Note : Les descriptions sont présentées dans la langue officielle dans laquelle elles ont été soumises.
wo 91/~880 2 ~ 8 3 9 ~ 8 Pcr/~s91/03~99
Benzoic Acid Derivatives For
Treating Leukotriene-related Diseases
Scope of the Tnvention
This invention relates to certain substituted pyridyl~
hydroxyethyl)benzoic acid derivatives and their ketone analo~s which
are useful for treating diseases associated with leukotrienes. These
compounds are particularly useful in treating diseases attributable to
~he hydroxyleukotrienes, especially LTB4 and LTB4-agonist active
substances .
Back~round of the Invention
The family of bioactive lipids known as the leukotrienes exert
pharmacological effects on respiratory, cardiovascular and
gastrointestinal systems. The leukotrienes are generally divided into
two sub-classes, the peptidoleukotrienes (leukotrienes C4, D4 and E1)
and the hydroxyleukotrienes (leukotriene B4). This invention is
primarily concerned with the hydroxyleukotrienes (LTB) bul is not
limited to this specific group of leukotrienes.
The peptidoleukotrienes are implicated with the biological
response associated with the "Slow Reacting Substance of
Anaphylaxis" (SRS-A). This response has been expressed in vivo as
prolonged bronchoconstriction, in cardiovascular effects such as
coronary artery vasoconstriction and numerous other biological
responses. The pharmacology of the peptidoleukotrienes include
smooth muscle contractions, myocardial depression, increased
2 ~ vascular permeability and enhanced mucous production.
By comparison, LTB4 exerts its biological effects through
stirnulation of leukocyte and lymphocyte functions. It stimulates
chemotaxis, chemokinesis and aggregation of polymorphonuclear
leukocytes (PMNs).
Leukotriene B4 (LTB4) was first described by Borgeat and
Samuelsson in 1979, and later shown by Corey and co-wor}cers to be
;5(S),12(R)-dihydroxy-(Z,E,E,Z)-6,8,10,14-eicosatetraenoic acid.
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2 0 8 ~ 2 ~
It is a product of the arachidonic acid cascade that results from the
enzymatic hydrolysis of LTA4. It has been found to be produced b\~
mast cells. polymorphonuclear leukocytes, monocytes and
macrophages. LTB4 has been shown to be a potent stimulus in vi~
5 for PMN leukocytes, causing increased chemotactic and chemokinetic
migration, adherence, aggregation, degranulation, superoxide
production and cytotoxicity. The effects of LTB4 are mediated through
distinct receptor sites on the leukocyte cell surface which exhibit a
high degree of stereospecificity. Pharmacological studies on human
l 0 blood PMN leukocytes indicate the presence of two classes of LTB4-
specific receptors that are separate from receptors specific for the
peptide chemotactic factors. Each of the sets of receptors appear to be
coupled to a separate set of PMN.leukocyte functions. Calcium
mobilization is involved in both mechanisms.
l 5 LTB4 has been established as an inflammatory mediator in ~iv~
It has also been associated with airway hyperresponsiveness in the
dog as well as being found in increased levels in lung lavages from
humans with severe pulmonary dysfunction. In addition, as with the
other leukotrienes, LTB4 has been implicated in inflammatory bowel
2 0 disease, rheumatoid arthritis, gout, and psoriasis They are critically
involved in mediating many types of cardiovascular, pulmonary,
dermatological, renal, allergic, and inflammatory diseases including
asthma, adult respiratory distress syndrome, cystic fibrosis, psoriasis,
and inflammatory bowel disease
2 5 By antagonizing the effects of LTB4, or other pharmacologically
active mediators at the end organ, for example airway smooth muscle,
the compounds and pharmaceutical compositions of the instant
invention are valuable in the treatment of diseases in subjects,
including human or animals, in which leukotrienes are a key factor.
3 0 SUMMARY OF T~E INVENT~ON
The compounds of this invention are represented by formula (I)
Rl)~LC}~2 -T~ R~ (I )
3 5 or a pharmaceutically acceptable salt or N-oxide thereof where
T is CO or CH(OH)
.
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f WO 91/18880 2 0 8 3 9 ~ 8 P(~ 591/03?~99
R is Cl to C20-aliphatic, unsubstituted or substituted phenyl C
to C l o -aliphatic where substituted phenyl has one or more radicals
selected from the group consisting of lower alkoxy, lower al~yl~
trihalomethyl, and halo, or R is Cl to C20-aliphatic-O-, or R is
5 unsubstituted or substituted phenyl Cl to Clo-aliphatic-O- where
substituted phenyl has one or more radicals selected from the group
consisting of lower alkoxy, lower alkyl, trihalomethyl, and halo;
R1 is -(C1 to Cs aliphatic)R3,-(C1 to C5 aliphatic)CHO, -(C1 to C5
aliphatic)CH20R7, R3, -CH20H or -CHO;
R2 and R3 are independently -COR4 where R4 is -OH, a
pharmaceutically acceptable ester-forming group -ORs, or -OX where
X is a pharmaceutically acceptable cation, or R4 is -N(R6)~ where R6 is
H, or an aliphatic group of 1 to 10 carbon atoms or a cycloalky]-
(CH2)n- group of 4 to 10 carbons where n is 0-3 or both R6 groups
15 form a ring having 4 to 6 carbons, or R2 is an amine, amide or
sulfonamide; and
R7 is hydrogen, C1 to C6-alkyl, or Cl to C6-acyl.
In another aspect, this invention covers pharmaceutical
compositions containing the instant compounds and a
2 0 pharmaceutically acceptable excipient
Treatment of diseases related to or caused by leukotrienes,
particularly LTB4, or related pharmacologically active mediators at
the end organ, are within the scope of this invention This treatment
can be effected by administering one or more of the compounds of
2 5 formula I alone or in combination with a pharmaceutically acceptable
excipient in an amount sufficient to prevent disease or treat it once it
has occurred
In yet another aspect, this invention relates to a method for
' making the compounds of this invention This aspect of the invention
3 0 is illustrated in the reaction schemes given below and in the examples
set forth in this specification.
DETAILED DESCRIPTION OF THE INVENT~ON
- The following definitions are used in describing this invention` and setting out what the inventors believe to be their invention
3~5 herein.
"Aliphatic" is intended to include saturated and unsaturated
radicals. This includes normal and branched chains, saturated or
mono or poly unsaturated chains where both double and triple bonds
may be present in any combination. The phrase "lower alkyl" means
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an alkyl group of I to 6 carbon atoms in any isomeric form, but
particularly the normal or linear form. "Lower alkoxy" means the
group lower alkyl-O-. "Halo" means fluoro, chloro, bromo or iodo.
"Acyl" means the radical having a terminal carbonyl carbon.
When reference is made to a substit1 ted phenyl ring, it is meant
that the ring can be substituted with one or more of the named
substituents as may be compatible with chemical synthesis. Multiple
substituents may be the same or different, such as where there are
three chloro groups, or a combination of chloro and alkyl groups and
further where this latter combination may have different alkyl
radicals in the chloro/alkyl substituent pattern.
The phrase "a pharmaceutically acceptable ester-forming group"
in R2 and R3 covers all esters which can be made from the acid
function(s) which may be present in these compounds. The resultant
esters will be ones which are acceptable in their application to a
pharmaceutical use. By that it is meant that the mono- or diesters
will retain the biological activity of the parent compound and will not
have an untoward or deleterious effect in their application and use in
treating diseases. Such esters are, for example, those formed with one
of the following radicals representing Rs: C1 to C1o alkyl, phenyl-C]-
C6 alkyl, cycloalkyl, aryl, arylalkyl, alkylaryl, alkylarylalkyl,
aminoalkyl, indanyl, pivaloyloxymethyl, acetoxymethyl,
propionyloxymethyl, glycyloxymethyl, phenylglycyloxymethyl~ or
thienylglycyloxymethyl. Aryl includes phenyl and naththyl, or
2 5 heteroaromatic radicals like furyl, thienyl, imidazolyl, triazolyl or
tetrazolyl. The most preferred ester-forming radicals are those where
Rs is alkyl, particularly alkyl of 1 to 10 carbons, [ie CH3-(CH2)n-
where n is 0-9], or phenyl-(CH2)n- where n is 0-4.
When R2 is referred to as being an amine, that includes the
3 0 radical -NH2 and mono- or dialkylate derivatives of this -NH2 radical.
Preferred alkylated amines are the mono- or disubstituted amines
having 1 to 6 carbons. When R2 is referred to as being an amide, that
includes all acylate derivatives of the NH2 radical. The preferred
amides are those having 1 to 6 carbons.
3 5 Where there is an acid group, amides may be formed. The most
preferred amides are those where -R6 is hydrogen or alkyl of I to 6
carbon atoms. Particularly preferred is the diethylamide.
The hydroxyl group of the 2-hydroxyethylene linking group
rnay be esterified. Lower alkyl acids of I to 6 carbon atoms may be
,
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WO 91/188X0 2 0 8 3 9 S 8 PC~/US91/03399
used to form such esters using standard reaction conditions. This
hydroxyl group also may be converted to an ether if so desired.
Again, such reactions are well known in the synthetic chemical arls.
Pharmaceutically acceptable salts of the instant compounds are
5 also intended to be covered by this invention. These salts will be ones
which are acceptable in their application to a pharmaceutical use. B~
that it is meant that the salt will retain the biological activity of the
parent compound and the salt will not have untoward or deleterious
effects in its application and use in treating diseases.
Pharmaceutically acceptable salts are prepared in a standard
manner, The parent compound in a suitable solvent is reacted with
an excess of an organic or inorganic acid, in the case of acid addition
salts of a basic moiety, or an excess of organic or inorganic base in the
case where R4 is OH, Representative acids are hydrochloric acid,
hydrobromic acid, sulfuric acid, phosphoric acid, acetic acid, maleic
acid, succinic acid or methanesulfonic acid, Cationic salts are readily
prepared from alkali metal bases such as sodium, potassium, calcium,
magnesium, zinc, copper or the like and ammonia, Organic bases
include the mono or disubstituted amines, ethylene diamine,
2 0 piperazine, amino acids, caffeine, tromethamine, other tris compounds
and the like,
Oxides of the pyridyl ring nitrogen may be prepared by means
known in the art and as illustrated herein. These are to be considered
part of the invention,
2 5 If by some combination of substituents, a chiral center is
created or another form of an isomeric-center is created in a
compound of this invention, all forms of such isomer(s) are intended
to be covered herein, Compounds with a chiral center may be
administered as a racemic mixture or the racemates may be separated
'` 3 0 and the individual enantiomer used alone,
As leukotriene antagonists, these compounds can be used in
treating a variety of diseases associated ~vith or attributing their
origin or affect to leukotrienes, particularly LTB4, Thus it is expected
that these compounds can be used to treat allergic diseases of a
3 5 pulmonary and non-pulmonary nature, For example these
compounds will be useful in antigen-induced anaphylaxis, 'f ney are
useful in treating asthma and allergic rhinitis, Ocular diseases such as
uveitis, and allergic conjunctivitis can also be treated with these
compounds .
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WO 91/188X0 2~ig~ ' PCr/~'S91/03399
The preferred compounds of this invention are those where R is
alkoxy, particularly alkoxy of 8 to 15 carbon atoms or substituted or
unsubstituted phenyl Cl to C1o-aliphatic-O-; R1 is -(C1 to Cs
aliphatic)R3 or -(Cl to C5 aliphatic)CH20R7, and R2 is -COOH or N(A)(B)
5 where A is ~, or alkyl of 1 to 6 carbons and B is ~, alkyl of 1 to 6
carbons, acyl of I to 6 carbons or -SO2R8 where R8 is -CF3, Cl to C6
alkyl or phenyl. The more preferred compounds of this invention are
those where R is alkoxy of 8 to 15 carbon atoms or alkoxy-substituted
phenyl-C~ to Cg-alkoxy; Rl is COR4, -CH2CH2COR3 or -CH=CH-COR3; and
10 R2 is -COOH or -NHSO2Rg, particularly where R2 is at the meta position
and R8 is -CF3.
The most preferred compounds are set out in Figure Il.
Figure II
R~ R2
_ ,
T R R1 -R2
H2IC10-- **HOOC~H-CH- m-COOH
..H~C-O-Ph-(CH2)~-O- ll ll
.
H21 Clo O HOOCCH2C~2-
_ H2]C10-O- **HOOCCH=CH p-COOH
* The methylene carbon is substituted on the pyridyl ring.
** Trans configuration.
2 0 Svnthesis
These compounds may be made by the star~ing materials,
intermediates and reagents and the synthetic steps set out in the
following reaction flow charts. These charts trace the path used to
.~ make these compounds and are based on the detailed chemistry set
2 5 out in the Examples recited below. These flow charts are intended to
act as a road map to guide one from known starting materials to the
desired products. These specific starting materials, intermediates and
` reagents are only given to illustrate the general case and are not
: ~ intended to limit the chemistry illustrated thereby. Reagents,
3 ~ intermediates, temperatures, solvents, reaction times, wor};-up
procedures all may be varied to accommodate differences and
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optimize the particular conditions for making a particular compound.
Such variations will be apparent to a chemist or will not require more
than minimal experimentation to optimize conditions and reagents for
a particular step.
These reaction schemes first illustrate how to make certain
portions of the R group which are not commercially available. then
illustrate a means to assemble the whole compound using the
materials frorn Reaction Scheme l or commercially available R-
forming groups.
l 0 The preparation of certain embodiments of R are given in
Scheme l.
Scheme l (a)
OH KAPA OH l-BU(Ph)~SiCI
(a)
l 5
H3CO-Ph-l
----(CH2)nOSi(Ph)2-t-Buph2 ~
(b) Pd [(Ph)3P]2 Cl2
, Pd-C
H3co~(cH2)nosi(ph)2-t-Buph2
(c)
H3co~(cH2)n~2osi(Ph)2~t-su Bu4NF
(d)
H3CO~(CH2)n+2-OH TsCI ~ H3Co~(CH2)n+2-OTs
(e) (f)
In those instances where an c~-yn- l -ol is not commercially
2 5 available, it can be prepared from a corresponding 3-yn-l -ol by
treating the alcohol with a strong base. An alkali metal amide may be
used. The alcohol is then protected in order to add the desired phenyl
group at the terminal triple bond. A silyl ether was formed in this
instance; it illustrates the general case. A halo-substituted-phenyl
30 adduct is used to add the phenyl group at the triple bond. The silyl
group is removed and the resulting alcohol converted to the tosylate,
- .
wo91/18880 2083n5~ PC:r/US91tO3399
or another group which is sufficiently reacti~, e so as to provide read~
formation of an ether later in the synthesis of these compound.
Scheme I(b) illustrates another method for malcing certain
alkoxy-substitutedphenylalkoxy R groups.
Scheme I (b)
H3CO~} (Ph)3P=CH(CH2)3C02- CH o~ H
(a)
1 0
TsCI
LiA l H4 ~,OH p y r .
(b)
CH30~-- , ,0T~
1 5 (c)
While the methoxyphenyl compound is illustrated here, this
series of steps and reagents may be used to make other
c~-(unsubstituted)phenylaliphatic or ~o-(substituted)phenylaliphatic
groups denoted by R. .The starting material, the benzaldehydes, are
2 0 commercially available or can be readily made by known methods
To make the acid, first an alkylsilazide is added to an inert
solvent under an inert atmosphere. Then the phosphonium salt is
added. This addition can be done at room temperature or
~` thereabouts. After a brief period of mixing, this mixture is usually a
2 5 suspension, the benzaldehyde is added slowly at about room
temperature. A slight molar excess of the phosphonium salt is
employed. After an additional brief period of stirring at about room
temperature, the reaction is quenched with water. The solution is
acidified and the acid (a) extracted with a suitable organic solvent.
3 0 Further standard separatory and purification procedures may be
employed as desired.
The alcohol is made by reducing the acid using a reducing agent.
Lithium alurninum hydride or similar reducing agents may be
employed and conditions may be varied as needed to effect the
3 5 reduction.
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The tosylate is prepared in an inert solvent employing
p-toluenesulfonylchloride and a base such as pyridine. Suitable
conditions include carrying out the reaction at room temperature or
thereabouts for a period of I to 5 hours. Other suitable leaving
5 groups similar in function to the tosylate may be prepared and will be
useful as a means for adding this R moiety to the pyridyl ring.
Compounds of formula I can then be synthesized by the
sequence of steps outlined in the following schemes.
Scheme 2
1 oxidation
HO~ 2 Rl/K~CO3 RO~3~ 2 m-io~obenzaldehyde
HO I~ N~ 3. hydrazine / 1 00C~ N 3. Pd(OAc)2 / dppf / CO
4. NiO2 N: N MeOH / DMSO / TEA
(2a)
RO~3,co M 1Br2/CH2C12 ~ ~ CO2Me
(2b) (2c)
1 5
1 (C6H5)3PCHCO2Me, RO~ OH
, HO2C~N ~CO2H
. (2d)
First 2,6-lutidine-a2,3-diol is oxidized to the 3-hydroxy-6-
methyl-2-pyridine carboxaldehyde. This aldehyde is then treated
2 0 with a I -halosubsti~uted group which adds to the 3-hydroxy group to
form an ether. This reaction is effected by base, for example a
carbonate such as K2CO2. Hydrazine hydrate is then used to form an
aminohydrazone. This reaction is carried out at an elevated
temperature. The reaction mixture is then cooled and treated with a
2 5 base before recovering the aminohydrazone. This hydrazone is then
- converted to a triazolo[l,S-a]pyridine(2a~ by means of NiO2 or another
oxidizing agent such as KFe(CN)6. If nickel peroxide is used, ~he
reaction can be effected at room temperature or thereabouts, though
it may require an extended reaction time. For the nickel peroxide
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process, an inert atmosphere is preferred, as are dry conditions.
Other oxidizing agents may require elevated temperatures~
The 2-hydroxyethyl product is then made by first preparin( i n
situ a reagent capable of extracting a proton from the triazolopyridine
5 c:ompound after which the triazolo compound is added followed by a
halobenzaldehyde. A useful base is lithium diisopropylamide. It is
preferable to prepare it at reduced temperatures, i.e. -40 to 0C or
thereabouts. After the triazolopyridine and benzaldehyde are added,
the reaction is al]owed to run its course at room temperature or
10 thereabouts. A carbonylation reaction is then carried out to introduce
a carboxyl group into the phenyl ring. This is effected by Pd(OAc)
and gaseous carbon monoxide in an appropriate solvent, preferabl~ at
an elevated temperature, i.e. 50-100C. This gives the carbomethox)~-
substituted phenyl compound (2b).
Treating the resulting triazolo compound with Br~ destroys the
triazole ring and brominates the resulting 2-position carbon on the
pyridine ring to afford the 2-(a,-dibromomethyl)pyridyl adduct.
This reaction is best carried out at reduced temperature, i.e. about 0C
and is complete in about an hour or so. Silver nitrate oxidation gives
aldehyde (2c). A Wittig reaction is then carried out to form the
carbomethoxyethylene group at position 2 on the pyridyl ring This
compound can be treated with a base to hydrolyze the esters, which is
then acidified if the free acid (2d) is desired.
Alternatively, the ethylene group at position 2 can be saturated
2 ~ by catalytic hydrogenation, then saponified using a base, which gives
the salt, or thereafter acidifying the soluton to obtain the free acid
(see Scheme 3 below). The acid can be converted to a
pharmaceutically acceptable salt or esterified by known means.
Amides can be made from the acids using known procedures.
3 0 Analogs of the compounds in Scheme 2 where Rl is an alkanoic
acid can be made by simply hydrogenating the unsaturated bonds in
that chain. Such process is illustrated in Scheme 3.
Scheme 3
H2~C~oO~ OH 1. H~ 21C10~,~, OH
MeO2C~ N ~co2Me 2- LiOH HO C--J` N ~CO2H
,.......................... . .
wo 91/1888() 2 0 ~ 3 ~ 5 8 PCr/US91/03399
11
Reducing the double bond is effected by catalytic mean using a
Iheavy metal catalyst and hydrogen gas. Mild conditions will suffice.
The illustra~ed esters can be hydrolyzed with base and further
converted to other forms of formula I from there or
5 transesterification can be used to convert to another ester.
Compounds where R I contains a terminal -OH group, or an ester
thereof, can be prepared by the series of steps given in Scheme 4.
Scheme 4
H~1C10~,~ OH 1 Br2
~ ~ 1 2 AgNO~ H21Cl0~ OH
N ,N ~ 3 (C6Hs)3pcHco2Me HO~
1 0 (4a)
1. Pd~OAc)2 / dppt / CO H21CloO~ OH
2 LiOH HO~ N l~`J~CO2H
(4b)
Starting material is derived from Scheme 2, then carried
through that set of steps, except that the R2 carbomethoxy function is
not introduced until after the Rl alcohol has been prepared. Also,
separately and apart from the steps in Scheme 2, the Rl
carbomethoxy group can be reduced to the alcohol using a reducing
2 0 agent such as diisobutylaluminum hydride (DIBAL) or a similar
reducing agent. Catalytic hydrogenation can be used to saturate the
ethylene group at position 2 on the pyridyl ring. A base can be used
to saponify the ester to obtain the acid salt, or that salt can be
acidified if the free acid is desired.
2 5 Each of the products containing an hydroxyl group in Schemes
2-4 can be oxidized to the corresponding ketone, that is where T is -
CH2C(O)-, by means of a mild oxidizing agent.
. -
~ Formu l ati on s
3 0 Pharmaceutical compositions of the present invention comprise
a pharmaceutical carrier or diluent and an amount of a compound of
the formula (I) or a pharmaceutically acceptable salt, such as an alkali
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metal salt thereof, sufficient to produce the inhibition of the effects of
leukotrienes.
When the pharmaceutical composition is employed in the form
of a solution or suspension, examples of appropriate pharmaceutical
5 carriers or diluents include: 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, lactose, kaolin and mannitol;
and for aerosol systems, dichlorodifluoromethane,
l 0 chlorotrifluoroethane and compressed carbon dioxide. Also, in
addition to the pharmaceutical carrier or diluent, 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
15 have a detrimental effect on the therapeutic action of the instant
compositions.
The nature of the composition and the pharmaceutical carrier or
diluent will, of course, depend upon the intended route of
administration, for example parenterally, topically, orally or by
2 0 inhalation.
In general, particularly for the prophylactic treatment of
asthma, the compositions will be in a form suitable for administration
by inhalation. Thus the compositions will comprise a suspension or
solution of the active ingredient in water for administration by means
2 5 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
3 0 diluent for administration from a powder inhalation device, In the
- above compositions, the amount of carrier or diluent will vary butpreferably will be the major proportion of a suspension or solution of
the active ingredient, When the diluent is a solid it may be present in
lesser, equal or greater amounts than the solid active ingredient.
3 ~ For parenteral administration the pharmaceutical composition
will be in the form of a sterile injectable liquid such as an ampule or
; an aqueous or nonaqueous liquid suspension.
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. wo 91/18X8~ 2 0 ~ 3 ~ ~ 8 Pcr/us9l/o33ss
For topical administration the pharmaceutical composition will
be in the form of a cream, ointment, liniment, lotion, pastes, and drops
suitable for administration to the eye, ear, or nose.
For oral administration the pharmaceutical composition will be
5 in the form of a tablet, capsule, powder, pellet, atroche, lozenge, syrup,
liquid, or emulsion.
Usually a compound of formula I is administered to a subject in
a composition comprising a nonto~ic amount sufficient to produce an
inhibition of the symptoms of a disease in which leukotrienes are a
10 factor. When employed in this manner, the dosage of the composition
is selected from the range of from 50 mg to 1000 mg of active
ingredient for each administration. For convenience, equal doses will
be administered l to 5 times daily with the daily dosage regimen
being selected from about 50 mg to about 5000 mg.
l 5 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
treating a disease mediated by LTB4 which comprises administering
2 0 to a subject a therapeutically effective amount of a compound of
formula I, preferably in the form of a pharmaceutical composition.
For example, inhibiting the symptoms of an allergic response resulting
from a mediator release by administration of an effective amount of a
compound of formula I is included within the scope of this disclosure.
2 5 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 symptoms is specifically required. However,
the method is also usefully carried out as continuous or prophylactic
- treatment. It is within the skill of the art to determine by routine
3 0 experimentation the effective dosage to be administered from the
dose range set forth above, taking into consideration such factors as
the degree of severity of the condition or disease being treated, and so
', forth.
Pharmaceutical compositions and their method of use also
3 5 include the combination of a compound of formula I with Hl blockers
where the combination contains sufficient amounts of both
compounds to treat antigen-induced respiratory anaphylaxis or
similar allergic reaction. Representative Hl blockers useful here
include cromolyn sodium, compounds from the ethanolamines
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WO 9l/l8~8n ' PCI/IJS91/03399
2083~ l4
(diphenhydramine), ethylenediamines (pyrilamine), the al~;ylamines
(chlorpheniramine), the piperazines (chlorcyclizine), and the
phenothiazines (promethazine). H ~ blockers such as 2-[4-(5-bromo-~-
methylpyrid-2-yl)butylamino]-S-[(6-methylpyrid-3-yl)metllyl]-4-
pyrimidone are particularly useful in this aspect of the invention.
Bioassays
The specificity of the antagonist activity of a number of the
compounds of this invention is demonstrated by relatively low levels
of antagonism toward agonists such as potassium chloride, carbachol,
1 0 histamine and PGF2-
The receptor binding affinity of the compounds used in the
method of this invention is measured by the ability of the compounds
to bind to [3H]-LTB4 binding sites .on human U937 cell membranes.
The LTB4 antagonists activity of the compounds used in the method of
1 5 this invention is measured by their ability to antagonize in ~ dose
dependent manner the LTB4 elicited calcium transient measured with
fura-2, the fluorescent calcium probe. The methods employed were
as follows.
U937 Cell Culture Conditions
2 0 U937 cells were obtained from Dr. John Bomalaski (Medical
College of PA) and Dr John Lee (SK&F, Dept. of Immunology) and
grown in RPMI-1640 medium supplemented with 10% (v/v) heat
inactivated fetal calf serum, in a humidified environment of 5% CO2,
95% air at 37C. Cells were grown both in T-flasks and in Spinner
culture. For differentiation of the U937 cells with DMSO to monocyte-
like cells, the cells were seeded at a concentration of 1 x 105 cells/ml
in the above medium with 1.3% DMSO and the incubation continued
for 4 days. The cells were generally at a density of 0.75-1.25 x 106
cells/ml and were harvested by centrifugation at 800 x g for 10 min .
3 0 Preparation of U937 Cell Membrane Enriched Fraction
Harvested U937 cells were washed with 50 mM Tris-HCI, pH 7.4
at 25C containing 1 mM EDTA (buffer A). Cells were resuspended in
buffer A at a concentration of 5 x 107 cells/ml and disrupted by
nitrogen cavitation with a Parr bomb at 750 psi for lO min at 0C.
The broken cell preparation was centrifuged at 1,000 x g for 1() min.
The supernatant was centrifuged at 50,000 x g for 30 min. The pellet
was washed twice with buffer A. The pellet was resuspended at
about 3 mg membrane protein/ml with 50mM Tris-HCl, pH 7.4 at 25C
and aliquots were rapidly frozen and stored at -70C.
WO 91/1X880 2 0 8 3 9 ~ 8 PCr/l!S9l /03399
15
Bindin~ of ~~Hl-LTB4 to U397 Membrane Receplors
13H]-LTB4 binding assays were performed at 25C, in 50 mM
Tris-HCI (pH 7.5) buffer containing lO mM CaC12, lO mM MgC12, [3H]-
LTB4, U937 cell membrane protein (standard conditions) in the
5 presence (or absence of varying concentrations of LTB4, or SK&F
compounds Each experimental point represents the means of
triplicate determinations, Total and non-specific binding of [3H]-LTB4
were determined in the absence or presence of 2 11 M of unlabeled
LTB4, respectively. Specific binding was calculated as the difference
10 between total and non-specific binding. The radioligand competition
experiments were performed, under standard conditions, using
approximately 0.2 nM [3H]-LTB4, 20-40 '~lg of U937 cell membrane
protein, increasing concentrations of LTB4 (O.l nM to lO nM) or other
competing ligands (O.l IlM to 30 ~LM) in a reaction volume of 0.2 ml
l 5 and incubated for 30 minutes at 25C. The unbound radioligand and
competing drugs were separated from the membrane bound ligand by
a vacuum filtration technique. The membrance bound radioactivity
on the filters was determined by liquid scintillation spectrometry.
Saturation binding experiments for U937 cells were performed,
2 0 under standard conditions, using approximately 15-50 llg of U937
- membrane protein and increasing concentrations of [3H]-LTB4 (0.02-
2.0 mM) in a reaction volume of 0.2 ml and incubation at 22~C, for 30
minutes. LTB4 (2 IlM) was included in a separate set of incubation
tubes to determine non-specific binding. The data from the saturation
2 5 binding experiments was subjected to computer assisted non-linear
least square curve fitting analysis and further analyzed by the
method of Scatchard.
Uptake of Fura-2 bv Differentiated U937 Cells
Harvested cells were resuspended at 2 x lO6 cells/ml in Krebs
30 Ringer Hensilet buffer containing 0.1% BSA (RIA grade), l.l mM
MgSO4, 1.0 mM CaCI2 and 5 mM HEPES (pH 7.4, buffer B). The
diacetomethoxy ester of fura-2 (fura-2/AM) was added to a final
concentration of 2 IlM and cells incubated in the dark for 30 minutes
at 37C. The cells were centrifuged at 800 x g for lO minutes and
3 5 resuspended at 2 x 106 cells/ml in fresh buffer B and incubated at
37C for 20 minutes to allow for complete hydrolysis of entrapped
ester. The cells were centrifuged at 800 x g for 10 minutes and
resuspended in cold fresh buffer 13 at 5 x 106 cells/ml. Cells were
. .
. :
,, . . .,i-
WO 91/1888() 2 ~ 8 3 n a 8 PCI/US91/0339~
16
maintained on ice in the dark until used for fluorescent
measuremen ts .
Fluorescent Measurements-Calcium Mobilization
The fluorescence of fura-2 containing U937 cells was measured
S with a fluorometer designed by the Johnson Foundation Biomedical
Instrumentation Group. Fluorometer is equipped with temperature
control and a magnetic stirrer under the cuvette holder. The wave
lengths are set at 339 nm for excitation and 499 nm for emission. All
experiments were performed at 37C with constant mixing.
U937 cells were diluted with fresh buffer to a concentration of l
x 106 cells/ml and maintained in the dark on ice. Aliquots (2 ml) of
the cell suspension were put into 4 ml cuvettes and the temperature
brought up to 37C, (maintained in 37C, water bath for 10 min).
Cuvettes were transferred to the fluorometer and fluorescence
15 measured for about one minute before addition of stimulants or
antagonists and followed for about 2 minutes post stirnulus. Agonists
and antagonists were added as 2 111 aliquots.
Antagonists were added first to the cells in the fluorometer in
order to detect potential agonist activity. Then after about one
2 0 minute 10 nM LTB4 (a near maximal effective concentration) was
added and the maximal Ca2+ mobilization [Ca2+]i was calculated using
the following formula:
[Ca2+Ji = 224{FmaX F}
F was the maximum relative fluorescence measurement of the sample
Fmax was determined by Iysing the cells with 10 1ll of 10% Triton X-
100 (final concentration 0.02%). After Fmax was determined 67 111 of
100 mM EDTA solution (pH 10) was added to totally chelate the Ca2t
30 and quench the fura-2 signal and obtain the Fmin. The [Ca2+]i level
for 10 nM LTB4 in the absence of an antagonist was 100% and basal
~Ca2+]i was 0%. The ICso concentration is the concentration of
antagonist which blocks 50% of the 10 nM LTB4 induced [Ca2+]i
mobilization The ECso for LTB4 induced increase in [Ca2+]i
3 5 mobilization was the concentration for half maximal increase. The K
for calcium mobilization was determined using the formula:
, ~ : . .
,
,
. . .- . . .
wo s1/l8R~o2 0 8 3 n ~ 8 Pcr/~s91/03399
. ,~,. . .
K _ ICso
I ~LTB4]
[EC501
With the experiments described, the LTB4 concen~ration was 10 nM
and the ECso was 2 nM.
Results for compounds tested by these methods are given in
Figure III.
Figure III
Bindin~. IC~o. (Kj). ~IM Ca-Mo~ilization
U-937 PMN U-937 P M N
Structure Membrane Whole Cell Whole cell ~Q~ % Agonist % Agonis
Ex 1 1.6(0.55) 0.77 0.60 3.8 o o
Ex 2 2.~(0.72) 0.74 - 3.4 o O
Ex 3 2.3(0.81) 1.6 - 2.9 o 20
Ex 4 8.8(3.1) 1.2 - 4.0 0 0
Examples
The following set of examples are given to illustrate how to
make and use the compounds of this invention. They are not
intended to circumscribe or otherwise limit the scope of this
invention. Reference is made to the claims for defining what is
reserved to the inventors by this document.
Example A
8-(4-MethoxYphenvl)octan-1 -(4-toluenesulfonate)
':
A(1 ) 7-OctYn-l -ol.
35% KH in mineral oil (27g, 240mmol) under an argon
atmosphere was washed with hexane and treated dropwise with 1,3-
; diaminopropane. The mixture was stirred at room temperature until
it became homogeneous. The flask was cooled to 0C and 3-octyn-1-ol
(lOg, 79mmol, Lancaster Synthesis) was slowly added. The reaction
~ 2 5 was then stirred at room temperature for l 8 hours. The reaction was
`~ quenched with H20 (SOmL) and the product was extracted into ether.
The organic layer was washed with 10% HCl (3XlSmL) and brine and
dried (MgS04). Evaporation gave the title product which was used
without further purification: 1H NMR (9OMHz, CDC13) ~ 3.65 (t, J=SHz,
30 2H, OCH2), 2.23 (m, 2H, CH2), 2.0 (m, IH, acetylenic), 1.7-1.2 (m, 8H,
(CH2)4); IR (neat) omax 3350, 2930, 2125 cm~l.
~'
. . .
:'
: . . , ., - -,
.
. ~ .
" .
wo 91/18880 2 ~ 8 3 ~ ~ ~ PCr/l_'S9l/03399~
18
A(2) 7-OctYn-l-t-butvldiphenylsilYl ether.
7-Octyn- I -ol (3.8g, 30mmol) was dissolved in dimethyl-
formamide (lOmL) and treated with t-butylchlorodiphenylsilane
S (10.2mL, 33mmol) and imidazole (3.65g, 45mmol) at 0C. The
reaction was stirred at 0C for 10 minutes and at room temperature
for 3 hours. Water was added and the product was extracted into
ethyl acetate. The ethyl acetate extract was washed with H2O and
brine and dried (Na2SO4). The solvent was evaporated and the
1 0 residue purified by flash column chromatography (silica, hexanes) to
give a yellow oil: IH NMR (250MHz, CDC13) ~ 7.7 (d, 4H, aryl), 7.4 (m,
6H, aryl), 3.63 (t, 2H, OCH2), 2.23 (m, 2H, CH2), 1.97 (t, lH, acetylenic),
1.6-1.3 (m, 8H, (CH2)4), 1.05 (s, 9H, t-butyl); IR~(film)~max 3321,
2940, 2125 cm~1.
1 5
At3~ 8-(4-MethoxvphenYl~-7-octvn-1-t-butYldiphenvlsilvl ether
To a flame-dried flask under an argon atmosphere was added
4-iodoanisole (5.34g, 22mmol) in triethylamine (SOmL) followed by
the addition of 7-octyn-1-t-butyldiphenylsilyl ether.(9.84g, 27mmol),
2 0 (Ph3P)2PdCl2 (350mg, 0.44mmol), and CuI (200mg, 0.88mmol). The
resulting mixture was heated at 50C for 4 hours. Upon cooling to
room temperature the reaction mixture was filtered and the solvent
evaporated. The residue was partitioned between ethyl acetate and
H 2 and the organic layer was collected and washed with brine and
2 5 dried (Na2SO4). The solvent was evaporated and the residue was
purified by flash column chromatography (silica, I % ethyl acetate in
hexanes) to give an oil: IH NMR (250MHz, CDC13) ~ 7 7 (d, 4H, aryl),
7.4 (m, SH, aryl), 7.35 (d, 2H, aryl), 6.8 (d, 2H, aryl), 3.8 (s, 3H, OCH3),
3.7 (t, 2H, OCH2), 2.4 (t, 2H, CH2), 1.7-1.3 (m, 8H, (CH2)4), 1.05 (s, 9H,
3 0 t-butyl).
A(4! 8-(4-MethoxyphenYl)octan-l-t-butYldiphenYlsilYl ether.
To 8-(4-methoxyphenyl)-7-octyn-1-t-butyldiphenylsilyl ether
(2.2g, 4.6mmol) in ethanol (1 OmL) and ethyl acetate (1 OmL) was
35 added 5% Pd/C (lOOmg). The mixture was subjected to 75 psi of H2
for 4 hours. The reaction was filtered through Celite and the solvent
evaporated to give an oil: -IH NMR (250MHz, CDC13) ~ 7.7 (d, 4H, aryl),
7.4 (m, 6H, aryl), 7.05 (d, 2H, aryl), 6.8 (d, 2H, aryl), 3.8 (s, 3H, OCH3),
. . .
WO 91/18XX(1 2 0 8 ~ 9 ~ 8 pcr/~!s91/o3399
1~
3.6 (t, 2~, OC~-12), 2.5 (t, 2~1, benzylic), 1.75-1.3 (m, 12~, (CH~)6). 1.0 (s~
9H, t-butyl ).
A(5! ~-(4-Methoxvphenvl)octan-l-ol.
8-(4-Methoxyphenyl)octan- I -t-butyldiphenylsilyl ether (2.2g,
4.6mmol) in tetrahydrofuran (20mL) was cooled to O~C and treated
with tetrabutylammonium fluoride (14mL, 14mmol, IM in
tetrahydrofuran). The cooling bath was removed and the reaction
was stirred at room temperature for 24 hours. The reaction was
I O diluted with ethyl acetate and was washed with H20 and brine and
dried (Na2S04) The solvent was evaporated and the residue was
purified by flash column chromatography (silica, 0-20% ethyl acetate
in hexanes) to give a white solid: 1H NMR (250MHz, CDC13) ~ 7.15 (d,
2H, aryl), 6.86 (d, 2H, aryl), 3.85 (s, 3H, OCH3), 3.68 (t, 2H, OCH2)~ 2.6
1 5 (t, 2H, benzylic), 1.75-1.3 (m, 12H, (CH2)6).
A(6! 8-(4-Methoxvphenvl)octan-1-(4-toluenesulfonate).
6-(4-Methoxyphenyl)octan-1-ol (5.9g, 25mmol) was dissolved
in dry CH2C12 (lOOmL) under an argon atmosphere and cooled to 0C.
2 0 To this was added pyridine (2,5mL, 30mmol) and 4-toluenesulfonyl
chloride (5.4g, 28mmol). The reaction was stirred at 0C for 20
minutes and at room temperature for 24 hours. The reaction soluIion
was washed with H20 and brine and dried (Na2S04). The solvent was
evaporated and the residue purified by flash column chromatography
25 (silica, 0-10% ethyl acetate in hexanes~ to give a white solid: IH NM~
(250MHz, CDC13) ~ 7.79 (d, 2H, aryl), 7.35 (d, 2H, aryl), 7.09 (d, 2~1,
aryl), 6.82 (d, 2H, aryl), 4.04 (s, 2H, OCH2), 3.8 (s, 3H, OCH3), 2.55 (t,
2H, benzylic), 2.46 (s, 3H, CH3), 1.75-1.15 (m, 12H, (CH~)6).
3 0 Example B
6-(4-Methoxyphenvl)hexan-1 -(4-toluenesulfonate)
B(1) 5-Hexyn- 1 -t-butyldiphenYlsilYI ether
5-Hexyn-l-ol (3g, 30mmol, Aldrich) was dissolved in
3 5 dimethylformamide (lOmL) and treated with
t-butylchlorodiphenylsilane (10.2mL, 33mmol) and imidazole (3.65g,
45mmol) at 0C. The reaction was stirred at 0C for 10 minutes and at
room temperature for 3 hours. Water was added and the producI was
extracted into ethyl acetate. The ethyl acetate extract was wash~d
:;
.
.
; . .
': .
W O 91/18X80 . . PC~r/~S91/0339920839~8 20 ~
with H20 and brine and dried (Na2S04). The solvent was evaporated
and the residue purified by flash column chromatography (silica,
hexanes) to give a yellow oil: IH NMR (250MHz, CDC13) ~ 7.7 (d, 4H,
aryl), 7.4 (m, 6H, aryl), 3.65 (t, 2H, OCH2), 2.2 (m, 2H, CH2), 1.9 (t, IH.
S acetylenic), 1.7 (m, 4H, CH2-CH2), l.OS (s, 9H, t-butyl).
B(2) 6-(4-Methoxvphenvl)-S-hexvn-l-t-butvldiphenvlsilvl ether.
To a flame-dried flask under an argon atmosphere was added
4-iodoanisole (5.34g, 22mmol) in triethylamine (50mL) followed by
1 0 the addition of S-hexyn-1-t-butyldiphenylsilyl et-her (8.83g, 27mmol),
(Ph3P)2PdC12 (350mg, 0.44mmol), and Cul (200mg, 0.88mmol). The
resulting mixture was heated at 50C for 4 hours. Upon cooling to
room temperature the reaction mixture was filtered and the solvent
evaporated. The residue was partitioned between ethyl acetate and
1 S H20 and the organic layer was collected and washed with brine and
dried (Na2S04). The solvent was evaporated and the residue was
purified by flash column chromatography (silica, I % ethyl acetate in
hexanes) to give an oil: IH NMR (250MHz, CDC13) ~ 7.7 (d, 4H, aryl),
7.4 (m, 6H, aryl), 7.35 (d, 2H, aryl), 6.8 (d, 2H, aryl), 3.8 (s, 3H, OCH3),
20 3.7 (t, 2H, OCH2), 2.4 (t, 2H, CH2), 1.7 (m, 4H, CH2-CH2), 1.05 (s, 9H,
t-butyl),
B(3! 6-(4-Methoxyphenyl!hexan-l-t-butv]diphenvlsilvl ether
To 6-(4-methoxyphenyl)-5-hexyn- 1 -t-butyldiphenylsilyl ether
2 5 (2.0g, 4.6mmol) in ethanol (lOmL) and ethyl acetate (lOmL) was
added 5% Pd/C (lOOmg), The mixture was subjected to 75 psi of H~
for 4 hours. The reaction was filtered through Celite and the solvent
evaporated to give an oil: 1H NMR (250MHz, CDC13) ~ 7.7 (d, 4H, aryl),
7.4 (m, 6H, aryl), 7.05 (d, 2H, aryl), 6.8 (d, 2H, aryl), 3.8 (s, 3H, OCH3),
3 0 3.6 (t, 2H, OCH2), 2.5 (t, 2H, benzylic), l.SS (m, 4H, CH2-CH2), 1.3 (m,
4H, CH2-CH2), 1.0 (s, 9H, t-butyl).
i
~
B(4! 6-(4-Methoxvphenvl!hexan-~-ol.
6-(4-Methoxyphenyl)hexan-l-t-butyldiphenylsilyl ether (2.0g,
3 S 4.6mmol) in tetrahydrofuran (20mL) was cooled to 0C and treated
with tetra~utylammonium fluoride (14mL, 14mmol, 1 M in
tetrahydrofuran). The cooling bath was removed and the reaction
. was stirred at room temperature for 24 hours. The reaction was
diluted with ethyl acetate and was washed with H ~O and brine and
. ~
- i : , . . . . . . .
, . . . .
.~ ;. ~ , -
,~? WO 91/18880 2 0 8 3 9 5 8 PCr/US91/03399
dried (Na2 S 0 4) . The solvent was evaporated and the residue was
purified by flash column chromatography (silica, 0-20% ethyl acetate
in hexanes) to give a white solid: IH NMR (250MHz, CDC13) ~ 7.05 (d,
2H, aryl), 6.8 (d, 2H, aryl), 3.8 (s, 3H, OCH3), 3.65 (t, 2H, OCH2), 2.55 (t,
S 2H, benzylic), 1.6 (m, 4H, CH2-CH2), 1.4 (m, 4H, CH2-CH2).
B(5! 6-(4-Methoxvphenyl)hexan- ] -(4-toluenesulfonate).
6-(4- Methoxyphenyl)hexan-l-ol (5.36g, 25mmol) was
dissolved in dry CH2C12 (lOOmL) under an argon atmosphere and
1 0 cooled to 0C. To this was added pyridine (2.5mL, 30mmol) and 4-
toluenesulfonyl chloride (5.4g, 28mmol). The reaction was stirred at
O C for 20 minutes and at room temperature for 24 hours. The
reaction solution was washed with H20 and brine and dried (Na2S04).
The solvent was evaporated and the residue purified by flash column
1 S chromatography (silica, 0-10% ethyl acetate in hexanes) to give a
white solid: 1H NMR (250MHz, CDC13) ~ 1.6-1.3 (m, 8H, (CH2)4), 2.4 (s,
3H, CH3), 2.5 (t, 2H, benzylic), 3.8 (s, 3H, OCH3), 4.0 (t, 2H, OCH2), 6.80
(d, 2H, aryl), 7.0 (d, 2H, aryl), 7.3 (d, 2H, aryl), 7.8 (d, 2H, aryl).
ExamPle C
E-6-(4-methoxvphenvl)-l -(4-toluenesulfonate)-S-hexene
:
C(1) E-4-Methoxyphenyl-5-hexenoic acid.
To a freshly prepared solution of lithium hexamethyldisilazide
2 S (64mmol) in tetrahydrofuran (30mL), under an argon atmosphere,
was added a suspension of (4- carboxybutyl)triphenylphosphonium
bromide (17.6g, 30mmol) in tetrahydrofuran (45mL) at room
: temperature. The reaction was stirred for l S minutes during which
time the orange-red color of the ylide developed. A solution of
: 3 0 4-anisaldehyde (4.5g, 30mmol) in tetrahydrofuran (30mL) was added
dropwise and stirring was continued for an additional 20 minutes.
The reaction was quenched with H20 (SOmL) and diluted with ether
(30mL). The aqueous layer was acidified to pH 1.0 with 3N HCI and
the product was extracted into ethyl aceta~e (3X50mL). The combined
organic layers were dried (MgS04) and the product was purified by
~lash column chromatography (silica, 1% methanol in CH2C12) to yield
the E-olefln as a solid: lH NMR (200MHz, Cr)C13) S 7.3 (d, 2H, aryl), 6.8
(d, 2H, aryl), 6.3 (d, lH, olefin), 6.0 (m, lH, olefin), 3.8 (s, 3H, OCH3),
2.3 (m, 4H, allylic CH2 and CH2C02), 1.8 (q, 2H, CH2).
- , . . . .. .. .
:.... .. - ; . :
wo gl/18XX0 2 o 8 3 ~ a 8 PCI`/I_'S91/03399
'~2
C(2) E-4-Methoxvphenyl-S-hexen- I -ol.
E-4-Methoxyphenyl-S-hexenoic acid ( 1.1 g, 5.0mmol~ in dry
ether ( I OmL) was slowly added to a suspension of LiAlH4 (240mg,
S 6.0mmol) in ether (lOmL) under an argon atmosphere. The reaction
mixture was refluxed for 45 minutes. Upon cooling to room
temperature the reaction was quenched with H20 (lOmL) followed by
6N H2S04 (7mL). Ethyl acetate (20mL) was added and the organic
layer was separa~ed and dried (MgS04); evaporation gave a white
crystalline solid: mp. 65-66C; lH NMR (200MHz, CDC13) ~ 7.2 (d, 2H,
aryl), 6.8 (d, 2H, aryl), 6.3 (d, lH, olefin), 6.1 (m, lH, olefin), 3.8 (s, 3H~
OCH3), 3.6 (t, 2H, OCH2), 2.2 (q, 2H, allylic), 1.5 (m, 4H, CH2- CH2);
Anal. Calcd. for C13H1802: C, 75.65; H, ~.80, found: C, 75.45; H, 8.95;
MS (CI): 207 (M+H).
C(3) E-6-(4-methoxvphenvl)-1-(4-toluenesulfonate)-5-hexene.
E-4-Methoxyphenyl-S-hexen-1-ol (1.6g, 7.0mmol) was
dissolved in dry CH2C12 (SOmL) under an argon atmosphere and
treated with 4-toluenesulfonyl chloride (7.0g, 36mmol) and pyridine
2 0 (3mL), The reaction solution was stirred at room temperature for 3.5
hours, Water (40mL) was added ~o the reaction and the organic layer
was separated and dried (MgS04). The product was purified by flash
column chromatography (silica, 10% ethyl acetate in hexane) to give
an oil: 1H NMR (200MHz, CDC13) ~ 7,8 (d, 2H, aryl), 7,3 (d, 2H, aryl),
7,2 (d, 2H, aryl), 6,8 (d, 2H, aryl), 6,2 (d, lH, olefin), 6.0 (m, lH, olefin),
s 4,1 (t, 2H, OCH2), 3.8 (s, 3H, OCH3), 2.4 (s, 3H, CH3), 2.1 (q, 2H, allylic ),
1.6 (m, 4H, CH2- CH2); MS (CI): 361 (M+H).
- ~ Example
3 0 2-(E-2-Carboxvethenyl!-3-decv]oxv-6-r2-(3-carboxvphenvl)-2-
hvdroxvlethylpvridine~ dilithium sa]t
, ~
~ 1 (a! 3-Hydroxv-6-methvl-2-pvridine carboxaldehvde.
- 2,6-Lutidine-a2,3-diol (1 .Og, 7.1 8mmol, Aldrich) was suspended
in dry CH2C12 (40mL) and treated with MnO2 (6.1g, 70mmol). The
reaction was stirred at room temperature for 6 hours. The reaction
mixture was filtered through a pad of Celite and the solvent was
removed in vacuo. The aldehyde was used directly in the next step
without further purification: IH NMR (250MHz, CDC13): ~ 10.65 (s, lH.
.. .. . . . . . .
,, ,
,
.
,
, ~- WO 91/18880 2 0 8 3 ~ ~ 8 PCI/US91/03399
. . " 2 3
OH), 10.30 (s, IH, CHO), 7.30 (dd, 2H, 4-pyridyl, S-pyridyl), 2.55 (s, 3H,
CH3)
I (b) 3-Decvloxv-6-methYl-2-pvridine carboxaldehvde.
S 3-Hydroxy-6-methyl-2-pyridine carboxaldehyde obtained
above was dissolved in dry dimethylformamide (1 OmL) and treated
with l-iododecane (2.1mL, 8.62mmol) and anhydrous K2C03 (3.0g,
21.7mmol) under an argon atmosphere. The reaction was heated a
90C for I hour with vigorous stirring. Upon cooling to room
1 0 temperature the reaction mixture was poured into ethyl acetate
(lOOmL); the ethyl acetate solution was washed with H20 (3X20mL)
and brine and dried (MgS04). The solvent was removed under
reduced pressure and the crude product was used directly in the next
step without further purification: IH NMR (250MHz, CDCI3): ~ 10.40
1 S (s, lH, CHO), 7.30 (dd, 2H, 4-pyridyl, S-pyridyl), 4.07 (t, 2H, OCH2),
2.6 (s, 3H, CH3), 1.85-0.90 (m, i9H, aliphatic).
1 (c! 3-DecyloxY-6-methYI-2-pYridine aminohvdrazone
3-decyloxy-6-methyl-2-pyridine carboxaldehyde (2.15g,
7.8mmol) was heated with hydrazine hydrate for 1 hour at 95C.
Upon cooling to room temperature 25% NaOH was added and the
mixture was extracted with ethyl acetate. The organic extract was
washed with H20 and brine and dried (Na2S04). The solvent was
evaporated to give an amorphous solid: 1H NMR (250MHz, CDCl3)
8.75 (broad singlet, 2H, NH2), 7.55 (s, lH, CH-N), 7.10 (d, lH, 5-
pyridyl), 6.95 (d, lH, 4-pyridyl), 3.95 (t, 2H, OCH2), 2.55 (s, 3H, CH3),
1.80-0.90 (m, 19H, aliphatic).
l (d! 4-Decyloxv-7 -methvl- 1,2,3-tri azolor l .S-alDvridine.
3 0 To a flame-dried flask under an argon atmosphere was added 3-
decyloxy-6-methyl-2-pyridine aminohydrazone (2.12g, 7.2mmol) in
dry benzene (30mL). To the resulting solution was added NiO2
(790mg, 8.7mmol). The resulting mixture was stirred at room
-'~ temperature for 72 hours and then filtered through Celite. The
3 5 solvent was evaporated and the residue purified by flash column
chromatography (silica, 10-15% ethyl acetate in hexanes) to give a
white solid: 1H NMR (250MHz, CDC13): ~ 8.2 (s, lH, CH-N), 6.68 (d, lH,
6-pyridyl), 6.4 (d, lH, S-pyridyl), 4.1 (t, 2H. OCH2), 2.8 (s. 3H, CH3),
,
. ~ . ., . - , ,.. - . -: .
.-' . :~ ' ,: .-c.,
"., .. ,. . . ~ . , , : :
: . . . . . .
.. . . . .
~: . ~ ., . - .
, . . . . . .
wo 91/18880 Pcr/~s9l/0339s
2083~:;)8 24 ~
1.90-0.90 (m, 19H~ aliphatic); Anal. Calcd. for Cl7Ha7N3: C, 70.5~; H,
9.40; N, 14.52, found: C. 70.60; H, 9.14; N, 14.47.
l (e) 1 -(3-~odophenvl)-2-(4-decvloxy-1,2 3-triazolo~ l .S-alpvridin-7-
yl)ethan- 1 -ol .
To a flame dried flask under an argon atmosphere was added
diisopropylamine (500mg, 4.9mmol) in dry ether ( I OmL). The
resulting solution was cooled to -40C (CH3CN/dry ice bath) and 2.5M
n-BuLi (1.97mL, 4.9 mmol) was added. The mixture was stirred at -
1 0 40C for 10 minutes followed by the dropwise addition of 4-decyloxy-
7-methyl-1,2,3-triazolo[l,S- a]pyridine (1.3g, 4.4mmol) in dry ether
(40mL) via addition funnel. The resulting red-brick colored mixture
was stirred at - 40C for 6 hours. 3-lodobenzaldehyde (1.15g,
4.9mmol) in ether (30mL) was added in one portion. A color change
1 S from deep-red to yellow was observed. The mixture was allowed to
warm to room temperature over a 2 hour period and then stirred at
room temperature for 12 hours. The resulting reaction mixture was
partitioned between ethyl acetate and H20 and the organic extract
was washed with H20, brine, and dried (Na2S04). The solvent was
2 0 evaporated and the residue purified by flash column chromatography
(silica, 10-30~b ethyl acetate in hexanes) to give the alcohol as a white
solid. A second component was isolated and identified as the
3-substituted triazolopyridine: NMR (250MHz, CDC13): ~ 8.2 (s, lH,
CH-N), 7.80 (s, lH, aryl), 7.59 (d, lH, aryl), 7.35 (d, lH, aryl), 7.07 (t,
lH, aryl), 6.65 (d, lH, 6-pyridyl), 6.4 (d, lH, 5-pyridyl), 5.36 (m, lH,
CH-O), 4.11 (t, 2H, OCH2), 3.64 (dd, lH, Py-CH), 3.45 (dd, lH, Py-CH'),
3.25 (d, lH, OH), 1.88-0.88 (m, 19H, aliphatic).
I(f) 1-(3-Carboxvmethvlphenvl)-2-(4-decvloxv-1~2.3-triazolo-
3 0 ~ l .S-alpvridine-7-vl )ethan - 1 -ol .
To a solution of 1-(3-iodophenyl)-2-(4-decyloxy-1,2,3-
triazolo[1,5-a]pyridine-7-yl)ethan-1-ol (500mg, 0.96mmol) in
dimethylsulfoxide (1 OmL) was added methanol (4mL), triethylamine
(0.3mL, 2.1mmol), Pd(OAc)2 (6.4mg, 0.029mmol), and bis
3 5 diphenylphosphinopropane (11.9mg, 0.029mmol). Carbon monoxide
was bubbled into the mixture for 4 minutes. The mixture was then
heated at 85C under positive carbon monoxide pressure for 6 hours.
The mixture was cooled to room temperature and parlitioned between
ethyl acetate and H20. The organic layer was washed with H20, brine,
. W O 91/188B0 2 ~ 8 3 ~ ~ 8 PC~r/US91/03399
and dried (Na2SO4). The solvent5was evaporated and the residue
purified by flash column chromatography (silica, 5-20% ethyl acetate
in hexanes) to give a white solid: IH NMR (250MHz, CDC13): ~ 8.2 (s,
lH, CH-N), 8.1 (s, lH, aryl), 7.95 (d, lH, aryl), 7.63 (d, lH, aryl), 7.4 (t,
lH, aryl), 6.65 (d, lH, 6-pyridyl), 6.4 (d, lH, 5-pyridyl), 5.45 (m, lH,
CH-O), 4.11 (t, 2H, OCH2), 3.9 (s, 3H, CO2CH3), 3.70 (dd, lH, Py-CH),
3.45 (dd, lH, Py-CH'), 3.25 (d, lH, OH), 1.90-0.88 (m, l9H, aliphatic);
Anal- Calcd- for C26H35N34: C, 68.85; H, 7.78; N, 9.26, found: C, 68.81;
H, 7.73; N, 9.31.
1 0
1(~) 3-Decvloxv-2-(a c~-dibromomethvl)-6-~2-(3-carboxymethv]-
phenvl)-2-hYdroxvlethvlpvridine.
1 -(3-Carboxymethylphenyl)-2-(4-decyloxy- 1,2,3-triazolo-
[1,5-a]pyridine-7-yl)ethan- 1 -ol (130mg, 0.28mmol) was dissolved in
1 5 CH2C12 (3mL) and cooled to 0C. To this was slowly added a solution
of Br2 (46mg, 0.28mmol) in CH2C12 (3mL); gas evolution was
observed and the reaction mixture was stirred at 0C for 1 hour. The
CH2C12 solution was washed with NaHCO3, H2O, and brine and dried
(Na2SO4), The solvent was evaporated to give a yellow oil: 1H NMR
20 (25~MHz, CDC13): ~ 8.1 (s, lH, aryl), 7.92 (d, lH, aryl), 7.63 (d, IH,
aryl), 7.4 (t, lH, aryl), 7.09 (d, lH, 3-pyridyl), 7.07 (s, lH, CHBr2), 7.0
(d, lH, 4-pyridyl), 6.08 (d, lH, OH), 5.25 (m, lH, CH- O), 4.05 (t, 2H,
OCH2), 3.9 (s, 3H, CO2CH3), 3.15 (m, 2H, Py-CH2), 1.90-0.88 (m, 19H,
aliphatic).
1 (h) 3-DecYloxv-6-~2-(3-carboxymethvlphenvl)-2-hydroxYlethYl-2
pyridine carboxaldehvde.
To a solution of 3-decyloxy-2-(a,~-dibromomethyl)-6-12-(3-
carboxymethylphenyl)-2-hydroxy]ethylpyridine (lSOmg, 0.26mmol)
3 0 in ethanol (3mL) was added AgNO3 (90mg, 0.56mmol) in H2O (lmL).
The resulting mixture was heated at reflux for 1 hour. The mixture
was cooled to room temperature and concentrated HCl (1 mL) was
.. added and the precipitated silver salt was removed by filtration. The
. filtrate was evaporated and the residue treated with saturated
''~A 35 NaHCO3. The product was extracted into ethyl acetate and was
washed with H2O and brine and dried (Na2SO4). The solvent was
evaporated and the residue purified by flash column chromatography
(silica, 10-30% ethyl acetate in hexanes) to give a yellow oil:
:-,
.,~. .
.. . .
- .. . . . . . . . .
.. , - ~ ., . . , ..
, . , : . ., , :
~ ~ ~ .: ; , - - : :
wo 9~/1888~ 2 0 8 3 ~ 5 8 - Pcr/us9l/o339s
26
lH NMR (250MHz, CDC13): ~ 10.4 (s, lH, CHO), 8.1 (s, lH, aryl),
7.92 (d, lH, aryl), 7.63 (d, lH, aryl), 7.4 (t, lH, aryl), 7.33 (d, lH, 3-
pyridyl), 7.25 (d, lH, 4-pyridyl), 5.25 (m, lH, CH-O), 5.0 (d, lH, OH),
4.1 (t, 2H, OCH2), 3.9 (s, 3H, CO2CH3), 3.15 (m, 2H, Py-CH2), 1.90- 0.88
S (m, 19H, aliphatic); MS (CI): 277 (M~H).
I (i~ 2-(E-2-Carboxvmethvlethenyl!-3-decvloxv-6-~2-(3-carboxY-
methvlphenvl)-2-hvdroxylethvlpvridine.
3-Decyloxy-6-[2-(3-carboxymethylphenyl)-2-hydroxy]ethyl-2-
1 0 pyridine carboxaldehyde (40mg, 0.09mmol) was dissolved in drybenzene (2mL) under an argon atmosphere. To this was added
methyl (triphenylphosphoranylidene)acetate (60mg, 0.18mmol) and
the resulting mixture was heated a~ 45C for 1 hour. Upon cooling to
room temperature the reaction was diluted with ethyl acetate and
lS was washed with H2O and brine and dried (Na2SO4). The solvent was
evaporated and the residue purified by flash column chromatography
(silica, 15-20% ethyl acetate in hexanes) to give a yellow oil:
1H NMR (250MHz, CDC13): ~ 8.1 (s, lH, aryl), 8.1 (d, J=16Hz, lH,
olefin), 7.9 (d, lH, aryl), 7.65 (d, lH, aryl), 7.4 (t, lH, aryl), 7.15 (d, lH,
20 S-pyridyl), 7.03 (d, lH, 4-pyridyl), 6.95 (d, J=16Hz, lH9 olefin), 5,65 (d,
lH, OH), 5.2 (m, lH, CH-O), 4.05 (t, 2H, OCH2), 3.9 (s, 3H, C02CH3), 3.8
(s, 3H, CO2CH3), 3.10 (m, 2H, Py-CH2), 1.90-0.88 (m, l9H, aliphatic);
MS (CI): 498 (M+H).
1 (j! 2-(E-2-carboxvethenvl!-3-decvloxy-6-r2-(3-carboxyphenvl!-2
hvdroxvle~hvlpvridine. dilithium salt.
2-(E-2-Carboxymethylethenyl)-3-decyloxy-6-[2-(3 -
carboxymethylphenyl)-2-hydroxy]ethylpyridine (22mg, 0.04mmol)
was dissolved in tetrahydrofuran, H2O, and methanol (0.SOmL each)
; 3 0 and treated with LiOH monohydrate (Smg, 0.2mmol). The reaction
- was stilTed at room temperature for 24 hours. The solvent was
'~ evaporated and the residue was dissolved in H2O and purified by
Reversed Phased MPLC (RP-18 silica, 10-40% MeOH in H2O). The
desired fractions were Iyophilized to give a colorless amorphous solid:
lH NMR (250MHz, CV30D): ~ 8.01 (s, lH, aryl), 7.80 (d,lH, aryl),
7.76 (d, J=16Hz, lH, olefin), 7.36 (d, lH, aryl), 7.30 (t, lH, aryl), 7.24
(d, lH, S- pyridyl), 7.07 (d,J=16Hz, lH, olefin), 7.01 (d, lH, 4-pyridyl),
S.11 (t, lH, CH-O), 4.0 (t, 2H, OCH2), 3.1 (m, 2H, Py-CH2), 1.83- 0.89
. . . . . . . .
' ~ ' . ,
: . . : , .
20839S~
~;wo 91/18880 PCr/US91/03399
27
~m, I 9H, aliphatic); FAB-MS: 474.3 (M-ll, monolithium salt). 468 (M-
H, free acid).
Example 2
2-(2-Carboxyethyl)-3-decyloxv-6-~2-(3-carboxYphenYl)-'7-
hydroxYlethYlpvridine. dilithium salt
2(a~ 2-(2-CarboxymethvlethYl~-3-decyloxY-6-~2-(3-carboxYmethvl-
12henvl)-2-hvdroxvlethylpvridine~
To 2-(E-2-carboxymethylethenyl)-3-decyloxy-6-[2-(3-
carboxymethylphenyl)-2-hydroxy]ethylpyridine (13mg, 0.02mmol) in
ethanol (3mL) was added 5% Pd/C (2mg). The mixture was subjected
to 5 psi of H2 for I hour. The mixture was filtered through Celite and
the solvent was evaporated to give an oil:
l 5 lH NMR (250MHz, CDC13): ~ 8.08 (s, lH, aryl), 7.9 (d, lH, aryl),
7.6 (d, lH, aryl), 7.4 (t, lH, aryl), 7,05 (d, lH, 5-pyridyl), 6.87 (d, lH,
4-pyridyl), 6.0 (broad singlet, lH, OH), 5.l5 (m, lH, CH-O), 4.01 (t, 2H,
OCH2), 3.9 (s, 3H, CO2CH3), 3.8 (s, 3H, CO2CH3), 3.2 (t, 2H, CH2), 3.05
(m, 2H, CH2)~ 2.8 (t, 2H, CH2), 1.83-0.88 (m, l9H, aliphatic).
2(b) 2-(2-Carboxvethvl)-3-decvloxY-6-~2-(3-carboxvphenvl)-2-
hvdroxvlethvlpvridine. dilithium salt.
2-(2-Carboxymethylethyl)-3-decyloxy-6-[2-(3-
carboxymeth ylphenyl) -2- hydroxy ] ethylpyrid i ne ( l Om g, 0. 0 l S mm ol )
2 5 was dissolved in tetrahydrofuran, H2O, and MeOH (0.5mL each) and
treated with LiOH monohydrate (2mg, 0. l Ommol). The reaction was
stirred at room temperature for 24 hours. The solvent was
'I evaporated and the residue was dissolved in H2O, filtered through a
nylon filter and purified by Reversed Phased MPLC (RP- l 8 silica, l 0-
40% methanol in H2O). The desired fractions were Iyophilized to give
a colorless amorphous solid: lH NMR (250MHz, CD30D): ~ 8.0 (s, lH,
aryl), 7.8 (d, lH, aryl), 7.32 (d, lH, aryl), 7.25 (t, lH, aryl), 7.l (d, lH,
5-pyridyl), 6.9 (d, lH, 4-pyridyl), 5.l (t, lH, CH-O), 4.0 (t, 2H, OCH2),
,' 3.1 (t, 2H, CH2), 3.05 (m, 2H, CH2), 2.5 (t, 2H, CH2), l.8-0.90 (m, 19H.
,' 35 aliphatic); FAB-MS: 484 (M+H).
,
: ,. ~ .: -- - .. ... ~ . ..
,
: . :.
;
. : : , , - .:
WO 91/188X02 ~ 8 3 ~ ~ 8 28 PCr/US91/0339~
Example 3
2-(E-3-Hvdroxypropenvl~-3-decyloxy-6-~2-(3-carboxyphenvl)-~-
hydroxYlethvlpYridine~ lithiurn salt.
~3(a! 3-Decyloxy-2-(o~ -dibromomethyl)-6-~2-(3-iodophenyl)-2-
hydroxvlethvlpvridine.
This compound was prepared from 1-(3-iodophenyl)-2-(4-
decyloxy- 1 ,2,3-triazolo[ 1 ,5-a]pyridin-7-yl)ethan- 1 -ol [Example I (d)]
according to the procedure described for 3-decyloxy-2-(a,o!-
1 0 dibromomethyl)-6^[2-(3-carboxymethylphenyl)-2-
hydroxy]ethylpyridine [Example 1 (f)].
3(b) 3-Decvloxv-6-~2-(3-iodophenvl)-2-hvdroxvlethYl-2-pvridine
carboxaldehvde.
This compound was prepared from 3-decyloxy-2-(a,a-
dibromomethyl)-6-[2-(3-iodophenyl)-2-hydroxy]ethylpyridine
according to the procedure described in Example 1 (g). It was
obtained as a white solid.
lH NMR (250MHz, CDC13): ~10.4 (s, lH, CHO), 7.8 (s, lH, aryl),
7.6 (d, lH, aryl), 7.4 (m, 2H, 3d pyridyl, aryl), 7.3 (d, lH, 4-pyridyl),
7.1 (t, lH, aryl), 5.1 (m, lH, CH-O), 4.95 (d, lH, OH), 4.1 (t, 2H, OCH2),
~` 3.1 (m, 2H, Pyd CH2), 1.80-0.90 (m, l9H, aliphatic).
. .
3(c) 2-(E-2-Carboxvmethvlethenvl)-3-decvloxv-6-~2-(3-iodophenvl)-
2 5 2-hvdroxvlethvlpvridine.
The captioned compound was prepared from 3-decyloxy-6-[2-
(3-iodophenyl)-2-hydroxy]ethyl-2-pyridine carboxaldehyde [Example
3(b)] according to the procedure described for in Examplel (h):
1H NMR (250MHz, CDC13): 8 8.1 (d, J=15.9H~, lH, olefin), 7.8 (s,
lH, aryl), 7.6 (d, lH, aryl), 7.4 (d, lH, aryl), 7.2 (d, lH, 5- pyridyl),
7.05 (m, 2H, 4-pyridyl, aryl), 6.95 (d, J=15.9Hz, lH, olefin), 5.6 (d, lH,
OH), 5.1 (m, lH, CH-O), 4.05 (t, 2H, OCH2), 3.8 (s, 3H, CO2CH3), 3.05 (m,
2H, Py-CH2), 1.85-0.90 (m, 19H, aliphatic).
3 5 3(d! 2-(E-3-Hvdroxypropenvl)-3-decvloxv-6-~2-(3-iodophenvl)-2-
hydroxylethvlpvridine.
2-(E-2-Carboxymethylethenyl)-3-decyloxy-6-~2-(3-
~- iodophenyl)-2-hydroxy]ethylpyridine (340mg, 0.60mmol) was
dissolved in dry CH2C12 (6mL) under an argon atmosphere and cooled
;."
'~
", -
.-
.
.
f ~ WO 91/18880 2 0 8 3 ~ 5 8 PCr/US91/03399
, .
29to 0C. DIBAL (I.SmL, 1.5mmol, IM in CH~C12) was added dropwise
and the reaction was maintained at 0C for 20 minutes. The reaction
was quenched with methanol (lOmL) and the solvent was ev~porated.
The residue was par~itioned between ethyl acetate and H~ O and the
5 organic layer was washed with H20 and brine and dried (Na~S04).
The solvent was evaporated and the residue purified by flash column
chromatography (silica, 10-30% ethyl acetate in hexanes) to give a
yellow oil.
1H NMR (250MHz, CDC13): ~7.8 (s, lH, aryl), 7.6 (d, IH, aryl), 7.4
10 (d, l H, aryl), 7. 1 0 (m, 5H, olefinic, 4-pyridyl, 3-pyridyl , aryl), 6.4
(broad singlet, lH, OH), 5.05 (m, lH, CH-O), 4.4 (d, 2H, allylic), 3.95 (t,
2H, OCH2), 3.0 (m, 2H, Py-CH2), 1.90-0.90 (m, l9H, aliphatic).
3(e) 2-(E-3-Hvdroxvpropenvl)-3-decv]oxy-6-~2-(3-carboxvmethvl-
l S phenyl!-2-hydroxvlethvlpvridine.
This compound was prepared from 2-(E-3-hydroxypropenyl)-3-
decyloxy-6-[2-(3-iodophenyl)-2-hydroxy]ethylpyridine according to
the procedure described in Example 1 (e).
1H NMR (250MHz, CDC13): ~ 8.15 (s, lH, aryl), 7.9 (d, lH, aryl),
20 7.65 (d, lH, aryl), 7.4 (t, lH, aryl), 7.10 (m, 4H, olefinic, 3-pyridyl, 4-
pyridyl), 6.4 (broad singlet, lH, OH), 5.2 (m, lH, CH-O), 4.4 (d, 2H,
allylic), 4.0 (t, 2H, OCH2), 4.9 (s, 3H, C02CH3), 3.1 (m, 2H, Py-CH2),
1.90-0.90 (m, 1 9H, aliphatic).
2 S 3(f) 2-(E-3-HYdroxvpropenvl)-3-decvloxv-6-~2-(3-carboxvphenyl)-
2-hvdroxylethvlpYridine, lithium salt.
This salt was prepared from 2-(E-3-hydroxypropenyl)-3-
;, decyloxy-6-[2-(3-carboxymethylphenyl)-2-hydroxy]ethylpyridine
lExample 3(e)] according to the procedure described for 2-(E-2-
3 0 carboxyethenyl)-3-decyloxy-6-12-(3-carboxyphenyl)-2-
hydroxy]ethylpyridine, dilithium salt [Example 1 (i)] .
H NMR (250MHz, CD30D): ~ 8.0 (s, lH, aryl), 7.8 (d, IH, aryl), 7.4
(d, lH, aryl), 7.3 (t, IH, aryl), 7.2 (d, lH, 3-pyridyl), 6.9 (m, 3H,
olefinic, 4-pyridyl), 5.1 (m, lH, CH-O), 4.3 (d, 2H, allylic), 4.0 (t, 2H,
35 OCH2), 3.1 (m, 2H, Py-CH2), 1.85-0.85 (m, 19H, aliphatic); FAB-MS:
(-ve), 460.3 (M-Li).
''
. .-. , - .
. ~. .. .
: ,
. .
.
:: . , : - .
wo 91/18X80 2 0 8 3 ~ 5 8 Pcr/us9l/033ss~
Example 4
2-(E-2-Carboxyethenyl~-3-~-(4-methoxyphenyl)hex~loxYl-6-~'~-(3-
carboxvphenyl!-2-hvdroxvlethvlpyridine~ dilithium salt
2-(E-2-Carboxyethenyl)-3-[6-(4-methoxyphenyl)hexyloxyl-6-
[2-(3-carboxyphenyl)-2-hydroxy]ethylpyridine, dilithium salt was
prepared according to the procedure described for 2-(E-2-
carboxyethenyl)-3-decyloxy-6-[2-(3-carboxyphenyl)-2-
hydroxy]ethylpyridine, dilithium salt recited in Examp]e 1, but
substituting 6-(4-methoxyphenyl)hexan-1-(4-toluenesulfonate)
I lExample B(5)] for l-iododecane.
4(a! 3-~6-(4-MethoxyphenYl)hexvloxvl-6-methvl-2-pvridine
carboxaldehvde: lH NMR (250MHz, CDC13): ~ 10.4 (s, lH, CHO), 7.3 (s,
2H, 4-pyridyl, 5-pyridyl), 7.05 (d, 2H, aryl), 6.8 (d, 2H, aryl), 4.1 (t,
1 5 2H, OCH~), 3.8 (s, 3H, OCH3), 2.6 (s, 3H, CH3), 2.6 (t, 2H, benzylic), 1.8-
1.35 (m, 8H, aliphatic); Anal. Calcd. for C20H25N3 1/8 H2O: C,
72.87; H, 7.72; N, 4.25, found: C, 72.75; H, 7.65; N, 4.10; MS (Cl): 328
(M+H)-
Following the procedures in Examples I (b) et seq, but
2 0 substituting the appropriate adducts here for those recited in Example
1, the following compounds were made:
4(b! 3-~6-(4-Methoxyphenyl!hexvloxYl-6-methvl-2-pvridine
aminohydrazone: 1H NMR (250MHz, CDCl3): ~ 8.2 (s, IH, CH-N), 7.15
(d, 2H, aryl), 7.1 (d, lH, 5-pyridyl), 7.0 (d, lH, 4-pyridyl), 6.8 (d, 2H,
aryl), 5.7 (b~oad singlet, 2H, NH2), 3.95 (t, 2H, OCH2), 3.8 (s, 3H, OCH3),
2.6 (s, 3H, CH3), 2.6 (t, 2H, benzylic), 1.8-1.35 (m, 8H, aliphatic).
4(c) 4-~6-(4-Methoxvphenvl!hexvloxvl-7-methvl-1~2.3-triazolo-~1~5-
alpvridine: IH NMR (250MHz, CDCl3): ~ 8.2 (s, IH, CH-N), 7.1 (d, 2H,
aryl), 6.8 (d, 2H, aryl), 6.65 (d, lH, 6-pyridyl), 6.4 (d, IH, 5-pyridyl),
4.1 (t, 2H, OCH2), 3.8 (s, 3H, OCH3), 2.8 (s, 3H, CH3), 2.63 (t, 2H,
benzylic), 1.90-1.35 (m, 8H, aliphatic).
3 5 4(d! 1 -(3-lodophenvl)-2- l-'6-(4-methoxvphenvl)hexvloxYl-l .2.3-
triazolo~].5-alpyridin-7-Ylleth;ln-l-ol: IH NMR (250MHz, CDC13): ~
8.2 (s, lH, CH-N), 7.8 (s, lH, aryl), 7.6 (d, IH, aryl), 7.37 (d, IH, aryl),
7.06 (t, IH, aryl), 7.05 (d, 2H, aryl), 6.8 (d, 2H, aryl), 6.65 (d, IH, 6-
pyridyl), 6.4 (d, lH, 5-pyridyl)~ 5.4 (m, IH, CH-O), 4.] (t, 2H, OCH2). 3.8
.
- ., :. . -
- , .
WO 91/ l X8X0 2 0 8 ~ 9 ~ 8 PC r/uss1 /03399
`.`~i 31
(s, 3H, OCH3), 3.7 (dd, lH, Py-CH), 3.5 (dd, lH, Py-CH'), 3.2 (d, lH, OH),
2.63 (t, 2H, benzylic), 1.90-1.35 (m, 8H, aliphatic); MS (Cl): 572 (M+H).
4(e~ 1-(3-Carboxvmethvlphenvl)-2-~4-~6-(4-methoxyphen~vl)-
hexvloxvl-1,2.3-triazolo~1,5-alpvridin-7-vllethan-l-ol: IH NMR
(250MHz, CDC13): ~ 8.2 (s, IH, CH-N), 8.11 (s, IH, aryl ), 7.95 (d, IH,
aryl), 7.6 (d, IH, aryl), 7.4 (t, IH, aryl), 7.1 (d, 2H, aryl), 6.8 (d, 2H,
aryl), 6.6 (d, IH, 6-pyridyl), 6.3 (d, IH, 5- pyridyl), 5.5 (m, lH, CH-O),
4.1 (t, 2H, OCH2), 3.9 (s, 3H, CO2CH3), 3.8 (s, 3H, OCH3), 3.7 (dd, lH, Py-
1 0 CH), 3.5 (dd, lH, Py- CH'), 3.2 (d, lH, OH), 2.55 (t, 2H, benzylic), 1.90-
1.40 (m, 8H, aliphatic); MS (CI): 504 (M+H).
4(f~ 3-~6-(4-Methoxvphenvl)hexvloxy,l-2-(a.o~-dibromomelhyl)-6-~2-
(3-carboxvmethYlphenyl)-2-hYdroxvlethYlpvridine: l H NMR
1 5 (250MHz, CDC13): ~ 8.15 (s, lH, aryl), 7.9 (d, lH, aryl), 7.65 (d, lH,
aryl), 7.4 (t, lH, aryl), 7.1 (m, 4H, 3-pyridyl, 4-pyridyl, aryl), 6.8 (d,
2H, aryl), 5.3 (m, IH, CH-O), 4.1 (t, 2H, OCH2), 3.95 (s, 3H, CO2CH3), 3.9
(s, lH, CHBr2), 3.8 (s, 3H, OCH3), 3.15 (m, 2H, Py-CH2), 2.55 (t, 2H,
benzylic), 1.85-1,40 (m, 8H, aliphatic).
4(~) 3-~6-(4-Methoxyphenvl)hexvloxvl-6-~2-(3-carboxvmethvl-
phenvl)-2-hvdroxY1ethyl-2-pvridine carboxaldehvde: 1 H NMR
(250MHz, CDC13): ~ 10.4 (s, lH, CHO), 8.1 (s, lH, aryl), 7.9 (d, lH, aryl),
7.65 (d, lH, aryl), 7.4 (t, lH, aryl), 7.35 (d, lH, 3-pyridyl), 7.25 (d, lH,
25 4-pyridyl), 7.1 (d, 2H, aryl), 6.8 (d, 2H, aryl), 5.4 (m, lH, CH-O), 5.0 (d,
lH, OH), 44.1 (t, 2H, OCH2), 3.95 (s, 3H, CO2CH3), 3.8 (s, 3H, OCH3), 3.2
(m, 2H, Py-CH2), 2.5 (t, 2H, benzylic), 1.90-1.40 (m, 8H, aliphatic); MS
(CI): 492 (M~H).
., .
3 0 4(h) 2-(E-2-Carboxvmethylethenvl)-3-~6-(4-methoxvphenvl)-
hexvloxyl-6-~2-(3-carboxy~methYlphenvl)-2-hYdroxvlethvlpYridine:
lH NMR (250MHz, CDC13): ~ 8.07 (s, lH, aryl), 8.05 (d, J=16Hz, IH,
olefin), 7.9 (d, lH, aryl), 7.65 (d, lH, aryl), 7.4 (t, lH, aryl), 7.1 (m, 4H,
4-pyridyl, 5-pyridyl, aryl), 6.95 (d, J=16Hz, IH, olefin), 6.8 (d, 2H,
` 35 aryl), 5.7 (d, lH, OH), 5.2 (m, IH, CH-O), 4.05 (t, 2H, OCH2), 3.9 (s, 3H,
CO2CH3), 3.8 (s, 3H, OCH3), 3.75 (s, 3H, CO2CH3), 3.15 (m, 2H, Py-CH2),
-' 2.55 (t, 2H, benzylic), 1.90-1.40 (m, 8H, aliphatic); Anal. Calcd. for:
C32H37NO7 9/8 H20: C, 67.68; H, 6.97; N, 2.47, found: C, 67.45; H,
` 6.63; N, 2.34; MS ~CI): 548 (M+H).
. ~ .
. .
wo 91/18880 2 0 8 3 9 S ~ 32 PCr/llssl/033g9~
4(i~ 2-(E-2-Carboxve~henvl)-3-~6-(4-methoxvphenvl~hexvloxYl-6-~2
,~3-car,boxyphenyl!-2-hydrQx~lethylpvridine~ dilithium salt:
IH NMR (2~0MHz, CD30D): ~ 8.05 (s, lH, aryl), 7.8 (d, IH, aryl)~ 7.75
(d, J=16Hz, IH, olefin), 7.35 (d, IH, aryl), 7.25 (t, lH, aryl), 7.2 (d,
.J=16Hz, IH, olefin);~7.0 (m, 4H, 4-pyridyl, 5- pyridyl, aryl), 6.75 (d, 2H,
aryl), 515 (t, lH, CH-O), 4.0 (t, 2H, OCH2), 3.7 (s, 3H, OCH3), 3.1 (m, 2H,
Py-CH2), 2.5 (t, 2H, benzylic), 1.80-1.35 (m, 8H, aliphatic); FAB-MS:
(+ve), 532.2 (M+H); (-ve), 524.4 (M-Li).
Example 5
Formulations for pharmaceutical use incorporating compounds
of the present invention can be prepared in various forms and with
numerous excipients. Examples of such formulations are given below.
Inhalant Formulation
A compound of formula I, I to 10 mg/ml, is dissolved in isotonic
saline and aerosolized from a nebulizer operating at an air flow
adjusted to deliver the desired amount of drug per use.
2 0 Tablets
Ingredients Per Tablet Per 10,000
Tablets
1. Active ingredient
(Cpd of Form. I) 40 mg 400 g
2 5 2. Corn Starch 20 mg 200 g
3. Alginic acid 20 mg 200 g
4. Sodium alginate 20 mg 200 g
5. Magnesium stearate 1.3 mg 13 g
101.3 mg 1013 g
Procedure for makin~ tablets:
Step 1 Blend ingredients No. 1, No. 2, No. 3 and No. 4 in a suitable
mixer/blender.
Step 2 Add sufficient water portion wise to the blend from Step I
35 with careful mixing after each addition. Such additions of water and
mixing until the mass is of a consistency to permit its conversion to
wet granules.
,
, ~ .
, .
W O 91/18880 2 0 8 3 9 5 8 PC~r/US91/03399
Step 3 The wet mass is converted to granules by passing it
through an oscillating granulator using a No. 8 mesh (2.38 mm)
screen .
S~ep 4 The wet granules are ~hen dried in an oven at 410F
5 (60C) until dry.
Step S The dry granules are lubricated with ingredient No. 5.
Step 6 The lubricated granules are compressed on a
suitable tablet press.
1 0 Suppositories:
Ingredients Per Supp. Per 1000 Supp.
1. Formula I compound 40.0 mg 40 g
Active ingredient
2. Polyethylene Glycol 1350.0 mg I ,350 g
1 5 1000
3 polyethylene glycol 450.0 m~ 450 ~e
; 4000 1840.0 mg I ,840 g
' ~rocedure:
Step 1. Melt ingredient No. 2 and No. 3 together and stir until
~ uniform
:l Step 2. Dissolve ingredient No. 1 in the molten mass from Step I
-' and stir until uniform.
Step 3. Pour the molten mass from Step 2 into suppository moulds
2 5 and chill.
Step 4. Remove the suppositories from moulds and wrap.
:~ .
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-
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