Note: Descriptions are shown in the official language in which they were submitted.
2, i
5/AOR12
- 1 - 17~85
TITLE OF T~E INVENTION
IMIDAZOLE COMPOUNDS AND THEIR USE AS TRANSGLUTAMINASE
IN~IBITORS
BACKGROUND OF TaE_l~Y~ lQ~
Transglutaminases, also known as
transamidases, are a family of enzymes which catalyze
the amide bond formation of the ~carboxamide group
of peptide glutamine residues with an ~-amino group
of peptide lysine residues.
A number o~ disease states have been
: associated with transglutaminase activ1ty. Thus, ~or
example, in acne lesions, transglutaminase activity
in sebaceous follicles has been report~d by DeYoung
et. aI., in J. Investigati~e Dermatology, 82, 275
(19~4). Also, the corni~ied cell envelope in acne
:: :
:
,~
.
:
:
: - , . . .
5/AORl2 -2- 17885
has been reported to be a result of transglutaminase
activity by Dalziel et. al., Br. J. E~p. Pathology,
65, 107-115 (1984).
Another dermatological disease, psoriasis,
is reported to be associated with exce~sive
transglutamina~e activity by ]3ernard et. al. British
Journal of Dermatology, 114, 279 ~1986).
Cataracts also have l)een reported to be
associated with elevated tran6glu~amina~e activities.
Factor XIIIa is a plasma transglutaminase
which is the activated form of Factor XIII also known
as fibrinase or Pibrin-stabilizing factor. It is
essential ~or normal hemostatis and is responsible
for the cro~s-linking o~ ~ibrin.
While the activity of thiæ enzyme may be
desirable and essential under most circumstances,
activity under certain other circumstances can be
hi~hly undesirable. Thus, excessive thrombosis, that
is the formation of clot within a blood vessel, gives
rise to thrombotic strokes, de~p vein thrombosis,
varian~ angina, myocardial infarction, and other
medical conditions which ~reguently re~ult in
necrosis of tiæsues and oftentimes in death of a
patient. Even i~ death does not occur, throm~otic
attacks are accompanied by damage to cells to which
circulation ha~ been prevented by thrombi ~ormation.
Removal o~ the thrombi by ly~i~ iæ e~sential and the
rate of lysis may be critical in ultimate patient
recovery.
Lysis may occur normally in hours or days by
~he action o.f a proteolytic enzyme, plasmin, which i3
present in plasma as the inactive precursor,
plasminogen, and which is activated ~y plasminogen
~; ~
: . .
.
- ' '; ' .: .
' ! , ' . . ~ . .
f,l i',J ' . ~
5/AORl2 -3- 17885
activators, such as (pro)urokinase, urokinase or
tissue plasminogen activator (tPA). Since the
occurrence of a thrombotic event calls for rapid
remedial action, administration of exogenous tissue
plasminogen activator or (pro)urokinase is the
current choice in thrombolytic or ~ibrinolytic
therapy. ~owever, a still further reduction in lysis
time is desirable to minimize cell injury.
Since Factor XIIIa is an enzyme responsible
for the .~inal event in the coagulation o~ blood,
lysis and maintaining the lytic state can be
facilitated by the presence of a Factor XIIIa
inhibitor. Moreover, the presence of a Fac~or XIIIa
inhibitor as in a prophylactic treatment where
thrombosis can be anticipated would inhibit hard clot
formation Thus, a Factor XIIIa inhibitor is use~ul
in inhibiting thrombosis, in treating thrombosis when
used with a plasminogen activator, a platelet
aggregation inhibitor or anticoagulant and in post
fibrinolytic therapy in maintainin~ the lytic state.
STATEMENT OF ~E INVENTION
A novel class o~ imidazole compounds has
been discovered which inhibits transglutamina3e
activity, particularly Factor XIXIa activity. The
invention also embraces composition and methods for
using the lmidazole compounds ag Factor ~IIIa
inhibitors in ~ibrinolytic or thrombolytic therapy.
For u8e as Factor XIIIa inhibitor~, the compounds may
be used alone or together with agents u~ed in
thromboIytic or fibrinolytic thera~y such as a
plasminogen activator, a pla~elet aggregation
inhibitor or an anticoagulant.
, ., . ~ . '
' . .
. .
J ~ r
5/AOR12 -4- 17885
D:E;TAILED DESCRIPTIO~ OF ~rHE INVENTION
The imidazole compounds of the present
invention are selected from the group consisting of:
(A) an imidazole represented by the formula;
R` O
Il
R~
R1 (I~
or its acid addition ~alt, and
(B) an imidazolium salt represented by the formula;
R4
2 5 N (~
S - C H2 - C - R
R~--¦ Q
Rl X ~ I I )
In the above and subsequent ~ormulas:
.
.. . : ,
.. . ' . - : ............................. .
;~ .
t ~ . ... J
5/AOR12 ~5-- 17885
R is hydrogen;
lower alkyl;
substituted lower alkyl wherein the
substituents are selected rom hydroxy,
lower alkoxy, pheno~y, phenylthio,
2-pyridinyl-N-oxide-thio, and halo;
cycloalkyl ~rom 3 to 6 carbon atoms;
benzyl;
substituted ben~yl wherein the substituents
are selected ~rom halo, hydroxy, lower
alkyl and lower alko~y;
phenyl;
substituted pheny~ containing 1 to 3
substituents selected ~rom hydroxy,
lower alkoxy, carbo(lower alkoxy),
carbamido, N-(lower alkyl)carbamido or
cyano;
pyridyl
pyrimidinyl; or
pyrazinyl;
Rl is lower alkyl;
substituted lower alkyl wherein the
substituent is earbalko~y or carbamido;
or
ArCnE2n- wherein Ar i~ phenyl, (lower alkyl)-
phenyl, (lower alkoxy)phenyl, or halo-
phenyl and n is 1-3;
R2 is nitro;
sarbo(lower alkoxy);
halo;
cyano;
pheny~;
.
S~ r~ 7~
5/AORl2 -6- 17885
substituted phenyl containing from 1 to 3
~ubstituent~ selected from lower alkyl,
lower alkoxy, halo, and hydro~y;
phenoxy;
phenylthio;
an amido group repre~ented by
NHCOQ whexein Q i8 lower alkyl,
-C~(N~2)C~2C6~5 or -NH(lower alkyl);
lo a hydroxyalkyl group represented by
R'
I
-C-R~' wherein R~ is hydrogen or R~
I
OH
wherein R~' is hydrogen, phenyl,
phenoxyphenyl, biphenylyl, (lower
alkyl)phenyl, lower alkyl and lower
cycloalkyl, or
R' and R" taken together is alkylene f~om 4
to 6 carbon atoms; or
: an ether-alkyl group represented by
-CH2-C~-C~2-0-R"" wherein
OR " '
R" ' is hydrogen, -CO-(lower al~yl),
-CO-phenyl, -CO-biphenylyl,
-CO-phenyl-O-phenyl and -CONH-phenyl,
and R"" is phenyl or lower alkyl;
: 30 R3 is ~ hydrogen; or when R2 i~ phenyl or
: æubstituted phenyl is optiona~ly the same as
; R2; or
.
~ : ~
: ~ :
' J
5/AOR12 -7- 17885
R2 and R3 taken together may ~be alkylene from 3 to 10
carbon atoms optionally substituted with
phenyl or spiroalkylene, or benzo;
R4 is lower alkyl, ArC~2~ wherein Ar iæ phenyl,
(lower alkyl)phenyl, (lower alkoxy)phenyl,
or halophenyl, and n is 1-3; and
X is an anion o a pharmaceutically acceptable
salt.
By the e~pression~ "lower alkyl" and "lower
alkoxy" as employed in the specification and claims
are meant radicals having from 1 to 6 carbon atoms..
By the e~pression "spiroalkylene" i~ meant
an al~ylene chain of from 3 to 6 carbon atoms, the
end carbons of which are attached to the ~ame carbon
of the nucleus.
By the expression "halo" i8 meant fluoro,
chloro, bromo and iodo.
Pharmaceutically acceptable ~alts ~uitable
as acid addi~ion salts as well as providing the anion
of the imidazolium salts are those from acids 3uch as
hydrochloric, hydrobromic, hydroiodic, p~osphoric,
sulfuric, trifluoroacetic, trichloroacetic, oxalic,
maleic, pyruvic, malonic, ~uccinic, citric, mandeli,
benzoic, cinn~mic, methanesul~onic, etha~esul~onic,
trifluoromethanesul~onic and the like, and include
other acids related to the pharmaceutically acceptable
~altR lis~ed in Journal of Pharmaceutical ~cience,
; 30 66, 2 (1977) and incorporated herein by reference.
The compounds, b~th those whirh are acid
addition salts o~ the compounds repreæented by
formula ~I) and thos~ quaternary ~altæ repre3ented hy
'
', . :
. .
: . .
~;~a ,~
5/AORl2 -8- 17885
formula (II) are solids soluble in polar ~olvents
such as water, methanol, ethanol and the like. The
imidazoles of formu~a (I) are E~oluble in non-polar
solvents such as ethyl acetate, methylene chloride,
ethylene dichloride, carbon te~:rachloride, and the
like.
The compounds of the present invention are
useful as transglutaminase inhibitors, particularly
lo as ~actor XIIIa inhibitors, and are adapted to be
employed in thrombolytic therapy. In such use, it is
administered to a thrombotic patient susceptible to
thrombotic attack either alone or in combination with
an antithrombotic agent.
Preferably, it i~ employed toðer with a
plasminogen activator, an enzyme w~ich converts
plasminogen to plasmin to increase the rate and
extent of lysis. Suitable activators include tissue
plasminogen activato~ (tPA), prourokinase (single
chain urokinase), urokinase (dual chain urokinase),
streptokinase and eminase (~uropean patent
application 028,489). T~e plasminogen activators may
be those isolated from natural sources or produced by
recombinant technology and include the genetically
engineered variants thereo~.
Also, it may be employed toge~her with
platelet aggregation inhibitors. Platelet
aggregation inhibitors may be drugs, naturally
occurring proteins or peptides or may be modified or
~emi-~ynthetic prot~ins or peptides. Established
dru~s which are platelet aggregation inhibitoxs
include aspirin and dipyridamole. Proteins or
polypeptides ~which are platelet aggregation
inhibitors have a certain peptide se~uence, most
, . . .
:
5/AOR12 ~9- 17885
frequently Arg-Gly-Asp. Some classes of natural
proteins having this propert~ are fibrinogen receptor
antagonists, thromboxane receptor antagonists,
thromboxane synthesis inhibitors, collagen receptor
antagonists and thrombin inhibitors. Among
especially useful polypepkides are those designated
'IEchistatin" and "Bit~statin" and having the amino
acid sequence:
X-Cys-R-R-R-Arg-Gly-Asp-R-R-R-R-R-Cys~Y where X is E
or an amino acid, Y is OH or an amino acid and each R
independently is an amino acid, described and claimed
in copending applications S.N. 184,649, filed April
22, 198B; S.N. 303,757, .~iled February 1, 1989; and
S.N. 307,642 filed February 7, 1989, all in the names
l~ of P.A. Friedman, et. al., the teachings of whlch are
incorporated by refexence.
Additionally, the imidazole compounds may be
employed for continued therapy after initial relief
from thrombotie attack thereby providing a more
2~ complete lysis and minimizing complications from
reocclusio~. Moxeover, the imidazole compounds may
be employed in post thrombosis therapy together with
anticoagulants such as heparin and coumarin drugs.
The preferred compounds for use as
transglutaminaæe inhibitors are the quaternary
imida~olium salts.
The compounds of the present in~ention which
are imidaæoles may have additional utility as
intermediates in one method o~ preparation of the
preferred imidazolium salts.
The imidazole~ (I) ueeful in ths present
inventiGn may be prepared aecording to the following
equation (1). (In a sub~e~uent table, thie method is
referred to as Method A).
~,r,~ ,4, " ~ ,
5/AORl2 -lO- 17885
R3 0
o-cH2-c-l~nln=
R2\ 1
(A) (~)
lo In the preparation o~ the imidazole o~
formula (I), the 2-mercaptoimidazole (A) starti~g
material, which may be prepared by known procedures
hereinafter detailed, i8 intimately contac~ed with
and caused to react with an acylmethyl ha~ide (B) in
the presence of a tertiary amine ~3 amine) in an
organic solvent at ambient temperature ~or time
sufficient for reaction to take place with the
formation of the desired imidazole o~ formula (I). ~:~
After completiorl o~ the reaction, the imidazole may
be recovered ~rom the reaction mi~ture by removing
the 30lvent by evaporation and purifying the residue
by conventional procedures.
Tertiary amine3 sui~able in the reaction
include triethylamine, trimethylamine, pyridine,
~2s picolines, co~lidinc~, and the like:.
Suitable~olvents ~or thç reactio~ include
acetone, methyl ethyl ketone, dimethylformamide,
dimethyl sulfoxide and the like.
In carrying out the reaction, a ~olution of
the acylmethyl~halide i~ added to:a solution of the
: 2-mercapto;midazo1e and tertiary amine a~d the
......... . ~ . .
, ~ : :
:, ~ ~ ' ' . '
,
5/AORl2 ~ 17885
mixture stirred at room temperature for several
hours, conveniently overnight. At the end of this
period, the æolvent is evapora.ted and the residue
partitioned between water and a water-immiscible
5 organic solvent ~uch as ethyl acetate. The organic
solution containing the imldazole is washed and
dried, the imidazole recovered from the dried
solution as residue, and therea~ter, puri~ied,
preferably by chromatography on silica gel using
lo methanol/chloro~orm as eluant.
The lmidazole then may be employed in the
therapeutic method of the present invention as such
or as an acid addition salt, or may be treated as an
intermediate and employed in the preparation of the
l~ imidazolium salts.
The acid addition salts may be prepared in a
conventional manner such as by intimately mixing the
imidazole and desired acid, preferably in a minimal
amount of polar solvent such as ethanol or by other
conventional procedures.
The imidazolium salts may be prepared
according ~o the ~ollowing equation (2).
2 5 R4
~ 2 ~ ~I R ~ 0
~ c) 1 1 z )
( II' )
, -
~p ~ ,. ,t f,
5/AOR12 -12- 17~85
wherein Z is a displaceable group of an active
quaternizing agent, and within the definitio~ o~ X.
The reaction is carried out by intimately contacting
the reactants in a solvent at ambient temperature for
a time sufficient ~or the reaction to take place with
the formation of an imidazolium salt (II'). The
imidazolium salt (II~) may be recovexed by conven
tional procedures and purified, if desired, or
converted to the ultimate imidazolium salt by use o~
an anion exchange resin:
ion
(2a~ (II') -~ > (II)
exchange
The quaternizing agent is preferably alkyl
~rifluoromethanesulfonate or other active agent. The
halide salts and many other salts are preferably
prepared ~rom the trifluoromethanesulfonate.
The reaction may be carried out for from as
little as about two hours to a wsel~ or so, depending
on the particular reactants.
In carrying out the reaction, methyl
2s trifluoromethane~ulfonate i8 added to a ~olution of
the appropria~e imidazole (I) in an aprotic organic
solvent ~uch as methylene chloride and the re~ulting
mixture ~tirred at ambient temp~rature ~or time
sufficient for ~ub~kantial completion of the
reaction. At the e~d of this period, the æolvent is
vaporized andl ~he re~idue crystallized to obtain the
trifluoromethanesulfonate salt or i8 converted into a
halide by ion-exchaDge chromatography, using
,
- ' . '
5/AOR12 -13- 17885
alkanol/water as eluant. The resulting imidazolium
salt is recovered from the eluate and purified, if
desired, by conventional procedures.
The imidazolium compound represented by
formula (II) may be prepared by an alternate
procedure in which a 1,3-di~ubstituted-imidaæoline-
2--thione is the starting material. This method may
be used when X i8 halogen and in such a case, a
1,3-disubstituted-imidazoline-2-thione is caused to
react with an acylmethyl halide according to the
following equation: (In a subsequent table, this
method is referred to as Method (B)).
1 ~
~3~ ,~=3 ~ halo-CH~-C-R ~ CH,-C-R
R1 R' halide
~A) ~B)
The thione starting material may be prepared
: as subseguently described or by any alt~rnate
procedure known to those skilled in the art. It may
: be prepared as a ~irst step and the alXanoylmethyl
halide added to the reaction ~ixture.
:
.:
... : ~ - ', I
.:' ,~ . ~: ': ' ' .
. , ' . , . . :, :
? f ~
5/AOR12 -14- 17885
In the ~oregoing reaction of the thione and
the acylmethyl halide, about equimolar amounts of the
reactants are employed and the reaction i~ carried
out in solution. Solvents ~uitable for carrying out
the reaction are acetone, methyl ethyl ketone and the
like. The reac~ion may be carried out between about
20O to about 50OC over a period of from about 4 to
about 24 hours. Conveniently, the reaction may be
carried out by stirring overnight. The reaction may
be catalyzed by the addition of a sma~l crystal o~
sodium iodide.
In carrying out the reaction, the acylmethyl
halide ~B) is added ~o a solution o~ the imidazoline-
2-thione (A) and the resulting mixture stirred
together for time sufficient to complete the reaction
with the formation of the desired imidazolium salt
which precipitates in ~he reaction mixture. The
imidazolium salt product may be recovered and
purified by conventional procedures.
The imidazolium 8alt8 in which ~~ is halide
may be converted to salts in which X~ is
trifluoromethanesulfonate or other anions by charging
an ion-exchange eolumn with the sodium salt of
trifluoromethanesulfonate or other deæired anion in a
conventional manner. Therea~ter, the imidazolium
halide i~ eharged on the column in a solvent such as
methanol and ~he desired imidazolium salt reco~ered
from the eluate by vaporizin~ o~f the ~olvent.
The usefulnes~ o~ the compounds as Factor
XIIIa inhibitoræ ~or e~hancing the rate of clot lysis
catalyzed by plasminogen activators may be
demonstrAted first by establishing the inhibitory
potencies of the compounds in a Factor XIIIa assay.
,.
.,
:
5/AOR12 -15- 17885
The Factor XIIIa inhibitor a~ay is based on
the incorporation o~ 14C-putrescine into casein
catalyzed by Factor XIIIa. The a~ay is carried out
employing the procedure described in Methods in
~nzymology, Vol. 45, Ch 1~.. pageæ 177-191 (1976) and
using Factor XIII (F XIII) isolated Prom human
plasma. The procedure is summarized briefly and
schematically illustrated as ~ollows:
F XIII
Thrombin
Ca*~
DTT
~ ,
F XIIIa
Control
or
inhibitor
25 ~4C-putrescine ~4C-pu~rescine
~ incorporated
+ F XIIIa - catalyæed into caæein
N,N-dimethylca~ein
~: Factor XIII as~ay mixture~ are prepared by
~ 30 adding ætepwise, appropriately prepared solu~ions of
: t~rombin and~dithiothre.itol (~TT~ to a ~i~ture
co~prising Fa-tor IIII at 140 mg/mL in glycerollwater
:.
5/AORl2 -16- 17885
and tris(hydro~Jmethyl)aminomethane hydrochloride
(Tris-HCl). To a portion of the mixture is added
calcium chloride as source of calcium ions required
for enzyme activity and to the remaining mixture is
added, instead of calcium ions, ethylenediaminetetra-
acetic acid (~DTA) which serves as a blank ~or
background.
A substrate mixture iæ prepared ~rom
lo 14C-putrescine and N,N-dimethylca~ein.
The assay tubes and control tubes are
charged with the substrate mixture and incubated at
37C ~or 20 minutes. Samples are withdrawn ~rom each
tube, spotted onto a ~ilter disk which is then
immersed in ice cold trichloroacetic acid ~olution to
precipitate the casein on the filter. The fil~er i8
then washed to remove unincorporated or ~ree
4C-putrescine and after drying is counted for
14C-putrescine incorporated to ca~ein from which
percent activity and/or inhibition can be calculated.
Imidazole compounds showing at least S0
percent activity at 2 X 10-5 M in the Factor XIIIa
assay are considered to be useful in in~ibiting hard
. clot ~ormation or especially in BUp~ ementing
fibrinolysis by plasminogen activator.
The imidazoles and imidazolium ~alts seen in
Table I are representatives of compound~ ha~ing IC50
at concentrations belo~ 2 X 10-5 M. ~lso seen in
Table I are the properties o~ the various compounds.
3~
.:
.
u~
~
co
oo
~ o
a~ ~ _ ~~ tl D O ~ O
O D
E o r ~ E I ~ I O , ~
~
O O V ~ U ~1 ~ U t ~ > U I r I
q I I I
~ î~
~ ~ v ~
a~ ~ [~ I ~ v u ~,~
c~l y ~ ~ ~ ~
C~ ~) U t l U U ~ V ~ ~ ) U V ~ U U U U
:~ ~ o
o 3
~: o, r~
: :
o
u~
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::: : : : :
~: ~ . : :
:: :
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u~
a:
o~
~ 8 ~ r n
æ~
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00
_I ~ ~ fk~r ~ o_~ uf--1--3 u~
~ ~ O c ,e c ~ ~ m a
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s~ r
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oo
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a~
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rb ~ x
c I~ ? ?
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: 0~ V , y y y y y ~ y
~ u m m m ,e ~e ,c ~ d
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: ~ 8 ~ -- ,~ S ~ 5
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u~
o~
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t~ q ~ q ~ q ~ d;
u 1 3
a ~ t
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a ~ ~ 5 rl ,o ,~
¢
,
;~
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5/AORl2 -23- 17885
For use in facilitating or æupplementing
fibrinolytic therapy, the imiclazole compound may be
administered in a pre- or post-lytic ætate alone or
in combination therapy. Preferably, it is used in a
combination therapy with a plasminogen activator,
with a platelet aggregation inhibitor or with natural
and synthetic anticoagulants.
The process for ~acilitating or
supplementing ~ibrinolytic therapy in prothrombic
o patients comprises administering a therapeutic dose
of an imidazole compound in an amount to provide
between 1.4-140 mg/kg/day while considering patient's
health, weight, age and other ~actors which influence
drug response. The drug may be admini~tered per os
or by injection, and if by injection, either by
single injection, multiple injections or continuous
infusion.
In the pre~erred process o~ the present
i~vention, the imidazole compound i8 administered
with a plasminogen activator in a combination
therapy. When combination therapy is employed, it is
preferable to administer the Factor XIIIa inhibitor
imidazole compound first in a single bolus and
. therea$ter to administer the plasminogen activator by
continuous in~usion. However, both may be
adminiætered simultaneously as a continuous
infusate Under certain circumætances it may be
desirable to administer the imidazole compound
subsequent to the administration of the plasminogen
activator. It is intended that the method of the
present in~e~tion embrace concurrent admi~iætsation
as well as æequential administfation, in any order.
- : ..
..
. :,- , .. .
.
;:
:, .
,~ .
C~J ,.~
~J ~J ~J ~ .3
5/AOR12 -24- 17885
When ~he Factor XIIIa inhibitor imidazole
compound and plasminogen activator are employed in a
combination therapy, it is most desirable to use the
plasminogen activator in the dose range of about 500
to 10,000 I.U./~g/minute for ~rom about 30 to 180
minutes and the imidazole compound in the range of 1
~g-100 ~g/kg/minute ~or a day (1440 minutes).
When ~he lmidazole compound i~ to be u~ed
with a platele~ aggregation inhibitor in combination
therapy, the dose range for platelet aggregation
inhibitor depends o~ the nature of the inhibitor.
When the platelet aggreg~tion inhibi~or is aspirin,
the aspirin may be employed at a dose o~ ~5-3~5 mg
twice a day. When the platelet aggregation inhibitor
compound is dipyridamole, the dipyridamole may be
employed at a dose o~ 25-100 mg ~our ~imes a day.
When the platelet aggregation inhibitor is a
semi-synthetic peptide such as "Echistatin" or
"Bitistatin", ~he peptide may be administered in a
dose range of 0.1 to 1 nanomole/kg/min. for from 30
to 180 minutes. In each ca~e, the imidazole compound
may be employed in the range o~ 100 ~glkg/min. ~or
a day. The administration may be carried out
simultaneously or sequentially in any order as in the
; 25 procedure for administration ~ith plasminogen
activators.
When the imidazole compound i~ to be used
with heparin, heparin may be admini6tered at doses of
4000 to 800Q uni~æ per 4 hours and the imidazole
; 30 compound in the range of 1 ~g - 100 ~g/kg/minute for
a day. When it i~ to be u~ed with coumarin dru~s
these drugs are administered orally at doses of
.
.
- ,
'' : ; ,.' '
.
.: . , ,
,~ : ' " ~
5/AOR12 -25- . 17885
10 to 15 mg/kg/day and the imidazole compound admin-
istered by in~usion at a rate of l ~g-lOO ~g/kg/minute
for a day.
Compositions to be employed in the practice
5 of the present invention whether parenteral, oral or
suppository compositions comprises an imidazole
compound in a pharmaceutically acceptable carrier.
Parenteral compositions comprise khe
imidazole compound in sterile physiologically
lo acceptable media such as physiolog;cal saline. Such
co~positions may also contain other ingredients for
purposes such as for aiding solubility or ~or
preservation or the like, said ingredients being
those acceptable ~or intravenous administration. The
compositions may be prepared as concentrate
compositions and lyophilized and then diluted to the
appropriate treating composition immediately prior to
administration~ A therapeutic composition as a unit
dose form may contain from lOO mg to 10 grams o~
imidazole compound. Compositions suitable in the
preferred practice o~ the present inventio~ of
co-admlnistering plasminogen activator and Factor
XIIa inhi~itor compound may contai~ (a) about 58
million I.U. o~ tissue plasminogerl activa~or (kPA) or
2s 1.5 million I.U. of ætreptokinase and (b~ ~rom 100 mg
to 10 grams of the imidazole compound,
Oral composition~ also may be prepared with
~he active ingredient in admi~ture with a
pharmaceutically acceptable carrier. Suitable
carriers ~or liquid compoæitions include water,
glycols, oils, alcohola, flavoring agents,
preservatives, coloring a~ents and the like; ~or
solid preparations, Qtarches, sugars, diluents,
- , ~ ,.
,,
,, .-. ~ .
::
.
~ f t ~, C',l ' ~ f;
5/AOR12 -26- 17885
granula~ing agents, lubricants, binders,
disintegrating agents and the like. Becau~e o~ their
ease in administration, tablets and capsules
represent the most advantageous oral dosage unit
form, in which case solid pha:rmaceutical carriers are
obviously employed.
Suppository composit:ions may be prepared
with ointments, jellies, carbowax, polyethylene
sorbitan monostearate, polyethylene glycol, cocoa
butter, and other conventional carriers.
The preparation of the imidazole compounds
suitable for inhibiting transglutaminase enzymes,
particularly Factor 2IlIa, and compositions ~uitable
for carrying out the process of the pre~ent invention
are illustrated by the following examples but are not
to be construed as limiting.
EXAMPL~I
Pr~çedur~
A. l-Methyl-5-phenyl-2[~2-oxo~opYl~thio]i~i~a
~ C H2- C- ~ ~3
C
6H~; I
: CH3
~::
:
': "-, ~ ,' ' ' - : ` '
'
,
5/AOR12 -27- 17885
To a 601ution of 4.76 g (0.025 mol) of
l-methyl-5-phenyl-imidazol-2-l:hiol and 3.15 g (0.031
mol) of triethylamine in 250 mL o~ acetone was added
2.55 g (0.28 mol) of chloroacetone in 250 mL of
acetone. ~he solution was ætirred at room temperature
for 16 hours. The acetone wa~ removed by evaporation
and the residue ~as partitioned betw~en ethyl acetate
and water. The ethyl acetate phaæe was wa~hed with
water, brine, and dried over magnesium æulfate. The
~olvent was evaporated ~rom the dried solution to
obtain as residue 6.16 grams of 1-methyl-5-phenyl-
2[(2-oxopropyl~thio]imidazole which was puri~ied by
~lash chromatography o~ silica gel using 2 percent
methanol in chloroform as eluant.
B. 1,3-Dimethyl-4-phenyl-2-[(2 o~opropyl)thio~-lH-
imidazolium ~hloride _ _ _
~o CH3
~ ~ ~ -CHz-C-CH3
C~
: 25 CH3 Cl~-
To a ~olution of 0.~5 gram (0.001 mol) of
the l-methyl-5-phenyl-2 r ( 2-oxopropyl)~hio~imidagole,
~ ~ 30 prepared aæ above de~cribed, in lO mL of methylene
: chloride ~as added~0.164 g (0.001 mol) o~ methyl
., - ;~
.. ~ , :
- . , ~
5/AOR12 -28- 17885
trifluoromethanesulfonate, and the solution was
stirred overnight at room temperature. The solvent
was evaporated and the residue was converted to the
chloride ion form by dissolvin,g in 20% methanol/water
and passing through Dowex-l (Cl~~ion e~change
column. The column was eluted with 20 percent
methanol in water and the eluate ~ubjected to r~duced
pressure to cvaporate the solvent and to obtain a
residue. The residue was ~tir:red overnight in ethyl
lo acetate, filtered and recrysta:llized ~rom
isopropanol/hexane to obtain purifi~d 1,3-dimethyl
4-phenyl-2-~(2-oxopropyl)thio]-1~-imidazolium
chloride, m.p. 143-146C.
Anal. Cald for C~H17ClN20S~1/6H~O:
C, 56.08; ~, 5.83; N, 9.34
C, 56.08; ~, 5.92; N, 9.28
~X~MPLE II
Proced~Q_~
A. l-Methyl-4,5-diphenyl-2[(2-oxopropyl)thio~-
imidaz~le
C6H~ \,N
/~N> S - CH2 - C- C~3
c6~
CH3
~ Ç? c~
5/AOR12 -29- 17885
l-~ethyl 4,5-diphenylimidazol-2-thiol was
~irst prepared as ollows: 10.0 grams (0.047 mol) of
benzoin and 4.25 grams methylthiourea were stirred
to~ether in 50 milliliters of hexanol-l and heated to
reflux and the refluxing continued for about 20 hours.
The mixture was allowed to cool whereupon crystals
formed in the reaction mixture. The cry~tals were
recovered by filtration and puri~ied on a ~ilica gel
column to obtain 0.68 gram o~ l-methyl-4,5-diphenyl-
imidazol-2-thiol, m.p. 280-283C.
The l-methyl-4,5~diphenylimidazol-2-thiol
(0.618 g, 0.00255 mol) was dissolved in 150
milliliters o~ acetone and to it was added with
stirring 0.236 gram (0.00255 mol) of chloroacetone
and 0.258 g (0.00255 mol) o~ triethylaminc. The
mixture was stirred overnight at room temperature.
Since the reaction was not complete a~ determined by
TLC, 0.25 gram of chloroacetone and 0.26 gram of
triethylamine was added and the mixture reflu~ed for
one hour. The mixture wa~ ~hen allowed to cool and
evaporated in vacuo to obtain a re~idue. The latter
was dissolved in e~hyl acetate, the ethyl acetate
solution was washed with water, 5% æodium hydroxide
and brine, the waQhed solution was dried (MgSO4), a~d
the dried æolution evapora~ed in Y~Ç~ to obtain a
~till impure product. The latter wa~ put over a
silica gel column and developed with 1 perc~nt
methanol in methylene chloride to recover ~rom the
elua~e after evapora~ing off the ~ol~ent, 0.561 ~ram
of 1 methyl-4,5-diphenyl-2~2-o~o-propyl)thio]-
imidazole. The produet after recry~tallization fromi~opropyl alcohol-hexane had a melting point of
~ .
5/AOR12 -30- 178~5
179-182C. Anal. Calcd. or C9H18N20S~Cl
C, 63.58; H, 5.34; N, 7.81
Found C, 63.56; ~, 5.37; N, 7.61
B. 1,3-Dimethyl-4,5-diphenyl-2~(2-oxopropyl)thio~
imidaæoli~Lm tri1uromethallesuli~onat~
H3
\,_N~
S - CH2 - C- CH3
C6H~ I CF3S03 (_)
CH3
0 . 561 gram (0 . 00174 mol) of the imidazole
prepared in part A was dissolv~d in 15 milliliters of
methylene chloride and while stirring at room
temperature there was added 0.31 g of me~hyl tri-
fluoromethanesulfona~e. The resulti~g mixture was
stirred one hour, then an additional 0.3 gram o~
methyl trifluoromethane~ulfonate waæ added and
stirred ~or 2 hour~. The mi~ture waæ coevaporated
with isopropanol ~o obtain crystal~ of ~he desired
imidazolium trifluoromethanesul~onate ~hich after
crystallization from isopropanol had a melting point
of 138-139~.
Anal. Calcd for C~lH21N2F304S2
C, 51.84; ~, 4.35, N, 5.76
Found: t', Sl.Sl; ~. 4.62, ~. 5.72
: ~ .
~ .~
.
'
g
5/AOR12 -31- 17~85
~ X~PLE III
A. l-Methyl-2[(2-oxopropyl)tlliQ]~nzi~ldazol~.
~>~ - C H2 - C - C H3
CH3
l-Methylbenzimidazole-2~thiol starting
material was prepared by the procedure o~ G.F. Duffin
et al., J. Chem. Soc. 3~1 (1956) ~rom 14.2 grams o~
N-methyl-nitro-aniline by reducin~ with zinc and
alkali in ethanol to obtain N-methyl-o-phenyl~ne-
diamine and intimately contacting the latter with
carbon disul~ide to obtain 6.1 grams of l-methyl-
benzimidazole-2-thiol, m.p. lg2-193C. :-
To 1.0 gram (0.0061 mol) of l-methyl-benz
imidazole-2-thiol in 50 milliliters o~ acetone was
: added 0.64 g (0.0069 mol) o~ chloroacetone; then 0.71
gram (0.007 mol) of triethylamine was added dropwise
: 25 ~ith stirring and the stirring eontinued overnight at
room temperature. The mixture was then evaporated in
va~U~ and the residue disæolved in 100 milliliters
:: o~ ethyl acetate. The ethyl acetate 801ution was
washed ~uccessively with water, 5 percent sodium
hydroxide and water:and dried sver MgSO~. Eth~l
acetate:was vaporized ~om the dried æolution to
, , - - , . . .
'~ 1,
..
, :
sJ
5/AOR12 -32- 17885
obtain as residue 1.4 ~rams of the thiol compound
which was further purified by recrystallizing from
butyl chloride, chromato~raph:ing over silica gel with
chloroform and recryætallizinp ~rom butyl chloride to
obtain 150 mg of the desired 1-methyl-2[(2-oxopropyl)-
thio]benzimidazole, m.p. 87-B8C.
Anal. Calcd. for C11~12N2S
C, 59.97; ~, 5.49; N, 12.72
Found: C, 59.83; E, 5.55; N, 1~.54
B. 1,3-Dimethyl-2-C(2-o~opropyl)thio]-lH-benzimida-
æolium ~rifluQ~Q~ethaneSulfon~e
0.340 gram (0.00154 mol) of the benzimidazole
above prepared was dissolved in 10 milliliters of
methylene chloride and to the resulting solution was
added 0.30 gram (0.00154 mol) of methyl trifluoro-
methanesulfonate and the mixture stirred for three
hours at room temperature. The mixture was then
subjected to reduced pressure to remove the solvent
and to obtain the imidazolium alt product as residue.
The product was crystallized from isopropyl alcohol
to obtain, after drying, a purified product melting
130-132C.
Anal. Calcd for Cl3~l5F~N2o4s2
; ~ C, 40.62; ~, 3.93; ~, 7.29
Found: C, 40.47; ~, 4.24; N, 7,?9
: 3
.
::
.
,:
.
J . , :' l J
5/AOR12 -33~ 17885
~ AMPLE IV
4-Chloro-1,3-dimethyl-2-~(2-oxopropyl)thio]-lH-
imidazolium chloride
CH3
~ CH~-C-CH3
Cl
C~I3 Cl
The startin~ material, 4-chloro-1,3-dimethyl-
imidazoline-2-thione was firs~ prepared by adding
dropwise and i~timately admixing 5.68 grams (0.04
mol) of methyl iodide to a ~olution of 4.0 grams
(0.0343 mol) of 5-chloro-1-methylimidazole in 25
milliliters o~ methylene chloride. The resulting
mixture was stirred overnight at room temperature
whereupon a crystalline product was found to have
formed. The latter was recovered ~th cold methylene
chloride to obtain 1,3-dimethyl-5-chloroimidazolium
chloride.
The imidazolium chloride thus obtained ~3.65
grams, 0.0141 mol), 0.45 gram (0.0141 gram atom)
sulfur, and 2.44 grams (0.0178) moles o~ potassium
carbonate and 25 mllliliters of anhydrous methanol
were stirred~vi~or~usly ouernight at room
temperature. Thereafter, the mixture waæ filtered
and the filtrate evaporated to drynesæ. The residue
. , ,
. . ~ .
- 2 ~ r~
5/AORl2 -34- 17885
was recrystallized from water to obta:in
4-chloro-1,3-dimethylimidazollne-2-thione which was
used in the preparation of the oxopropylthio-
imidazolium salt.
To a solution of 1.00 gram (0.0062 mol) o~
4-chloro-1,3-dimethyl-imidazol.ine-2-thione in 180 mL
of acetone was added 0.S69 g (0.0062 mol) of chloro-
acetone and a ~mal.l crystal of sodium iod~de. The
mixture was ~tirred overnight at room temperature
lo whereupon a product precipitated. The latter wa8
recovered by filtration and recrystallized from
isopropyl alcohol-hexane to obtain purified 4-chloro-
1,3-dimethyl-2-~2-oxopropyl)thioJ-lH-imidazolium
chloride, m.p. 134-136C.
Anal. Calcd, for C8H12ClN2OS:
C, 37.65; ~1 4.74; N, 10.95
Found: C, 37.65; H, 4.58; N, 11.03
EXAMPLE V
1,3-Dimethyl-4-(methylaminocarbonylamino)~2-~(2-oxo-
propyl)thio~ -i.m dazolium_~hlorid~
CH3
CH2-CCH3
CX3NHC- CH3 Cl~
o
: 30
~: :
~; :
:
. . , , . , . . . ,. . . - -
. : . - . :~ , : .
..
. .
~: ' " ;. : . ,
.
5/AOR12 35- 17885
To a solution of 3.22 ~rams (0.0225 mol) of
1,3-dimethyl-4-amino 4-imidazoline-2-thione (prepared
from methylaminoacetonitrile and methyl i~othiocyanate
by the method of T. Kinoshita et al, Bull. Chem. Soc.
Japan., 5~, 442 (1980)) in 12 mL of pyridine under an
atmosphere of nitrogen was added with stirring 2.61
grams (0.0457 mol) of methyl i~ocyanate. The mixture
became e~othermic. Stirring was continued overnight
whereupon a solid precipitate formed in the reaction
mixture. The latt0r wa6 collected by filtration and
washed with isopropyl alcohol to obtain 2.11 grams of
1,3-dimethyl 4-(methylaminocarbonylamino)-imidazo-
line-2-thione, m.p. 200-201C.
A solution of 0.50 gram (0.0025 mol) of the
thione intermediate in 60 mL of acetone was intimately
mixed with 0.28 gram of chloroacetone. A small
crystal of potassium iodide wa~ added and the solution
was stirred for three days whereupon a white solid
formed in the reactio~ mi~ture. The latter was
~0 collected a recrystallized from ethanol to obtain
purified 1,3-dimethyl-4 (methylaminocarbonylamino)-2-
~(2-oxopropyl)thio]-1~-imidazolium chloride, m,p.
~77-179C.
~nal. Calcd for CloH17ClN4O25:
C, 41.02, H, 5.~5; N, 19.14
Found: C, 40.86; H, 5.80; N, 18.87
_ ..
. .
-
.:
.
,
,
5/AOR12 -36- 17885
EXAMPLE ~I
1~3-Dimethyl-4-(acetylamino)-2-c(2-oxopropyl)thio~-l~I
imidazolium chlQrid Q
c~3
~[N\>~ C~a - CCH3
C H3 C ~ HN I )
0.50 gram (0.0027 mol) o~ 1,3-dimethyl-4-
(acetylamino)-imidazoline-2-thione (prepared by
l~ acetylating by conventional means the 1,3-dimethyl-
4-amino-4-imidazoline-2-thione described in ~xample
V) and 0.28 gram (0.003 mol) o.~ chloroacetone were
mixed together and refluxed for t~o hours after which
time a solution 0.428 gram of sodium iodid0 in 5
milliliters of acetone was added whereupon a reaction
took place with the immediate formation of a
precipitate o~ sodium chloride by-p~oduct. The
latter was removed by ~iltration and the filtrate
evaporated to dry~ess to obtain a residue. The
residue ~as di~solved in acetone and from ~he
~olution was reco~ered a ~olid which a~ter
purification on a Dowe~-l (Cl ) column and
: crysta~lization from iæopropyl alcohol was obtained
the desired 1,3-dimethyl-4~(acetylamino)-2
~(2-oxopropyl)-thio3-lH-imidazolium chloride, m.p.
:171-173C. ~ :
~. - . . . :: .
.
.
. - . ~
., . :
~ 3
5/AOR12 -37- 17885
Anal. Calcd, for CloHl~ClN3O2';:
C, 43.24; X, 5.81; N~ 15.13
Found: C, 43.21; H, 5.68i N, 15.09
5EXAM~LE VII
1 t 3-Dimethyl-4,5-bi~(methoxyphenyl)-2-~(2~oxopropyl)-
~thio~-lx-imidazQLium chlorld~
10CH3
(p)CH3OC~H ~ I ~ O
~S - C H2 - C - C H3
( p) cH3c)c6H~
CH3 Cl ~
1 ~
1l3-Dimethyl-4,5-bis(p-methoxyphenyl)-
imidazolin-2-thione was fir~t prepared by heating
together 5.45 grams (0.02 mol) of anlsoin and 2.08
grams (0.0~ mol) of N,N-dimethylthiourea in 20
milliliters o~ l-hexanol under a~ atmosphere of
nitro~en.
: In an operatlon carried out in a manner
æimilar to the preceeding examples, 0.185 gram o~
chloroacetone was added to a ~olution o~ 0.68 gram
~0. 002 mole? Of the imidazolidinethione in 20 milli-
liters o~ ac~tone and ~h0 mixture allowed first to
stand at room temperature, then refluxed for 3
~ ~ hours. Therea~ter,;a:godium iodide crystal was
:~ added, 0.0~5 gram (0.002 mole) chloroacetone added
:
,
.
:
~ t ~
5/AOR12 -38- 17885
and the mixture heated to obtain the desired
1,3-dimethyl-4,5-bis(p-methoxyphenyl)-2[(2-
oxopropyl~-thio]-l~-imidazolium chloride, which after
a series o~ recrystallizations was obtained as a
product melting 179-180C.
Anal. Calcd for C22H25ClN203S
C, 61.03; H, 5.~2; N, 6.47
Found: C, 60.80; H, 5.61; N, 6.64
EXAMPLE YIII
1,3-Dimethyl-4,5-decamethylene-2-t(2-oxo~
~ropvl2thiQ~ d~zolium chlo~ide
l- (CH2 ~ CH2-CcH3
(~)
CH3 c
1,3-Dimethyl 4,5~decamethylene-imidazolin-2-
thione was prepared by heating together at re~lux
temperature 2.4 gram (0.012 mol) of 2-hydroxy-
cyclododecanone and 1.47 grams (0.014 mol) of .
l-methyl-2-thiourea in 10 milliliters of hexanol.
After heating for about 20 hours, the mixture was
cooled, isopropanol added and chilled to obtain the
imidazolin-2-thione.
.
:
5/AOR12 -39- 17885
In a manner similar to that previously
described, 0.553 gram (0.002 mole) o~ the thione
above prepared and 0.185 gram (0.002 mole) o~
chloroacetone were stirred together in 10 milliliters
of acetone at room temperature; then ~odium iodide
crys~al was added and ~he mixture heated to obtain
the desired 1,3-dimethyl-4,5-decamethylene-2-[(2-
oxopropyl)thio]-l~-imidazolium chloride, m.p. 73-78C.
Anal. Calcd for C18H31ClN20S:
C, 60.~2; H, 8.71; N, 7.81
Found C, 60.13; ~, 8.96; N, 8.01
~XAMPLE IX
1,3-Dimethyl-4(~(L)-phenylalanyl]amino~-2-~(2-oxo-
1~ propyl!~hio]-lH--imidazoliu~ chloride hydrochlQ~ide
~H3
~ H -CH-gNH ~ N ~ -CH -CCH3
NH2-HCl CH Cl`
1,3-Dimethyl-4(E(L>-N~(t-butyloxycarbonyl)-
phenylalanyl]amino)-4-~midazoline-2-thione o~ the
following ~ormula
,.
5/AOR12 -40~ 17885
CH3
~ ~2-CH_C~
INH2 CH3
CCC(C~3)3
was first prepared: To a solution o~ 2~8 g (0.020~
mol) of l-hydroxybenztriaæole and 3.99 g (0.0208 mol)
of l-(3-dimethylaminopropyl)-3-ethylcarbodiimide
hydrochloride in 30 mL o~ dimethylformamide wag added
1~ 5.05 g (O.019 mol) of ~-butyloxycarbonylphenylalanine.
~ To thiæ stirred solution wa~ added dxopwi6e over 15
mi~utes, a ~olution of 2.47 g (0.0173 mol) of 1,3 di-
methyl-4 amino-4-imidazoline-2-thion@ (prepared ~rom
: methylaminoaceto~itrile and methyl i~ocyanate as
described in Bull. Chem. Soc. Japan ~, 442 (1980))~
The p~ o~ the solution was adjusted to approxi~ately
8.5 by the addition of 7 mL of trie~hylamine. The
mixture was stirred ~or 3 hours at room temperature
under a nitrogen atmospherei and then was poured onto
500 mL of water. The a~ueous soIution waæ e~tract0d
with ~ive 75 mL portion~ oP ethyl acetate. The
combin~d extract~ were wa~hed with ~ive 100 mL
portions o~ water, dried over MgSO4, filtered, and
: : ~he et~l acetate r~moved under reduced presæure.
The residue was recrystallized ~rom ethanol to obtain
4.15 g ~61%) of 1,3-di~ethy~-4(~(L~-N-(t-butylo~y
carbonyl)phenylalanyl~amino)-4-imidazoline-2-thione
as a white, cry talline material, m.p. 168-169~C.
'
- ; .,~.................... ...
., ; , , ~
, .
G,~ ' `J'
5/AOR12 -41- 17885
1,3-Dimethyl-4-(t(L~-phenylalanyl]amino)-2-
[(2-oxopropyl)thio]imidazolium chloride hydrochloride
was then prepared: To a æolution of 0.050 g (0.00128
mol) of ~he ~hione above prepared in 12 mL of acetone
5 was added 0.12 g (0.0013 mol) of chloroacetone, and
the ~olution wa~ stirred 3 days. Additional
chloroacetone (0.5 mL) waæ added at this time and the
~olution was stirred and refluxed for 24 hours. The
solution was concentrat~d undler vacuum and the
residue was stirred wi~h 5 mL o~ ethanol saturated
with HCl (gas) ~or 6 hours. The solution was
concentrated under vacuum and th~ residue was
cryætallized from isopropyl alcohol ether to obtaln
the desired 1,3-dimethyl-4-([(L)-phenylalanyl]
l- amino)~ (2-oxopropyl~thio~ l~-imidazolium chloride
hydrochloride; m.p. indeterminate at 90-100C
(softens, then ~oams).
Anal. Calcd for C17H~3ClN4O2S~Cl-2H20:
C, 44.83; H, 6.20; N, 12.30
Found: C, 44.36; ~, 5.63; N, 12.02
: 25
~ ~ 30
:
::
;: ~
:
:
.
,
'
'
2~ ,.a~
5/AOR12 ~42- 17885
EXA~L~
l-Benzyl-3-methyl-2-~(2-oxopropyl)thio]-lH-imidazolium
.~hlori~
C~13 o
C HZ - C - C H~,
(~)
~2 (~)
l-Benzyl-3-methylimidazoline-2-thione was
1~ first prepared as follows: 8.g3 grams o~ methyl
iodide was added slowly with stirri~g to 10.0 grams
of benzylimidazole in 25 milliliters of methylene
chloride. The mixture was stirred for three hours
and then an additlonal 2.0 grams of methyl iodide was
23 added. The mixture was then evaporated to dryness in
vacuo to obtain 18.97 grams of an re~idue of
l-benzyl-3-methylimidazol;um iodide.
The qua~ernary salt thus obtained was then
stirred tog~ther at room temperature with 10.9~ gxams
of anhydrou~ potas~ium carbonat~, 2.03 grams of
powdered sulfur and 100 milliliter~ of anhydrous
methanol ~o~ about 48 hours. Thereafter, the mixture
was filtered through a pad of celite and washed with
200 milliliters o~ methylene chloride. The ~iltrate
was evaporated to drynes~ ae~o and the residue
.
.:
.
r
~1 b~ S .; ~ J
5/AOR12 -43~ 17885
boiled with 100 milliliters of water. The resldue
was recrystallized from 100 millil~ters of absolute
methanol to obtain purified l-benzyl-3-methyl-
imidazoline-2-thione.
1-Benzyl-3-methyl-2-~(2-oxopropyl)thio~
imidazolium chloride was prepa~d a3 ~ollo~s: 0.5
gram of chloroacetone was added with ~tirring to a
~olution o~ 1.0 gram of the thione above prepared in
20 milliliters of acetone. A very small crystal of
sodium iodide was added and the mixture stirred
overnight at room temperature. A ~mall amount o~
c.rystalline material p~ecipitated. The mixturc was
then heated at reflu~ temperature ~or 18 hours to
obtain a crystalllne product. The latter was
recovered, washed with acetone and recry~tallized
from a 10 milliliter isopropyl alcohol-~0 milliliter
ether mixture to obtain purified l-benzyl-3-methyl-
2~[(2-oxo-propyl)thio]-l~-imidazolium chloride, m.p.
119-1~1C.
2~ Anal. Calcd for Cl~Hl7ClN2OS00.25~70:
C, 55.21; E, 5.76; N, 9.16
Found: C, 55.39; H, 5.59; ~, 9.14
2s 1-(3-Metho~y-3-oxopropyl)-3-methyl-2[(2 o~opropyl)-
: thiQ~-lH-imLdaæQliu~ chloride
CH3
[I~N~-CH~-C-C~13
Cl (-~)
I H~ lo~
CHa- C- OCH3
::
:
'
', ~
s~ ( I '3 -~
'"''~
5/AOR12 -44- 17885
In operations carried out in a manner
similar to that described in Example I, 1-(3-methoxy-
8-oxopropyl)imidazol-2-thiol was reacted first with
chloroacetone and triethylamine to obtain 1-(3-
methoxy-3-oxopropyl)-3-methyl-2-~(2-oxopropyl)thio-lH-
imidazole, which thcn was rc~.cted with methyl tri-
~luoromethanesulfonate to obtain 1-(3-methoxy-3-oxo-
propyl)-3 methyl-2-~(2-oxopropyl)thio]~ imidaæolium
trifluoromethanesulfonate, which in turn was pa~sed
through a Dowex-l (Cl~~ ion exchange column to obtain
1-(3-methoxy-3-oxopropyl)-3-methyl-2-~(2-o~o-
propylthio)-lH-imidazolium chloride having m.p.
121-124C.
Anal. Calcd ~or C~ 17ClN23S
1~ C, 45.12; H, 5.85; N, 9.57
Found: C, 45.24; ~, 5.76; N, 9.45
EXAMPLE XII
1-(3-Amino-3-oxopropyl)-3-methyl-2[(2-oxopropyl)thioJ-
2~ l~-imi~azolium Ghloride
C ~ ~ -C~ _C-C}b
l~2 o
C~2-C- ~2
.
,~
5/AOR12 -45~ 17885
In operations carried out in a manner
similar to that described in Example I, 1-(3-amino-
3-oxopropyl~imidazol-2-thiol was reacted fir~t with
chloroacetone in the preæence of triethylamine to
obtain 1-(3-amino-3-oxopropyl)-3-methyl-2~2-oxo-
propyl)thio-lH-imidazole, which then was reacted with
methyl trifluoromethanesu~fonate ~o obtain 1-~3-amino-
3-oxopropyl)-3-methyl 2[(2-oxopropyl)thio-lH-
imidazolium trifluoromethanesulfonate which in turn
was passed through a Dowex 1 (Cl-) ion e~change
column to obtain 1-(3-amino-3-oxopropyl) 3-methyl-
2~(2-oxopropyl)thio-lH-imida~olium chloride, m.p.
124-125C.
Anal. Calcd for C10~16ClN302S:
C, 43.24; ~, 5.81; N, 15.13
Eound: C, 42.96; H, 5.80; N, 14.95
EXA~E ~III
1,3-Dimethyl-4,5,6~7-tetrahydro-2-[(2-o~o-
2~ R~Q~Yl)thi~ çn~imidaz~lium ChlQrid~ _
CH3
I ~ O
~ ~ -C~2-C-CH3
CH~ Cl(-)
- . .. .
.
.
.
5/AOR12 ~46-- 17885
1,3-Dimethyl-~4,5,6,7-tetrahydro-2 thiono-
benzim;dazole~ was first prepared by heating together
at reflux temperature 11.41 grams (10.0 mmols) of
2-hydro~ycyclohexanone and 10.42 grams (0.014 mol) of
N,N'-dimethylthiourea in 10 milliliters of hexanol
under argon atmosphere ~or 24 hours. The mixture was
subjected to reduced pressure to remove the
volatiles, isopropanol then was added and the
resulting mixture chilled to obtain a yellow ~olid
which after filtration and drying amounted to 10.0
grams o.f the imidazolin-2-thione.
A solution of 4.00 grams (22.0 mmol) of the
thione abo~e prepared in 110 mL of acetone was
stirred at 25C while 2.0 grams ~22.0 mmol) of
chloroacetone was added. A~ter 18 hours an
additional 2.0 grams of chloroacetone was stirring
continued. This was repeated with three more
portions o~ chloroacetone. The resultin~ mi2ture was
concentrated to obtain a solid which ~as then
2~ tritura~ed with ethyl acetate and recrystallized ~rom
isopropyl alcohol and ethyl acetate to obtain 4.00
grams of the desired 1,3-dimethyl-4,5,6,7-
tetrahydro-2-[(2-oxopropyl)thio]-1~-benzimidazolium
chloride, m.p. 149~151C.
XY
In operations carried out in a manner
similar to that de~cribed in E~ample~ VIII-X, the
compounds in Table II are prepared:
(1) 1-~2-(p-Propylphenyl)ethyl]-2[(~-oxopropyl)-
thio]-lH-imidazole;
.
. .
.. : . ' ' ' ~'-.' .
:-
.3, ~
5/AOR12 -47-- ~ 17885
1-[2(p-Propylphenyl)ethyl~-3-methyl-2-[(2-oxo-
propyl)thio]-l~-imidazolium trifluoromethane-
sulfonate;
1-[2(p-Propylphenyl)ethyl]-3-methyl~2C(2-oxo-
propyl)thio3-lH-imidazolium chloride.
(2) 1-t3-(p-Chlorophenyl)propyl]-~-[(2-oxopropyl)
thio]-l~-imidazole;
1-[3-(p-Chlorophenyl)propyl]-2-[(2-oxopropyl)-
thio]-lH-imidazolium trifluoromethanesulfonate;
1-[3-(p-Chlorophenyl)propyl~-2-[(2-oxopropyl)-
thio]-lH-imidazolium chloride.
(3) l-Isopropyl-2-[~2-oxopropyl)thio-1~-imidazole;
1,3-Diisopropyl-2-~(2-oxopropyl)thio)~
imidazolium trlfluoromethanesul~onate;
1,3-Dii~opropyl-2-[(2-oxopropyl~thio]-lH-
imidazolium chloride.
. ' - ' '
~ fi; ~ f.~
5/AOR12 -48- 17885
~X~ LE ~VI
In operations carried out in a manner
similar to that described in Example I, the compounds
in Table II are prepared:
T~3 bl ~ I ~
-CH~CH~CH~ -C}13 -Cl H -CH3 CF,903'
-CH,CH30H -CH~ -CN H -C}~ CF,90J'
-CHlDCHgC~b -CH3 -ND~ H -CH~ CF~803
-C4,CH~COOCHJ -CH3 -CO~ H -C~b CF19C13'
l 0 -CH~CONH~ CH;~CHl -OC~}13 H -CH~CH1 Cl'
-CH~CN -C1H7 _C"N~C1~ P) H -C3HT CF980
-C~l~C~H~ -C}l~ -COOC}I~ H -CH~ Cl'
-Co4 -C~tCHJ)I -CoHo ~ -CNtCH3~9 CEr~503'
CO~40CN9tP) -CH~ -(CN~)~- -CH3 CF~903
-COH ~CHJtO) -CN~ -C~ II -CH3 CF~303
-CoNIOHtp) -CH~C~ -CA}I3 C3H~ -CHgC~b Cl'
-Co~l~COCCH~ -CH9 ~J~ -CH1 Cl~
-C~H~CN -CH1 -8Co~S~ N -CN~ Cl-
- CoH,CONHCH3 - CH~ - CO1~3Br~ p) H - CH3 Cl'
-~CNa)~CH, -C~ Co}13 -Cl H -CH3 CF~601-
-CoH~CONH, -C~CONNI -CN H -CH3 CF,803'
-CH,C~Cl - CIH~CC~C}13 - Nl~l H - CHg CF7803
-CH~C~ c~3 -CoN~OCH~p) H -CN9 CF~80~'
-CH~C~ r~ p) -CH3 ¢~ -C}l~ CF~503
-CH~C~ O~tto) -CH3 -~C~9)~- -C~ CF~903
-CH,CoH,OCH, -CH3 ~(CNa)3~ -C~b CF~803'
~ -CHg -C~bON ~ _CNJ CF3809'
-C}l, -Cl ~ -c~ C~,SO,'
-CH3 -c6~ H -CM~ CF~803-
3~ .
. ~
: , ~ ~, . . .
~ ~;{ ~1 h
5/AOR12 -49- 17885
EXAMPLE %VII
In operations carried out in a manner
similar to the foregoing e~amples, the ~ollowing
compounds are prepared:
TA~3LE I I I
R R ~ l t
-C~13 -C~13 -NIICOC3H7 H -CH3 Cl-
- CIH~- C~13 - CH( NH;,~ C}~C~ }I - C~13 Cl -
C~13 - CH3 - NHCH3 H - CH3 CF3SO3
-CH3 -CH3 -CHC~H3 H -CH3 CF3SO3-
0~1
- H - CH3 - CHCo H3 OCH3( p) H - CH3 Cl -
OH
-CH3 -CH3 -CHCoH~C~(p) H -CH3 CF3SQ3-
OH
-CH3 -C}~3 -7(C0~5)2 H -CH3 Cl-
OH
-H -CH3 -CHCo~C}b(P) H -CH3 CF3SO3-
2 0 OH ~_~
- CH3 - CH3 - CH~_) H - CH3 Cl-
OH
-CH3 -CH3 -~2)9 H -CH3 CF3SO3-
OH
-H -CH3 -CH2-CH-CH2OC~ H -CH3 Cl-
2 5 COC~13
-H -C}13 -C~zCJlCH2OCo}15 H -CH3 Cl-
OCI,H9
_~ -C~3 -CH2-CH-C~OCH3 H -CH3 CF3SO3-
OC~C~,H,,OC~,Y~( p)
O
:
. .
.
. .
, : ~ ' ' . . '
~' , ~ '~ ' ; '
,'
'~ .
5/AOR12 -50 17885
~AMPLE~
The following compounds are prepared by
intimately mixing the imidazole with ethanolic
hydrogen chloxide, letting the mixture stand at
ambient temperature to allow the cry~tals of the salt
to ~orm and then recovering by filtration.
l-Methyl 5-phenyl-2~(2-oxopropyl)thio]-
imidazole hydrochloride.
1-Methyl-4,5-diphenyl-2[(2-oxopropyl)thio~-
imidazole hydrochloride.
5-Chloro-l-methyl-2-~(2~o20propyl)thio]-
imidazole hydrochloride.
l-Methyl-5-~methylaminocarbonylamino) 2-
~(2-oxopropyl)thioJimidazole hydrochloride.
~AMpLE~
In a similar manner the ~ollowing æalts are
prepared by mixing the imidazole with an ethanolic
solution of the aeid corr~sponding to the salt
desired:
1 Ethyl-4,5~diphenyl-2[(2-oxopropyl)thio]-
imidazole hydrogen maleate.
2s 5-ChIoro-l-methyl-2~(2-oxopropyl)thio~- :
lmidaæole hydrogen citrate.
:
: ~o
`
:: ~: :
~: '
~, - - - -
,
.
:
5/AOR12 ~51- 17885
E~A~¢E~
1.3.4.~--Tçtrameth~l=2-~L~IoL~or~=~=g~
propyl-thi~]-l~-imidazoli~m ~loride _ _
l-Chloro-3-phenoxy-propan-2 one was prepared
by the method of R. Longonl, P. Bernt~æon, N. ~ild,
and M. Hesse, ~lev. C~im. Aeta, 60, 103 (1977) and/or
K.J. Steven60n and L.B. Smillie, Canadian Journal o~
Biochemistry, 4~, 1357 (1968). In the preparation,
8.0 grams pheno~yacetyl chloride wa~ added dropwise
to a cooled (ice bath) æolution o~ about 3.0 grams of
diazomethane in 60 milliliter~ of ether to obtain an
intermediate l-diazo-3-phenoxypropan-2-one. Hydrogen
chloride was bubbled into the resulting mixture
whereupon nitrogen evolution occurred with the
formation of l-chloro-3-pheno~y-propan-2-one which
was reco~ered by conventional procedures.
To a solution of 0.50g (0.0032 mol) of
1,3,4,5-tetramethylimidazoline-2-thione in 25 mL of
acetone was added 0.59g ~0.0032 mol) of 1-chloro-3-
phenoxy-propan-2-one and the solutlon was ~tirred
overnight at room temperature. The product that
crystallized was collected by ~iltration.
Recrystallization from i~o-propyl-alcohol-~ther ~ave
1,3,4,5-tetram~thyl-2-[(3-phenoxy-2-o~o-propyl)thio)~-
lR-imidazolium chloride, m.p. 156-157.
Anal. Calcd for Cl~H21N202S: C, 56.37; ~, 6.21;
N, 8.22.
Found: C, 55.98; ~, 6.24; N, 8.05
:, :
~ , .
. ~ .. . .
2 ~ r,~) ~"
5/AOR12 -52- 17885
~XAMPLE ~XI
BenzYl-3-methyl-2- r ( 3-p~Qxy-2-oxopropyl ~
thiol-lH-imidazoli~m chlori~e
The above-identified compound, m.p.
110-112C was prspared in substantially the ~ame
manner as described in Example XX from l-chloro-3-
phenoxy-propan 2-one and 1-be!nzyl-3~methyl-imida-
zolium-2-thione.
EXAMPLE X~II
1~3-Dlmethyl~ -{[~ y~ldinyl-N-~ide ._
thio~-2-oxoprop~l]thio~ k~L~Lu~ ~hL~cLIe
0.51 g (0.002 mol) of 1,3-dimethyl-2-~(3- -
chloro-2-oxopropyl)thio]-lH-imidazolium chloride was
lS added to a solutio~ of 0.30 g (0.002 mol) of 2-
mercaptopyridine N-oxide æodium 6alt hydrate in 6 mL
of absolute ethanol. The resulting mixture was
.stirred at room temperature for 7.5 hours wher~upon a
white pr~cipitat:e formed. It waæ remoYed by
filtration and recry~tallized ~rom isopropyl alcohol-
ethanol to obtain 1,3-dimethyl-2-{[3-(2 pyridinyl-N-
oxide thio-2-oxopropyl]thio}-1~-imidazGlium chloride,
m.p. 194 (dec).
Anal. Calcd for C13~16ClN3O2S2: C, 40.94; E, 4.23;
N, 11.02;
Found: C, 41.01; ~, 4.13; N, 11.19;
.
,
:
"
5/AOR12 -53- 17885
EXAMPLE XXI I I
One liter o a parent:eral composition
comprising one of the foregoirlg compounds may be
prepaxed from the following formulation:
Grams
Imidazolium salt 5.0
Polysorbate 80 2.0
Sodium chloride 9.0
10 Sodium carboxymethylcellulose10.00
Methyl paraben 1.8
Propyl paraben 0.2
Water, USP g.s. to 1 liter
The parabens, sodium chloride and earboxy-
methylcellulose are dissolved in one-hal~ the total
volume of water by heating to 95C to obtain a
solution which is then filtered and autoclaved. The
polysorbate is dissolved in o~e-third of the total
volume of water, and the resulting solution also
filtered and autoclaved. Sterile aetive ingredient
- is added to the second ~olution and the mixture
passed through a sterile colloid mill to obtain a
suspension o~ active ingredient. The ~irst solution
is added to the suspen~ion with ~tirring then U.S.P.
~ater added to 1 liter. Sterile vials are filled
with the 6uspension while stirring.
3 0
~ ,
~ ~ f ? ~
J ' i )
5/AOR12 -54- 17885
EXAMPL~ IV
Oral ~omposition
5000 compressed tablets, each containing as
active ingredient 100 milligrams of one o the
5 foregoing compounds, may be prepared ~rom the
following formation:
~ra~
Imidazolium salt SOO
Starch 700
Dibasic calcium phosphate hydrous 5000
Calcium stearate 25
The ingredients are finely powdered, mixed
1~ well, and then granulated with 10 percen~ starch
paste. The granulation is dried and compre~sed into
tablets using starch as a disintegrant and calcium
stearate as lubricant.
Prepara~ion of ~he Startin~ Material~
A. ~-Mercaptoimldazole
The 2-mercaptoimidazoles may be obtained by
a reaction between an appropriate acyloin and
mono-substituted urea according to the following
equation:
:
:
:
:: :
.
,
:
,: .
5/AOR12 -55- 17885
R3~c~o S R ~ N
,CHOH R,N~ICNH2 ~ ~ H
R2 R2 R~
The reaction may be carried out by ~using
the reactants or by refluxing the component~ in
hexanol-l as more ~ully described by Nuhn, P. et.
al., J. fur praktische Chemie, 312, 90 (1970~ for the
fusion method and by Kjellin, G. et. al., Acta
Chemica Scandinavica, 23, 2879 (1969) for the method
where the ~~hydro~yketones and N-alkylthioureas are
re~luxed in l-he~anol with a water separator. The
teachingæ of the foregoing articles on the
preparation of the star~ing 2-mercaptoimidazoles are
incorporated by refercnce.
23 The acyloins may be prepared in a~y manner
within the knowledge of those skilled in the art.
B. 1.3-DiRub~ uted-imida~Qline-2-~h~Qa~
1,3-di~ub~tituted imidazoline-2-thione may
be obtained by the reaction bet~een an a-hydroxy-
ketone and di-~ubstituted thiourea according to the
equation
; :
,
-
.
" . J
5/AOR12 ~56- 17885
R~\ ~ S R4
C ~R4NHCNHRl ~~CN)~ S
R2,JCHOH R2
R1
The reactants may be intimately contacted in
the manner above described ~or the preparation o~ the
mercaptoimidazoles.
23
: ~5
:
~:~ 30
:: :
~ :
: : ; ~
. .
- ~
- ., ' :. . : . : ~ .
