S: C, 63.57; H, g.94. Found:
C, 63.56, H, 9.82.

Preparation of 4-hydroxy-5-[(2-amino-2-carboxy-ethyl)thio]-6(Z)-nonadecenoic acid [Pormula (II) wherein m ~ is l; Rl is amino and R2 is hydroxyl] as the 4~R),5(S) isomer and the 4(R),S(S) isomer (a) 4~ oxy-S-[(2-trifluoroacetamido-2-carbomethox~- -ethyl)thio]-6(Z)-nonadecenoic acid, meth~l ester and y-lactone To the epoxide, methyl-4,5-epoxy-6(Z)-non-adecenoate (6) (0.9 g, 0.00277 mol) under argon at room temperature was added dropwise a solution of 1.2 g (0.00519 mol) of 2-trifluoroacetamido-2-carbomethoxy-ethylmercaptan in triethylamine (0.88 ml)/methanol (lS
ml). The reaction mixture was stirred at room temperature for 30 hours, concentrated in vacuo and then filtered through a silica gel bed, eluting the product mixture with chloroform. The chloroform wash was concentrated to give ~; a mixture of the above-noted methyl ester and y-lactone.

, .~ ~

1~3~9~7 ~ - 25 -1 (b) 4-Hydroxy-5-[(2-trifluoroacetamido-2-carbomethoxy-ethyl)thio]-6(Z)-nonadecenoic acid, ~-lactone The crude mixture (2 g) of the methyl ester and y-lactone from Example 8(a) in 75 ml of toluene was heated to 80C in the presence of 27 mg of p-toluenesulfonic acid for 20 minutes. The raaction mixture was then concentrated in vacuo and the residue was taken up in methylene chloride, washed with 5% sodium bicarbonate solution, dried with brine and anhydrous sodium sulfate 10 and then concentrated in vacuo. The resul~ing crude oil was applied to the Water's Preparative HPLC (2 Prepak silica columns, eluting with 25% ethyl acetate in hexane) to give the desired products as the 4(R),5(S) isomer and the 4(S),5(R) isomer. Each individual diastereomer was 15 purified further on a silica "flash" column, eluting with 30% ethyl acetate in hexane.
(c) 4(R~-Hydr~ 5(S)-[(2-amino-2-carboxyeth l)thio]-6(Z)-nonadecenoic acid, sodium salt A partial suspension of 4(R),5(S) isomer of t,he ~ (-lactone of Example 8(b) (0.3 g, 0.00057 mol) in aqueous sodium hydroxide (0.205 g, 0.004 mol in 13 ml water) was stirred for 24 hours at room temperature. The pH of the reaction mixture was then adjusted to 3.5 with concentrated hydrochloric acid to give the desired 25 product: mp 168-170C. Anal. calcd. for C22H41NO5S~
lNa (-lH): C, 58.12; H~ 8.87; N, 3.08. Found: C, 57.99;
H, 8.85 N, 3.70.
(d) 4(S~-Hvdroxv-5(R)-[(2-amino-2-carboxvethvl)thio]-6(Z)-nonadecenoic acid, sodium salt The reaction was carried out as described in Example 8(c), to giYe the desired product: mp 159-161C.
Anal. Calcd. for C22H41 O5S .5 C, 56.69; H, 8.54; N, 3.01. Found: C, 56.40; H, 8.19; N, 3.33.

,: ' .
: - - ;

:

~2329i.7 Preparation of 5[-(2-amino-3-carboxamidomethyl-amino-3-oxopropyl)thio~-4-hydroxy-6(Z)-nonadecenoic acid [Formula (II) wherein m is l; Rl is amino and R2 is -NHCH2~NH2]
(a) 4(R)-Hydroxy-5(S)-[(2-trifluoroacetamido-3-carbamo~l-met_ylamino-3-oxopro~y~)thio]-6(Z)-nonadecenoic acid, ~-lactone 4(R)-hYdroxy-5(s)-[(2-trifluoroacetamido-3-carbamoylmethylamino-3-oxoprop~l)thio]-6(5)-nonadecen-.

amide A mixture of lactone methyl ester prepared according to Example l(g) (0.41 g, 0.7 mmol) in 10 ml of dimethoxyethane, 10 ~1 of methanol, and 20 ml of concentrated ammonium hydroxide solution was stirred at ~~
0C for 2 hours. The mixture was neutralized in the coldwith concentrated hydrochloric acid to pH 7.5. The crude product was partitioned into methylene chloride. The aqueous phase was further extracted 2 x 40 ml methylene chloride. The combined extracts were washed with saturated sodium chloride solution, dried over sodium sulfate, a~d evaporated to give oil residue. Flash column chromatography (silica gel, 1.5" x 6", 3% CH30H/CHC13, 25 ml fraction) gave the desired products. The 6(2)-nonadecenamide had the following properties: mp 25 143-5C; Anal. calcd. for C26H45F3N405 53.59; H, 7.78; N, 9.61. Found: C, 53.86; H, 7.52; N, 9.83.
(b) 4(R)-Hydr~y-5(S)-[(2-amino-3 carbamoylmeth~lamino-3-oxopropylthio]-6(Z~-nonadecenoic acid A mixture of 120 mg (0.2 mmol) of the y-lactone of Example 9(a) in 5 ml of 0.2 M sodium hydroxide solution was stirred at room temperature for 18 hours. The mixture was acidified to pH 3 by adding a concentrated hydro-chloric acid solution in ice-bath~ The resulting precipitates were filtered and washed quic~ly with cold water and dried at 56C for 24 hours to give the desired ~, , .

~3~7 1 product, mp. 143 -5C; Anal. calcd.: C, 57.51; H, 9.35;
N, 8.38; Found C, 57.50; H, 9.42; N, 6.72.

~ ;
Preparation of 3-hydroxy-4-[(2-carboxye~hyl)thio]-5(Z)-octadecenoic acid [Formula (III) wherein m is l; X i5 hydroxyl, Rl is hydrogen and R2 is hydroxyl]
(a) He~Ptadec-2(E)~4(z)-dienyl tetrahydrop~ranyl ether_ lO(a)(1) He~tadec-2(E),4(E)-dienyl tetrahydropvranyl ether ._ lO(a)(2) Tridecyltriphenyl phosphonium bromide (189 g, a. 36 mole) was dissolved in 900 ml of tetrahydrofuran and cooled to 0C in an ice-salt bath while stirring under argon. A 2.2 N solution of n-butyllithium in hexane (250 ml, 0.36 mole) was added dropwise over a period of 30 ; minutes. The mixture was stirred for an additional 20 minutes and then cooled to -70C in a dry ice-acetone bath~ The 4-hydroxybut-2(E)-ene-l-al tetrahydropyranyl ether (51 g, 0.3 mole) in 2~5 ml o tetrahydrofuran was added dropwise over a period of 35 minutes and the mixture stirred for an additional hour at -70C. The mixture was then poured into 6.25 liters of ether and stirred for 20 minutes. The resulting mixture was filtered through glass fiber filter paper. The filtrate was evaporated and the residue ~riturated wi~h hexane, filtered, evaporated, and flash chromatographed to give a ~3:1 mixture of lO(a)(l):
lO(a~(2).
(b) He~adec-2(E)_,4(Z)-dlen-l-ol lO(b)(l) Heptadec-2(E?!4(E)-dlen-l-ol lO(b)(2) The mixture of compounds lO(a)(l) and lO(a)(2) ; (80 g t 0.24 mole) was dissolved in 3 liters o methanol and the pyridinium p-toluenesulfonate (3 g, 0.012 mole) was added to the mixture stirring under argon at room temperature. The progress of the reaction was monitored by tlc. When the reaction was complete the solvent was ; evaporated anu the residue flash chromatographed on 500 :
:, , ~ '' . ' ~

~;~3~9~

1 grams of silica gel eluted with 10% ethyl acetate in hexane to give 52 grams (87%) o a ~3:1 mixture of lO(b)(l):lO(b)(2). Separation of lO(b)(l) from lO(b)(2) was accomplished by careful chromatography on silica gel.
Compound lO(b)(l) mp 34-37C. Compound lO(b)(2) mp 51-55C.
(c~ Trans-2~3-epoxy-heptadec-4(z)-ene-l-ol lO(c) Compound lO(b)(l) (2.52 9, 10 mmol) was dissolved in 100 ml o~ methylene chloride stirring at room temperature under argon. A 0.5 N solution of sodium bicarbonate (30 ml~ was added. The 85% m-chloroperben~oic acid l2.03 g, io mmol) was added slowly in small portions. The mixture was stirred for 1.5 hours after the addition was complete. The phases were separàted and the aqueous phase washed with methylene chloride. The combined organic phases were dried over anhydrous sodium sulfate filtered and evaporated. The residue was flash chromatographed on 100 grams of silica gel eluted with 10-20% ethyl acetate-hexane to give compound lO(c).
~d) Methvl_3-tl,2-dihydroxyheptadec-4(Z)-enYl)thio-propionate lO(d) Compound lO(c) (7.2 g, 26.9 mmol) was dissolved in 40.2 ml of methanol containing 2% triethylamine. This solution was stirred at room temperature under argon and a solution of methyl 3-mercaptopropionate (4.92 ml, 44.4 mmol) and triethylamine (11.16 ml, 80.2 mmol) in 40.2 ml of methanol was added dropwise over a period of 15 minutes. The mixture was stirred for S hours at room temperature and then placed in the refrigerator overnight. The solvents were evaporated and the residue flash chromatographed on 500 grams of silica gel eluted wi~h 10-S0~ ethyl acetate in hexane to give compound lO(d), mp. 33-36.
(e) 2-[(2-CarbomethoxYethyl)thio~hexadec-3(z)-enal lO~e) Compound lO(d) ~2 9, S.15 mmol) was dissolved in - 10 ml of diethyl ether and stirred in a room temperature :

:

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~L~3~

1 water bath. A saturated solution (100 ml) of periodic acid in diethyl ether was added in a single portion. The resulting mixture was stirred for two minutes and then immediately flash chromatographed on 150 g of silica gel with 10~ ethylaceta~e in hexane to give compound lO(e).
(f) Methvl 3-hYdroxY-4-[(2-carbomethoxvethvl)thio]octadec-5(Z)-enoate lO(f) A dry flask sealed with a septum and maintained under an argon atmosphere was charged with 4.5 ml of hexane and cooled in an ice bath. A 2~2 M solution of n-BuLi (1.03 ml, 2.25 mmol) was added followed by the dropwise addition of diisopropyl amine (0.315 ml, 2.25 mmol). The solution was stirred at 0C for 10 minutes and then cooled to -78C in a dry ice-acetone bath for 15 -15 minutes. A solution of methyl acetate (0.18 ml, 2~25 mmol) in 1.5 ml hexane was added over a period of 1 minute and the mixture stirred at -78C for an additional minute. The mixture at this time was almost clear.
Compound lO(e) (750 mg, 2~1 mmol) in 1.5 ml of hexane was added over a period of 1 minute resulting in a clear yellow solution ~hich was stirred at -78C for an additional 15 minutes. The reaction mixture was then flash chromatographed on 100 grams of silica gel eluted with 15% ethyl acetate in hexane to give compound lO(f)o (9) 3-HydroxY-4-[(2-carboxyethyl)thio]-5(Z)-octadecenoic acid Compound lO(f) (0.2 g, 0.46 mmol) was dissolved in 5 ml of methanol and stirred under an argon atmosphere at 0C. A lN solution of sodium hydroxide (2 ml, 2 mmol) was added dropwise over a period of 0.5 minute. The ice ` bath was removed and the reaction allowed to warm to room temperatuse for ~ hours. Most of the methanol was ; evaporated and the aqueous residue was cooled in an ice bath and acidified with dilute hydrochloric acid. The aqueous phase was extracted twice with diethyl ether. The combined ether extracts were dried over anhydrous sodium ~3;~9~7 1 sulfate, filtered, and evaporated to give crude product.
This was recrystallized from diethyl ether-hexane to give the desired compound, mp 88-97C. Anal. Calcd. C: 62.65;
H: 9.51; S: 7.96; Found C: 62.32, H: 9.38; and S: 8.10.
The following compounds are prepared by the general method o Example 10 by employing the appropriate thiol containing compound for methyl-3-mercaptoproprionate:
3-hydroxy-4-[(carboxymethyl)thio]-5(Z)-octa-decenoic acid;
3-hydroxy-4-[(3-carboxypropyl)thio]-5(Z)-octa-decenoic acid;
3-hydroxy-4-[(3-carboxym~thylamino-3-oxopropyl)-thio]-5(Z)-octadecenoic acid;
3-hydroxy-4-[(2-amino-3-carboxymethylamino-3-oxo-15 propyl)thio]-5(Z)-octadecenoic acid;
3-hydroxy-4-[[(aminocarbonyl)ethyl]thio3-5(Z)-octadecenoic acid; and 3-hydroxy-4-[(~-amino-2-carboxyethyl)thio]-S(Z)-octadecenoic acid.

Preparation of 5-[(2-carboxyethyl)thio]-6(Z)-non-adecenoic acid [Formula (IV) wherein n is 25 m is 1; p is 11; Rl is hydrogen and R2 is hydroxyl]
(a) 2-Bromo-5^carbomethoxypentanoic acid ll(a) To a solution of 30 gm (0.187 M) of 5-carbomethoxypentanoic acid in 250 ml of chloroform was added 90 gm of thionyl chloride. The mixture was refluxed for 2 hours, cooled and the solvent removed under vacuumO
The residue was redissolved in 200 ml of carbon 30 tetrachloride, 40 gm (0.225 M) of N-bromosuccinimide was added, the mixture heated to reflux, eight drops of a HBr/acetic acid mixture added, and the reaction refluxed ~or 3 hours. After cooling, the mixture was fil~ered and evaporated to dryness. The residue was dissolved in 250 ml of acetone to which was added dropwise 200 ml of a freshly prepared 5% sodium bicarbonate solution. After ;

~ .

~2329 l stirring for 15 minutes during which time the pH was constantly adjusted to p~ 9 with sodium bicarbonate the mixture was acidified with dilute HCl, to a pH l.0, concentrated in vacuum, and extracted with chloroform.
The organic phase was extracted with dilute bicarbonate solution, acidified and reextracted with chloroform. The extract was dried over Mg504, filtered and evaporated to dryness to afford the desired compound as a viscous oil.
(b) 5-Carbomethoxy-2-~(methoxY~carbonylpropyl)thio]-pentanoic acid ll(b) A mixture of 42 gm (0.175 M) of ll(a), 50.68 g(0.42 M) oE methyl-3-mercaptoproprionate and 147 ml (1.05 M) of triethylamine in 250 ml of methanol was refluxed for two hours, cooled and concentrated in vacuum. The residue 15 was di~solved in ethyl acetate and extracted with 5%
sodium bicarbonate. The aqueous extract was acidified with 3N HC1, extracted with chloroform, the extract dried over MgSO4, filtered a~d evaporated to dryness to afford the desired compound.
(c) 5-Carbomethoxy-2[(methoxycarbonylpropyl)thio]-pentanol ll(c?
A solution of ll(b) (40 g, 0.142 M) in THF (200 ml~ was placed in a methanol-ice (-10C) bath. The contents were permitted to cool for 20 minutes at which time 12.57 g (0.156 M) of borane-methyl sulfide complex was added slowly over a period of 50 minutes, followed by refluxing for 30 minutes. Acetic acid (lO ml) was then added and excess T~F evaporated. The reaction mixture was dissolved in ethyl acetate (300 ml), washed with 5% sodium bicarbonate, dried over MgSO4 and filtered. Evaporation afforded the crude alcohol.
(d~ S Carbomethoxy-2[(methoxycarbonylpropyl)thiol-pentanal ll(d) Methylene chloride (lO ml.) and Me2SO (1.78 g, 0.23 M) were combined and cooled to -65C. TFAA (1.73 g, 0.015 M) was added dropwise to the cold solution, at which ` ~329~7 1 time a white precipitate formed. After S minutes at -65C, a solution of ll(c) (2.0 g, 7.58 mmol) in CH2C12 (S ml) was added dropwise while maintaining the reaction mixture at -65C. The mi~ture was then stirred at -65C for 30 minutes, followed by addition of TEA (2.30 g, 0.023 M) dropwise. A temperature below -60C was maintained until addition of TEA was complete. The bath was then removed and the reaction permitted to warm to room temperature. The mixture was diluted with CH2C12, washed with 3N HCL, H20, twice with 5~
NAHC03 and once with brine. The organic phase was then dried over MgS04~ filtered and evaporated to dryness to afford the crude aldehyde.
(e) MethYl-5-[(2-carbomethoxyethyl)thio]-6(Z)-non- -lS adecanoate ll(e) A mixture of tridecyltriphenyl phosphonium bromide (1.76 g, 3.35 M) and THF (20 ml) was permitted to stir for 5 minutes at room temperature. The solution was then cooled to -68~C and stirred for an additional 10 minutes. At which time N-butyllithium (3.05 mmol) was slowly added while maintaining a -65C température.
Following stirring for 10 minutes at -68C the reaction mixture was stirred for an additional 10 minutes at -10C. Upon recooling to -68C, (0.918 g, 3.50 mmol) of ll(d) was dissolved in THF (10 ml) and slowly added. The reaction was stirred for one hour at -68C followed by removal of the bath in order to warm the mixture to room temperature. THF was then evaporated and the residue dissolved in ethyl acetate, which was washed with 3N HCL, water and twice with 5% NAHC03. The extract was dried over MgS04, filtered and evaporated to dryness. The product was 1ash chromatographed with 91~ he~ane: 9%
EtOAc to provide the nonadecanoate.
(f) Potassium carbonate (0.81 g, 5.84 mmol) was dissolved in H20 (10 ml), to which was added methanol ~ :
: , ~ ~,., , `~ ' . .

:~3~ 7 .

1 (5 ml). The solution was stirred for 2 minutes at room temperature, followed by the addition of ll(e) (100 mg;
0.234 mmol) in MeOH (24 ml). The mixture was permitted to stir overnight at room temperature, after which time excess MeOH was evaporated and the remaining aqueous phase was then washed wi~h ethyl acetate, acidified to a pH =
1.0 with 3N HCL and extracted twice with ethyl acetate, the extract was dried over MgSO4, filtered and evaporated to an oil. The product was recrystallized from ether-petroleum ether and cooled for one hour to afford the nonadecanoic acid as a white crystalline solid (mp 53-54C).
The following compounds are prepared by the general method of Example 11 by employing the appropriate thiol containing compound for methyl-3-mercaptoproprionate:
5-[(carboxymethyl)thio]-6(Z)-nonadecenoic acid;
5-~3-carboxypropyl)thio]-6(2)-nonadecenoic acid;
5-[(3-carboxymethylamino-3-oxopropyl)thio]-6(Z)-nonadecenoic acid;
5-[(2-amino-3-carboxymethylamino-3-oxopropyl)thio]-6(Z)-nonadecenoic acid;
5-[[(aminocarbonyl)ethyl]thio]-6(Z)-nonadecenoic acid; and S-[(2-amino-2-carboxyethyl)thio]-6(Z)-nonadecenoic acid.

Preparation of 5-[(2-carboxyethyl)thio]-4-hydroxy-6~2)-nonadecenamide, [Formula (V) where m is 1, Rl is hydrogen and R2 is hydroxyl]
The above named compound was prepared by treatment of the compound of Example 2 with 1% solution of trifluoroacetic anhydride in CH2C12 for 2 hours at room temperature, followed by treatment with NH3 in methanol solution at 0C for 0.5 hour and room temperature overnigh~. Product was obtained as a crystalline solid mp 82-85C; Anal. Calcd. for C22H41NO4S 0.3 M NH3:

~23~917 1 C, 62.77; H, 9.89; N, 4.33. Found: C, 62.67, 62.98; H, 9.92, 9.78; N, 3.98, 4.00.

Preparation of 5-[(3-carboxymethylamino-3-oxopropyl)thio]-4-hydroxy~6(Z)-nonadecenamide [Formula (V) where m is 1, Rl is amino and R2 is -NHCH2CO2H]
A mixture of 0.9 g (1.4 mmol) of compounds of Example l(g) in 100 ml of 3.8~ anhydrous ammonia in ethanol solution was stirred at room temperature for 2 ~ays. The reaction mixture was concentrated to dryness.
The residue was azeotroped with methylene chloride to give 0.9 g of off-white powder. The hydroscopic material was dissolved in 10% NaOH solution (5 ml) and chromatographed on a 3-cm-by-9-cm column of XAD-7 resin. After washing with ~2 (100 ml), the column was eluted with aqueous methanol solution (1:1) taking 20 ml per fraction.
Fraction 13 was collected to give the desired product as a white amorphous powder, mp. 195-8C. Anal.C~lcdlfor C24H4~N3O5S Na(H) 2~2O: C, 52.82; H, 8.87; N,

7.70. Found: C, 52.65; H, 8.15; N, 7.72.

Preparation of 4-hydroxy-5-[(2-carboxyethyl)thio]-(Z)-hepadecenoic acid [Formula (VI) wherein m is 1, R
is hydrogen and R2 is hydroxyl]
The above named compound was prepared by the general method of Example 2 using 4,5-epoxy-6(Z)-hepta-decenoic acid methyl ester, which was obtained by employing the general method of Example l(e) using undecyltriphenyl phosphonium bromide. The product obtained had a melting point of 7B.5-80C. Anal. calcd~:
C, 61.82; H, 9.34; S, 8.25. Found C, 61.82; H, 9.10; S,

8.42.
The following compounds are prepared by the general method of Example 14 by employing the appropriate thiol containing compound for methyl-3-mercaptoproprionate:

., .

1~3~9~7 1 4-hydroxy-5-[(carboxymethyl)thio]-6(z)-hepta-decenoic acid;
4-hydroxy-5-[(3-carboxypropyl)thio]-6(Z)-hepta-decenoic acid;
4-hydroxy-5-[(3-carboxymethylamino-3-oxopropyl)-thio~-6(Z)-heptadecenoic acid;
4-hydroxy-S-[(2-amino~3-carboxymethylamino-3-oxo-propyl)thio]-6(Z)-heptadecenoic acid;
4-hydroxy-S-~[(aminocarbonyl)ethyl]thio]-6(Z)-heptadecenoic acid; and 4-hydroxy-5-[(2-amino-2-carboxyethyl)thio~-6(Z)-heptadecenoic acid.

Preparation of 4-Hydroxy-S-[(2-carboxyethyl)thio-6(Z)-heneicosenoic acid [Formula (VII) wherein m is 1, R1 is hydrogen and R2 is hydroxyl]
The above named compound was prepared by the general method of Example 2 using 4,5-epoxy-6(Z)-heneicosenoic acid methyl ester, which was obtained by employing the general method of Example l(e) usi~g pentadecyltriphenyl phosphonium bromide. The product obtained had a melting point 59-61C. Anal,calcd.:
C, 64.82; H, 9.97; S, 7.21. Found C, 65.16; H, 10.07;
S, 7.36.
The following compounds are prepared by the general method of Example 15 by employing the appropriate thio containing compound for methyl-3-mercaptoproprionate:
: 4-hydroxy-5-[(carboxymethyl)thio]-6(Z)-heneico-senoic acid;
4-hydroxy-5-[(3-carboxypropyl)thio]-6(Z)-heneico-senoic acid;
4-hydroxy-5-[(3-carboxymethylamino-3-oxopropyl)-;~ thio] 6(Z)-heneicosenoic acid;
4-hydroxy-S-[(2-amino-3-carboxymethylamino-3-oxo-propyl)thio]-6(Z~-heneicosenoic acid;

' .

~2329~7 1 4-hydroxy-5-[[(aminocarbonyl)e~hyl]thio]-6(Z)-heneicosenoic acid; and 4-hydroxy~5-[(2-amino-2-carboxyethyl)thio~-6(Z)-heneicosenoic acid.

Preparation of 4-[(2-carboxyethyl)thio~-5(Z)-octa-de enoic acid [Formula (III) wherein m is l; X is hydrogen; Rl is hydrogen and R2 is hydroxyl~
(a) Methyl-4-bromo-4-carboxybutanoate 16(a) Methyl-4-(chloroformyl)butyrate (30.0 g, 0.182 M) was charged to a flask, along with 200 ml of CC14. To this solution N-bromosuccinimide (40.0 9, 0.219 ~) was added. The reaction mixture was heated to reflux and seven drops of 47% aqueous HBr was added, reflux conditions continued for approximately 2 hours. Following cooling to room temperature, the reaction was filtered and evaporated to dryness. The residue was then placed in the refrigerator overnight. The residue was dissolved in 250 ml of acetone, to which is added saturated sodium 20 .bicarbonate during which time the ph was adjusted to nine. With dilute HCl the mixture was then acidified to a pH = 1.0, concentrated and extracted with chloroform. The extract was dried over MgSO4, filtered and evaporated to dryness to afford the desired compound as an oil.
(b) Meth~1-4-[(2-carbomethox~ethyl)thio]-4-carbox~
butanoate 16(b) To a stirred solution of 29.0 g (0.24 M) of methyl-3-mercaptoproprionate and 61.0 g (0.60 M) of triethylamine in 200 ml of methanol was added 22.5 (0.10 M) of 16(a) under argon. After stirring at room ~ temperature for ten minutes, ~he reaction was heated to ; re~lux for one and a half hours, cooled to room temperature and concentrated in a vacuum. The residue was then diluted with ethyl acetate, extracted with 5% sodum bicarbonate, acidified to pH 1.6 with dilute ~Cl and reextracted into chloroform several times. Organic , -~ , ~3~9 :lL7 1 extracts were then combined, dried over MgSO4, filtered and evaporated to dryness to afford the desired compound.
(c) Methyl 4~_(2-carbomethoxyethyl)thio]-5-hydroxy ~entanoate 16(c) A solution of 16(b) (21.4 g; 0.08 M) in THF
(175 ml) was cooled to -10C in a methanol-ice bath. The contents were permitted to cool for 10 minutes at which time borane methyl sulfide complex (7.2 g; 0.09 M) was added slowly. Following addition reaction was warmed to room temperature and then refluxed for approximately 15 minutes. After cooling acetic acid (6 ml) was added and execess THF evaporated. The reaction mixture was dissolved in ethyl acetate, washed with 5~ sodium bicarbonate, dried over MgSO4, filtered, evaporated to dryness to afford the crude alcohol. This material was then flash chromatographed using a 60~ ethyl acetate in hexane system to yield a purer product.
(d) Methyl-4-[(2-carbomethoxyethYl)thio]-4-formYl butanoate 16(d) Methylene chloride (20 ml) and Me2SO (1.14 ml;
O.016M) were combined and cooled to -65C. TFAA (O.91 ml;
0.012 M) was added dropwise to the cold solution at which time a white precipitate formed. After- 5 minutes a solution of 16(c) (2.0 g; 0.008 M) in methylene chLoride (3 ml) was added dropwise while maintaining the temperature at -65C. The reaction mixture was then stirred for thirty minutes followed by addition of T~A
(3~1 ml; 0.022M) dropwise. The reaction mixture was stirred for an additional thirty minutes at -65C, at which it was diluted with methylene chloride, washed with dilute HCl, water and then twice with 5% sodium bicarbonate. The organic phase was then dried over MgSO4, filtered and evaporated to dryness to afford the crude aIdehyde. This material was flash chromatographed using 50% ethyl acetate in hexane resulting in purer product.

..
' -~3~9~7 1 (e) Methyl-4-[(2-carbomethoxyethyl)thio]-5(Z)-octa-decenoate 16(e) A mixture of tridecyltriphenyl phosphonium bromide (1.27 g; 2.4 mmol) and THF (20 ml) was allowed to stir for 3 minutes at room temperature. The solution was then cooled to -68C, at which time n-butyllithium (1.05 ml; 2.4 mmol) was added slowly while maintaining a reaction temperature less than -65C. Following stirring at -68C the reaction was permitted to warm to -10C for approximately 10 minutes. Upon recooling to -68C, (0.05 g; 2.0 mmol) of 16~d) was dissolved in TH~ (10 ml) and slowly added. The reaction was stirred for one hour at -68C followed by removal of the bath. Product was then triturated into hexane and also into ether, hexane and ether fractions were combined, evaporated to dryness and the remaining residue was flash chromatographed affording the desired octadecenoate.
(f) 4-[(2-Carboxyeth~l)thio]-S(Z)-octadecenoic acid (319 mg; 0;826 mmol) of 16(e) was dissolved in methanol (7.5 ml) and permitted to stir under argon at 0C. A lN solution o~ sodium hydroxide (3.1 ml; 3.08 mmol) was added dropwise. The ice bath was then removed and the reaction allowed to warm to room temperature for approximately three hours. After which it was placed in the refrigerator and stored overnight. Most of the methanol was then evaporated and the aqueous residue was cooled in an ice bath followed by acidification with dilute HCL. The aqueous phase was extracted twice with diethyl ether. The ether extracts were then combined and dried over magnesium sulfate, filtered and evaporated to give crude product. This was then recrystallized from diethyl ether-hexane to afford the final product, mp 81C;
Anal. Calcd C: 65.24; H: 9.91; S: 8.29; Found C: 65.50;
~: 10.05; S: 8.61.
The following compounds are prepared by the general method of Example 16 by employing the appropriate , 1 thiol containing compound for methyl-3-mercaptoproprionate:
4-[(carboxymethyl~thio]-5(Z3-octadecenoic acid;
4-[(3-carboxypropyl~thio]-5(Z)-octadecenoic acid;
4-[(3-carboxymethylamino-3-oxopropyl~thio]-5(Z)-S octadecenoic acid;
4-[(2-amino-3~carboxymethylamino-3-oxopropyl)thio]-5(Z)-octadecenoic acid;
4-[[(aminocarbonyl)ethyl]thio]-5(Z)-octadecenoic a~id; and 4-[(2-amino-2-carboxyethyl)thio]-5(Z)-octadecenoic acid.

As a specific embodiment of a composition of this invention, an active ingredient, such as the compound of Example l(i), is dissolved in sterile water at a concentration of 0.5 percent and aerosolized from a nebulizer operating at an air flow adjusted to deliver the desired aerosolized weight of drug~

As an additional specific embodiment of a composition of this invention, an active ingredient, such s as the compound of Example 2, is admixed w-th mannitol at a concentration of 1.0 percent and administered from a powder inhalation device adjusted to deliver the desired ; 25 weight of drug.

' :

` ' '

Claims (14)

The embodiments of the invention in which an exclusive property or privilege is claimed are defined as follows:

1. A process for preparing the compounds represented by the formula (I) (I) wherein n is 1 or 2 m is 0, 1 or 2; p is 9, 10, 11, 12 or 13; X is hydrogen or hydroxyl; R is hydroxyl or amino R1 is hydrogen, amino or ; R2 is hydroxyl, amino, -NHCH2CO2H, or -NHCH2CONH2 with the proviso that when m is 0, R1 is hydrogen, or a pharmaceutically acceptable salt thereof which comprises:
(A) when X is hydroxyl and n is 2, reacting wherein COR3 is COR of a radical such as CO2-alkyl easily convertible into an acid moiety with , wherein R1' and R2' are respectively R1 and R2 or a radical such as an alkyl ester of trifluoromethylacetamide easily convertible into R1 or R2;
(B) when X is hydroxyl and n is 1, reacting with CH3COR3; or (C) when x is hydrogen, reacting with CH3(CH2)pCH=P?3;
and then optionally converting R1', R2' and R3 into R1, R2 and R and optionally forming the pharmaceutically acceptable salt of the compounds.

2. A process for preparing the compounds of the formula (I) (I) wherein n is 2 and X is hydroxyl which comprises reacting with wherein p, m, R, R1, R2, R1', R2' and R3 are as defined in claim 1.

3. A process for preparing 5-[(2-amino-3-carboxymethylamino-3-oxopropyl)thio]-4-hydroxy-6(Z)-nonadecenoic acid which comprises reacting methyl 4,5-epoxy-6(Z)-nonadecenoate with 2-trifluoro-acetamido-3-carbomethoxymethylamino-3-oxopropylmercaptan and converting the resultant compound with aqueous base to the desired product.

4. A process for preparing 4-hydroxy-5-[(2-carboxyethyl)thio]-6(Z)-nonadecenoic acid which comprises reacting methyl 4,5-epoxy-6(Z)-non-adecenoate with methyl-3-mercaptopropionate and converting the resultant compound with aqueous base to the desired product.

5. A process for preparing the compounds of the formula (I) (I) wherein n is 1 and X is hydroxyl and m, p, R, R1 and R2 are as defined in claim 1, which comprises reacting with CH3COR3, wherein m, p, R3, R1' and R2' are as defined in claim 1, and then optionally converting R1', R2' and R3 into R1, R2 and R respectively by treatment with an aqueous base.

6. A process for preparing 3-hydroxy-4-[(2-carboxyethyl)thio]-5(Z)-octadecenoic acid which comprises reacting 2-[(2-carboxyethyl)thio]-hexadec-3(Z)-enal with methyl acetate and converting the resultant compound with aqueous base to the desired product.

7. A process for preparing the compounds of the formula (I) (I) wherein X is hydrogen and m, n, p, R, R1 and R2 are as defined in claim 1, which comprises reacting with CH3(CH2)pCH=P? wherein m, n, p, R3, R1' and R2' are as defined in claim 1.

8. A process for preparing 4-[(2-carboxyethyl)thio]-5(Z)-octadecenoic acid which comprises reacting methyl-4-[(2-carbomethoxyethyl)thio]-4-formylbutanoate with tridecyltriphenylphosphonium ylid and converting the resulting compound in aqueous base to the desired product.

9. A compound of the formula (I) (I) wherein n is 1 or 2; m is 0, 1 or 2; p is 9, 10, 11, 12 or 13; X is hydrogen or hydroxyl; R is hydroxyl or amino;

R1 is hydrogen, amino or ; R2 is hydroxyl, amino -NHCH2CO2H, or -NHCH2CONH2 with the proviso that when m is 0, R1 is hydrogen or a pharmaceutically acceptable salt thereof whenever prepared by the process of Claims 1 or 2, or by any obvious chemical equivalent thereof.

10. A compound of the formula (I) (I) wherein n is 1 or 2 m is 0, 1 or 2 p is 9, 10, 11, 12 or 13; X is hydrogen or hydroxyl; R is hydroxyl or amino;

R1 is hydrogen, amino or ; R2 is hydroxyl, amino -NHCH2CO2H, or -NHCH2CONH2 with the proviso that when m is 0, R1 is hydrogen or a pharmaceutically acceptable salt thereof whenever prepared by the process of Claims 5 or 7 or by any obvious chemical equivalent thereof.

11. 5-[(2-Amino-3-carboxymethylamino-3-oxo-propyl)thiol-4-hydroxy-6(Z)-nonadecenoic acid whenever prepared by the process of Claim 3 or any obvious chemical equivalent thereof.

12. 4-Hydroxy-5-[(2-carboxyethyl)thio]-6(Z)-nonadecenoic acid whenever prepared by the process of Claim 4 or any obvious chemical equivalent thereof.

13, 3-Hydroxy-4-[(2-carboxyethyl)thio]-5(Z)-octa-decenoic acid whenever prepared by the process of Claim 6 or any obvious chemical equivalent thereof.

14. 4[(2-Carboxyethyl)thio]-5(Z)-octadecenoic acid whenever prepared by the process of Claim 8 or any obvious chemical equivalent thereof.