OA13293A - Method for producing (3-oxo-2,3-dihydro-1H-isoindol-1-yl) acetylguanidine derivatives. - Google Patents

Abstract

The invention relates to methods for producing (3-oxo-2,3-dihydro-1H-isoindol-1-yl) acetylguanidine derivatives of formula (I) using 3-hydroxy-2,3-dihydro-1 H-isoindol-1-one derivatives or 3-(2-carbamoyl-phenyl) acrylic acid ester derivatives as intermediate stages, to a method for the resolution of racemates and to intermediate products of the inventive method.

Description

1

The présent invention relates to processes for preparing (3-oxo-2,3-dihydro-1 H-isoindol-1-yl)acetyiguanidine dérivatives via 3-hydroxy-2,3-dihydro-1H-isoindol-1-onedérivatives or 3-(2-carbamoylphenyl)acrylic ester dérivatives as intermediates, to aprocess for optical resolution, and also to intermediates of the process according tothe invention. (3-Oxo-2,3-dihydro-1H-isoindol-1-yl)acetylguanidine dérivatives of the formula IO nh2

R1 R2 are NHE1 inhibitors and are described in PCT/EP03/05279. However, the synthèsesdescribed there lead to racemic regioisomer mixtures, which entails costly andinconvénient séparation processes and reduces the yield of the desired compound.Hitherto, it has only been possible to obtain the isomers by a costly and inconvénientchromatographie séparation on chiral supports. However, the substance throughputis restricted in chromatographie séparations.

There is therefore a great interest in finding regioselective préparation processes for(3-oxo-2,3-dihydro-1H-isoindol-1-yl)acetylguanidine dérivatives and processes forrecovering the enantiomers. The improved, regioselective préparation of the racemic(3-oxo-2,3-dihydro-1 H-fsoindol-1-yl)acetyiguanidine dérivatives succeeds by twoindependent routes which are shown in scheme 1 and scheme 3. The resolution ofthe racemates succeeds by crystallization as the salts of 2,3-O-acylated D- orL-tartaric acids, as shown in scheme 5. Gentle base-catalyzed racemization of the ineach case undesired enantiomer makes possible substantial conversion of theracemate to the desired enantiomer. The processes mentioned enable the simplepréparation of enantiomerically enriched or enantiomerically pure (3-oxo-2,3-dihydro-1H-isoindol-1-yl)acetylguanidine dérivatives. The novel processes now makepossible the simple préparation of large amounts of substance of the compounds of 13293 the formula I on the industrial scale.

The présent invention thus relates to a process for preparing compounds of theformula I

whereR1 and R2 are each independently hydrogen, F, Cl, trifluoromethoxy, 2,2,2-trifluoroethoxy, trifluoromethyl, 2,2,2-trifîuoroethyi or alkyl having 1,2,3 10 or 4 carbon atoms; R3 is Alk-R4 or trifluoromethyl;

Alk is alkyl having 1,2, 3 or 4 carbon atoms; R4 is hydrogen, trifluoromethyl or cycloalkyl having 3, 4, 5, 6 or 7 carbon atoms; 15 and salts thereof; which comprises, as shown in scheme 1,

Scheme 1 1 3293 a) formylating the amide of the formula IV and then cyclizing to the compound of theformula VI, b) reacting the compound of the formula VI with an alkoxycarbonylmethylene-triphenylphosphorane, with a 1-alkoxy-1-trimethylsiloxyethylene or with a trialkyl 5 phosphonoacetate to give the compound of the formula VII, and c) reacting the compound of the formula VII with guanidine to give the compound ofthe formula I, where, in the compounds of the formulae IV, VI and VII, R1 to R3 are each as defined in formula I and 10 R5 is alkoxy having 1,2, 3 or 4 carbon atoms;and salts thereof.

The invention also provides a process for preparing compounds of the formula Iwhere 15 R1 and R2 are each independently hydrogen, F, Cl, trifluoromethoxy, 2,2,2-trifluoroethoxy, trifluoromethyl, 2,2,2-trifluoroethyl oralkyl having 1,2, 3or 4 carbon atoms; R3 is Alk-R4 or trifluoromethyl; 20 Alk is alkyl having 1,2, 3 or 4 carbon atoms;

I R4 is hydrogen, trifluoromethyl or cycloalkyl having 3, 4, 5, 6 or7 carbon atoms; and salts thereof; wherein, as shown in scheme 2, 1 3293

Scheme 2 a) the compound of the formula II is reacted with the amine of the formula III to give5 the amide of the formula IV, b) the amide of the formula IV is formylated at the ortho-position to the amidefunction to give the formyl amide of the formula V, c) the formyl amide of the formula V is cyclized to the compound of the formula VI, d) the compound of the formula VI is reacted with an 10 alkoxycarbonylmethylenetriphenylphosphorane, with a 1-alkoxy-1-trimethylsiloxy-ethylene or with a trialkyl phosphonoacetate to give the compound of the formula VIIand e) the compound of the formula VII is reacted with guanidine to give the compoundof the formula I, 15 where, in the compounds of the formulae II, III, IV, V, VI and VII, R1 to R3 are each as defined in formula I, R5 is alkoxy having 1,2, 3 or 4 carbon atoms andX is Cl, Br, OH or alkoxy having 1,2, 3 or 4 carbon atoms;and salts thereof. 20 13293

The compound of-the formula II is typically reacted, in an inert solvent such as anether, hydrocarbon or halogenated hydrocarbon, for example dichloromethane, at atempérature between -30°C and the boiling point of the solvent, preferably at RT,with an amine of the formula III, if appropriate in the presence of an activating agent,to give the amide of the formula IV.

The ortho-formylation may, for example, be carried out by initially charging an alkyl-metal compound, for example an alkyllithium compound, preferably t-BuLi, with acomplex ligand, preferably TMEDA, in an inert solvent such as an ether orhydrocarbon, for example THF, at a température between -100°C and 0°C,preferably between -80°C and -50°C. Then, the amide of the formula IV is addedand deprotonation is effected over a period between 10 minutes and 10 hours,preferably between 10 minutes and 60 minutes, at a température between -100°Cand 0°C, preferably between -80°C and -50°C. Subsequently, a formylating agent,preferably DMF, is added and reaction with the anion is effected at a températurebetween -100°C and 40°C, preferably between -80°C and room température.Préférence is given to leaving the solution to corne to RT after addition of the DMFover a period of from 10 minutes to 3 hours, for example within 30 minutes. Amide ofthe formula V formed as an intermediate generally cyclizes directly to the isoindoloneof the formula VI.

The isoindolone of the formula VI is reacted with a (Ci-C4)-alkoxycarbonyl-methylenetriphenylphosphorane in an inert solvent such as an ether, hydrocarbon orhalogenated hydrocarbon, for example toluene, at a température between 0°C andthe boiling point of the solvent, preferably between 20°C and the boiling point of thesolvent, or with a tri(C^ -C4)-alkyl phosphonoacetate in the presence of a base, forexample sodium hydride, in an inert solvent such as an ether, hydrocarbon orhalogenated hydrocarbon, for example 1,2-dimethoxyethane, at a températurebetween 0°C and the boiling point of the solvent, preferably between 20°C and theboiling point of the solvent. Alternatively, the isoindolone of the formula VI is reactedwith a 1-(C-i-C4)-alkyloxy-1-trimethylsiloxyethylene in the presence of a Lewis acid, for example titanium(IV) chloride or trimethylsiiyl triflate, in an inert solvent such asan ether, hydrocarbon or halogenated hydrocarbon, for example dichloromethane, at 1 3293 a température between -80°C and the boiling point of the solvent, preferably at atempérature between -80°C and 20°C (Synth. Commun. 1987,17, 1).

The ester of the formula VII may be reacted by commonly known processes withguanidine to give the acylguanidine of the formula I. The reaction is preferablyeffected in the manner known to those skilled in the art in a protic or aprotic, polarbut inert organic solvent. For example, in the reaction of the methyl ester (formulaVII; R5=OCH3) with guanidine, useful solvents hâve been found to be methanol, isopropanol or THF at températures of from 20°C up to the boiling température ofthese solvents. In most reactions of compounds of the formula VII with salt-freeguanidine, operation is effected, for example, in aprotic, inert solvents, for exampleethers such as THF, dimethoxyethane or dioxane. However, water may also be usedwhen use is made of a base, for example NaOH, as a solvent in the reaction ofcompounds of the formula VII with guanidine. In the reaction of compounds of theform,ula VII with salts of guanidine, for example guanidine hydrochloride, the reactionis typically effected in the presence of a base, for example potassium tert-butoxide,sodium methoxide or sodium ethoxide, in an inert solvent such as dimethylformamide, NMP, 2-propanol, at a température between 20°C and theboiling point of the solvent.

In addition to the carboxyiic esters of the formula VII, it is also possible to use furtheractivated acid dérivatives in the reaction with guanidine, for example carbonylchlorides, carboxyiic thioesters or carboxyiic anhydrides. An activation of thecarboxyiic acid with, for example DCC can also be effected. The activated aciddérivatives can be prepared in the manner known to those skilled in the art directlyfrom the parent carboxyiic esters of the formula VII or from the correspondingcarboxyiic acids which can be obtained from the esters by customary hydrolysismethods. A sériés of suitable methods for preparing activated carboxyiic aciddérivatives are specified with citation of source literature in J. March, AdvancedOrganic Chemistry, Third Edition (John Wiley & Sons, 1985, p. 350).

The process steps described in scheme 1 and 2 may each independently be effectedcontinuously or batchwise. A workup of the reaction mixture may be effected at any 1 3 293 of the process steps. The workup and, if desired, the purification of the products iseffected by the customary methods such as extraction, pH séparation,chromatography or crystallization and the customary dryings. 5 The starting compounds of the formulae II and III are commercially available or canbe prepared according to or in a similar manner to the processes described in theliterature and familiar to those skilled in the art.

Also claimed is a process for preparing compounds of the formula I

10 whereR1 and R2 are each independently hydrogen, F, Cl, trifluoromethoxy, 2,2,2-trifluoroethoxy, trifluoromethyl, 2,2,2-trifluoroethyl oralkyl having 1, 2, 3 15 or 4 carbon atoms; R3 is Alk-R4 or trifluoromethyl;

Alk is alkyl having 1,2, 3 or 4 carbon atoms; R4 is hydrogen, trifluoromethyl or cycloalkyl having 3, 4, 5, 6 or 7 carbon atoms; 20 and salts thereof; which comprises, as shown in scheme 3,

H2NR3 iii

Scheme 3

IX 1 3293 a) reacting the amine of the formula IX via a diazonium sait with an alkyl acrylate togive the cinnamic acid dérivative of the formula XI, b) reacting the compound of the formula XI with the amine of the formula III and withguanidine to give the acylguanidine of the formula I, where, in the compounds of the formulae III, IX and XI, R1 to R3 are each as defined in formula I and R6 is alkoxy having 1,2, 3 or 4 carbon atoms; and salts thereof.

The présent invention also relates to a process for preparing the compounds of theformula I

where R1 and R2 are each independently hydrogen, F, Cl, trifluoromethoxy, 2,2,2-trifluoroethoxy, trifluoromethyl, 2,2,2-trifluoroethyl or alkyl having 1,2, 3or 4 carbon atoms; R3 isAlk-R4 or trifluoromethyl;

Alk is alkyl having 1,2, 3 or 4 carbon atoms; R4 is hydrogen, trifluoromethyl or cycloalkyl having 3, 4, 5, 6 or 7 carbon atoms; and salts thereof; wherein, as shown in scheme 4, 1 3293

XIV

Scheme 4 a) the nitro compounds of the formula VIII is converted to the amine of theformula IX, 5 b) the amine of the formula IX is converted to the diazonium sait of the formula X, c) the diazonium sait of the formula X is reacted with an alkyl acrylate to give thecinnamic acid dérivative of the formula XI, d) the compound of the formula XI is converted to the amide of the formula XII and e) the compound of the formula XII is converted to the acylguanidine of the formula I,10 either by converting the compound of the formula XII in the presence of a base to the isoindolone dérivative of the formula XIII and subsequently by reaction withguanidine with activation to give the acylguanidine of the formula I (alternative A), or,after formation of the isoindolone dérivative of the formula XIII, in the presence of a 10 1 3293 base, from the compound of the formula XII, by converting the compound of theformula XIII to the ester of the formula XIV and subsequently by reacting withguanidine to give the acylguanidine of the formula I (alternative B), orby converting the compound of the formula XII in the presence of a strong base to 5 the ester of the formula XIV and subsequently by reacting with guanidine to theacylguanidine of the formula I (alternative C), or by directly reacting the compound of the formula XII with guanidine in the presenceof a base with simultaneously proceeding guanylation and cyclization to give theisoindolone of the formula I (alternative D), 10 where, in the compounds of the formulae VIII, IX, X, XI, XII, XIII and XIV, R1 to R3 are each as defined in formula I and R6 and R7 are each independently alkoxy having 1,2, 3 or 4 carbon atoms;and salts thereof. . 15 The nitro compounds of the formula VIII may be reduced by known methods (forexample described in “Houben-Weyl, Methoden derorganischen Chemie", VolumeXI/1, Nitrogen compounds II, Georg Thieme Verlag Stuttgart, 1957, p. 360ff)to theaniline of the formula IX. Preference is given to catalytic hydrogénation, for exampleusing Pd/C, for example using 5% Pd/C or 10% Pd/C, in a solvent, for example an 20 alcohol, preferably éthanol, under a hydrogen atmosphère of from 1 bar to 200 barpressure, preferably from 1 bar to 10 bar of pressure.

The subséquent diazotization of the aniline of the formula IX is effected in an inertsolvent, preferably éthanol, in the presence of an acid whose anion does not 25 substitute the diazonium ion itself, for example HBF4 or HPFg, preferably HBF4, or,for example, H2SO4 and in the presence of a nitrite, preferably NaNC>2, at a température between -30°C and the boiling point of the solvent, preferably between0°C and 30°C. 30 The diazonium sait of the formula X is preferably reacted directly with a (C4-C4)-alkylacrylate, preferably ethyl acrylate, in the presence of a palladium catalyst, preferablyPd(OAc)2, at a température between 0°C and the boiling point of the solvent,preferably between 45°C to 55°C, to give the cinnam'rc acid dérivative of the 1 3293 formula XI.

The benzoic acid function of the compound of the formula XI may be converted tothe amide of the formula XII by methods known to those skilied in the art, preferablyvia the acid chloride or with the aid of DCC. This réaction may also be conducted insuch a way that the amide of the formula XII is cyclized in the reaction mixturedirectly to the ester of the formula XIV, i.e. the reaction of the compound of theformula XI to give the ester of the formula XIV is carried out in one step. This may bedone either under the basic reaction conditions of amide formation or the cyclizationmay be brought about by adding a base, for example triethylamine, Hünig’s base orpotassium tert-butoxide. A further alternative consists in converting the compound ofthe formula XI directly to the compound of the formula I by successively carrying outamide formation, cyclization and guanidation in the same reaction vessel, in whichcase the Feaction may be effected without isolating intermediates.

For the further conversion of the compound of the formula XII to the acylguanidine ofthe formula I there are 4 alternatives: ,

Alternative A: The conversion of the amide of the formula XII is preferably effectedusing aqueous alkali solution, preferably aqueous NaOH solution, in a solvent suchas an alcohof, preferably methanol or éthanol, at a température between -30°C andthe boiling point of the solvent, preferably at RT. Both the hydrolysis of the esterfunction and the cyclization to the isoindoione dérivative of the formula XIII takeplace. The compound of the formula XIII is activated for acylation by commonlyknown processes (and as described for scheme 1), for example using the acidchloride or with DCC, and the acylguanidine of the formula I is obtained.

Alternative B: As in alternative A, the carboxylic acid of the formula XIII issynthesized. Subsequently, standard processes for ester préparation, preferablyusing SOCI2 in an alcohol such as methanol or éthanol, are used to préparé, for example, the methyl or ethyl ester of the formula XIV. The ester of the formula XIV issubsequently converted to the acylguanidine of the formula l as described forscheme 1. 12 1 3293

Alternative C: The conversion of the amide of the formula XII is effected in a solution of a strong base, preferably methoxide ort-butoxide in an alcohol such as methanol or éthanol, and the methyl or ethyl ester of the formula XIV is obtained. The conversion of the ester of the formula XIV to the acylguanidine of the formula I is effected as described for scheme 1.

Alternative D: The amide of the formula XII is converted under customary conditionsfor the acylation of guanidine. The solvent used is an inert solvent such as an ether,hydrocarbon or halogenated hydrocarbon, preferably DMF. Typically, a guanidiniumsait is initially reacted with a strong base, preferably KOtBu, which releases the freeguanidine. The mixture is added to the solution of the compound of the formula XII ina solvent such as an alcohol, ether, hydrocarbon or halogenated hydrocarbon, forexample DMF, NMP or 2-propanol, In the course of the addition, the guanylation andthe cyclization to the isoindolone of the formula I occurs simultaneously. In onevariant, the compound of the formula XI is cyclized to the compound of the formulaXIV and then converted in situ to the compound of the formula I successiveiy in timeusing a catalytic amount of a strong base, for example potassium tert-butoxide orsodium methoxide or sodium ethoxide, in a solvent, for example DMF, NMP or 2-propanol.

Preference is given to alternative D in which the conversion of the benzoic aciddérivative of the formula XI is carried out in a one-pot process to give acylguanidineof the formula I.

The process steps described in scheme 2 may be effected continuously orbatchwise. A workup of the reaction mixture may be effected after any of the processsteps. The workup and, if desired, the purification of the products is effected by thecustomary methods such as extraction, pH séparation, chromatography orcrystallization and the customary dryings.

The starting compounds of the formulae III and VIII are commercially available orcan be prepared according to or in a similar manner to the processes described inthe literature and known to those skilled in the art. 13 1 3293

The invention also provides compounds of the formula XII

where 5 R1 and R2 are each independently hydrogen, F, Cl, trifluoromethoxy, 2,2,2-trifluoroethoxy, trifluoromethyl, 2,2,2-trifluoroethyl or alkyl having 1,2, 3or 4 carbon atoms; R3 is Alk-R4 or trifluoromethyl; 10 Alk is alkyl having 1,2, 3 or 4 carbon atoms; R4 is hydrogen, trifluoromethyl or cycloalkyl having 3,4, 5, 6 or. 7 carbon atoms; R6 is alkoxy having 1, 2, 3 or 4 carbon atoms;and salts thereof. 15

Also claimed is the use of the compounds of the formula XII as a syntheticintermediate.

Compounds of the formula I in enantiomerically enriched or in enantiomerically pure20 form may advantageously be prepared by a novel optical resolution process which likewise forms part of the subject matter of the présent invention. To this end, theracemates of the compounds of the formula I are crystallized as salts of 2,3-0-acyiated D- or L-tartaric acid, in the course of which the enantiomers are enriched inthe crystal or in the mother liquor. Subsequently, the free bases are released again 25 from the salts.

The présent invention thus relates to a process for isolating compounds of theformula la and Ib 14 1 3293

whereR1 and R2 are each independently hydrogen, F, Cl, trifluoromethoxy, 5 2,2,2-trifIuoroethoxy, trifluoromethyl, 2,2,2-trifluoroethyl or alkyl having 1,2, 3 or 4 carbon atoms; R3 is AIk-R4 or trifluoromethyl;

Alk is alkyl having 1,2, 3 or 4 carbon atoms; R4 is hydrogen, trifluoromethyl or cycloalkyl having 3, 4, 5, 6 or 10 7 carbon atoms; and salts thereof; which comprises, as shown in scheme 5,

la lb

Scheme 5 15 a) converting the compound of the formula I to salts of a 2,3-O-acylated D- or 15 13 293 L-tartaric acid and.obtaining the two salts of the formulae XVa and XVb separatelyby crystallization, and b) releasing the free bases of the formulae la and Ib from the two salts of the formulae XVa and XVb respectively, where, in the compounds of the formulae I, XVa and XVb, R1 to R3 are each as defined in the formulae la and IbR* is O R8 O R8 HO.

O

CO2H R8 R8 R8 is alkyl having 1,2, 3, 4, 5 or 6 carbon atoms or phenyl which is unsubstituted orsubstituted by 1, 2 or 3 substituents from the group of F, Cl, Br, I, alkyl having 1,2, 3or 4 carbon atoms or alkoxy having 1,2, 3 or 4 carbon atoms.

Also claimed is the above-described process in which the undesired enantiomer ofthe formula la or Ib is racemized again.

The racemate of the compound of the formula I is crystallized with a tartaric aciddérivative R*, for example Ο,Ο’-dibenzoyl-D-tartaric acid, O,O’-dibenzoyl-L-tartaricacid, O,O’-di(4-methylbenzoyl)-L-tartaric acid, O,O’-di(4-methylbenzoyl)-D-tartaricacid, O,O’-di(4-methoxybenzoyl)-L-tartaric acid or O,O'-di(4-methoxybenzoyl)-D-tartaric acid, preferably with O,O’-dibenzoyl-L-tartaric acid or Ο,Ο’-dibenzoyl-D-tartaric acid, in a suitable solvent, for example in an ether, e.g. diethyl ether,diisopropyl ether, dimethoxyethane, tetrahydrofuran ordioxane, in a halogenatedhydrocarbon, e.g. dichloromethane, trichloromethane, tetrachloromethane, 1,2-dichloromethane or trichloroethylene, in an alcohol, e.g. methanol, éthanol,n-propanol, 2-propanol, butanol, in an ester, e.g. ethyl acetate or butyl acetate, inwater, or in mixtures of solvents, preferably in 2-propanol, dimethoxyethane or ethylacetate, at a température between -10°C and the boiling point of the solvent,preferably at from 0°C to 40°C. In one variant of the process, mixtures of two or more 16 13293 2,3-O-acylated D- or L-tartaric acids of the same configuration which bear differentacyl groups are used for the séparation.

The sait formation from the compound of the formula I and the tartaric acid dérivativeR* can be effected using équivalent amounts, i.e. 0.5 mol of the tartaric aciddérivative R* which contains two carboxylic acid groups may be used per mole of thecompound of the formula I. However, the compound of the formula I may also becrystallized with less than 0.5 mol équivalent of the 2,3-O-acylated D- or L-tartaricacid, for example with from 0.25 mol to 0.5 mol of tartaric acid dérivative R* per moleof the compound of the formula I, in particular with from 0.25 mol to 0.3 mol oftartaric acid dérivative R* per mole of the compound of the formula I. The desiredenantiomer then crystallizes out in the form of the sait of the formula XVa or XVb andthe undesired enantiomer is for the most part présent in the mother liquor in the formof the enantiomers of the formula lb or la and not in the form of the sait of theformula XVa or XVb. The enantiomeric purity of the salts of the formulae XVa andXVb may be increased by repeâted crystallization or by stirring of the first crystalswith fresh solvent at elevated température and subséquent cooling.

After séparation of the two salts of the formulae XVa and XVb and séparation of thesait of the formula XVa or XVb from the undesired enantiomer lb or la, theenantiomerically enriched compounds of the formulae la and lb are subsequentlytypically released from the salts by addition of an auxiliary base, for example anamine, e.g. triethylamine, an inorganic base such as NaHCO3, Na2CO3 or aqueoussolutions thereof. It is customary to work in a suitable solvent, for example in anether, e.g. diethyl ether, diisopropyl ether, dimethoxyethane, tetrahydrofuran ordioxane, in a halogenated hydrocarbon, e.g. diçhloromethane, trichloromethane,tetrachloromethane, 1,2-dichloroethane or trichloroethylene, in an alcohol, e.g.methanol, éthanol, n-propanol, 2-propanol or butanol, in an ester, e.g. ethyl acetateor butyl acetate, or in water or in solvent mixtures, preferably in ethyl acetate,2-propanol, diçhloromethane or water or mixtures thereof, in which case the reactionmixture may hâve one or more phases, at a température between -10°C and theboiling point of the solvent, preferably at from 10°C to 40°C. This may be done, forexample, in such a way that the sait is dissoived in aqueous NaHCOs solution and 17 13293 the enantiomer of the formula la or Ib is then extracted using an organic solvent, forexample ethyl acetate.

The enantiomer la or Ib which is undesired in each case may be converted back tothe racemate of the formula I by a racemization process and is thus available foranother optical résolution step. In this case, the undesired enantiomer is preferablytreated in a solvent such as an alcohol, e.g. 2-propanol, at a température between-10°C and the boiling point of the solvent, preferably at from 0°C to 40°C, with smallamounts of a base, for example KOH, the reaction mixture is neutralized and theracemate is isolated after aqueous-extractive workup. This process may be carriedout by suitable sélection of the amount of base and reaction température in such away that virtually exclusively racemization and no Chemical change in the substanceoccurs.

The présent invention also provides compounds of the formulae XVa and XVb

R1 and R2 are each independently hydrogen, F, Cl, trifluoromethoxy,2,2,2-trifluoroethoxy, trifluoromethyl, 2,2,2-trifluoroethyl or alkyl having 1, 2, 3or 4 carbon atoms; R3 is Alk-R4 or trifluoromethyl;

Alk is alkyl having 1,2, 3 or 4 carbon atoms; R4 is hydrogen, trifluoromethyl or cycloalkyl having 3, 4, 5, 6 or 7 carbon atoms; R* is 18

13293

R8 R8 R8 is alkyl having 1,2, 3, 4, 5 or 6 carbon atoms or phenyl which is unsubstituted orsubstituted by 1,2 or 3 substituents from the group of F, Cl, Br, I, alkyl having 1,2, 3or 4 carbon atoms or alkoxy having 1, 2, 3 or 4 carbon atoms.

When the above-described compounds, for example the compounds of the formulaeI, la, Ib, VII, XIII, XIV, XVa orXVb, contain one or more centers of asymmetry, theymay each independently hâve either S or R configuration, unless stated otherwise.The compounds may be présent in the form of optical isomers, of diastereomers, ofracemates or of mixtures thereof, unless they are more precisely defined. On doublebonds, either E orZ configuration may be présent, unless stated otherwise. Theprésent invention encompasses ail tautomeric forms of the above-describedcompounds, for example of the compounds of the formulae I, la, Ib, XVa and XVb.

Alkyl radicals may be straight-chain or branched. This is also true when they bearsubstituents or occur as substituents of other radicals, for example in fluoroalkylradicals or alkoxy radicals. Examples of alkyl radicals are methyl, ethyl, n-propyl,isopropyl (= 1-methylethyl), n-butyl, isobutyl (= 2-methylpropyl), sec-butyl(= 1-methylpropyl), tert-butyl (= 1,1-dimethylethyl), n-pentyl, isopentyl, tert-pentyl,neopentyl and hexyl. Preferred alkyl radicals are methyl, ethyl, n-propyl andisopropyl, more preferably methyl or ethyl. In alkyl radicals, one or more, for exampie1,2, 3, 4 or 5, hydrogen atoms may be substituted by fluorine atoms. Examples ofsuch fluoroalkyl radicals are trifluoromethyl, 2,2,2-trifluoroethyl and pentafluoroethyl,preferably trifluoromethyl or 2,2,2-trifluoroethyl. Substituted alkyl radicals may besubstituted in any positions.

Examples of cycloalkyl radicals are cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl orcycloheptyl. 19 1 3293

Phenyl radicals may be unsubstituted or mono- or polysubstituted, for examplemono-, di- or trisubstituted, by identical or different radicals. When a phenyl radical issubstituted, it preferably bears one or two identical or different substituents. Inmonosubstituted phenyl radicals, the substituent may be be disposed in the 5 2-position, the 3-position or the 4-position. Disubstituted phenyl may be substitutedin the 2,3-position, 2,4-position, 2,5-position, 2,6-position, 3,4-position or3,5-position. In trisubstituted phenyl radicals, the substituents may be disposed in the 2.3.4- position, 2,3,5-position, 2,4,5-position, 2,4,6-position, 2,3,6-position or 3.4.5- position. 10,

The above-described compounds, for example the compounds of the formulae I, laand Ib, may be used in the process according to the invention in the form of theirsalts and/or Îsolated in the form of their salts. Salts may be obtained by thecustomary methods, for example by reacting with acids or bases in a solvent, or by 15 anion exchange or cation exchange from other salts. Useful acid addition salts, forexample of the compounds of the formulae I, la and Ib, are, for example, halides, inparticularhydrochlorides, hydrobromides, lactates, sulfates, citrates, tartrates,acétates, phosphates, methylsulfonates, benzenesulfonates, p-toluenesulfonates,adipates, fumarates, gluconates, glutamates, glycerolphosphates, maleates, 20 benzoates, oxalates and pamoates and trifluoroacetates. In the case of the préparation of active ingrédients, preference is given to physiologically tolerated saltsand pharmaceutically acceptable salts. Examples include salts of compounds of theformulae I, la and Ib with fumaric acid, in particular salts which contain 1 mole offumaric acid per mole of the compound of the formula I, la or Ib, and are thus 25 hydrogenfumarates or hemifumarates. Advantageous properties such as crystallinity,stability, particularly low hygroscopicity, a low tendency to racemization and goodsolubility are features especially, for example, of (S)-N-{2-[3-oxo-2-(2,2,2-trifluoroethyl)-6-trifluoromethyl-2,3-dihydro-1H-isoindol-1-yl]acetyl}guanidinehydrogenfumarate hydrate of the formula XVI which likewise forms part of the 30 subject matter of the présent compound in ail its tautomeric forms. 20 13293

When the compounds contain an acid group, they may form salts with bases, forexample alkali métal salts, preferably sodium salts or potassium salts, or ammoniumsalts, for example salts with ammonia or organic amines or amino acids. Compounds 5 which contain a basic group and an acid group may also be présent in the form of azwitterion.

One embodiment of the présent invention relates to compounds in which R1 and R2are not both hydrogen, in particularto compounds in which R1 is hydrogen and R2 is 10 fluorine, chlorine or trifluoromethyl, especially trifluoromethyl. In compounds in whichR1 is hydrogen, the R2 substituent is preferably disposed in the para-position of thebenzene ring relative to the C=O group in the isoindolone System.

The Alk group is preferably alkyl having 1,2 or 3 carbon atoms, in particular having 1 15 or 2 carbon àtoms, especially having 1 carbon atom. R4 is preferably trifluoromethylor cycloalkyl having 3, 5 or 6 carbon atoms, in particular 3 carbon atoms, morepreferably trifluoromethyl. One embodiment of the présent invention relates tocompounds in which R3 is trifluoromethyl or 2,2,2-trifluoroethyl, in particular2,2,2-trifluoroethyl. 20 A spécial embodiment of the présent invention relates to the préparation of N-{2-[3-oxo-2-(2,2,2-trifluoroethyl)-6-trifluoromethyl-2,3-dihydro-lH-isoindol-1-yl]âcetyl}-guanidine and its enantiomeric forms and salts thereof. 25 X is preferably chlorine or methoxy, in particular chlorine. R5 is preferably methoxyor ethoxy, in particular ethoxy. R6 is preferably methoxy or ethoxy, in particularethoxy. R7 is preferably methoxy or ethoxy, in particular ethoxy.

In one embodiment of the présent invention, R8 is phenyl which is unsubstituted or 21 1 3293 substituted by 1, 2 or 3 substitutents from the group of F, Cl, alkyl having 1,2, 3 or 4carbon atoms, or alkoxy having 1,2, 3 or 4 carbon atoms, in particular unsubstitutedphenyl.

The compounds of the formulae I, la, Ib, XVa and XVb and their pharmaceuticallytolerated salts are substituted acylguanidines and inhibit the cellular sodium-protonantiporter (Na+/H+ exchanger, NHE), in particular the subtype NHE-1.

Owing to the NHE inhibitory properties, the compounds of the formulae I, la, Ib, XVa,XVb and XVI and/or their pharmaceutically acceptable salts are suitable for theprévention and treatment of disorders which are caused by activation of or byactivated NHE, and also of disorders which hâve the NHE-related damage as asecondary cause.

The compounds of the formulae I, la, Ib, XVa and XVb may also be used for thetreatment and prévention of disorders in which the NHE is only partially inhibited, forexample by using a lower dose.

Since NHE inhibitors act predominantiy via their influence on cellular pH régulation,they can generally be advantageously combined with other compounds whichregulate the intracellular pH value, useful combination partners being inhibitors of theenzyme group of the carbonic anhydrases, inhibitors of the Systems transportingbicarbonate ions, such as of the sodium bicarbonate cotransporter (NBC) or of thesodium-dependent chloride-bicarbonate exchanger (NCBE), and also NHE inhibitorswith inhibitory action on other NHE subtypes, because they can reinforce ormodulate the pharmacologically relevant pH-regulating effects of the NHE inhibitorsdescribed here.

The use of the compounds of the formulae I, la, Ib, XVa, XVb or XVI relates to theprévention and to the treatment of acute and chronic disorders in veterinary and inhuman medicine. For instance, the inventive inhibitors of the NHE are suitable fortreating disorders which are induced by ischemia and by reperfusion.

As a resuit of their pharmacological properties, the compounds described here aresuitable as antiarrhythmic médicaments. Their cardioprotective component makes 22 1 3293 NHE inhibitors outstandingly suitable for infarction prophylaxis and infarctiontreatment, and also for the treatment of angina pectoris, in which cases they alsopreventively inhibit or greatly reduce the pathophysiological processes whenischemia-induced damage arises, in particular when ischemia-induced cardiacarrhythmias are triggered. Owing to their protective actions against pathologicalhypoxie and ischémie situations, the compounds of the formuiae I, la, Ib, XVa, XVband XVI used in accordance with. the invention and/or pharmaceuticaily acceptablesalts thereof, as a resuit of inhibition of the cellular Na+/H+ exchange mechanism,may be used as médicaments for the treatment of ail acute or chronic damageinduced by ischemia or disorders induced primarily or secondarily thereby.

This also relates to their use as médicaments for surgical interventions. Thus, thecompounds may be used in organ transplants, in which case the compounds may beused for the protection of the organs in the donor before and during the removal, toprotect removed organs, for example in the course of treatment with or their storagein physiological bath liquids, and also in the course of transfer to the récipientorganism.

The inventive compounds are likewise valuable médicaments having a protectiveaction in the performance of angioplastie surgical interventions, for example on theheart and also on peripheral organs and vessels.

The inventive compounds may also be used when performing bypass operations, forexample in bypass operations on coronary vessels and in Coronary Artery BypassGraft (CABG).

Depending on their action against ischemia-induced damage, the inventivecompounds of the formula I can even be used for resuscitation after a cardiac arrest.

The inventive compounds are of interest for médicaments against life-threateningarrhythmias. Ventricular fibrillation is terminated and the physiological sinus rhythmof the heart is restored. 23 1 3293

Since NHE1 inhibitors of human tissue and organs, especially the heart, protecteffectively not only against damage caused by ischemia and reperfusion but alsoagainst the cytotoxic action of médicaments like those finding use in particular incancer therapy and the therapy of autoimmune diseases, combined administrationwith NHE inhibitors is suitable for inhibiting the cytotoxic, especially cardiotoxic, sideeffects of the compounds mentioned. The réduction in the cytotoxic effects,especially the cardiotoxicity, resulting from comedication with NHE1 inhibitors alsomakes it possible to increase the dose of the cytotoxic therapeutic agents and/or toprolong the médication with such médicaments. The therapeutic benefit of such acytotoxic therapy can be considerably increased by the combination with NHEinhibitors.

Moreover, NHE1 inhibitors can be used in the event of heart-damaging overproduction of thyroid hormones, thyrotoxicosis, orwhen thyroid hormones aresupplied externally. The compounds of the formulae I, la, tb, XVa, XVb and XVI.and/or the pharmaceutically acceptable salts thereof are thus suitable for improvingtherapy with cardiotoxic médicaments.

In accordance with their protective effect against ischemia-induced damage, theinventive compounds are also suitable as médicaments for the treatment ofischemias of the nervous System, especially of the central nervous System, and theyare suitable, for example, for the treatment of stroke or of cérébral edema. NHE inhibitors are also suitable for thé therapy and prophylaxis of diseases anddisorders which are induced by hyperexcitability of the central nervous system, inparticular for the treatment of epileptic disorders, centrally induced clonie and toniespasms, States of psychological dépréssion, anxiety disorders and psychoses. Inthese cases, it is possible to use the NHE inhibitors described here alone or incombination with other substances having antiepiieptic activity or antipsychotic activeingrédients, or carbonic anhydrase inhibitors, for example with acetazoiamide, andwith other inhibitors of NHE or of the sodium-dependent chloride-bicarbonateexchanger (NCBE). 24 13293

In addition, NHE irihibitors are likewise suitable for the treatment of types of shock,for example of allergie, cardiogenic, hypovolémie and bacterial shock.

The compounds of the formulae I, la, Ib, XVa, XVb and XVI and/orthe pharmaceutically acceptable salts thereof may likewise be used for the préventionand treatment of thrombotic disorders, since they, as NHE inhibitors, are themselvesable to inhibit platelet aggregation. In addition, they are able to inhibit or prevent theexcessive release, occurring after ischemia and reperfusion, of mediators ofinflammation and coagulation, especially of von Willebrand factor and ofthrombogenic selectin proteins. It is thus possible to reduce and eliminate thepathogenic action of significant thrombogenic factors. The NHE inhibitors of theprésent invention can therefore be combined with other anticoagulant and/orthrombolytic active ingrédients, for example recombinant or natural tissueplasminogen activator, streptokinase, urokinase, acetylsalicylic acid, thrombinantagonists, factor Xa antagonists, médicinal substances with fibrinolytic activity,thromboxane receptor antagonists, phosphodiesterase inhibitors, factor Vllaantagonists, clopidogrel, ticlopidine etc. It is particularly favorable to use the présentNHE inhibitors in combination with NCBE inhibitors and/or with inhibitors of carbonicanhydrase, for example with acetazolamide.

In addition, NHE inhibitors feature a strong inhibitory effect on the prolifération ofcells, for example fibroblast cell prolifération and the prolifération of smooth vascularmuscle cells. The compounds of the formulae I, la, Ib, XVa, XVb and XVI and/orthepharmaceutically acceptable salts thereof are therefore useful as valuabletherapeutic agents for diseases in which prolifération represents a primary orsecondary cause, and can therefore be used as antiatherosclerotics, agents forchronic rénal failure, cancers.

It has been possible to show that cell migration is inhibited by NHE inhibitors. Thecompounds of the formulae I, la, Ib, XVa, XVb and XVI and/orthe pharmaceuticallyacceptable salts thereof are therefore suitable as valuable therapeutic agents fordiseases in which cell migration represents a primary or secondary cause, forexample, cancers with a pronounced tendency to metastasis. 25 Ί 3293 ΝΗΕ inhibitors further feature a retardation or prévention of fibrotic disorders. Theyare thus suitable as excellent agents for the treatment of cardiac fibroses, and ofpulmonary fibrosis, hepatic fibrosis, rénal fibrosis and other fibrotic disorders. Theycan thus be used for the treatment of organ hypertrophies and hyperplasias, forexample of the heart and the prostate. They are therefore suitable for the préventionand treatment of heart failure (congestive heart failure = CHF) and for the treatmentand prévention of prostate hyperplasia or prostate hypertrophy.

Since there is significant élévation in NHE in essential hypertensives, thecompounds of the formulae I, la, Ib, XVa, XVb and XVI and/orthe pharmaceuticallyacceptable salts thereof are suitable for the prévention and treatment of high bloodpressure and for the treatment of cardlovascular disorders. In these cases they canbe used alone orwith a suitable combination and formulation partner for thetreatment of high blood pressure and of cardiovasculan disorders. For example, oneor more diuretics with a thiazide-like action, loop diuretics, aldostérone andpseudoaldosterone antagonists, such as hydrochlorothiazide, indapamide,polythiazide, furosemide, piretanide, torasemide, bumetanide, amiloride, triamterene,spironolactone or eplerone, may be combined. The NHE inhibitors of the présentinvention may also be used in combination with calcium antagonists such asverapamil, diltiazem, amlodipine or nifedipine, and with ACE inhibitors, for exampleramipril, enalapril, lisinopril, fosinopril or captopril. Further favorable combinationpartners are also ^-blockers such as metoprolol, albuterol etc., antagonists of theangïotensin receptor and its receptor subtypes such as losartan, irbesartan,valsartan, omapatrilat, gemopatrilat, endothelin antagonists, renin inhibitors,adenosine receptor agonists, inhibitors and activators of potassium channels such asglibenclamide, glimepiride, diazoxide, cromakalim, minoxidil and dérivatives thereof,activators of the mitochondrial ATP-sensitive potassium channel (mitoK(ATP)channel), inhibitors of Kv1.5 etc.

It has been shown that NHE1 inhibitors hâve a significant antiinflammatory effect andcan thus be used as antiinflammatory drugs. The inhibition of the release ofmediators of inflammation is noteworthy in this connection. The compounds maythus be used alone or in combination with an antiinflammatory drug in theprévention or treatment of chronic and acute inflammatory disorders. The 26 13293 combination partners used advantageously are stéroïdal and non-steroidalantiinflammatory drugs. The inventive compounds may also be used for theprévention or treatment of diseases which are caused by protozoa, such as in theevent of malaria or coccidiosis in poultry.

It has also been found that NHE inhibitors exhibit a bénéficiai effect on sérumlipoproteins. It is generally acknowledged that blood fat levels which are too high,known as hyperlipoproteinemias, constitute an essential risk factor for thedevelopment of arteriosclerotic vascular lésions, especially coronary heart disease.The réduction of elevated sérum lipoproteins is therefore of exceptional importancefor the prophylaxrs and the régression of atherosclerotic lésions. In addition to theréduction in total sérum cholestérol, it is particularly important to reduce theproportion of spécifie atherogenic lipid fractions in this total cholestérol, in particularof the low density lipoproteins (LDL) and of the very low density lipoproteins (VLDL),since these lipid fractions constitute an atherogenic risk factor. By contrast, aprotective function against coronary heart disease is ascribed to the high densitylipoproteins. Accordingly, hypolipidémies should be capable of reducing not only totalcholestérol but also in particular the VLDL and LDL sérum cholestérol fractions. Ithas now been found that NHE1 inhibitors exhibit valuable therapeutically utilizableproperties in relation to influencing the sérum lipid levels. For instance, theysignificantly reduce the elevated sérum concentrations of LDL and VLDL, as can beobserved, for example, as a resuit of increased dietary intake of a cholestérol- andlipid-rich diet or in cases of pathological metabolic alterations, for examplegenetically related hyperlipidemias. They can therefore be used for the prophylaxisand for the régression of atherosclerotic lésions by eliminating a causal risk factor.These include not only the. primary hyperlipidemias but also certain secondaryhyperlipidemias, as occur, for example, in the event of diabètes. In addition, NHEinhibitors lead to a marked réduction in the infarctions induced by metabolicabnormalities and in particular to a significant réduction in the induced infarction sizeand the severity thereof.

The inventive compounds of the formulae I, la, Ib, XVa, XVb and XVI thereforeadvantageously find use for preparing a médicament for the treatment ofhypercholesterolemia; for preparing a médicament for the prévention of 27 13293 atherogenesis; for preparing a médicament for the prévention and treatment ofatherosclerosis, for preparing a médicament for the prévention and treatment ofdiseases whiçh are induced by elevated cholestérol levels, for preparing amédicament for the prévention and treatment of diseases induced by endothélialdysfunction, for preparing a médicament for the prévention and treatment ofatherosclerosis-induced hypertension, for preparing a médicament for the préventionand treatment of atherosclerosis-induced thromboses, for preparing a médicamentfor the prévention and treatment of hypercholesterolemia-induced and endothélialdysfunction-induced ischémie damage and post-ischemic reperfusion damage, forpreparing a médicament for the prévention and treatment of hypercholesterolemia-induced and endothélial dysfunction-induced cardiac hypertrophies andcardiomyopathies and of congestive heart failure (CHF), for preparing a médicamentfor the prévention and treatment of hypercholesterolemia-induced and endothélialdysfunction-induced coronary vasospasms and myocardial infarctions, for preparinga médicament for the treatment of the disorders mentioned in combinations withhypotensive substances, preferably with angiotensin converting enzyme (ACE)inhibitors and angiotensin receptor antagonists. A combination of an NHE inhibitorwith an active ingrédient lowering the blood fat levels, preferably with an HMG-CoAreductase inhibitor (for example lovastatin or pravastatin), the latter bringing abouthypolipidémie action and thus increasing the hypolipidémie properties of the NHEinhibitor, is found to be a favorable combination with enhanced action and reduceduse of active ingrédients.

For instance, NHE inhibitors lead to effective protection against endothélial damageof various origins. This protection of the vessels against the syndrome of endothélialdysfunction makes compounds of the formulae I, la, Ib, XVa, XVb and XVI and/orthepharmaceutically acceptable salts thereof valuable médicaments for the préventionand for the treatment of coronary vasospasms, peripheral vascular diseases, inparticular intermittent claudication, atherogenesis and atherosclerosis, left ventricularhypertrophy and dilated cardiomyopathy and thrombotic disorders.

It has also been found that NHE inhibitors are suitable in the treatmenf of non-insulin-dependent diabètes (NIDDM), in the course of which the insulin résistance isrestrained. In this case, it may be bénéficiai to enhance antidiabetic activity andquality of action of the inventive compounds by combining them with a biguanide 28 1 3293 such as metformin, with an antidiabetic sulfonylurea such as glyburide, glimepiride,tolbutamide etc., with a glucosidase inhibitor, with a PPAR agonist such asrosiglitazone, piogtitazone etc., with an insulin product of different administrationform, with a DB4 inhibitor, with an insulin sensitizer or with meglitinide.

In addition to the acute antidiabetic effects, NHE inhibitors counteract thedevelopment of late complications of diabètes and can therefore be used asmédicaments for the prévention and treatment of late damage from diabètes, suchas diabetic nephropathy, diabetic neuro'pathy, diabetic retinopathy, diabeticcardiomyopathy and other disorders occurring as a conséquence of diabètes. Theycan in this connection be advantageously combined with the antidiabeticmédicaments just described under NIDDM treatment. The combination with abénéficiai dosage form of insulin might be particularly important in this connection. NHE inhibitors exhibit, in addition to the protective effects against acute ischémieevents and the subséquent equally acutely stressing reperfusion events, also directtherapeutically utilizable actions against diseases and disorders of the entiremammalian organism which are associated with the manifestations of the chronicallyprogressive aging process and which occur independently of acute hypoperfusionStates and under normal, non-ischemic conditions. These pathological, age-relatedmanifestations induced overthe long aging period, such as illness, invalidity anddeath, which can now be made amenable to treatment with NHE inhibitors, arediseases and disorders which are essentially caused by age-related changes in vitalorgans and the function thereof and become increasingly important in the agingorganism.

Disorders which are connected with an age-related functional impairment or withage-related manifestations ofwearon organs are, for example, the inadéquateresponse and reactivity of the blood vessels to contraction and relaxation reactions.This age-related décliné in the reactivity of vessels to constricting and relaxingstimuli, which are an essential process of the cardiovascular System and thus of lifeand health, can be significantly eliminated or reduced by NHE inhibitors. Oneimportant function and a measure of the maintenance of the reactivity of vessels isthe blockade or retardation of the age-related progression in endothélial dysfunction,which can be eliminated highly significantly by NHE inhibitors. NHE inhibitors are 29 1 3293 thus outstandingly.suitable for the treatment and prévention of the age-relatedprogression in endothélial dysfunction, especially of intermittent claudication.

An example ofanother variable characterizing the aging process isthe décliné in thecontractability of the heart and the décliné in the adaptation of the heart to a requiredpumping output of the heart. This diminished efficiency of the heart as aconséquence of the aging process is in most cases connected with a dysfunction ofthe heart which is caused inter alia by déposition of connective tissue in themyocardial tissue. This déposition of connective tissue is characterîzed by anincrease in the weight of the heart, by an enlargement of the heart and by restrictedcardiac function. It is surprising that it has been possible to virtually completely inhibitsuch aging of the heart organ. NHE inhibitors are thus outstandingly suitable for thetreatment and prévention of heart failure, of congestive îieart failure (CHF). .

Inhibition of prolifération allows not only forms of cancer which hâve alreadyoccurred to be cured, but also réduction and highly significant retardation of the age-related incidence of cancer through NHE inhibitors. A particularly noteworthy findingis that the disorders, occurring as a resuit of aging, of ail organs and not only certaintypes of cancer are suppressed or occur with a highly significant delay. NHEinhibitors are thus outstandingly suitable for the treatment and, in particular, theprévention of age-related types of cancer.

Using NHE inhibitors, a delay, shifted highly significantly in time, is found in theoccurrence of age-related disorders of ail the organs investigated, including theheart, vessels, liver etc., and also a highly significant delay in cancer of the elderly.Moreover, there is also surprisingly a prolongation of life to an extent which has todate been achievable by no other group of médicaments or by any natural products.This unique effect of NHE inhibitors also enables, in addition to the sole use of theactive ingrédients on humans and animais, these NHE inhibitors to be combined withother activity principles, measures, substances and natural products which are usedin gerontology and which are based on a different mechanism of action. Suchclasses of active ingrédients used in gerontological therapy are: in particular vitaminsand substances with antioxidant activity. Since there is a corrélation between calorieload or food intake and the aging process, it is possible to combine with dietary 30 1 3 2 9 3 measures, for example with appetite suppressants. It is equally possible to consider a combination with hypotensive médicaments such as with ACE inhibitors, angiotensin receptor antagonists, diuretics, Ca+2 antagonists etc. or with metabolism- normalizing médicaments such as cholesterol-lowering agents. NHE inhibitors are thus outstandingly suitable for the prévention of age-related tissuechanges and for prolonging life while retaining a high quality of life.

The inventive compounds are effective inhibitors of the cellular sodium-protonantiporter (Na/H exchanger) which in numerous disorders (essential hypertension,atherosclerosis, diabètes etc.) fs also increased in cells which are readily amenableto measurements, for example in érythrocytes, thrombocytes or leukocytes. Thecompounds used in accordance with the invention are therefore suitable asoutstanding and simple scientific tools, for example in their use as diagnostic agentsfor determining and distinguishing different types of hypertension, but also ofatherosclerosis, diabètes and the late complications of diabètes, proliférativedisorders etc.

Also claimed is a medicine for human, veterinary or phytoprotective use which,togetherwith pharmaceutically acceptable carriers and excipients, comprises aneffective amount of one or more compounds of the formulae XVa, XVb and XVIand/or pharmaceutically acceptable salts thereof, alone or in combination with otherpharmacological active ingrédients or médicaments. Médicaments which comprise acompound of the formulae I, la, Ib, XVa, XVb and XVI and/or the pharmaceuticallyacceptable salts thereof can be administered, for example, orally, parenterally,;intravenously, rectally, percutaneously or by inhalation, the preferred administrationbeing dépendent on the particular characteristics of the disorder. The compounds ofthe formulae I, la, Ib, XVa, XVb and XVI may be used alone or together withpharmaçeutical excipients, both in veterinary medicine and in human medicine. Themédicaments generally comprise active ingrédients of the formulae I, la, Ib, XVa, XVb and XVI and/or the pharmaceutically acceptable salts thereof in an amount offrom 0.01 mg to 1 g per dose unit.

Which excipients are suitable for the desired pharmaceutical formulation are familiarto those skilled in the art on the basis of their expert knowledge, in addition to 31 1 3293 solvents, gel formers, suppository bases, tablet excipients, and other activeingrédient carriers, it is possible to use, for example, antioxidants, dispersants,emulsifiers, antifoams, flavorings, preservatives, solubilizers or.colors.

For an oral administration form, the active compounds are mixed with additivessuitable therefor, such as carriers, stabilizers or inert diluents, and converted byconventional methods to suitable dosage forms such as tablets, coated tablets, hardgelatin capsules, aqueous, alcoholic or oily solutions. Examples of inert carrierswhich can be used are gum arabic, magnesia, magnésium carbonate, potassiumphosphate, lactose, glucose or starch, especially corn starch. Thé formulation maybe effected either in the form of a dry granule or a wet granule. Examples of suitableoily carriers or solvents are vegetable or animal oils such as sunflower oil or fish liveroil.

For subcutaneous, intramuscular or intravenous administration, the activecompounds used, if desired with the substances customary therefor, such assolubilizers, emulsifiers or other excipients, are brought into solution, suspension orémulsion. Examples of useful solvents are: water, physiological saline or alcohols,e.g. éthanol, propanol, glycerol, as well as sugar solutions such as glucose ormannitol solutions, or else a mixture of the different solvents mentioned.

Suitable pharmaceutical formulations for administration in the form of aérosols orsprays are, for example, solutions, suspensions or émulsions of the active ingrédientof the formulae I, la, Ib, XVa, XVb and XVI and/or the pharmaceutically acceptablesalts thereof in a pharmaceutically acceptable solvent, in particular éthanol or water,or a mixture of such solvents. The formulation may, if required, also contain otherpharmaceutical excipients such as surfactants, emulsifiers and stabilizers, and apropellant gas. Such a formulation normally contains the active ingrédient in aconcentration of about 0.1 to 10%, in particular of about 0.3 to 3% by weight.

The dose of the active ingrédient of the formulae I, la, Ib, XVa, XVb and XVI to beadministered, and the frequency of administration, dépend upon the potency andduration of action of the compounds used; additionally also on the nature and 32 1 3293 severity of the disorderto be treated and pn the sex, âge, weight and individualresponsiveness of the mammal to be treated.

On average, the daily dose of a compound of the formulae I, la, Ib, XVa, XVb andXVI and/or the pharmaceutically acceptable salts thereof for a patient weighing about75 kg is at least 0.001 mg/kg, preferably 0.01 mg/kg, up to at most 10 mg/kg,preferably 1 mg/kg, of body weight. In the event of acute épisodes of the disorder, forexample immediately after suffering a myocardial infarction, higher and especiallymore frequent dosages may also be necessary, for example up to 4 single doses aday. Especially in the case of i.v. administration, for instance for a patient withinfarction in the intensive care unit, for example up to 700. mg per day may benecessary, and the inventive compounds may be administered by infusion.

List of abbreviations: DCC dicyclohexyl carbodiimide DIP diisopropyl ether TLC thin-layer chromatography DMF N,N-dimethylformamide EA ethyl acetate eq. équivalent Et3N triethylamine Et2O diethyl ether EtOH éthanol h hour(s) HEP n-heptane HOAc acetic acid KOtBu potassium 2-methyl-2-propoxide MeOH methanol min minute(s) mp melting point MTB tert-butyl methyl ether NMP 1-methylpyrrolidin-2-one Pd(OAc)2 palladium(ll) acetate 33 1 3293

RT rt tBu

THF

5 TMEDA room température rétention time tert-butyl tetrahydrofuran N,N,N',N'-tetramethylethane-1,2-diamine

The rétention times (rt) reported hereinbelow relate to HPLC analyses having thefollowing parameters: 10 Method A:

Stationary phase: Waters Symmetry C8 (5μ) 3.9x150mm

Mobile Phase: isocratic CH3CN/0.1% aqueo.us CF3CO2H 35:65; λ=220 nm; 1 ml/min. 15 Method B:

Stationary phase: Waters Symmetry C8 (5μ) 3.9x150mm

Mobile Phase: isocratic CH3CN/0.1% aqueous CF3CO2H 40:60; λ=230 nm; 1 ml/min. 20 Method C:

Stationary phase: Waters Symmetry C8 (5μ) 3.9x150mm

Mobile Phase: isocratic CH3CN/0.1% aqueous CF3CO2H 50:50; À=220nm; 1 ml/min. 25 Example 1 a) N-(2,2,2-Trifluoroethyl)-4-trifluoromethylbenzamide

5.0 g (24 mmol) of 4-trifluoromethylbenzoyl chloride and 5.0 ml (36 mmol) oftriethylamine were dissolved in 50 ml of CH2CI2 and 2.4 g (24 mmol) of 2,2,2- 34 1 3293 trifluoroethylamin&were slowly added dropwise at RT. The mixture was stirred at RTfor 4 h, then the volatile constituents were removed under reduced pressure. Theresidue was taken up using 100 ml of MTB and washed initially with 30 ml of asaturated aqueous Na2CC>3 solution and then with 30 ml of a saturated aqueousNaHSO4 solution. Drying was effected over MgSO.4 and 6.1 g (94%) of a colorlessresin were obtained which crystallized when left to stand; mp: 117°C..

Rf (DIP) = 0.50 MS (El): 271 (M+1)+ b) (R,S)-3-Hydroxy-2-(2,2,2-trifluoroethyl)-5-trifluoromethyl-2,3-dihydroisoindol-1-one 0.37 ml (2.4 mmol) ofTMEDA and 1.4 ml (2.3 mmol) of a 1.5 M solution oft-BuLi inn-pentane were dissolved at -75°C in 2 ml of THF (anhydrous) and a solution of0.30 g (1.1 mmol) of N-(2,2,2-trifIuoroethyl)-4-trifluoromethylbenzamide in 2 ml ofTHF was added dropwise at -75°C. The mixture was stirred at -75°C for 3 h, then0.43 ml (5.5 mmol) of DMF was added dropwise and the mixture was warmed to RTover 30 minutes. The reaction mixture was poured onto 100 mi of a saturatedaqueous NaHCC>3 solution and extracted 3 times with 30 ml each time of EA. Dryingwas effected over MgSC>4 and the solvent was removed under reduced pressure.Chromatography on silica gel using DIP afforded 80 mg of (R,S)-3-hydroxy-2-(2,2,2-trifluoroethyl)-5-trifluoromethyl-2,3-dihydroisoindol-1-one in addition to 110 mg ofmixture with starting material. This mixture was separated again by reversed phaseHPLC (conditions see below) and a further 40 mg of (R,S)-3-Hydroxy-2-(2,2,2-trifluoroethyl)-5-trifluoromethyl-2,3-dihydroisoindol-1-one were obtained; overall yield30%. HPLC: gradient, run time 20 min

Eluent: 0.1% aqueous CF3CO2H, acetonitrile (Chromasolv); fl ow rate: 30 ml/minColumn: Waters XterraTM MS C-|8 5 pm, 30x100 mm 35 1 3293

Gradient 0 - 2.5 min 10% acetonitrile 3.0 min 25% acetonitrile 14.0 min 75% acetonitrile 15.0 min 95% acetonitrile 17.5 min 10% acetonitrile Rf (DIP) = 0.50 MS (El): 299 (M+1)+ c) Ethyl (RS)-[3-oxo-2-(2,2,2-trifluoroethyl)-6-trifIuoromethyl-2,3-dihydro-1 H-isoindol-1 -yljacetate

Under argon, ethyl (diethoxyphosphoryl)acetate (135 mg, 0.6 mmol) was dissolved inanhydrous dimethoxyethane (10 ml). 17.6 mg of NaH (60% in oil) were added at RTto this solution which was stirred at RT for 10 min. Afterward, a solution of 120 mg(0.04 mmol) of (RS)-3-hydroxy-2-(2,2,2-trifluoroethyl)-5-trifluoromethyl-2,3-dihydroisoindol-1-one in anhydrous dimethoxyethane (5 ml) was added and themixture was subsequently stirred at reflux for 2 h. The reaction solution was left tocool; the reaction solution was then poured onto 50 ml of 5% sodium hydrogencarbonate solution and extracted twice with 20 ml each time of ethylacetate; the organic phase was dried over MgSÜ4 and concentrated under reducedpressure, and the residue was purified by chromatography on silica gel using DIP asthe eluent. 90 mg (61%) of ethyl (RS)-[3-oxo-2-(2,2,2-trifIuoroethyl)-6-trifluoromethyl-2,3-dihydro-1 H-isoindol-1-yljacetate were obtained as a colorless oil whichcrystallized from heptane as a beige solid.

Rf(DIP)=0.31

The NMR spectrum was identical to the material prepared in example 4. 36 1 3293 d) (R,S)-N-{2-[3-Oxo-2-(2,2,2-trifluoroethyl)-6-trifluoromethyl-2,3-dihydro-H-isoindol-1-yl]acetyl}guanidine

As described in example 2g), ethyi (RS)-[3-oxo-2-(2,2,2-trifluoroethyl)-6-trifluorornethyl-2,3-dihydro-1 H-isoindol-1-yl]acetate may be reacted with guanidine.

Example 2 10 a) 2-Nitro-4-trifluoromethylbenzoic acid

11.97 g of 4 trifluoromethylbenzoic acid (63 mmol) were added slowly in portions atRT to 48 ml of HNO3 (100%). The mixture was subsequently heated to reflux for 1 h,then cooled to RT and poured onto about 600 g of ice. The mixture was stirred for 15 1 h, then the precipitate was filtered off and washed with 1 I of water. The filtrate was extracted with 300 ml of CH2CI2, and the organic phase was combined with theprecipitate and dried over Na2SO4. The solvent was removed under reducedpressure and the residue was recrystallized by dissolving in 1 I of DIP at 68°C,adding 2 I of HEP at this température and finally cooling the solution slowly to RT. 20 The crystallized product was washed with 1 I of HEP and dried under reducedpressure to obtain 7.1 g (48%), mp 136°C-138°C. b) 2-Amino-4-trifluoromethylbenzoic acid 37 1 3293

250 g of 2-nitro-4-trif]uoromethylbenzoic acid (1.06 mol) were dissolved in 1 I ofEtOH and 7.5 g of Pd/C (5%) were added. The mixture was hydrogenated under 1- 2.5 bar of hydrogen pressure. During the hydrogen uptake, the température rose 5 temporarily from 10°C to 104°C. After 2 h, the hydrogen uptake was complété.Subsequently, the catalyst was filtered off and the solvent was removed underreduced pressure to obtain 215 g (99%) of a pale yellow solid, mp 174-176°C. c) 2-((E)-2-Ethoxycarbonylvinyl)-4-trifluoromethylbenzoic acid

520 mg of NaNC>2 (7.6 mmol) were dissolved in 2 ml of water and added dropwise at 0°C to a solution of 1.3 g of 2-amino-4-trifluoromethylbenzoic acid (6.5 mmol) in 2.6 ml of a 48% aqueous HBF4 solution and 30 ml of éthanol. The mixture was then stirred at 0°C for 10 minutes, then warmed to RT. A further 0.3 ml of a 48% aqueous 15 HBF4 solution was then added, then 30 ml of éthanol, 0.9 g of ethyl acrylate (9.0 mmol) and 26.9 mg of Pd (OAc)2 (0.12 mmol). Subsequently, the mixture was stirred at 50-60°C for 1 h. The solvent was then removed under reduced pressure,and the residue taken up with 25 ml of EA and washed initially with 25 ml of a 1Naqueous HCl solution then with 25 ml of a saturated aqueous NaCI solution. The 20 organic phase was dried over Na2SO4 and the solvent removed under reducedpressure. The residue was suspended in 25 ml of heptane and the precipitatedproduct was filtered off. Yield: 1.3 g (69%). of a pale brownish solid. An analyticalsample was purified by crystailization from heptane/ethyl acetate. 38 1 3293

The NMR spectrum was identical to the material prepared in example 3a. d) Ethyl (E)-3-[2-(2,2,2-trifluoroethylcarbamoyl)-5-trifluoromethylphenyl]acrylate

5 1.3 g of 2-(2-ethoxycarbonylvinyl)-4-trifluoromethylbenzoic acid (4.5 mmol) and 453 mg of 2,2,2-trifluoroethylamine (4.5 mmol) were dissolved in 5 ml of DMF and0.93 g of DCC was added. The mixture was stirred at RT for 4 h. The urea by-product was removed by filtration and then the solvent was removed under reducedpressure. The residue was recrystallized from DIP to obtain 1.6 g (96%) of white 10 crystals.

The NMR spectrum was identical to the material prepared in example 3b. e) (RS)-[3-Oxo-2-(2,2,2-trifluoroethyl)-6-trifluoromethyl-2,3-dihydro-1H-isoindol-1-yljacetic acid

2.2 g of ethyl (E)-3-[2-(2,2,2-trifluoroethyicarbamoyl)-5-trifIuoromethylphenyl]acrylate(5.9 mmol) were dissolved in 10 ml of methanol and 1.5 ml of a 5 M aqueous NaOHsolution (7.5 mmol) wére added. The mixture was stirred at RT for 18 h and then setto pH = 7 using aqueous HCl solution. The solvents were removed under reduced 20 pressure and the residue was suspended in 10 ml of water. This suspension was setto pH = 2 using a 2N aqueous HCl solution and extracted 3 times with 10 mleachtime of EA. Drying was effected over Na2SO4 and the solvent was removed underreduced pressure. The residue was crystallized with diethyl ether/DIP, mp: 39 1 3293 202-204°C.

Yield: 1.8 g (89%). 1H NMR (400 MHz, CDCI3): δ = 3.07 (dd, ^=17 Hz, J2=6 Hz, 1 H), 3.23 (dd, J1=17Hz, J2= 5 Hz, 1 H), 4.27 (nri, 1 H), 4.58 (m, 1 H), 5.08 (t, J=5 Hz, 1 H), 7.91 (d, J=8 5 Hz, 1 H), 7.96 (d, J=8 Hz, 1 H), 8.12 (s, 1 H), 12.50 (bs, 1 H) ppm.

Combustion analysis: C13H9F6NO3 (341.2): cale. C 45.76 H 2.66 N4.10; foundC 45.71 H 2.43 N 4.11. f) Ethyl (RS)-[3-oxo-2-(2,2,2-triflUoroethyl)-6-trifluoromethyl-2,3-dihydro-1 H-isoindol-10 1 -yljâcetate

2.6 ml of SOCI2 (35 mmol) were dissolved in 20 ml of éthanol and 3.4 g of (R,S)-[3-oxo-2-(2,2,2-trifluoroethyl)-6-trifluoromethyl-2,3-dihydro-1 H-isoindol-1 -yljacetic acid(10 mmol) were added at -10°C, The mixture was stirred at RT for 18 h and the 15 volatile constituents were subsequently removed under reduced pressure. The residue was chromatographed on silica gel using 3:1 HEP/EA. Yield: 3.0 g (81%) ofa colorless oil which crystallized from heptane as a beige solid.

The NMR spectrum was identical to the material prepared in example 4. 20 g) (RS)-N-{2-[3-Oxo-2-(2,2,2-trifluoroethyl)-6-trifluoromethyl-2,3-dihydro-H-isoindol-1 -yl]acetyl}guanidine

Guanidine hydrochloride (11.5 g, 120 mmol) was dissolved in NMP (45 ml) and 40 1 3293 KOtBu (11.2 g, 1Q0 mmol) was added with stirring, and the mixture was left to stir atRT for 1.5 h and filtered. The filtrate was added dropwise at RT with stirring to asolution of ethyl (RS)-[3-oxo-2-(2,2,2-trifluoroethyl)-6-trifluoromethyl-2,3-dihydro-1H-isoindol-1-yl]acetate (7.38 g, 20 mmol) in NMP (12 ml) and left to stir at RT for a 5 further 60 min. Subsequently, ice-water (270 ml) was added, the mixture was set topH 7 using 2N HCl, ethyl acetate (60 ml) was added and the pH was subsequentlyadjusted to 8-8.5 by adding NaHCO3 solution. The mixture was stirred vigorously atRT for 1 h and the precipitate formed was filtered off with suction and washed withwater. 7.06 g (83%) of (R,S)-N-{2-[3-oxo-2-(2,2,2-trifluoroethyl)-6-trifIuoromethyl-2,3- 10 dihydro-H-isoindol-1-yl]acetyl}guanidine, inclusion compound with 0.5 équivalent ofethyl acetate, were obtained as fight yellow crystals, mp. 160-161 °C with graduaiheating, escape of ethyl acetate from approximately 90°C.

Rf (ethyl acetate/methanol) = 0.45 1H NMR (400 MHz, CDCI3): δ = 2.54 (dd, Ji=8 Hz, J2=16 Hz, 1 H), 3.09 (dd, ^=4 Hz, 15 J2=16 Hz, 1 H), 4.25 (m, 1 H), 4.64 (m, 1 H), 5.18 (m, 1 H), 6,65 (bs, 2 H), 7.75 (bs, 2 H), 7.88 (d, J=8 Hz, 1 H), 7.95 (d, J=8 Hz, 1 H), 8.02 (s, 1 H) ppm.C14H12F6N4O2 · % C4H8O2 (426.33): cale. C 45.08, H 3.78, N 13.14; found C 45.07, H3.79, N 13.01. 20 Example 3 a) 2-((E)-2-Ethoxycarbonylvinyl)-4-trifluoromethylbenzoic acid (variant ofexample 2c) r» n

F

F

F 658 ml of a 48-50% aqueous HBF4 solution were added at RT to 339 g of 2-amino- 25 4-trifluoromethylbenzoic acid (1,65 mol) in 6.8 I of EtOH (anhydrous). The température rose from 21 °C to 26°C. The mixture was then cooled to 0°C and asolution of 125 g of NaNC>2 in 500 ml of water was added dropwise between 0°C and 41 1 3293

5°C over 17 minutes. The initially pale yellow solution became initially an orange-redsuspension and fïnally a light yellow suspension. The progress of the reaction wasmonitored by-HPLC (method B; 2-amino-4-trifluorornethylbenzoic acid rt = 6.4 min;2-carboxy-5-trifluoromethylbenzenediazoniüm sait intermediate = 1.1 min. Within 30minutes, the conversion to the 2-carboxy-5-trifluoromethylbenzenediazonium saitwas >99% complété. The mixture was then added to 231 g of ethyl acrylate(2.31 mol), 11.1 g of Pd(OAc)2 (49 mmol) and 6.8 I of éthanol (anhydrous) and theréaction mixture was heated to 49-5ΓΟ. A uniform évolution of nitrogen increasingwith increasing température was observed. The reaction was monitored by HPLC(method B; 2-((E)-2-ethoxycarbonylvinyl)-4-trifluoromethylbenzoic acid rt = 16.4 min).After 45 min, the degree of conversion was above 99%. The mixture was thencooled to RT and the solvent removed under reduced pressure. The residue wastaken up in 3 I of EA and filtered off. The filtrate was then washed initially 3 timeswith 2.1 I each time of an aqueous HCl solution, then with 1 I of a saturated aqueousNaCI solution. Drying was effected over Na2SC>4 and the solvent was removedunder reduced pressure to obtain 449 g of a light brown solid. Taking into account animpurity (4-trifluoromethylbenzoic acid; 6.3%) and solvent residues (EA; 4%), a yieldof 83% was obtained. An analytical sample was purified by crystallization fromheptane/ethyl acetate. Mp.: 132-133°C 1H NMR (400 MHz, CDCI3): δ = 1.36 (t, J=7 Hz, 3 H), 4.31 (q, J=7 Hz, 2 H), 6.41 (d,J=16 Hz, 1 H), 7.72 (d, J=8 Hz, 1 H), 7.86 (s, 1 H), 8.21 (d, J=8 Hz, 1 H), 8.51 (d,J=16 Hz, 1 H), 8.5-9.5 (bs, 1 H) ppm.

Combustion analysis: C-13H11F3O4 (288.23): cale. C 54.17, H 3.85; found C 54.24, H 3.74. b) Ethyl (E)-3-[2-(2,2,2-trifIuoroethylcarbamoyl)-5-trifluoromethylphenyl]acrylate

F

F

F

NH

F 42 1 3293 315 g of oxalyl chloride (2.48 mol) were added at a température between 15 and18°C over 24 min to a mixture of 650 g of 2-((E)-2-ethoxycarbonylvinyl)-4-trifluoromethylbenzoic acid (2.25 mol), 33 ml of DMF and 7.8 I of CH2CI2. During theaddition, gas évolution was observed. The mixture was stirred at RT for 1 h and thencooled to 5°C, and 285 g of Et3N (2.81 mol) was then added at a températurebetween 5°C and 10°C over a period of 27 min. The mixture was stirred at 5°C for afurther 10 min, then 279 g of 2,2,2-trifluoroethylamine (2.81 mol) were added at atempérature between 9°C and 20°C over a period of 27 min. The mixture was stirredat RT for 10 min, in the course of which a thïck precipitate precipitated out, and, toimprove the stirrabiiity of the mixture, an additional 1 I of CH2CI2 was added. The reaction was monitored by HPLC (method C; 2-((E)-2-ethoxycarbonylvinyl)-4-trifluoromethylbenzoic acid rt. = 5.9 min; ethyl (E)-3-[2-(2,2,2-trifluoroethylcarbamoyl)- 5-trifIuoromethylphenyl]acrylate rt = 13.2 min). After stirring at RT fora further50 min, the reaction was complété. Volatile constituents of the reaction mixture werethen removed under reduced pressure, and the residue was taken up with 12 I of EAand washed 3 times with 2.5 I each time of water, then twice with 2.5 I each time of asaturated aqueous NaHCO3 solution and finally with 1.5 I of a saturated aqueousNaCI solution. Drying was effected over MgSO4, and the solvent was removed under reduced pressure to obtain 802 g of ethyl (E)-3-[2-(2,2,2-trifIuoroethylcarbamoyl)-5-trifluoromethylphenyljacrylate as a brown solid. This crude substance was combinedwith the crude product of another batch (177 g) and dissolved at 60-70°C in 3 I ofEA, and 14 I of HEP were added at this température in 1 I portions. The mixture wasthen heated to 80°C and stirred at this température for 1.5 h. This mixture was thenadded to 5.6 I of HEP at 70°C and the mixture was then cooled to RT with stirringover a period of 5 h. The product was then filtered off, washed with 3 I of HEP anddried under air to obtain 689 g of ethyl (E)43-[2-(2,2,2-trifluoroethylcarbamoyl)-5- "trifluoromethylphenyl]acrylate (67%) as a light brown solid. Mp; 161,5-162°C. 1h NMR (400 MHz, CDC!3): δ = 1.33 (t, J=7 Hz, 3 H), 4.05 (m, 2 H), 4.26 (q, J=7 Hz, 2 H), 6.19 (bs, 1 H), 6.46 (d, J=16 Hz, 1 H), 7.63 (d, J=8 Hz, 1 H), 7.68 (d, J=8 Hz, 1 H), 7.87 (s, 1 H), 7.90 (d, J=16 Hz, 1 H) ppm.

Combustion analysis; C15H13F5NO3 (369.27);calc. C 48.79, H 3.55, N 3.79; foundC 48.93, H 3.51, N 3.92. 43 1 3293 c) (R,S)-N-{2-[3-Oxo-2-(2,2,2-trifluoroethyl)-6-trifluoromethyl-2,3-dihydro-H-isoindoI-1-yl]acetyl}guanidine o NH. 386 g of ethyl (E)-3-[2-(2,2,2-trifluoroethylcarbamoyl)-5-trîfluoromethylphenyl]acrylate(1.05 mol) was suspended in 600 ml of DMF and added at a température between5°C and 15°C in portions slowly to 4.7 g of KOtBu (42 mmol). The cyclization fo theisoindolone was monitored byTLC (HEP/EA = 2:1; ethyl acrylate: Rf = 0.32;isoindolone: Rf = 0.41). After one hour, the reaction was complété. In the meantime,587 g of KOtBu were suspended in 2.2 I of DMF and 600 g of guanidinium chloridewere added at a température of between 20°C and 25°C. The mixture was stirred at25°C for 1 h and then the KCI was filtered off. The filtrate comprising the releasedguanidine was then added to the reaction mixture comprising the isoindolone andstirred at RT for 2 h. The conversion to the acylguanidine was monitored via HPLC(method B; wavelength 230 nm and 254 nm; isoindolone: rt = 15.1 min;acylguanidine: rt = 2.9 min). Subsequently, the reaction mixture was poured onto 141of ice-water, set to pH 8.5-9.0 using aqueous HCL solution and extracted 4 timeswith 3 I each time of EA. The mixture was then washed 3 times with 3 I each timë ofa saturated aqueous NaCI solution and dried over Na2SC>4, and the solvent wasremoved under reduced pressure. 329 g (82%) of a brown solid were obtained. Theproduct was combined with 3 other batches of the same préparation process; totalamount 842 g. These 842 g (2.2 mol) were digested in 2 I of EA and 5 I of Et2O at30°C for 2 h. The solid was then filtered off, washed twice with 2 I each time withEt2Q and dried under reduced pressure. 693 g (82% recoveiy) of an almost whitesolid were obtained. The compound crystallized from 2-propanol as the inclusioncompound with 0.5 équivalent of 2-propanol.

The NMR spectrum was identical to the S-enantiomer prepared in example 5b. 44 1 3293

Example 4

Ethyl (RS)-(2-(2,2,2-trifluoroethyl)-3-oxo-6-trifluoromethyl-2,3-dihydro-1 H-isoindol-1 -yl)acetate by one-pot reaction starting from 2-((E)-2-ethoxycarbonyivinyl)-4- 5 trifluoromethylbenzoic acid

SOCI2 (1.98 g, 27.2 mmol) was added at room température to a suspension of2-((E)-2-ethoxycarbonylvinyl)-4-trifluoromethylbenzoic acid (2.9 g, 10.1 mmol) intoluene (30 ml). The mixture was stirred at room température for 5 min and then 10 heated to 105°C (bath température) within 30 min. At about 70°C, gas évolutioncommenced. The mixture was stirred at 105°C for 3 h, then cooled to roomtempérature, filtered with suction through a kieselguhr layer (2.5 χ 0.5 cm) andwashed with toluene, and the filtrate was concentrated by évaporation underreduced pressure. The acid chloride was obtained in the form of a red-brown oil 15 (3.34 g). 2,2,2-Trifluoroethylamine (1.2 g, 12.1 mmol) and triethylamine (2.58 g, 25.3 mmol) was dissolved at 5°C in dichloromethane (15 ml), and the acid chloride,dissolved in dichloromethane (20 ml), was added dropwise with ice cooling at such arate that the température was kept between 5°C and 10°C. The ice bath was thenremoved and the excess oftrifluoroethylamine and a portion of the dichloromethane 20 distilled off under géntle vacuum. Subsequently, the mixture was heated to boilingunder reflux for 10 h. After it had been cooled, the mixture was diluted withdichloromethane (50 ml) and extracted twice with aqueous 2N HCl solution (50 mieach time), and the combined organic phases were washed with water (100 ml),dried over Na2SO4 and concentrated under reduced pressure. Ethyl (RS )-(2-(2,2,2- 25 trifluoroethyl)-3-oxo-6-trifluoromethyl-2,3-dihydro-1 H-isoindol-1-yl)acetate (3,51 g,94%) was obtained as a dark brown oil which was purified by crystallization fromn-heptane. Mp: 54.5-55.5°C. 1H NMR (400 MHz, CDCI3): δ = 1.15 (t, J=7 Hz, 3 H), 2.85 (dd, ^=6 Hz, J2=16 Hz, 45 1 3293 1 H), 3.01 (dd, J-] =5 Hz, J2=16 Hz), 1 H), 3.83 (m, 1 H), 4.12 (q, J=7 Hz, 2 H), 4.73(m, 1 H), 5.17 (t, J=6 Hz, 1 H), 7.80 (m, 2 H), 8.01 (d, J=8 Hz, 1 H) ppm.

Combustion analysis: C15H-13F5NO3 (369.27): cale. C 48.79, H 3.55, N 3.79;foundC 48.54, H 3.49, N 3.79.

Example 5 a) (S)-N-{2-[3-Oxo-2-(2,2,2-trifluoroethyl)-6-trifluoromethyl-2,3-dihydro-1 H-isoindol-1 -yl]acetyl}guanidine, O,O'-dibenzoyl-L-tartaric acid sait

10 (RS)-N-{2-[3-Oxo-2-(2,2,2-trifluoroethyl)-6-trifluoromethyl-2,3-dihydro-1 H-isoindol-1 -yl]acetyl}guariidine (inclusion compound with ethyl acetate, content 87.06% by NMR,44 g, 100 mmol) and O,O'-dibenzoyl-L-tartaric acid (11.2 g, 31 mmo!) were initiallycharged as solids and 2-propanol (500 ml) was added dropwise with stirring. Thesolids initially dissolved fully, bear a white solid precipitated out. After 30 min, the 15 mixture was heated to 70°C. This again gave an almost clear solution. This was leftto cool to room température within 4 h and subsequently stirred at this températureovernight. Afterward, the mixture was stirred at 10°C for 4 h and subsequentlyfiltered with suction. The residue was washed twice with 2-propanol (100 ml eachtime) and dried under air. 28.05 g of (S)-N-{2-[3-oxo-2-(2,2,2-trifluoroethyl)-6- 20 trifluoromethyl-2,3-dihydro-1 H-isoindol-1 -yl]acety!}guanidine, O.O'-dibenzoyl-L-tartaric acid sait (74% yield based on the (S)-enantiomer), in enantiomeric purity82% ee by HPLC (Chiracel OD/21,250 χ 4.6 mm, 50:5:2 n-heptane/ethanol/methanol, 1 ml/min, 30°C) were obtained as colorless crystals. 20 g (14.6 mmol) of these crystals were initially charged and 2-propanol (400 ml) . 25 was added dropwise. The mixture was heated to 80°C with stirring and then left to 46 1 3293 cool gradually to room température. The mixture was stirred at this température for afurther 2 h and then filtered with suction, and the residue was washed twice with2-propanoi (50 mleach time) and dried underair. 16.3 g (100% yield based on the(S)-enantiomer) of (S)-N-{2-[3-oxo-2-(2,2,2-trifluoroethyl)-6-trifluoromethyl-2,3- 5 dihydro-1 H-isoindol-1-yl]acetyl}guanidine, Ο,Ο'-dibenzoyl-L-tartaric acid sait wereobtained as colorless crystals, rhp: 192-193°C, enantiomeric purity >97% ee byHPLC (conditions as above).

Combustion analysis: C14H12F6N4O2 · C^gH^Og (561.43): cale. C 49.21, H3.41, N 9.98; found C 49.17, H 3.30, N 9.97. 10 b) (S)-N-{2-[3-Oxo-2-(2,2,2-trifluoroethyl)-6-trifluoromethyl-2,3-dihydro-1H-isoindol-1-yl]-acetyl}guanidine

o F 15 (S)-N-{2-[3-Oxo-2-(2,2,2-trifluoroethyl)-6-trifluoromethyl-2,3-dihydro-1 H-isoind ol-1 -yl]-acetyljguanidine, O,O'-dibenzoyl-L-tartaric acid sait (113 mg, 0,20 mmol) wasdissolved in a mixture of water (1 ml) and ethyl acetate (5 ml) and a solution ofNaHCOg (50 mg) in water (7.5 ml) was added. The mixture was left to stir at roomtempérature for 16 h and then extracted three times with ethyt acetate (5 ml each 20 time). The combined organic phases were shaken once with a solution of NaHCOg(50 mg) in water (20 ml) and then with pure water (20 ml), dried over Na2SÜ4 and concentrated by évaporation under reduced pressure. 75 mg (97%) of ,(S)-N-{2-[3-oxo-2-(2,2,2-trifluoroethyl)-6-trifluoromethyl-2,3-dihydro-1H-isoindol-1-yl]-acetyl}-guanidine were obtained. The product crystallized from 2-propanol as an inclusion 25 compound with 0.5 équivalent of 2-propanol, mp: 80-82°C. The substance may berecrystallized from ethyl acetate and crystallizes with 0.5 équivalent of ethyl acetate,mp: 121.5-122°C.

Enantiomeric purity > 97% by HPLC (Chiracel OD/21,250 · 4.6 mm, 4:1 n-heptane/2-propanol, 1 ml/min, 30°C). 1 3293 1H NMR (400 MHz, CDCI3): δ = 2.54 (dd, J 1=8 Hz, J2=16 Hz, 1 H), 3.09 (dd, Jt=4Hz, J2=16 Hz, 1 H), 4.25 (m, 1 H), 4.64 (m, 1 H), 5.18 (m, 1 H), 6.65 (bs, 2 H), 7.75(bs, 2 H), 7.88 (d, J=8 Hz, 1 H), 7.95 (d, J=8 Hz, 1 H), 8.02 (s, 1Ή) ppm.Combustion analysis (inclusion compound with 0.5 équivalent of 2-propanol):C-14H12F6N4O2 · 1/2 C3H8O (412.32): cale. C45.15, H 3.91, N 13.59; foundC 45.23, H 4.27, N 13.10.

Example 6 (RS)-N-{2-[3-Oxo-2-(2,2,2-trifluoroethyl)-6-trifluoromethyl-2,3-dihydro-1H-isoindol-1-yl]acetyl}guanidine by racemization of (R)-N-{2-[3-oxo-2-(2,2,2-trifluoroethyl)-6-trifluoromethyl-2,3-dihydro-1 H-isoindol-1 -yl]acetyl}guanidine

(R)-N-{2-[3-Oxo-2-(2,2,2-trifluoroethyl)-6-trifluoromethyl-2,3-dihydro-1H-isoindol-1-yl]acetyl}guanidine (inclusion compound with 0.5 équivalent of 2-propanol(M=412.3), 43 g, 104 mmol; obtained by concentrating the mother liquor which wasobtained in the concentration of (S)-N-{2-[3-oxo-2-(2,2,2-trifluoroethyl)-6-trifluoromethyl-2,3-dihydro-1 H-isoindol-1 -yl]acetyl}guanidine, O,O'-dibenzoyl-L-tartaric acid sait in example 5a, and treatment with NaHCO3 as described inexa.mple 5b)) was dissolved in 2-propanol (1.8 I) and a solution of KOH, (85 percent,660 mg, 10 mmol) in 2-propanol (400 ml) was added at room température withstirring. The mixture was stirred at room température for 24 h and then acidifiedusing glacial acetic acid (720 mg, 1.5 ml) and concentrated by évaporation underreduced pressure at a max. bath température of 40°C, and the residue waspartitioned between water (500 ml) and ethyl acetate (400 ml). The aqueous phasewas extracted twice with ethyl acetate (300 ml each time). The combined organicphases were shaken with a solution of NaHCO3 (10 g) in water (500 ml) and thenonce again with pure water. The organic phase was dried over Na2SO4 andconcentrated by évaporation under reduced pressure at a max. bath température of40°C. 39.8 g of (RS)-N-{2-[3-oxo-2-(2,2,2-trifluoroethyl)-6-trifluoromethyl-2,3-dihydro- 48 1 3293 1 H-isoindol-1 -yl]acetyl}guanidine, content 87% by NMR, inclusion compound with0.5 équivalent of ethyl acetate, were obtained as pale yellow crystals, yield: 87%.Mp: 164-166°C on gentle heating, escape of ethyl acetate from approximately 10Û°C. 5 Enantiomeric ratio 49:51 by HPLC (Chirace! OD/21,250 x 4.6 mm, 4:1 n-heptane/2-propanol, 1 mL/min, 30°C).

Example Ί (S)-N-{2-[3-Oxo-2-(2,2,2-trifluoroethyl)-6-trifluoromethyl-2,3-dihydro-1 H-isoindol-1 -yl]10 acetyl}guanidine hydrogenfumarate hydrate

O NH

OH (S)-N-{2-[3-Oxo-2-(2,2,2-trifluoroethyl)-6-trifluoromethyl-2,3-dihydro-1 H-isoindol-1 -yl]acetyljguanidine [inclusion compound with 0.5 équivalent of 2-propanol, (M=412.3),110 g, 266 mmol] was dissolved in dimethoxyethane (2 I) and admixed with fumaric 15 acid solution (0.5M in 9:1 dimethoxyethane/water, 512 ml) and the clear solutionformed was concentrated by évaporation under reduced pressure. The residue wastaken up with dichloromethane (2 I) and the mixture concentrated again byévaporation under reduced pressure. The residue was suspended in water (1.51),filtered with suction at room température and dried under air at room température 20 overnight. 125.9 g (95%) of (S)-N-{2-[3-oxo-2-(2,2,2-trifluoroethyl)-6-trifIuoromethyl-2,3-dihydro-1 H-isoindol-1-yl]acetyl}guanidine hydrogenfumarate hydrate wereobtained as colorless crystals, mp. 210°C. Détermination of the NHE inhibition

The inhibitory concentration IC5Q for NHE-1 inhibition was determined as follows:

The NHE-1 inhibition IC50 was determined in an FLIPR assay by measurement ofthe recovery in pHj in transfected cell lines which express human NHE-1. 25 49 1 3293

The assay was carried out in an FLIPR (fluorescent imaging plate reader) with blackwalled 96-well microtiter plates having clear bases. The transfected cell linesexpressing the different NHE subtypes (the parental cell line LAP-1 shows no 5 endogenous NHE activity as a conséquence of mutagenesis and subséquentsélection) were seeded the preceding day at a density of ~25 000 cells/well. Thegrowth medium for the transfected cells (Iscove +10% fêtai calf sérum) additionallycontained G418 as a sélection antibiotic in orderto ensure the presence of thetransfected sequences. 10

The actual assay started with the removal of the growth medium and addition of100 μΙ of loading buffer per well (5 μΜ BCECF-AM [2‘,7‘-bis(carboxyethyl)-5- (and-6-)carboxyfluorescein, acetoxymethyl ester] in 20 mM NH4CI, 115 mM choline chloride, 1 mM MgCl2, 1 mM CaCl2, 5 mM KCI, 20 mM HEPES, 5 mM glucose; 15 pH 7.4 [adjusted with KOH]). The cells were then incubated at 37°C for 20 minutes.

This incubation led to loading of the cells with the fluorescent dye whosefluorescence intensity dépends on pHi, and with NH4CI which makes the cellsslightly alkaline.

The nonfluorescent dye precursor BCECF-AM is, as the ester, membrane-20 permeable. Inside the cell, esterases release the actual BCECF dye which is membrane-impermeable.

Afterthis incubation for 20 minutes, the loading buffer which contained NH4CI and free BCECF-AM was removed by washing three times in a cell washer (Tecan25 Columbus) with in each case 400 pl of washing buffer (133.8 mM choline chloride, 4.7 mM KCI, 1.25 mM MgCI2, 1.25 mM CaCI2, 0.97 mM K2HPO4, 0.23 mM KH2PO4 5 mM HEPES, 5 mM glucose; pH 7.4 [adjusted with KOH]). The residual volumeremaining in the wells was 90 μΙ (50-125 μΙ possible). This washing step removedthe free BCECF-AM and led, as a conséquence of the removal of the external NH4+ 30 ions, to intracellular acidification (~ pHj 6.3-6.4). 50 1 3293

Since the equilibrium of intracellular NH4+ with NH3 and H+ was disturbed by theremoval of the extracellular NH4+ and by the subséquent instantaneous passage of the NH3 through the cell membrane, the washing process led to H+ remaining inside the cells, which was the cause of the intracellular acidification. This may eventuallylead to cell death if it persists long enough. It was important at this point that thewashing buffer was sodium-free (<1 mM), since extracellular sodium ions would leadto instantaneous recovery in the pHj as a resuit of the activity of the cloned NHEisoforms.

It was likewise important for ail buffers used (loading buffer, washing buffer, recoverybuffer) not to contain any HCO3 ions, since the presence of bicarbonate would leadto activation of interfering bicarbonate-dependent pHj regulatory Systems which areprésent in the parental LAP-1 cell line.

The microtiter plates with the acidified cells were then (up to 20 minutes aftertheacidification) transferred to the FLIPR. In the FLIPR, the intracellular fluorescent dyewas excited by light having a wavelength of 488 nm which was generated by anargon laser, and the measured parameters (laser power, illumination time andaperture of the CCD caméra incorporated in the FLIPR) were selected such that theaverage fluorescence signal per well was between 30 000 and 35 000 relativefluorescence units.

The actual measurement in the FLIPR started with an image being taken by the CCDcaméra every two seconds under software control. After ten seconds, the recovery ofthe intracellular pH was initiated by adding 90 pl of recovery buffer (133.8 mM NaCI, 4.7 mM KCI, 1.25 mM MgCI2, 1.25 mM CaCI2, 0.97 mM K2HPO4, 0.23 mM KH2PO4> 10 mM HEPES, 5 mM glucose; pH 7.4 [adjusted with NaOH]) by means of the96-well pipettor incorporated in the FLIPR.

The positive Controls (100% NHE activity) used were wells to which pure recoverybuffer had been added, while négative Controls (0% NHE activity) received washingbuffer. Recovery buffer with twice the concentration of test substance was added to 51 13293 ail other wells. Measurement in the FLIPR ended after 60 measurements (twominutes).

The raw data were exported into the ActivityBase program. This program firstly5 calculated the NHE activities for each tested substance concentration and, from these, the IC50 values for the substances. Since the course of pHj reçovery was notlinear overthe entire experiment, but ratherfell at the end owing to decreasing NHEactivity at higher pHj values, it was important to select, for évaluation of the measurement, the part in which the increase in fluorescence of the positive contrais10 was linear. -

Substance NHE1 inhibition IC50 [nM] (S)-N-{2-{3-Oxo-2-(2,2,2-trifluoroethyl)-6-trifluoromethyl-2,3-dihydro-1 H-isoindol-1 - yl]acetyl}guanidine 0.3

Claims (18)

1. 52 1 3293 What is claimed is:

   1. A process for preparing compounds of the formula I

   5 whereR1 and R2 are each independently hydrogen, F, Cl, trifluoromethoxy, 2,2,2-trifluoroethoxy, trifluoromethyl, 2,2,2-trifluoroethyl oralkyl having 1,2, 3or 4 carbon atoms; 10 R3 is Alk-R4 or trifluoromethyl; Alk is alkyl having 1,2, 3 or 4 carbon atoms; R4 is hydrogen, trifluoromethyl or cycloalkyl having 3, 4, 5, 6 or 7 carbon atoms; and salts thereof; 15 which comprises

   a) formylating the amide of the formula IV and then cyclizing to the compound of theformula VI, 53 13293 b) reacting the compound of the formula VI with an alkoxycarbonylmethylene-triphenylphosphorane, with a 1-alkoxy-1-trimethylsiloxyethylene or with a trialkylphosphonoacetate to give the compound of the formula VII, and c) reacting the compound of the formula VII with guanidine to give the compound of5 the formulai, where, in the compounds of the formulae IV, VI and VII, R1 to R3 are each as defined in formula I andR5 is alkoxy having 1, 2, 3 or 4 carbon atoms;and salts thereof. 10
2. 2. The process for preparing compounds of the formula I as claimed in claim 1 where R1 and R2 are each independently hydrogen, F, Cl, trifluoromethoxy, 15 2,2,2-trifluoroethoxy, trifluoromethyl, 2,2,2-trifluoroethyl or alkyl having 1,2, 3 or 4 carbon atoms; R3 is Alk-R4 or trifluoromethyl; Alk R4 is alkyl having 1, 2, 3 or 4 carbon atoms; is hydrogen, trifluoromethyl or cycloalkyl having 3, 4, 5, 6 or7 carbon atoms; and salts thereof;wherein 20 54 1 3293

   a) the compound of the formula II is reacted with the amine of the formula III to givethe amide of the formula IV, 5 b) the amide of the formula IV is formylated at the ortho-position to the amidefunction to give the formyl amide of the formula V, c) the formyl amide of the formula V is cyclized to the compound of the formula VI, d) the compound of the formula VI is reacted with analkoxycarbonylmethylenetriphenylphosphorane, with a 1-alkoxy- 1 -trimethylsiloxyethylene or with a trialkyl phosphonoacetate to give the compound ofthe formula VII and e) the compound of the formula VII is reacted with guanidine to give the compoundof the formula I, where, in the compounds of the formulae II, III, IV, V, VI and VII, 15 R1 to R3 are each as defined in formula I, R5 is alkoxy having 1,2, 3 or 4 carbon atoms andX is Cl, Br, OH or alkoxy having 1,2, 3 or 4 carbon atoms;and salts thereof.
3. 3. The process as claimed in claim 1 and/or 2, in which the process steps are 55 13293 each independently conducted continuously or batchwise.
4. 4. The process as claimed in one or more of daims 1,2 or 3, wherein thecompound of the formula I is defined as N-{2-[3-oxo-2-(2,2,2-trifluoroethyl)- 6-trifluoromethyl-2,3-dihydro-1H-isoindol-1-yl]acetyl}guanidine, and pharmaceuticallyacceptable salts thereof.
5. 5. A process for preparing compounds of the formula I

   10 where R1 and R2 are each independently hydrogen, F, Cl, trifluoromethoxy, 2,2,2-trifluoroethoxy, trifluoromethyl, 2,2,2-trifluoroethyl or alkyl having 1,2, 3or 4 carbon atoms; 15 R3 is Alk-R4 or trifluoromethyl; AlkR4 and salts thereof; 20 which comprises is alkyl having 1,2, 3 or 4 carbon atoms; is hydrogen, trifluoromethyl or cycloalkyl having 3, 4, 5, 6 or 7 carbon atoms;

   IX

   XI R6 H2NR3

   56 1 3293 a) reacting the amine ofthe formula IX via a diazonium sait with an alkyl acrylate to give the cinnamic acid dérivative ofthe formula XI, ' b) reacting the compound ofthe formula XI with the amine ofthe formula III and withguanidine to give the acylguanidine ofthe formula I, 5 where, in the compounds of the formulae III, IX and XI, R1 to R3 are each as défined in formula I andR6 is alkoxy having 1,2, 3 or 4 carbon atoms; and salts thereof. · '
6. 6. The process for preparing compounds of the formula I as claimed in claim 5 whereR1 and R2 are each independently hydrogen, F, Cl, trifluoromethoxy, 2,2,2-trifluoroethoxy, trifluoromethyl, 2,2,2-trifluoroethyl or alkyl having 1,2, 3 15 or 4 carbon atoms; R3 is Alk-R4 or trifluoromethyl; Alk is alkyl having 1,2, 3 or 4 carbon atoms; R4 is hydrogen, trifluoromethyl or cycloalkyl having 3, 4, 5, 6 or 7 carbon atoms; 20 and salts thereof;wherein 57 1 3293

   R2 Ο XIV a) the nitro compounds of the formula VIII is converted to the amine of theformula IX, 5 b) the amine of the formula IX is converted to the diazonium sait of the formula X, c) the diazonium sait of the formula X is reacted with an alkyl acrylate to give thecinnamic acid dérivative of the formula XI, d) the compound of the formula XI is converted to the amide of the formula XII and e) the compound of the formula XII is converted to the acylguanidine of the formula I,10 either by converting the compound of the formula XII in the presence of a base to the isoindolone dérivative of the formula XIII and subsequently by reaction withguanidine with activation to give the acylguanidine of the formula I (alternative A), or,after formation of the isoindolone dérivative of the formula XIII, in the presence of a 58 13293 base, from the compound of the formula XII, by converting the compound of theformula XIII to the ester of the formula XIV and subsequently by reacting withguanidine to give the acylguanidine of the formula I (alternative B), orby converting the compound of the formula XII in the presence of a strong base tothe ester of the formula XIV and subsequently by reacting with guanidine to theacylguanidine of the formula I (alternative O), or by directly reacting the compound of the formula XII with guanidine in the presenceof a base with simultaneously proceeding guanylation and cyclization to give theisoindolone of the formula I (alternative D), where, in the compounds of the formulae VIII, IX, X, XI, XII, XIII and XIV, R1 to R3 are each as defined in formula I and R6 and R7 are each independently alkoxy having 1,2, 3 or 4 carbon atoms;and salts thereof.
7. 7. The process as claimed in claim 6, wherein, in process step e), alternative D isused.
8. 8. The process as claimed in claim 6 and/or 7, in which process steps d) and e)are carried out in a one-pot process.
9. 9. The process as claimed in claim 5 and/or 6, in which the process steps areeach independently conducted continuously or batchwise.
10. 10. The process as claimed in one or more of daims 5 to 9, wherein the compoundof the formula l is defined as N-{2-[3-oxo-2-(2,2,2-trifluoroethyl)-6-trifluoromethyl-2,3-dihydro-1H-isoindol-1-yl]acetyl}guanidine, and pharmaceutically acceptable saltsthereof.
11. 11. A compound of the formula Xi I 59 1 3293

    where R1 and R2 ~ 5 are each independently hydrogen, F, Cl, trifluoromethoxy, 2,2,2-trifluorôethoxy, trifluoromethyl, 2,2,2-trifluoroethyl oralkyl having 1,2, 3or 4 carbon atoms; R3 is Alk-R4 or trifluoromethyl; Alk is alkyl having 1,2, 3 or 4 carbon atoms; 10 R4 is hydrogen, trifluoromethyl or cycloalkyl having 3,4, 5, 6 or 7 carbon atoms; R6 is alkoxy having 1,2, 3 or 4 carbon atoms;and salts thereof. 15 12. A compound ofthe formula XII as claimed in claim 11 for use as a synthetic intermediate.
12. 13. A process for isolating compounds of the formula la and Ib

    whereR1 and R2 20 60 13293 are each independently hydrogen, F, Cl, trifluoromethoxy, 2,2,2-trifluoroethoxy, trifluoromethyl, 2,2,2-trifluoroethyl oralkyl having 1,2, 3or 4 carbon atoms; R3 is Alk-R4 or trifluoromethyl; Alk is alkyl having 1, 2, 3 or 4 carbon atoms; R4 is hydrogen, trifluoromethyl or cycloalkyl having 3, 4, 5, 6 or 7 carbon atoms; and salts thereof;which comprises 10

    la lt> a) converting the compound of the formula I to salts of a 2,3-O-acylated D- orL-tartaric acid and obtaining the two salts of the formulae XVa and XVb separatelybycrystallization, and 15 b) releasing the free bases of the formulae la and Ib from the two salts of theformulae XVa and XVb respectively,where, in the compounds of the formulae I, XVa and XVb, R1 to R3 are each as defined in the formulae la and Ib R* is 61 1 3293

    R8 Ο

    R8 R8 is alkyl having 1,2, 3, 4, 5 or 6 carbon atoms or phenyl which is unsubstituted orsubstituted by 1,2 or 3 substituents from the group of F, Cl, Br, I, alkyl having 1,2, 3 5 or 4 carbon atoms or alkoxy having 1,2, 3 or 4 carbon atoms.
13. 14. The process as claimed in claim 13, wherein the undesired enantiomer of theformulae la or Ib is racemized again.
14. 15. The process as claimed in claim 13 and/or 14, wherein the compounds of the formulae la and Ib are (R)-N-{2-[3-oxo-2-(2,2,2-trifluoroethyI)-6-trifluoromethyl-2,3-dihydro-1 H-isoindol-1 -yl]acetyl}guanidine and (S)-N-{2-[3-oxo-2-(2,2,2-trifluoroethyl)- 6-trifluoromethyl-2,3-dihydro-1 H-isoindol-1-yl]acetyl}guanidine. 15 16. A compound of the formula XVa and/or XVb

    whereR1 and R2 20 are each independentiy hydrogen, F, Cl, trifluoromethoxy, 2,2,2-trifluoroethoxy, trifluoromethyl, 2,2,2-trifluoroethyl or alkyl having 1,2, 3or 4 carbon atoms; R3 is Alk-R4 or trifluoromethyl; 62 1 3293 Alk ' is alkyl having ï,2, 3 or 4 carbon atoms; R4 is hydrogen, trifluoromethyl or cycloalkyl having 3, 4, 5, 6 or 7 carbon atoms; R* is

    or HO,C. O à O R8^CO,H 5 R8 R8 R8 is alkyl having 1,2, 3, 4, 5 or 6 carbon atoms or phenyl which is unsubstituted orsubstituted by 1,2 or 3 substituents from the group of F, Cl, Br, I, alkyl having 1,2, 3or 4 carbon atoms or alkoxy having 1,2, 3 or 4 carbon atoms. 10 17. (S)-N-{2-[3-oxo-2-(2,2,2-trifluoroethyl)-6-trifluoromethyl-2,3-dihydro-1 H- isoindol-1-yl]acetyl}guanidine hydrogenfumarate hydrate of the formula XVI

    15 18. A compound of the formulae XVa or XVb as claimed in claim 16 or of the formula XVI as claimed in claim 17 for use as a médicament. 19. 19. The use of a compound of the formulae XVa and/or XVb as claimed in claim 16 or of the formula XVI as claimed in claim 17, alone or in combination with other20 médicaments or active ingrédients, for producing a médicament for the treatment or prophylaxis of acute or chronic damage, disorders or indirect sequelae of organs andtissues caused by ischémie or by reperfusion events, for the treatment or prophylaxisof arrhythmias, of life-threatening cardiac ventricular fibrillation, of myocardialinfarction, of angina pectoris, for the treatment or prophylaxis of ischémie States of 25 the heart, of ischémie States of the peripheral and centra! nervous System or of 63 1 3293 stroke or of ischémie States of peripheral organs and tissues, for the treatment orprophylaxis of States of shoek, of diseases in which cellular prolifération represents aprimary or secondary cause, of cancer, of metastasis, of prostate hypertrophy and ofprostate hyperplasia, of atherosclerosis or of disturbances of lipid metabolism, ofhigh blood pressure, of essential hypertension, of disorders of the central nervoussystem, of disorders resulting from overexcitability of the CNS, epilepsy or centrallyinduced convulsions, of disorders of the central nervous system, especially ofanxiety States, dépréssions or psychoses, for the treatment or prophylaxis of non-insulin-dependent diabètes mellitus (NIDDM) or late damage from diabètes, of thromboses, of disorders resulting from endothélial dysfunction, of intermittent » claudicating, for the treatment or prophylaxis of fibrotic disorders of internai organs,fibrotic disorders of the liver, fibrotic disorders of the kidney, fibrotic disorders ofvessels and fibrotic disorders of the heart, for the treatment or prophylaxis of heartfailure or of congestive heart failure, of acute or chronic inflammatory disorders, ofdisorders caused by protozoa, of malaria or of coccidiosis in poultry, and for use forsurgical operations and organ transplantations, for preserving and storing transplantsfor surgical procedures, for use in bypass operations, for use in resuscitation aftercardiac arrest, for preventing age-related tissue change, for producing a médicamentdirected against aging or for prolonging life, for the treatment and réduction ofcardiotoxic effects in thyrotoxicosis or for producing a diagnostic aid.
15. 20. A medicine for human, veterinary and/or phytoprotective use, comprising aneffective amount of a compound of the formulae XVa or XVb as claimed in claim 16together with pharmaceutically acceptable excipients and additives.
16. 21. A medicine for human, veterinary or phytoprotective use, comprising aneffective amount of a compound of the formulae XVa or XVb as claimed in claim 16together with pharmaceutically acceptable excipients and additives in combinationwith other pharmacological active ingrédients or médicaments.
17. 22. A medicine for human, veterinary and/or phytoprotective use, comprising aneffective amount of a compound of the formula XVI as claimed in claim 17 togetherwith pharmaceutically acceptable excipients and additives. 13293
18. 23. A medicine for human, veterinary or phytoprotective use, comprising an effective amount of the compound of the formula XVI as claimed in claim 17 together with pharmaceutically acceptable excipients and additives in combination with other pharmacological active ingrédients or médicaments. 64