CA2022116A1 - Imidazole compounds and their use as transglutaminase inhibitors - Google Patents

Abstract

TITLE OF THE INVENTION
IMIDAZOLE COMPOUNDS AND THEIR USE AS TRANSGLUTAMINASE
INHIBITORS

ABSTRACT OF THE DISCLOSURE
Imidazole compounds including imidazoles and imidazolium salts, and their use as transglutaminase inhibitors are disclosed.

Description

2, i - 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 :: :

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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 ~; ~

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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.

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J ~ r 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;

2 5 N (~

R~--¦ Q
Rl X ~ I I ) In the above and subsequent ~ormulas:

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.. . : , .. . ' . - : ............................. .
;~ .

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 .

~ : ~

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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 to&ether 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 , . . .

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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 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 ~ ~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 , - ' . ' 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.

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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.

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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.

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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.

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~J ~J ~J ~ .3 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 .

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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, - , ~ ,.

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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 ~::
:

': "-, ~ ,' ' ' - : ` ' ' , 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 ., - ;~

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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~

~ Ç? c~

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~

\,_N~

C6H~ I CF3S03 (_) 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 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

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 ~ 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~

CX3NHC- CH3 Cl~
o : 30 ~: :

~; :

:
. . , , . , . . . ,. . . - -. : . - . :~ , : .
..
. .
~: ' " ;. : . , .

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 _ ..

. .
-.:

.
, , 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 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~

(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 ~

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.

.

:

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 ~ ~2-CH_C~

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' 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 [I~N~-CH~-C-C~13 Cl (-~) I H~ lo~
CHa- C- OCH3 ::

:
' ', ~

s~ ( I '3 -~
'"''~

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~ _ 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.~

~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 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

:

. .

.
. .

, : ~ ' ' . . ' ~' , ~ '~ ' ; ' ,' '~ .

~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,~) ~"

~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;

.

, :

"

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.

~ , ~ ~ f ? ~
J ' i ) 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:

:
:
:

:: :

.
, :

,: .

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 The reactants may be intimately contacted in the manner above described ~or the preparation o~ the mercaptoimidazoles.

: ~5 :

~:~ 30 :: :

~ :
: : ; ~

. .
- ~

- ., ' :. . : . : ~ .

Claims (8)

1. An imidazole compound selected from the group consisting of:

(A) an imidazole having the formula;

or its acid addition salt, and (B) an imidazolium salt having the formula;

wherein in the above formulas, R is hydrogen;
lower alkyl;
substituted lower alkyl wherein the substituents are selected from hydroxy, lower alkoxy, phenoxy, phenylthio,

2-pyridinyl-N-oxide thio, and halo;
cycloalkyl from 3 to 6 carbon atoms;
benzyl;
substituted benzyl wherein the substituents are selected from halo, hydroxy, and lower alkoxy;
phenyl;
substituted phenyl containing 1 to 3 substituents selected from hydroxy, lower alkoxy, carbo(lower alkoxy), carbamido, N-(lower alkyl)carbamido or cyano;
pyridyl;
pyrimidinyl; or pyrazinyl;
R1 is lower alkyl, substituted lower alkyl wherein the substituent is carbalkoxy or carbamido;
or ArCnH2n-wherein Ar is phenyl, (lower alkyl)phenyl, (lower alkoxy)phenyl, or halopenyl and n is 1-3;
R2 is nitro;
carbo(lower alkoxy);
halo;
cyano;
phenyl;

5/AORl2 -59- 17885 substituted phenyl containing from 1 to 3 substituents selected from lower alkyl, lower alkoxy, halo and hydroxy;
phenoxy;
phenylthio:
an amido group represented by -NHCOQ wherein Q is lower alkyl, -CH(NH2)CH2C6H5 or -NH(lower alkyl);
a hydroxyalkyl group represented by wherein R' is hydrogen or R"

wherein R" is hydrogen, phenyl, phenoxyphenyl, biphenylyl, (lower alkyl)phenyl, lower alkyl and lower cycloalkyl, or R' and R" taken together is alkylene from 4 to 6 carbon atoms; or an ether-alkyl group represented by wherein R''' is hydrogen, -CO-(lower alkyl), -CO-phenyl, -CO-biphenylyl, -CO-phenyl-O-phenyl and -CONH-phenyl, and R"" is phenyl or lower alkyl;
R3 is hydrogen, or when R2 is phenyl or substituted phenyl is optionally the same as R2; or R2 and R3 taken together is alkylene from 3 to 10 carbon atoms optionally substituted with phenyl or spiroalkylene, or benzo;

R4 is lower alkyl or ArCnH2n wherein Ar is phenyl, (lower alkyl)phenyl, (lower alkoxy)phenyl, or halophenyl and n is 1-3; and X is a negative radical of a pharmaceutically acceptable salt;
wherein "lower" is from 1 to 6 carbon atoms, inclusive.

2. A compound according to Claim 1 which is an imidazolium salt.

3. A compound according to Claim 2 where R2 and R3 taken together is alkylene from 3 to 10 carbon atoms.

4. A compound according to Claim 3 where R
is lower alkyl or substituted lower alkyl.

5. A compound according to Claim 3 which is 1,3-dimethyl-4,5,6,7 tetrahydro-2[(2-oxopropyl) thio]-1H-imidazolium chloride.

6. A composition suitable for thrombolytic therapy in inhibiting or combatting thrombosis or for supplementing fibrinolytic therapy comprising an amount of an imidazole compound of Claim 1 in a pharmaceutically acceptable carrier.

7. A composition according to Claim 6 in unit dose form wherein the imidazole compound is present in amount of 100 mg to 10 grams.

8. A composition suitable for thrombolytic therapy in unit dosage form comprising (a) a plasminogen activtor selected from tPA in an amount of about 58 million I.U. and streptokinase in an amount of 1.5 million I.U. and (b) an imidazole compound according to Claim 1 in an amount of from 100 mg to 10 grams in a pharmaceutically acceptable carrier.