Note: Descriptions are shown in the official language in which they were submitted.
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 -
1 1
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;
0
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
-- 3 --
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
-- 4 --
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;
CH3
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
-- 5 --
~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
-- 6 --
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
-- 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
-- 8 --
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
-- 10 --
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
:
:, :,
:: - :, -,
,.. ~ - .~
- 11 -
(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:
Image (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
- 12 -
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
- 13 -
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
- 14 -
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
-- 15 --
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~
- 16 -
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
- 17 -
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~
- 18 -
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
-- 19 --
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
- 20 -
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.
. :
-~ ;
. ~ .
32917
- 21 -
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
- 22 -
1 EXAMPLE 2
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.
EXAMPLE 3
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
: ~'
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,
.
- :1232~7
- 23 -
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.
XAMPLE 4
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
NCH2C02H
~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.
EXAMPLE 6
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~
- 24 -
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.
EXAMPLE 7
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.
EXAMPLE 8
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
-- 26 --
1 EXAMPLE 9
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
- 27 -
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.
EXAMPLE 10
~ ;
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~
- 28 -
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
:
:
j; , .
:;'' ~'' ~
~L~3~
- 29 -
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
- 30 -
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.
EXAMPLE 11
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
- 31 -
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
- 32 -
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
- 33 -
.
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.
EXAMPLE 12
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
- 34 -
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.
EXAMPLE 13
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.
EXAMPLE 14
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
- 35 -
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.
EXAMPLE 15
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
- 36 -
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.
EXAMPLE 16
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
- 37 -
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
- 38 -
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
,
- 39 -
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.
EXAMPLE 17
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~
EXAMPLE 18
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.
'
:
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