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(11) Patent Numbe" KE 417   

(45) Date of grant: 03/11/2010   

(51) Int.CL7: C 07C 233/66, C 07D 209/46

(21) Application Number: KElP/ 2004/ 000373

(22) Filing Date: 19/11/2004

(30) Priority data: 10356717.8  02/12/2003  DE

(86) PCT data PCT/EP04/013153 19/11/2004 W0/2005/054195 16/06/2005

(73)0wner:SANOFI-AVENTIS DEUTSCHLAND GMBH of 65929 Frankfurt am Main, Germany

(72) Inventor: SCHUBERT, Gerrit ofFeldbergstrasse 2a, 65779 KelkheimGermany; RIEKE- ZAPP, Joerg of Arndtstrasse 50, 60325 FrankfurtGermany; KEIL, Johannes, of Johannesallee 12, 65925 Frankfurt.Germany; KLEEMANN, Heinz- Werner of Mainstrasse 29,65474 BischofsheimGermany; HANNA, Reda of 1043 Treeline Drive, Allentown, PA 18103U.S.A.; HUANG, Bao-Guo oflO Shields Lane, Bridgewater, NJ 08807U.S.A.; WIJ, Xiao-Dong of 58 Bond St., Bridgewater, NJ 08807U.S.A. and GOURAUD, Yves of 87 Avenue des Pervenches, F-93370 Montfermeil, France.

(74) Agent/address for correspondence: Kaplan & Stratton Advocates, P.O. Box 40111-00100, Nairobi

(54) Title: METHOD FOR PRODUCING (3- OX0-2,3-DIHYDRO- 1H- ISOINDOL-1-YL) ACETYLGUANIDE DERNATIVES

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

Method for producing (3-<>xo-2,3-{lihydro-1H-isoindol-1-yl)acetylguanidine denvatives

The present invention relates to processes for prepanng (3-<:>xo-2,3-{lihydro-1H-isoindol-1-yl)acetylguanidine denvatives via 3-hydroxy-2,3-dihydro-1 H-isoindol-1-one

5    derivatives or 3-(2-carbamoylphenyl)acrylic ester derivatives as intermediates, to a process for optical resolution, and also to intermediates of the process according to the invention.

(3-0xo-2,3-dihydro-1 H-isoindol-1-yl)acetylguanidine denvatives of the formula 1

~ ;)::H, I

R1~N-R3

R2    0

10

are NHE1 inhibitors and are described in PCT/EP03/05279. However, the syntheses descnbed there lead to racemic regioisomer mixtures, which entails costly and inconvenient separation processes and reduces the yield of the desired compound. Hitherto, it has only been possible to obtain the isomers by a costly and inconvenient

15    chromatographic separation on chiral supports. However, the substance throughput is restricted in chromatographic separations.

There is therefore a great interest in finding regioselective preparation processes for

(3-oxo-2,3-dihydro-1H-isoindol-1 -yl)acetylguanidine derivatives and processes for

20    recovering the enantiomers. The improved, regioselective preparation of the racemic (3-oxo-2,3-dihydro-1 H-isoindol-1-yl)acetylguanidine derivatives succeeds by two independent routes which are shown in scheme 1 and scheme 3. The resolution of

the racemates succeeds by crystallization as the salts of 2,3-0-acylated D- or L-tartaric acids, as shown in scheme 5. Gentle base-catalyzed racemization of the in

25    each case undesired enantiomer makes possible substantial conversion of the racemate to the desired enantiomer. The processes mentioned enable the simple preparation of enantiomencally enriched or enantiomerically pure (3-oxo-2,3-dihydro-1H-isoindol-1-yl)acetylguanidine derivatives. The novel processes now make possible the simple preparation of large amounts of substance of the compounds of


the fonnula I on the industrial scale.

The present invention thus relates to a process for preparing compounds of the formula I

-    ~NJ:~H, I

R1~N-R3

R2    0

5

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, 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,

R1 -- . f) - ( - R3

R2    0

IV
- !R5

R1~N-R3

R2    0

VII

Scheme 1

a)    formylating the amide of the formula IV and then cyclizing to the compound at the formula VI,

b)    reacting the compound of the formula VI with an alkoxycarbonylmethylene-triphenylphosphorane, w~h 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 of the 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 I

where

15    R1 and R2

are each independen~y hydrogen, F, Cl, trifluoromethoxy, 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;

20    Alk    is alky! 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 2,
 


4

    +        _("1ft   
R1 - f) - {            R1    ~N-R3   
                       
R2II0        Ill        R2    0   
                    IV   
                       
                       
-0f'CH~            OH   
    R1~N-R3   
R1~-R3       
R2    v    0    R2        0   
                    VI   
,-J':               
R2        0               
        VII               

Scheme 2

a) the compound of the fonmula II is reacted with the amine of the fonmula Ill to give

5    the amide of the formula IV,

b)    the amide of the fonmula IV is fonmylated at the ortho-position to the amide function to give the fonmyl amide of the formula V,

c)    the fonmyl amide of the fonmula Vis cyclized to the compound of the formula VI,

d)    the compound of the fonmula 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 fonmula VII
and

e) the compound of the fonmula VII is reacted with guanidine to give the compound

of the formula I,

15    where, in the compounds ofthefonmulae II, Ill, IV, V, VI and VII, R1 to R3 are each as defined in fonmula I,
R5 is alkoxy having 1, 2, 3 or 4 carbon atoms and

X is Cl, Br, OH or alkoxy having 1, 2, 3 or 4 carbon atoms;

and salts thereof.

20

The compound of.the formula II is typically reacted, in an inert solvent such as an ether, hydrocarbon or halogenated hydrocarbon, for example dichloromethane, at a temperature between -30'Cand the boiling point of the solvent, preferably at RT, with an amine of the formula Ill, if appropriate in the presence of an activating agent,

5    to give the amide of the formula IV.

The ortho-formylation may, for example, be carried out by inijially charging an alkyl-metal compound, for example an alkyllithium compound, preferably t-Buli, with a complex ligand, preferably TMEDA, in an inert solvent such as an ether or

10 hydrocarbon, for example THF, at a temperature between -100'Cand O'C, preferably between ~o•c and -50'C.Then, the amide of the formula IV is added and deprotonation is effected over a period between 10 minutes and 10 hours, preferably between 10 minutes and 60 minutes, at a temperature between -1 OO'C and O'C,preferably between ~o•c and -50'C.Subsequently, a formylating agent,

15    preferably DMF, is added and reaction with the anion is effected at a temperature between -100'Cand 40'C,preferably between -BO'Cand room temperature. Preference is given to leaving the solution to come to RT after addition of the DMF over a period of from 10 minutes to 3 hours, for example within 30 minutes. Amide of the formula V formed as an intermediate generally cyclizes directly to the isoindolone
20    of the formula VI.

The isoindolone of the formula VI is reacted with a (C1-C4)-alkoxycarbonyl-

methylenetriphenylphosphorane in an inert solvent such as an ether, hydrocarbon or

halogenated hydrocarbon, for example toluene, at a temperature between O'Cand

25    the boiling point of the solvent, preferably between 20'Cand the boiling point of the solvent, or with a tri(C1-C4)-alkyl phosphonoacetate in the presence of a base, for example sodium hydride, in an inert solvent such as an ether , hydrocarbon or halogenated hydrocarbon, for example 1 ,2-<limethoxyethane, at a temperature between O'Cand the boiling point of the solvent, preferably between 20'Cand the
30    boiling point of the solvent. Alternatively, the isoindolone of the formula VI is reacted with a 1-(C1-C4)-alkyloxy-1-trimethylsiloxyethylene in the presence of a Lewis acid, tor example titanium(IV) chloride or trimethylsilyl triflate, in an inert solvent such as an ether, hydrocarbon or halogenated hydrocarbon, for example dichloromethane, at

a temperature between -sooc and the boiling point of the solvent, preferably at a temperature between -aooc and 20°C (Synth. Commun. 1987, 17, 1).

The ester of the formula VII may be reacted by commonly known processes with

5    guanidine to give the acylguanidine of the formula I. The reaction is preferably effected in the manner known to those skilled in the art in a protic or aprotic, polar but inert organic solvent. For example, in the reaction of the methyl ester (formula VII; R6=0CH3) with guanidine, useful solvents have been found to be methanol,

isopropanol or THF at temperatures of from 20oc up to the boiling temperature of

10    these solvents. In most reactions of compounds of the formula VII with salt-free guanidine, operation is effected, for example, in aprotic, inert solvents, for example
ethers such as THF, dimethoxyethane or dioxane. However, water may also be used when use is made of a base, for example NaOH, as a solvent in the reaction of compounds of the formula VII with guanidine. In the reaction of compounds of the

15    formula VII with salts of guanidine, for example guanidine hydrochloride, the reaction is 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 temperature between 20°C and the boiling point of the solvent.

20

In addition to the carboxylic esters of the formula VII, it is also possible to use further activated acid derivatives in the reaction with guanidine, for example carbonyl chlorides, carboxylic thioesters or carboxylic anhydrides. An activation of the carboxylic acid with, for example DCC can also be effected. The activated acid
25    derivatives can be prepared in the manner known to those skilled in the art directly from the parent carboxylic esters of the formula VII or from the corresponding carboxylic acids which can be obtained from the esters by customary hydrolysis methods. A series of suitable methods for preparing activated carboxylic acid derivatives are specified with citation of source literature in J. March, Advanced

30    Organic Chemistry, Third Edition (John Wiley & Sons, 1985, p. 350).

The process steps described in scheme 1 and 2 may each independently be effected continuously or batchwise. A workup of the reaction mixture may be effected at any
 

7

of the process steps. The workup and, if desired, the purification of the products is effected by the customary methods such as extraction, pH separation, chromatography or crystallization and the customary dryings.

5    The starting compounds of the formulae II and Ill are commercially available or can be prepared according to or in a similar manner to the processes described in the literature and familiar to those skilled in the art.

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

~ ~NJ::H, I

R1~N-R3

R2    0

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, 3

15    or4 calbon 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 calbon atoms;

20    and salts thereof;

which comprises, as shown in scheme 3,

R1-Y;~ _,~~    H2NR3        ON~H,   
    Ill    R1    R3NH2   
R2    OH    R2OH        R2    0   
IX                       
        XI               
Scheme 3    *

a)    reacting the amine of the formula IX via a diazonium salt with an alkyl acrylate to give the cinnamic acid derivative of the formula XI,

b)    reacting the compound of the formula XI with the amine of the formula Ill and with 5 guanidine to give the acylguanidine of the formula I,

where, in the compounds of the formulae Ill, 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.

10

The present invention also relates to a process for preparing the compounds of the formula I
~ !NJ::H, I

R1~N-R3

R2    0

where

15    R1 and R2

are each independently hydrogen, F, Cl, trifiuoromethoxy, 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;

20    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,

25
 

9

---CYNH2
R1--~o

R2    IX    OH

~ !R6    A,B    ~/OH
R1~0    R1~N-R3

R2  XII   HN,R3    R2    XIII  O

~rR7
B,C    R1~N-R3

R2    0

XIV

Scheme 4

a) the nitro compounds of the fonnula VIII is converted to the amine of the

fonnula IX,

5    b) the amine of the fonnula IX is converted to the diazonium salt of the fonnula X,

c)    the diazonium salt of the fonnula X is reacted wtth an alkyl acrylate to give the cinnamic acid derivative ofthe fonnula XI,
d)    the compound of the fonnula XI is converted to the amide of the fonnula XII and

e)    the compound of the fonnula XII is converted to the acylguanidine of the fonnula I,

10    etther by converting the compound ofthe formula XII in the presence of a base to the isoindolone derivative of the fonnula XIII and subsequently by reaction with guanidine with activation to give the acylguanidine of the fonnula I (alternative A), or, after fonnation of the isoindolone derivative of the fonnula XIII, in the presence of a
 

10

base, from the compound of the formula XII, by converting the compound of the formula XIII to the ester of the formula XIV and subsequently by reacting with guanidine to give the acylguanidine of the formula I (alternative B), or

by 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 the acylguanidine of the formula I (alternative C), or

by directly reacting the compound of the formula XII with guanidine in the presence of a base with simultaneously proceeding guanylation and cyclization to give the

isoindolone 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 (for example described in "Houben-Weyl, Methoden der organischen Chemie", Volume Xl/1, Nitrogen compounds II, Georg Thieme Ve~ag Stuttgart, 1957, p. 360ft) to the aniline of the formula IX. Preference is given to catalytic hydrogenation, for example using Pd/C, for example using 5% Pd/C or 10% Pd/C, in a solvent, for example an

20    alcohol, preferably ethanol, under a hydrogen atmosphere of from 1 bar to 200 bar pressure, preferably from 1 bar to 10 bar of pressure.

The subsequent diazotization of the aniline of the formula IX is effected in an inert

solvent, preferably ethanol, in the presence of an acid whose anion does not

25    substitute the diazonium ion itself, for example HBF4 or HPF5, preferably HBF4, or, for example, H2SO, and in the presence of a nitrite, preferably NaN02. at a temperature between -3o•c and the boiling point of the solvent, preferably between o•cand 3o•c.

30    The diazonium salt of the formula X is preferably reacted directly with a (C1-C4)-alkyl acrylate, preferably ethyl acrylate, in the presence of a palladium catalyst, preferably Pd(OAc)2. at a temperature between o•c and the boiling point of the solvent, preferably between 45°C to 55°C, to give the cinnamic acid derivative of the

fonnula XI.

The benzoic acid function of the compound of the formula XI may be converted to the amide of the fonnula XII by methods known to those skilled in the art, preferably

5    via the acid chloride or with the aid of DCC. This reaction may also be conducted in such a way that the amide of the formula XII is cyclized in the reaction mixture directly to the ester of the fonnula XIV, i.e. the reaction of the compound of the fonnula XI to give the ester of the fonnula XIV is carried out in one step. This may be

done either under the basic reaction condttions of amide fonnation or the cyclization

10    may be brought about by adding a base, for example triethylamine, HOnig'sbase or potassium tert-butoxide. A further alternative consists in converting the compound of the fonnula XI directly to the compound of the formula I by successively carrying out amide formation, cyclization and guanidation in the same reaction vessel, in which case the reaction may be effected without isolating intennediates.

15

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

Alternative A: The conversion of the amide of the formula XII is preferably effected

20    using aqueous alkali solution, preferably aqueous NaOH solution, in a solvent such as an alcohol, preferably methanol or ethanol, at a temperature between -30°C and the boiling point of the solvent, preferably at RT. Both the hydrolysis of the ester
function and the cyclization to the isoindolone derivative of the fonnula XIII take

place. The compound of the fonnula XIII is activated for acylation by commonly

25    known processes (and as described for scheme 1), for example using the acid chloride or with DCC, and the acylguanidine of the formula I is obtained.

Alternative B: As in alternative A, the carboxylic acid of the fonnula XIII is

synthesized. Subsequently, standard processes for ester preparation, preferably

30    using SOCI2 in an alcohol such as methanol or ethanol, are used to prepare, for example, the methyl or ethyl ester of the fonnula XIV. The ester of the fonnula XIV is subsequently converted to the acylguanidine of the fonnula I as described for scheme 1.
 

12


Alternative C: The conversion of the amide of the formula XII is effected in a solution of a strong base, preferably methoxide or t-butoxide in an alcohol such as methanol or ethanol, 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 conditions for the acylation of guanidine. The solvent used is an inert solvent such as an ether,

10    hydrocarbon or halogenated hydrocarbon, preferably DMF. Typically, a guanidinium salt is initially reacted with a strong base, preferably KOtBu, which releases the free guanidine. The mixture is added to the solution of the compound of the formula XII in

a solvent such as an alcohol, ether, hydrocarbon or halogenated hydrocarbon, for

example DMF, NMP or 2-propanol. In the course of the addition, the guanylation and

15    the cyclization to the isoindolone of the formula I occurs simultaneously. In one variant, the compound of the formula XI is cyclized to the compound of the formula XIV and then converted in sttu to the compound of the formula I successively in time using a catalytic amount of a strong base, for example potassium tert-butoxide or sodium methoxide or sodium ethoxide, in a solvent, for example DMF, NMP or

20    2-propanol.

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

25

The process steps described in scheme 2 may be effected continuously or batchwise. A workup of the reaction mixture may be effected after any of the process steps. The workup and, if desired, the purification of the products is effected by the customary methods such as extraction, pH separation, chromatography or

30    crystallization and the customary dryings.

The starting compounds of the formulae Ill and VIII are commercially available or can be prepared according to or in a similar manner to the processes described in the literature and known to those skilled in the art.

The invention also provides compounds of the formula XII

~ !R6

R1~0

R2    XII   HN,R3

where

5    R1 and R2

are each independently hydrogen, F, Cl, trifiuoromethoxy, 2,2,2-trifiuoroethoxy, trlfiuoromethyl, 2,2,2-trifluoroethyl or alkyl having 1, 2, 3 or 4 carbon atoms;

R3    is Alk-R4 or trifiuoromethyl;

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 synthetic intermediate.

Compounds of the formula I in enantiomerically enriched or in enantiomerically pure

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

The present invention thus relates to a process for isolating compounds of the formula Ia and lb
 

14

~ ~N~~H
"--t:r{-~ ,:

R2    0

where

R1 and R2

are each independently hydrogen, F, Cl, trifluoromethoxy,

5    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    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,

,,    lb


Scheme 5

15    a) converting the compound of the fonmula I to salts of a 2,3-0-acylated D- or
 


15

L-tartalic acid and. obtaining the two salts of the formulae Y0/a and Y0/b separately by crystallization. and

b) releasing the free bases of the formulae Ia and lb from the two salts of the formulae Y0/a and Y0/b respectively,

5    where, in the compounds of the formulae I, Y0/a and Y0/b, R1 to R3 are each as defined in the formulae Ia and lb
R'11 is

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

15    the formula Ia or lb is racemized again.

The racemate of the compound of the formula I is crystallized with a tartaric acid

derivative R", for example 0,0'-<Jibenzoyi-D-tartaricacid, 0,0'-<Jibenzoyi-L-tartaric acid, O,O'-<Ji(4-methylbenzoyi)-L-tartaricacid, O,O'-di(4-methylbenzoyi)-D-tartalic

20    acid, O,O'-di(4-methoxybenzoyi}-L-tartalicacid or O,O'-di(4-methoxybenzoyi)-D-tartalic acid, preferably with 0,0'-dibenzoyi-L-tartaricacid or 0,0'-dibenzoyi-D-tartaric acid, in a suitable solvent, for example in an ether, e.g. diethyl ether, diisopropyl ether, dimethoxyethane, tetrahydrofuran or dioxane, in a halogenated

hydrocarbon, e.g. dichloromethane, trichloromethane, tetrachloromethane,

25    1,2-<Jichloromethane or trichloroethylene, in an alcohol, e.g. methanol, ethanol, n-propanol, 2-propanol, butanol, in an ester, e.g. ethyl acetate or butyl acetate, in water, or in mixtures of solvents, preferably in 2-propanol, dimethoxyethane or ethyl

acetate, at a temperature between -10°C and the boiling point of the solvent, preferably at from ooc to 40°C. In one valiant of the process, mixtures of two or more
 


16

2,3-0-acylated D-.or L-tartaric acids of the same configuration which bear different acyl groups are used for the separation.

The salt formation from the compound of the fonnula I and the tartaric acid derivative

5    R" can be effected using equivalent amounts, i.e. 0.5 mol of the tartaric acid derivative R" which contains two carboxylic acid groups may be used per mole of the compound of the formula 1. However, the compound of the forrnula I may also be crystallized with less than 0.5 mol equivalent of the 2,3-0-acylated D- or L-tartaric
acid, for example with from 0.25 mol to 0.5 mol of tartaric acid derivative R'per mole

10    of the compound of the formula I, in particular with from 0.25 mol to 0.3 mol of tartaric acid derivative R' per mole of the compound of the tonnula I. The desired enantiomer then crystallizes out in the form of the salt of the formula XVa or XVb and
the undesired enantiomer is for the most part present in the mother liquor in the form

of the enantiomers of the formula lb or Ia and not in the form of the salt of the

15    formula XV a or XVb. The enantiomeric purity of the salts of the formulae XVa and XVb may be increased by repeated crystallization or by stirring of the first crystals with fresh solvent at elevated temperature and subsequent cooling.

After separation of the two salts of the formulae XV a and XVb and separation of the

20    salt of the formula XVa or XVb from the undesired enantiomer lb or Ia, the enantiomerically enriched compounds of the formulae Ia and lb are subsequently typically released from the salts by addition of an auxiliary base, for example an
amine, e.g. triethylamine, an inorganic base such as NaHCOa, Na2C03 or aqueous

solutions thereof. It is customary to work in a suitable solvent, for example in an

25    ether, e.g. diethyl ether, diisopropyl ether, dimethoxyethane, tetrahydrofuran or dioxane, in a halogenated hydrocarbon, e.g. dichloromethane, trichloromethane, tetrachloromethane, 1,2-dichloroethane or trichloroethylene, in an alcohol, e.g. methanol, ethanol, n-propanol, 2-propanol or butanol, in an ester, e.g. ethyl acetate or butyl acetate, or in water or in solvent mixtures, preferably in ethyl acetate,

30    2-propanol, dichloromethane or water or mixtures thereof, in which case the reaction

mixture may have one or more phases, at a temperature between -tooc and the boiling point of the solvent, preferably at from 1ooc to 40°C. This may be done, for example, in such a way that the salt is dissolved in aqueous NaHC03 solution and
 

17

the enantiomer of.the formula Ia or lb is then extracted using an organic solvent, for example ethyl acetate.

The enantiomer Ia or lb which is undesired in each case may be converted back to

5    the racemate of the formula I by a racemization process and is thus available for another optical resolution step. In this case, the undesired enantiomer is preferably treated in a solvent such as an alcohol, e.g. 2-propanol, at a temperature between -10'Cand the boiling point of the solvent, preferably at from O'Cto 40'C,with small

amounts of a base, for example KOH, the reaction mixture is neutralized and the

10    racemate is isolated after aqueous-extractive workup. This process may be carried out by suitable selection of the amount of base and reaction temperature in such a way that virtually exclusively racemization and no chemical change in the substance occurs.

15    The present invention also provides compounds of the formulae ~a and ~b

xva

where

R1 and R2

20    are each independently hydrogen, F, Cl, trifluoromethoxy, 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    is alkyl having 1, 2, 3 or 4 carbon atoms;

25 R4 is hydrogen, trifluoromethyl or cycloalkyl having 3, 4, 5, 6 or 7 carbon atoms;

R* is

RB is alkyl having 1, 2, 3, 4, 5 or 6 carbon atoms or phenyl which is unsubstituted or substituted by 1, 2 or 3 substituents from the group ofF, Cl, Br, I, alkyl having 1, 2, 3 or 4 carbon atoms or alkoxy having 1, 2, 3 or 4 carbon atoms.
5

When the above-described compounds, for example the compounds of the fonmulae I, Ia, lb, VII, XIII, XIV, XVa or XVb, contain one or more centers of asymmetry, they may each independently have either S or R configuration, unless stated otherwise. The compounds may be present in the fonm of optical isomers, of diastereomers, of

10    racemates or of mixtures thereof, unless they are more precisely defined. On double bonds, either E or Z configuration may be present, unless stated otherwise. The present invention encompasses all tautomeric fonms of the above-described compounds, for example of the compounds of the fonmulae I, Ia, lb, XV a and XVb.

15    Alkyl radicals may be straight-chain or branched. This is also true when they bear substituents or occur as substituents of other radicals, for example in fluoroalkyl radicals 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,

20    neopentyl and hexyl. Preferred alkyl radicals are methyl, ethyl, n-propyl and isopropyl, more preferably methyl or ethyl. In alkyl radicals, one or more, for example 1, 2, 3, 4 or 5, hydrogen atoms may be substituted by fluorine atoms. Examples of such fluoroalkyl radicals are trifluoromethyl, 2,2,2-trifluoroethyl and pentafluoroethyl,
preferably trifluoromethyl or 2,2,2-trifluoroethyl. Substituted alkyl radicals may be

25    substituted in any positions.

Examples of cycloalkyl radicals are cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl or cycloheptyl.
 

19

Phenyl radicals may be unsubstituted or mono- or polysubstttuted, for example mono-, di- or trisubstituted, by identical or different radicals. When a phenyl radical is substituted, it preferably bears one or two identical or different substituents. In monosubstituted phenyl radicals, the substituent may be be disposed in the

5    2-position, the 3-position or the 4-position. Disubstituted phenyl may be substituted in the 2,3-position, 2,4-position, 2,5-position, 2,6-position, 3,4-position or 3,5-position. In trisubstituted phenyl radicals, the substituents may be disposed in the 2,3,4-position, 2,3,5-posttion, 2,4,5-position, 2,4,6-posttion, 2,3,6-position or

3,4,5-position.

10

The above-described compounds, for example the compounds ofthe fonmulae I, Ia and lb. may be used in the process according to the invention in the form of their salts and/or isolated in the fonm of their salts. Salts may be obtained by the customary 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, for example of the compounds of the fonmulae I, Ia and lb. are, for example, halides, in particular hydrochlorides, hydrobromides, lactates, sulfates, citrates, tartrates, acetates, phosphates, methylsulfonates, benzenesu~onates, p-toluenesulfonates, adipates, fumarates, gluconates, glutamates, glycerolphosphates, maleates,

20    benzoates, oxalates and pamoates and trifluoroacetates. In the case of the preparation of active ingredients, preference is given to physiologically tolerated salts and phanmaceutically acceptable salts. Examples include salts of compounds of the fonmulae I, Ia and lb with fumaric acid, in particular salts which contain 1 mole of fumaric acid per mole of the compound of the fonmula I, Ia or lb. and are thus

25    hydrogenfumarates or hemifumarates. Advantageous properties such as crystallinity, stability, particularly low hygroscopictty, a low tendency to racemization and good solubility are features especially, for example, of (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 fonmula XVI which likewise fonms part of the

30    subject matter of the present compound in all its tautomeric fonms.
 

20

0NH2       
FF~NANH2    o   
F~N-,J    X  HO~OH XVI   
~ (f    XHO0   
    2   
0   F       

When the compounds contain an acid group, they may form salts with bases, for example alkali metal salts. preferably sodium salts or potassium salts, or ammonium salts. 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 present in the form of a zwitterion.

One embodiment of the present invention relates to compounds in which R1 and R2

are not both hydrogen, in particular to compounds in which R1 is hydrogen and R2 is

0    fluorine. chlorine or trifluoromethyl, especially triftuoromethyl. In compounds in which R1 is hydrogen, the R2 substituent is preferably disposed in the para-position of the benzene 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

5    or 2 carbon atoms, especially having 1 carbon atom. R4 is preferably triftuoromethyl or cycloalkyl having 3, 5 or 6 carbon atoms. in particular 3 carbon atoms, more preferably trifluoromethyl. One embodiment of the present invention relates to compounds in which R3 is trifluoromethyl or 2,2,2-trifluoroethyl, in particular 2.2,2-trifluoroethyl.
10

A special embodiment of the present invention relates to the preparation of N-{2-[3-oxo-2-(2,2,2-trifluoroethyiHi-trifluoromethyl-2,3-<lihydro-1 H-isoindol-1-yl]acetyl}-guanidine and its enantiomeric forms and salts thereof.

25    X is preferably chlorine or methoxy, in particular chlorine. R5 is preferably methoxy or ethoxy, in particular ethoxy. R6 is preferably methoxy or ethoxy, in particular ethoxy. R7 is preferably methoxy or ethoxy, in particular ethoxy.

In one embodiment of the present invention, RSis phenyl which is unsubstituted or
 

21

substHuted by 1, 2or 3 substitutents from the group ofF, Cl, alkyl having 1, 2, 3 or 4 carbon atoms, or alkoxy having 1, 2, 3 or 4 carbon atoms, in particular unsubsmuted phenyl.

5    The compounds of the formulae I, Ia, lb, XCII a and XC~~b and their pharmaceutically tolerated salts are substituted acylguanidines and inhibit the cellular sodium-proton antiporter (Na•/H• exchanger, NHE), in particular the subtype NHE-1.

Owing to the NHE inhibitory properties, the compounds of the formulae I, Ia, lb, XC~~ a,

10    XCIIb and XCIII and/or their pharmaceutically acceptable salts are suitable for the prevention and treatment of disorders which are caused by activation of or by activated NHE, and also of disorders which have the NHE-related damage as a secondary cause.
The compounds of the formulae I, Ia, lb, XCIIa and XCIIb may also be used for the

15    treatment and prevention of disorders in which the NHE is only partially inhibited, for example by using a lower dose.

Since NHE inhibitors act predominanUy via their influence on cellular pH regulation,

they can generally be advantageously combined with other compounds which

20    regulate the intracellular pH value, useful combination partners being inhibitors of the enzyme group of the carbonic anhydrases, inhibitors of the systems transporting bicarbonate ions, such as of the sodium bicarbonate cotransporter (NBC) or of the sodium-dependent chloride-bicarbonate exchanger (NCBE), and also NHE inhibitors

with inhibitory action on other NHE subtypes, because they can reinforce or

25    modulate the pharmacologically relevant pH-regulating effects of the NHE inhibHors described here.

The use of the compounds of the formulae I, Ia, lb, XCII a, XCIIb or XCIII relates to the

prevention and to the treatment of acute and chronic disorders in veterinary and in

30    human medicine. For instance, the inventive inhibitors of the NHE are suHable for treating disorders which are induced by ischemia and by reperfusion.

As a result of their pharmacological properties, the compounds described here are suHable as antiarrhythmic medicaments. Their cardioprotective component makes
 

22

NHE inhibitors outstandingly suitable for infarction prophylaxis and infarction treatment, and also for the treatment of angina pectoris, in which cases they also preventively inhibit or greatly reduce the pathophysiological processes when ischemia-induced damage arises, in particular when ischemia-induced cardiac

5    arrhythmias are triggered. Owing to their protective actions against pathological hypoxic and ischemic situations, the compounds of the formulae I, Ia, lb, )Wa, )Wb and )WI used in accordance with the invention and/or pharmaceutically acceptable

salts thereof. as a result of inhibtlion of the cellular Na •tH• exchange mechanism, may be used as medicaments for the treatment of all acute or chronic damage
1o    induced by ischemia or disorders induced primarily or secondarily thereby.

This also relates to their use as medicaments for surgical interventions. Thus, the compounds may be used in organ transplants, in which case the compounds may be used for the protection of the organs in the donor before and during the removal, to

15    protect removed organs, for example in the course of treatment with or their storage in physiological bath liquids. and also in the course of transfer to the recipient organism.

The inventive compounds are likewise valuable medicaments having a protective

20    action in the performance of angioplastic surgical interventions, for example on the heart and also on peripheral organs and vessels.

The inventive compounds may also be used when performing bypass operations, for example in bypass operations on coronary vessels and in Coronary Artery Bypass
25    Graft (CABG).

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

30    The inventive compounds are of interest for medicaments against life-threatening arrhythmias. Ventricular fibrillation is terminated and the physiological sinus rhythm of the heart is restored.
 

23

Since NHE1 inhibitors of human tissue and organs, especially the heart, protect effectively not only against damage caused by ischemia and reperfusion but also against the cytotoxic action of medicaments like those finding use in particular in cancer therapy and the therapy of autoimmune diseases, combined administration

5    with NHE inhibitors is suitable for inhibiting the cytotoxic, especially cardiotoxic, side effects of the compounds mentioned. The reduction in the cytotoxic effects, especially the cardiotoxicity, resulting from comedication with NHE1 inhibitors also makes it possible to increase the dose of the cytotoxic therapeutic agents and/or to

prolong the medication with such medicaments. The therapeutic benefrt of such a

10    cytotoxic therapy can be considerably increased by the combination with NHE inhibitors.

Moreover, NHEI inhibitors can be used in the event of heart-damaging

overproduction of thyroid hormones, thyrotoxicosis, or when thyroid hormones are

15    supplied externally. The compounds of the formulae l,la,lb, >Wa, >Wb and >WI and/or the pharmaceutically acceptable salts thereof are thus suitable for improving therapy with cardiotoxic medicaments.

In accordance with their protective effect against ischemia-induced damage, the

20    inventive compounds are also suitable as medicaments for the treatment of ischemias of the nervous system, especially of the central nervous system, and they are suitable, for example, for the treatment of stroke or of cerebral edema.

NHE inhibitors are also suitable for the therapy and prophylaxis of diseases and

25    disorders which are induced by hyperexcitability of the central nervous system, in particular for the treatment of epileptic disorders, centrally induced clonic and tonic spasms, states of psychological depression, anxiety disorders and psychoses. In these cases, it is possible to use the NHE inhibitors described here alone or in
combination with other substances having antiepileptic activity or antipsychotic active

30    ingredients, or carbonic anhydrase inhibitors, for example with acetazolamide, and with other inhibitors of NHE or of the sodium-<lependent chloride-bicarbonate exchanger (NCBE).
 

24

In addition, NHE inhibttors are likewise suHable for the treatment of types of shock, for example of allergic, cardiogenic, hypovolemic and bacterial shock.

The compounds of the formulae I, Ia, lb. 'INa, YNb and '1011and/or the

5    pharmaceutically acceptable salts thereof may likewise be used for the prevention and treatment of thrombotic disorders, since they, as NHE inhibitors, are themselves able to inhibit platelet aggregation. In addition, they are able to inhibit or prevent the excessive release, occurring after ischemia and reperfusion, of mediators of

inflammation and coagulation, especially of von Willebrand factor and of

10    thrombogenic selectin proteins. It is thus possible to reduce and eliminate the pathogenic action of significant thrombogenic factors. The NHE inhibitors of the present invention can therefore be combined with other anticoagulant and/or thrombolytic active ingredients, for example recombinant or natural tissue plasminogen activator, streptokinase, urokinase, acetylsalicylic acid, thrombin
15    antagonists, factor Xa antagonists, medicinal substances with fibrinolytic activity, thromboxane receptor antagonists, phosphodiesterase inhibttors, factor VIla antagonists, clopidogrel, ticlopidine etc. It is particularly favorable to use the present NHE inhibttors in combination with NCBE inhibitors and/or with inhibitors of carbonic anhydrase, for example with acetazolamide.
20

In addttion, NHE inhibitors feature a strong inhibitory effect on the proliferation of cells, for example fibroblast cell proliferation and the proliferation of smooth vascular muscle cells. The compounds of the formulae I, Ia, lb, 'INa, YNb and '1011 and/or the pharmaceutically acceptable salts thereof are therefore useful as valuable
25    therapeutic agents for diseases in which proliferation represents a primary or secondary cause, and can therefore be used as antiatherosclerotics, agents for chronic renal failure, cancers.

It has been possible to show that cell migration is inhibited by NHE inhibttors. The

30    compounds of the formulae I, Ia, lb, 'INa, YNb and '1011 and/or the pharmaceutically acceptable salts thereof are therefore suitable as valuable therapeutic agents for diseases in which cell migration represents a primary or secondary cause, for example, cancers with a pronounced tendency to metastasis.
 

25

NHE inhibitors further feature a retardation or prevention of fibrotic disorders. They are thus suitable as excellent agents for the treatment of cardiac fibroses, and of pulmonary fibrosis, hepatic fibrosis, renal fibrosis and other fibrotic disorders. They can thus be used for the treatment of organ hypertrophies and hyperplasias, for

5    example of the heart and the prostate. They are therefore suitable for the prevention and treatment of heart failure (congestive heart failure= CHF) and for the treatment and prevention of prostate hyperplasia or prostate hypertrophy.

Since there is significant elevation in NHE in essential hypertensives, the

10    compounds of the formulae I, Ia, lb, X:Va, X:Vb and X:VI and/or the pharmaceutically acceptable salts thereof are suitable for the prevention and treatment of high blood pressure and for the treatment of cardiovascular disorders. In these cases they can
be used alone or with a suitable combination and formulation partner for the

treatment of high blood pressure and of cardiovascular disorders. For example, one

15    or more diuretics with a thiazide-like action, loop diuretics, aldosterone and pseudoaldosterone antagonists, such as hydrochlorothiazide, indapamide, polythiazide, furosemide, piretanide, torasemide, bumetanide, amiloride, triamterene, spironolactone or eplerone, may be combined. The NHE inhibitors of the present
invention may also be used in combination with calcium antagonists such as

20    verapamil, diltiazem, amlodipine or nifedipine, and with ACE inhibitors, for example ramipril, enalapril, lisinopril, fosinopril or captopril. Further favorable combination partners are alsoP-blockers such as metoprolol, albuterol etc., antagonists of the angiotensin receptor and its receptor subtypes such as losartan, irbesartan, valsartan, omapatrilat, gemopatrilat, endothelin antagonists, renin inhibitors,

25    adenosine receptor agonists, inhibitors and activators of potassium channels such as glibenclamide, glimepiride, diazoxide, cromakalim, minoxidil and derivatives thereof, activators of the mitochondrial AlP-sensitive potassium channel (mitoK(ATP} channel), inhibitors of Kv1.5 etc.

30    It has been shown that NHEI inhibitors have a significant antiinflammatory effect and can thus be used as antiinflammatory drugs. The inhibition of the release of mediators of inflammation is noteworthy in this connection. The compounds may thus be used alone or in combination with an antiinflammatory drug in the prevention or treatment of chronic and acute inflammatory disorders. The
 

26

combination partners used advantageously are steroidal and non-steroidal antiinflammatory drugs. The inventive compounds may also be used for the prevention or treatment of diseases which are caused by protozoa, such as in the event of malaria or coccidiosis in poultry.
5

It has also been found that NHE inhibitors exhibit a beneficial effect on serum lipoproteins. It is generally acknowledged that blood fat levels which are too high, known as hyperlipoproteinemias, constitute an essential risk factor for the development of arteriosclerotic vascular lesions, especially coronary heart disease.

10    The reduction of elevated serum lipoproteins is therefore of exceptional importance for the prophylaxis and the regression of atherosclerotic lesions. In addition to the reduction in total serum cholesterol, ~ is particularly important to reduce the
proportion of specific atherogenic lipid fractions in this total cholesterol, in particular

of the low density lipoproteins (LDL) and of the very low density lipoproteins (VLDL),

15    since these lipid fractions constitute an atherogenic risk factor. By contrast, a protective function against coronary heart disease is ascribed to the high density lipoproteins. Accordingly, hypolipidemics should be capable of reducing not only total cholesterol but also in particular the VLDL and LDL serum cholesterol fractions. It
has now been found that NHE1 inhib~ors exhibit valuable therapeutically utilizable

20    properties in relation to influencing the serum lipid levels. For instance, they significantly reduce the elevated serum concentrations of LDL and VLDL, as can be observed, for example, as a result of increased dietary intake of a cholesterol- and lipid-rich diet or in cases of pathological metabolic alterations, for example

genetically related hyperlipidemias. They can therefore be used for the prophylaxis

25    and for the regression of atherosclerotic lesions by eliminating a causal risk factor. These include not only the primary hyperlipidemias but also certain secondary hyperlipidemias, as occur, for example, in the event of diabetes. In add~ion, NHE inhibitors lead to a marked reduction in the infarctions induced by metabolic
abnormalities and in particular to a significant reduction in the induced infarction size

30    and the severity thereof.

The inventive compounds of the formulae I, Ia, lb, XI/ a, XVb and XVI therefore advantageously find use for preparing a medicament for the treatment of hypercholesterolemia; for preparing a medicament for the prevention of
 

27

atherogenesis; for. preparing a medicament for the prevention and treatment of atherosclerosis, for preparing a medicament for the prevention and treatment of diseases which are induced by elevated cholesterol levels, for preparing a medicament for the prevention and treatment of diseases induced by endothelial

5    dysfunction, for preparing a medicament for the prevention and treatment of atherosclerosis-induced hypertension, for preparing a medicament for the prevention

and treatment of atherosclerosis-induced thromboses, for preparing a medicament for the prevention and treatment of hypercholesterolemia-induced and endothelial dysfunction-induced ischemic damage and post-ischemic reperfusion damage, for

10    preparing a medicament for the prevention and treatment of hypercholesterolemia-induced and endothelial dysfunction-induced cardiac hypertrophies and cardiomyopathies and of congestive heart failure (CHF), for preparing a medicament for the prevention and treatment of hypercholesterolemia-induced and endothelial dysfunction-induced coronary vasospasms and myocardial infarctions, for preparing
15    a medicament for the treatment of the disorders mentioned in combinations with hypotensive substances, preferably with angiotensin converting enzyme (ACE) inhibitors and angiotensin receptor antagonists. A combination of an NHE inhibitor

with an active ingredient lowering the blood fat levels, preferably wtth an HMG-CoA

reductase inhibitor (for example lovastatin or pravastatin), the latter bringing about

20    hypolipidemic action and thus increasing the hypolipidemic properties of the NHE inhibttor, is found to be a favorable combination with enhanced action and reduced use of active ingredients.

For instance, NHE inhibitors lead to effective protection against endothelial damage

of various origins. This protection of the vessels against the syndrome of endothelial

25    dysfunction makes compounds of the formulae I, Ia, lb, XI/ a, Xl/b and XI/I and/or the pharmaceutically acceptable salts thereof valuable medicaments for the prevention and for the treatment of coronary vasospasms, peripheral vascular diseases, in particular intermittent claudication, atherogenesis and atherosclerosis, left ventricular hypertrophy and dilated cardiomyopathy and thrombotic disorders.

30

It has also been found that NHE inhibitors are suitable in the treatment of non-insulin-dependent diabetes (NIDDM), in the course of which the insulin resistance is restrained. In this case, tt may be beneficial to enhance antidiabetic activity and quality of action of the inventive compounds by combining them with a biguanide
 

28

such as metformin, with an antidiabetic sulfonylurea such as glyburide, glimepiride, tolbutamide etc., wtth a glucosidase inhibttor, wtth a PPAR agonist such as rosiglitazone, pioglitazone etc., with an insulin product of different administration form, with a DB4 inhibttor, wtth an insulin sensttizer or wtth meglttinide.

5    In addition to the acute antidiabetic effects, NHE inhibitors counteract the development of late complications of diabetes and can therefore be used as medicaments for the prevention and treatment of late damage from diabetes, such as diabetic nephropathy, diabetic neuropathy, diabetic retinopathy, diabetic

cardiomyopathy and other disorders occurring as a consequence of diabetes. They

10    can in this connection be advantageously combined with the antidiabetic medicaments just described under NIDDM treatment. The combination with a beneficial dosage form of insulin might be particularly important in this connection.

NHE inhibitors exhibit, in addiTion to the protective effects against acute ischemic

15    events and the subsequent equally acutely stressing reperfusion events, also direct therapeutically utilizable actions against diseases and disorders of the entire mammalian organism which are associated with the manifestations of the chronically progressive aging process and which occur independently of acute hypoperfusion states and under normal, non-ischemic conditions. These pathological, age-related

20    manifestations induced over the long aging period, such as illness, invalidity and death, which can now be made amenable to treatment wtth NHE inhibitors, are diseases and disorders which are essentially caused by age-related changes in vital organs and the function thereof and become increasingly important in the aging

organism.

25    Disorders which are connected with an age-related functional impairment or with age-related manifestations of wear on organs are, for example, the inadequate response and reactivity of the blood vessels to contraction and relaxation reactions. This age-related decline in the reactivtty of vessels to constricting and relaxing
stimuli, which are an essential process of the cardiovascular system and thus of life

30    and health, can be significantly eliminated or reduced by NHE inhibttors. One important function and a measure of the maintenance of the reactivity of vessels is the blockade or retardation of the age-related progression in endothelial dysfunction, which can be eliminated highly significantly by NHE inhibitors. NHE inhibitors are
 

29

thus outstandingly. suitable for the treatment and prevention of the age-related progression in endothelial dysfunction, especially of intenmittent claudication.

An example of another variable characterizing the aging process is the decline in the

5    contractability of the heart and the decline in the adaptation of the heart to a required pumping output of the heart. This diminished efficiency of the heart as a consequence of the aging process is in most cases connected w~h a dysfunction of the heart which is caused inter alia by deposition of connective tissue in the

myocardial tissue. This deposition of connective tissue is characterized by an

10    increase in the weight of the heart, by an enlargement of the heart and by restricted cardiac function. It is surprising that tt has been possible to virtually completely inhibit such aging of the heart organ. NHE inhibitors are thus outstandingly suitable for the treatment and prevention of heart failure, of congestive heart failure (CHF).

15    Inhibition of proliferation allows not only fonms of cancer which have already occurred to be cured, but also reduction and highly significant retardation of the age-related incidence of cancer through NHE inhibitors. A particularly noteworthy finding

is that the disorders, occurring as a result of aging, of all organs and not only certain

types of cancer are suppressed or occur with a highly significant delay. NHE

20    inhibttors are thus outstandingly suitable for the treatment and, in particular, the prevention of age-related types of cancer.

Using NHE inhibitors, a delay, shifted highly significantly in time, is found in the

occurrence of age-related disorders of all the organs investigated, including the

25    heart, vessels, liver etc., and also a highly significant delay in cancer of the elderty. Moreover, there is also surprisingly a prolongation of life to an extent which has to date been achievable by no other group of medicaments or by any natural products. This unique effect of NHE inhibttors also enables, in addition to the sole use of the

active ingredients on humans and animals, these NHE inhibttors to be combined with

30    other activity principles, measures, substances and natural products which are used in gerontology and which are based on a different mechanism of action. Such classes of active ingredients used in gerontological therapy are: in particular vttamins and substances with antioxidant activity. Since there is a correlation between caloric load or food intake and the aging process, tt is possible to combine with dietary
 

30

measures, for example with appetite suppressants. II is equally possible to consider a combination with hypotensive medicaments such as with ACE inhibttors, angiotensin receptor antagonists, diuretics, ca• 2 antagonists etc. or with metabolism-normalizing medicaments such as cholesterol-lowering agents.

5    NHE inhibitors are thus outstandingly suitable for the prevention of age-related tissue changes and for prolonging life while retaining a high quality of life.

The inventive compounds are effective inhibitors of the cellular sodium-proton

antiporter (Na/H exchanger) which in numerous disorders (essential hypertension,

10    atherosclerosis, diabetes etc.) is also increased in cells which are readily amenable to measurements, for example in erythrocytes, thrombocytes or leukocytes. The compounds used in accordance with the invention are therefore suitable as outstanding and simple scientific tools, for example in their use as diagnostic agents

for determining and distinguishing different types of hypertension, but also of

15    atherosclerosis, diabetes and the late complications of diabetes, proliferative disorders etc.

Also claimed is a medicine for human, veterinary or phytoprotective use which,

together with pharmaceutically acceptable carriers and excipients, comprises an

20    effective amount of one or more compounds of the formulae Y0/a, Y0/b and Y0/l and/or pharmaceutically acceptable salts thereof, alone or in combination with other pharmacological active ingredients or medicaments. Medicaments which comprise a
compound of the formulae I, Ia, lb, Y0/a, Y0/b and Y0/i and/or the pharmaceutically

acceptable salts thereof can be administered, for example, orally, parenterally,

25    intravenously, rectally, percutaneously or by inhalation, the preferred administration being dependent on the particular characteristics of the disorder. The compounds of the formulae I, Ia, lb, Y0/a, Y0/b and Y0/l may be used alone or together with pharmaceutical excipients, both in veterinary medicine and in human medicine. The

medicaments generally comprise active ingredients of the formulae I, Ia, lb, Y0/a,

30    Y0/b and Y0/l and/or the pharmaceutically acceptable salts thereof in an amount of from 0.01 mg to 1 g per dose unit.

Which excipients are suttable for the desired pharmaceutical formulation are familiar to those skilled in the art on the basis of their expert knowledge. In addition to
 

31

solvents, gel formers, suppository bases, tablet excipients, and other active ingredient carriers, it is possible to use, for example, antioxidants, dispersants, emulsifiers, antifoams, flavorings, preservatives, solubilizers or colors.

5    For an oral administration form, the active compounds are mixed with additives suitable therefor, such as carriers, stabilizers or inert diluents, and converted by conventional methods to suitable dosage forms such as tablets, coated tablets, hard gelatin capsules, aqueous, alcoholic or oily solutions. Examples of inert carriers

which can be used are gum arabic, magnesia, magnesium carbonate, potassium

10    phosphate, lactose, glucose or starch, especially corn starch. The formulation may be effected either in the form of a dry granule or a wet granule. Examples of suitable oily carriers or solvents are vegetable or animal oils such as sunflower oil or fish liver oil.

15    For subcutaneous, intramuscular or intravenous administration, the active compounds used, if desired with the substances customary therefor, such as solubilizers, emulsifiers or other excipients, are brought into solution, suspension or emulsion. Examples of useful solvents are: water, physiological saline or alcohols,

e.g. ethanol, propanol, glycerol, as well as sugar solutions such as glucose or

20    mannitol solutions, or else a mixture of the different solvents mentioned. Suitable pharmaceutical formulations for administration in the form of aerosols or
sprays are, for example, solutions, suspensions or emulsions of the active ingredient

of the formulae I, Ia, lb. ~a. ~b and ~I and/or the pharmaceutically acceptable

25    salts thereof in a pharmaceutically acceptable solvent, in particular ethanol or water, or a mixture of such solvents. The formulation may, if required, also contain other pharmaceutical excipients such as surfactants, emulsifiers and stabilizers, and a propellant gas. Such a formulation normally contains the active ingredient in a concentration of about 0.1to 10%, in particular of about 0.3to 3% by weight.

30

The dose of the active ingredient of the formulae I, Ia, lb, ~a. ~band XVI to be administered, and the frequency of administration, depend upon the potency and duration of action of the compounds used; additionally also on the nature and
 

32

severity of the disorder to be treated and on the sex, age, weight and individual responsiveness of the mammal to be treated.

On average, the daily dose of a compound of the formulae I, Ia, lb. ~a. ~band

5    ~I and/or the phanmaceutically acceptable salts thereof for a patient weighing about 75 kg is at least 0.001 mglkg, preferably O.Q1 mg/kg, up to at most 10 mg/kg, preferably 1 mglkg, of body weight. In the event of acute episodes of the disorder, for
example immediately after suffering a myocardial infarction, higher and especially more frequent dosages may also be necessary, for example up to 4 single doses a day. Especially in the case of i.v. administration, for instance for a patient with infarction in the intensive care unit, for example up to 700 mg per day may be necessary, and the inventive compounds may be administered by infusion.
 

List of abbreviations:

15    DCC DIP TLC DMF EA

20    eq.

Et3N

Et20 EtOH h

25    HEP HOAc KOtBu MeOH

min

30    mp MTB NMP
Pd(OAc)2
 


dicyclohexyl carbodiimide diisopropyl ether thin-layer chromatography N,N-<limethylformamide ethyl acetate

equivalent triethylamine diethyl ether ethanol hour(s) n-heptane acetic acid

potassium 2-methyl-2-propoxide methanol

minute(s) melting point

tert-butyl methyl ether 1-methylpyrrolidin-2-one palladium(ll) acetate
 

33

RT    room temperature

rt    retention time

tBu    tert-butyl

THF    tetrahydrofuran

5    TMEDA    N,N ,N',N'-tetramethylethane-1,2-<liamine

The retention times (rt) reported hereinbelow relate to HPLC analyses having the following parameters:

10    Method A:

Stationary phase:   Waters Symmetry CB (5~) 3.9x150mm

Mobile Phase;    isocratic CH3CN/0.1% aqueous CF3C02H 35:65; 1.=220 nm;

1 ml/min.

15    Method B:

stationary phase:   Waters Symmetry CB (5~) 3.9x150mm

Mobile Phase:    isocratic CH3CN/0.1% aqueous CF3C02H 40:60; 1.=230 nm;

1 ml/min.

20    Method C:

Stationary phase:   Waters Symmetry CB (5~) 3.9x150mm

Mobile Phase:    isocratic CH3CN/0.1% aqueous CF3C02H 50:50; 1.=220nm;

1 ml/min.

25    Example 1

a) N-(2,2,2-Trtfluoroethy1}4-trifluoromethylbenzamide

F
;~ H     FF

~NJF

0

5.0 g (24 mmol) of 4-trifluoromethylbenzoyl chloride and 5.0 ml (36 mmol) of triethylamine were dissolved in 50 ml of CH2CI2 and 2.4 g (24 mmol) of 2,2,2-
 

34

trifluoroethylamine. were slowly added dropwise at RT. The mixture was stirred at RT for 4 h, then the volatile constituents were removed under reduced pressure. The residue was taken up using 100 ml of MTB and washed initially with 30 ml of a saturated aqueous Na2C03 solution and then with 30 ml of a saturated aqueous

5    NaHS04 solution. Drying was effected over MgS04 and 6.1 g (94%) of a colorless resin were obtained which crystallized when left to stand; mp: 11 ?"C.

Rf (DIP} = 0.50    MS (EI): 271 (M+1)+

b) (R,S}-3-Hydroxy-2-(2,2,2-trifluoroethyl)-5-trifluoromethyl-2,3-dihydroisoindoi-1-<Jne

F~FOH
F    I """'  N
~    \    F
of;
10

0.37 ml (2.4 mmol) of TMEDA and 1.4 ml (2.3 mmol) of a 1.5 M solution of t-Buli in n-pentane were dissolved at -75oc in 2 ml ofTHF (anhydrous) and a solution of 0.30 g (1.1 mmol) of N-(2,2,2-trifluoroethyl}-4-trifluoromethylbenzamide in 2 ml of THF was added dropwise at -75°C. The mixture was stirred at -75°C for 3 h, then

15    0.43 ml (5.5 mmol) of DMF was added dropwise and the mixture was warmed toRT over 30 minutes. The reaction mixture was poured onto 100 ml of a saturated aqueous NaHC03 solution and extracted 3 times with 30 ml each time of EA. Drying was effected over MgS04 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-

20    trifluoroethyl}-5-trifluoromethyl-2,3-dihydroisoindol-1-one in addition to 110 mg of mixture wtth starting material. This mixture was separated again by reversed phase HPLC (conditions see below) and a further 40 mg of (R,S)-3-Hydroxy-2-(2,2,2-trifluoroethyl}-5-trifluoromethyl-2,3-<lihydroisoindol-1-one were obtained; overall yield
30%.

25

HPLC: gradient, run time 20 min

Eluent: 0.1% aqueous CF3C02H. acetonitrile (Chromasolv}; flow rate: 30 mUmin

Column: Waters XterraTM MS C18 5 ~m. 30x1 00 mm
 

35


Gradient:

0-2.5 min 10% acetonitrile 3.0min 25% acetonitrile
5    14.0 min 75% acetonitrile 15.0min 95% acetonitrile 17.5 min 10% acetonitrile

Rf (DIP} = 0.50    MS (EI): 299 (M+1)+

10

c) Ethyl (RS}-[3-oxo-2-(2,2,2-trifluoroethyl)-6-trifluoromethyl-2,3-dihydro-1H-isoindol-1-yl]acetate
,t'~ ~

''CQ~F
0    F

Under argon, ethyl (diethoxyphosphoryl)acetate (135 mg, 0.6 mmol} was dissolved in

15    anhydrous dimethoxyethane (1 0 ml). 17.6 mg of NaH (60% in oil} were added at RT to 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 the

mixture was subsequently stirred at reflux for 2 h. The reaction solution was left to

20    cool; the reaction solution was then poured onto 50 ml of 5% sodium hydrogencarbonate solution and extracted twice with 20 ml each time of ethyl acetate; the organic phase was dried over MgS04 and concentrated under reduced pressure, and the residue was purified by chromatography on silica gel using DIP as

the eluent. 90 mg (61%) of ethyl (RS)-[3-oxo-2-(2,2,2-trifluoroethyl}-6-trifluoromethyl-

25    2,3•dihydro-1H-isoindol-1-yl]acetatewere obtained as a colorless oil which crystallized from heptane as a beige solid.

Rf (DIP}= 0.31

The NMR spectrum was identical to the material prepared in example 4.
 

36


d) (R,S}-N-{2-[3-0xo-2-(2,2,2-trifluoroethyl}-6-trifluoromethyl-2,3-<lihydro-H-isoindol-1-yl]acetyl}guanidine

0    X'
N    NH2

F    F
\j--F
0    F

As described in example 2g), ethyl (RS}-[3-oxo-2-(2,2,2-trifluoroethyl}-6-trifluoromethyl-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 at RT to 48 ml of HN03 (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 CH2Ciz, and the organic phase was combined with the precipitate and dried over Na2S04. The solvent was removed under reduced

pressure and the residue was recrystallized by dissolving in 1 I of DIP at 68°C,

adding 2 1 of HEP at this temperature and finally cooling the solution slowly to RT.

20    The crystallized product was washed with 1 I of HEP and dried under reduced pressure to obtain 7.1 g (48%), mp 136°C-138°C.

b) 2-Amino-4-trifluoromethylbenzoic acid
 
250 g of 2-nttro-4-trifluoromethylbenzoic acid (1.06 mol) were dissolved in 1 1 of EtOH 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 temperature rose

5    temporarily from 10'Cto 104'CAfter. 2 h, the hydrogen uptake was complete. Subsequently, the catalyst was filtered off and the solvent was removed under reduced pressure to obtain 215 g (99%) of a pale yellow solid, mp 174-176'C.

c) 2-((E)-2-Ethoxycarbonylviny1}4-trifluoromethylbenzoic acid


520 mg of NaN02 (7 .6 mmol) were dissolved in 2 ml of water and added dropwise at O'Cto a solution of 1.3 g of 2-amino-4-trifluornmethylbenzoic acid (6.5 mmol) in
2.6 ml of a 48% aqueous HBF4 solution and 30 ml of ethanol. The mixture was then stirred at O'Cfor 10 minutes, then warmed toRT. A further 0.3 ml of a 48% aqueous
15    HBF 4 solution was then added, then 30 ml of ethanol, 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'Cfor 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 1N

aqueous HCI solution then with 25 ml of a saturated aqueous NaCI solution. The

20    organic phase was dried over Na2S04 and the solvent removed under reduced pressure. The residue was suspended in 25 ml of heptane and the precipitated pn:>duct was filtered off. Yield: 1.3 g (69%) of a pale brownish solid. An analytical sample was purified by crystallization from heptane/ethyl acetate.
 

38

The NMR spectrum was identical to the material prepared in example 3a.
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 and 0.93 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 reduced pressure. 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-Dxo-2-(2,2,2-trifluomethyl)-6-trifluoromethyl-2,3-dihydro-1 H-isoindol-1-yl]acetic acid
:~~'
0    F  F
15

2.2 g of ethyl (E}-3-[2-(2,2,2-trifluoroethylcarbamoyl}-5-trifluommethylphenyl]acrylate

(5.9 mmol} were dissolved in 10 ml of methanol and 1.5 ml of a 5 M aqueous NaDH

solution (7.5 mmol) were added. The mixture was stirred at RT for 18 hand then set

to pH = 7 using aqueous HCI solution. The solvents were removed under reduced

20    pressure and the residue was suspended in 10 ml of water. This suspension was set to pH = 2 using a 2N aqueous HCI solution and extracted 3 times with 10 ml each time of EA. Drying was effected over Na2SD4 and the solvent was removed under

reduced pressure. The residue was crystallized with diethyl ether/DIP, mp:
 

39

202-204°C.

Yield: 1.8 g (89%).

1H NMR (400 MHz, CDCI,): o = 3.07 {dd, J,=17 Hz, J2=6 Hz, 1 H), 3.23 {dd, J,=17 Hz, J2= 5Hz. 1 H). 4.27 (m, 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: C13H9FsN03 (341.2): calc. C45.76 H 2.66 N4.10; found C45.71 H2.43N4.11.

f)    Ethyl (RS )-[3-oxo-2-(2,2,2-trifluoroethyl)-6-trlfluoromethyl-2,3-dihydro-1 H-isoindol-

10    1-yl]acetate

:~~'
0    F  F

2.6 ml of SOCI2 (35 mmol) were dissolved in 20 ml of ethanol and 3.4 g of (R,S)-[3-oxo-2-(2,2,2-trifluoroethyl)-6-trifluoromethyl-2,3-dihydro-1 H-isoindol-1-yl]acetic acid (10 mmol) were added at -10°C. The mixture was stirred at RT for 18 hand 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 %) of a 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-[3-0xo-2-(2,2,2-trifluoroethyl)-6-trifluoromethyl-2,3-dihydro-H-isoindol-1-yl]acetyl}guanidine

0    !'
N    NH2

F    '!---F

0    F

Guanidine hydrochloride (11.5 g, 120 mmol) was dissolved in NMP (45 ml) and
 

40

KOtBu (11.2 g, 100 mmol) was added wtth stirring, and the mixture was left to stir at RT for 1.5 hand filtered. The filtrate was added dropwise at RT with stirring to a solution of ethyl (RS)-[3-oxo-2-(2,2,2-trifluoroethyl)-6-trifluoromethyl-2,3-dihydro-1 H-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 to pH 7 using 2N HCI, ethyl acetate (60 ml) was added and the pH was subsequently adjusted to 8-8.5 by adding NaHCO, solution. The mixture was stirred vigorously at RT for 1 h and the precipitate formed was filtered off wtth suction and washed with

water. 7.06 g (83%) of (R,S)-N-{2-[3-oxo-2-(2,2,2-trifluoroethyl)-6-trifluoromethyl-2,3-

10    dihydro-H-isoindol-1-yl]acetyl)guanidine, inclusion compound with 0.5 equivalent of ethyl acetate, were obtained as light yellow crystals, mp. 160-161•c with gradual heating, escape of ethyl acetate from approximately 9o•c.

Rf (ethyl acetate/methanol)= 0.45

1H NMR (400 MHz, CDCI,): o= 2.54 (dd, J,=8 Hz, J2=16 Hz, 1 H), 3.09 (dd, J1=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.

C14H12F6N40 2 • y, C4H80 2 (426.33): calc. c 45.08, H 3.78, N 13.14; found c 45.07, H 3.79, N 13.01.

20    Example 3

a) 2-((E)-2-Ethoxycarbonylvinyl)-4-trifluoromethylbenzoic acid (variant of

example 2c)

658 ml of a 48-50% aqueous HBF 4 solution were added at RT to 339 g of 2-amino-

25    4-trifluoromethylbenzoic acid (1,65 mol) in 6.81 of EtOH (anhydrous). The temperature rose from 21•c to 26•c.The mixture was then cooled to o•cand a solution of 125 g of NaN02 in 500 ml of water was added dropwise between o•cand
 

41

soc over 17 minutes. The initially pale yellow solution became initially an orange-red suspension and finally a light yellow suspension. The progress of the reaction was monitored by HPLC (method B; 2-amino-4-trifluoromethylbenzoic acid rt = 6.4 min; 2-carboxy-5-trifluoromethylbenzenediazonium salt intermediate= 1.1 min. Within 30

5    minutes, the conversion to the 2-carboxy-5-trlfluoromethylbenzenediazonium salt was >99% complete. The mixture was then added to 231 g of ethyl acrylate

(2.31 mol), 11.1 g of Pd(OAc)z (49 mmol) and 6.81 of ethanol (anhydrous) and the reaction mixture was heated to 49-51 oc. A uniform evolution of nitrogen increasing

with increasing temperature was observed. The reaction was monitored by HPLC

10    (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 then cooled toRT and the solvent removed under reduced pressure. The residue was

taken up in 3 I of EA and filtered off. The filtrate was then washed initially 3 times

with 2.1 I each time of an aqueous HCI solution, then with 1 I of a saturated aqueous

15    NaCI solution. Drying was effected over Na2S04 and the solvent was removed under reduced pressure to obtain 449 g of a light brown solid. Taking into account an impurity (4-trifluoromethylbenzoic acid; 6.3%) and solvent residues (EA; 4%), a yield of 83% was obtained. An analytical sample was purified by crystallization from
heptane/ethyl acetate. Mp.: 132-133°C

20    'HNMR (400 MHz, CDC!,): 8 = 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: C13H11F304 (288.23): calc. C 54.17, H 3.85; found C 54.24,

H 3.74.

25

b) Ethyl (E}-3-[2-(2,2,2-trifluoroethylcarbamoyl)-5-trifluoromethylphenyl]acrylate
 

42

315 g of oxalyl chloride (2.48 mol) were added at a temperature between 15 and 18'Cover 24 min to a mixture of 650 g of 2-((E)-2-ethoxycarbonylviny1}4-trifluoromethylbenzoic acid (2.25 mol), 33 ml of DMF and 7.8 I of CH2CI2. During the addition, gas evolution was observed. The mixture was stirred at RT for 1 h and then
5    cooled to S'C,and 285 g of Et3N (2.81 mol) was then added at a temperature between S'Cand 1O'Cover a period of 27 min. The mixture was stirred at 5'Cfor a further 10 min, then 279 g of 2,2,2-trifluoroethylamine (2.81 mol) were added at a
temperature between 9'Cand 20'Cover a period of 27 min. The mixture was stirred

at RT for 10 min, in the course of which a thick precipitate precipitated out, and, to

10    improve the stirrability of the mixture, an additional! 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-trifluoromethylphenyl]acrylate rt = 13.2 min). After stirring at RT for a further
50 min, the reaction was complete. Volatile constituents of the reaction mixture were

15    then removed under reduced pressure, and the residue was taken up with 12 I of EA and washed 3 times with 2.5 I each time of water, then twice with 2.5 I each time of a saturated aqueous NaHC03 solution and finally wtth 1.5 I of a saturated aqueous NaCI solution. Drying was effected over MgS04, and the solvent was removed under

reduced pressure to obtain 802 g of ethyl (E}-3-[2-(2,2,2-trifluoroethylcarbamoyl)-5-

20    trifluoromethylphenyl]acrylate as a brown solid. This crude substance was combined with the crude product of another batch (177 g) and dissolved at 60-70'Cin 31 of EA, and 14 I of HEP were added at this temperature in 1 I portions. The mixture was then heated to 80'Cand stirred at this temperature for 1.5 h. This mixture was then
added to 5.6 I of HEP at 70'Cand the mixture was then cooled to RT with stirring

25    over a period of 5 h. The product was then filtered off, washed with 31 of HEP and dried under air to obtain 689 g of ethyl (E}-3-[2-{2,2,2-trifluoroethylcarbamoyl}-5-trifluoromethylphenyl]acrylate (67%) as a light brown solid. Mp: 161.5-162'C.

1 H NMR (400 MHz, CDCI3): o = 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,

30    1 H), 7.87 (s, 1 H), 7.90 (d, J=16 Hz, 1 H) ppm.

Combustion analysis: CtsH13F5N03 (369.27):calc. C 48.79, H 3.55, N 3.79; found C 48.93, H 3.51, N 3.92.
 

43


c) (R,S)-N-{2-[3-0xo-2-(2,2,2-trifluoroethyl}-6-trifluoromethyl-2,3-dihydro-H-isoindol-1-yl]acetyl}guanidine

386 g of ethyl (E)-3-[2-(2,2,2-trifluoroethylcarbamoyl}-5-trifluoromethylphenyl]acrylate (1.05 mol) was suspended in 600 ml of DMF and added at a temperature between 5'Cand 15'Cin portions slowly to 4. 7 g of KO!Bu (42 mmol). The cyclization to the

10    isoindolone was monitored by TLC (HEP/EA =2:1; ethyl acrylate: Rf =0 .32; isoindolone: Rf = 0.41 ). After one hour, the reaction was complete. In the meantime,

587 g of KO!Bu were suspended in 2.2 I of DMF and 600 g of guanidinium chloride

were added at a temperature of between 20'Cand 25'C.The mixture was stirred at

25'Cfor 1 h and then the KCI was filtered off. The filtrate comprising the released

15    guanidine was then added to the reaction mixture comprising the isoindolone and

stirred 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 14 I

of ice-water, set to pH .8.5-9.0 using aqueous HCL solution and extracted 4 times

20    with 3 1 each time of EA. The mixture was then washed 3 times with 3 I each time of a saturated aqueous NaCI solution and dried over Na2S04, and the solvent was removed under reduced pressure. 329 g (82%) of a brown solid were obtained. The product was combined with 3 other batches of the same preparation process; total amount 842 g. These 842 g (2.2 mol) were digested in 2 I of EA and 5 I of Et20 at

25    30'Cfor 2 h. The solid was then filtered off, washed twice with 2 I each time with Et2o and dried under reduced pressure. 693 g (82% recovery) of an almost white solid were obtained. The compound crystallized from 2-propanol as the inclusion compound with 0.5 equivalent of 2-propanol.

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

44


Example 4

Ethyl (RS }-(2-(2,2,2-tritluoroelhyt}-3-oxo-6-lriftuoromethyl-2,3-dihydro-1 H-isoindol-1-yl)acetate by one-pot reaction starling from 2-((E)-2-ethoxycarbonylvinyl}-4-

5    tritluoromethylbenzoic acid


SOCI2 (1.98 g, 27.2 mmol) was added at room temperature to a suspension of 2-({E)-2-ethoxycarbonylvinyl)-4-trifluoromethylbenzoic acid (2.9 g, 10.1 mmol) in toluene (30 ml). The mixture was stirred at room temperature for 5 min and then

10    heated to 105'C(bath temperature) within 30 min. At about 70'C,gas evolution commenced. The mixture was stirred at 1OS'Cfor 3 h, then cooled to room temperature, filtered with suction through a kieselguhr layer (2.5 x 0.5 em) and washed with toluene, and the filtrate was concentrated by evaporation under reduced 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 S'Cin dichloromethane (15 ml), and the acid chloride,

dissolved in dichloromethane (20 ml}, was added dropwise with ice cooling at such a rate that the temperature was kept between S'Cand 10'C.The ice bath was then removed and the excess of trifluoroethylamine and a portion of the dichloromethane

20    distilled off under gentle vacuum. Subsequently, the mixture was heated to boiling under reflux for 10 h. After it had been cooled, the mixture was diluted with dichloromethane (50 ml) and extracted twice with aqueous 2N HCI solution (50 ml each time), and the combined organic phases were washed with water (1 00 ml), dried over Na2S04 and concentrated under reduced pressure. Ethyl (RS)-(2-(2,2,2-

25    trifluoroethyl}-3-oxo-6-trifluoromethyt-2,3-dihydro-1 H-isoindol-1-yt)acetate (3,51 g, 94%) was obtained as a dark brown oil which was purified by crystallization from n-heptane. Mp: 54.5-55.5'C.

1H NMR (400 MHz, CDC/g): 8 = 1.15 (t, J=7 Hz, 3 H), 2.85 (dd, J1=6 Hz, J2=16 Hz,
 

45

1 H), 3.01 (dd, J1=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: C15H13F5N03 (369.27): calc. C 48.79, H 3.55, N 3.79; found C 48.54, H 3.49, N 3.79.
5

Example 5

a) (S}-N-{2-[3-0xo-2-(2,2,2-trifluoroethyl)-6-trifluoromethyl-2,3-{lihydro-1 H-isoindol-1-yl]acetyl)guanidine, 0,0'-{libenzoyl-l-tartaricacid salt

10    (RS )-N-{2-[3-0xo-2-(2,2,2-trifluoroethyl}-6-trifluoromethyl-2,3-dihydro-1 H-isoindol-1-yl]acetyl)guanidine (inclusion compound wtth ethyl acetate, content 87.06% by NMR, 44 g, 100 mmol) and 0,0'-{libenzoyi-L-tartaricacid (11.2 g, 31 mmol) were initially charged as solids and 2-propanol (500 ml) was added dropwise with stirring. The
solids 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 left to cool to room temperature within 4 h and subsequently stirred at this temperature overnight. Afterward, the mixture was stirred at 10"C for 4 hand subsequently filtered with suction. The residue was washed twice with 2-propanol (100 ml each

time) and dried under air. 28.05 g of (S)-N-{2-[3-<Jxo-2-(2,2,2-trifluoroethyl}-6-20 trifluoromethyl-2,3-dihydro-1 H-isoindol-1-yl]acetyl}guanidine, 0,0 •- dibenzoyi-L-

tartaric acid salt (74% yield based on the (S)•<:>nantiomer), in enantiomeric purity 82% ee by HPLC (Chiracel OD/21, 250 • 4.6 mm, 50:5:2 n-heptanelethanollmethanol, 1 mllmin, 30"C) were obtained as colorless crystals. 20 g (14.6 mmol) of these crystals were inttially charged and 2-propanol (400 ml)
25    was added dropwise. The mixture was heated to 80"C with stirring and then /eft to
 

46

cool gradually to room temperature. The mixture was stirred at this temperature for a further 2 h and then filtered with suction, and the residue was washed twice with 2-propanol (50 ml each time) and dried under air. 16.3 g (tOO% 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, 0,0'-{libenzoyi-L-tartaric acid salt were obtained as colorless crystals, mp: 192-193°C, enantiomeric purity >97% ee by HPLC (conditions as above).
Combustion analysis: C14H12F5N402 • Y. C1sH140s (561.43): calc. C 49.21, H

3.41, N 9.98: found C 49.17, H 3.30, N 9.97.

10

b) (S)-N-{2-[3-0xo-2-(2,2,2-trifluoroethyl)-6-trifluoromethyl-2 ,3-dihydro-1 H-isoindol-1-yl]-acetyl}guanidine

15    (S)-N-{2-(3-0xo-2-{2,2,2-trifluoroethyl)-6-trifluoromethyl-2,3-dihydro-1 H-isoindol-1-yl]-acetyl}guanidine, 0,0'-{libenzoyi-L-tartaric acid salt (113 mg, 0,20 mmol) was dissolved in a mixture of water (1 ml) and ethyl acetate (5 ml) and a solution of NaHC03 (50 mg) in water (7 .5 ml) was added. The mixture was left to stir at room

temperature for 16 h and then extracted three times with ethyl acetate (5 ml each

20    time). The combined organic phases were shaken once with a solution of NaHC03 (50 mg) in water (20 ml) and then w~h pure water (20 ml), dried over Na2S04 and concentrated by evaporation under reduced pressure. 75 mg (97%) of (S)-N-{2-[3-oxo-2-(2,2,2-trifluoroethyl)-6-trifluoromethyl-2,3-dihydro-1 H-isoindol-1-yl]-acetyl}-
guanidine were obtained. The product crystallized from 2-propanol as an inclusion

25    compound with 0.5 equivalent of 2-propanol, mp: 80-82°C. The substance may be recrystallized from ethyl acetate and crystallizes with 0.5 equivalent 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 mllmin, 30°C).
 

47

1H NMR (400 MHz, CDCI3): o= 2.54 (dd, J1=8 Hz, J2=16 Hz, 1 H), 3.09 (dd, J1=4 Hz, 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.
Combustion analysis (inclusion compound with 0.5 equivalent of 2-propanol}:

5    c1 4H12F5N402 • Y. C3H50 (412.32): calc. C 45.15, H 3.91, N 13.59; found C 45.23, H 4.27, N 13.10.

Example 6 (RS}-N-{2-[3-0xo-2-(2,2,2-trifluoroethyl}-6-trifluoromethyl-2,3-<lihydro-1 H-isoindol-

10    1-yl]acetyl}guanidine by racemization of (R}-N-{2-[3-<>xo-2-(2,2,2-trifluoroethyl}-6-trifluoromethyl-2,3-dihydro-1 H-isoindol-1-yl]acetyl)guanidine
0    NH2
1-F ~ rNANfl, F'"CLiN}-F

0    F

(R}-N-{2-[3-0xo-2-(2,2,2-trifluoroethyl}-6-trifluoromethyl-2,3-<lihydro-1H-isoindol-

1-yl]acetyl)guanidine (inclusion compound with 0.5 equivalent of 2-propanol

15    (M=412.3), 43 g, 104 mmol; obtained by concentrating the mother liquor which was obtained 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, 0,0"- dibenzoyi-L-tartaric acid salt in example 5a, and treatment with NaHC03 as described in

example 5b}} was dissolved in 2-propanol (1.81) and a solution of KOH, (85 percent,

20    660 mg, 10 mmol) in 2-propanol (400 ml) was added at room temperature with stirring. The mixture was stirred at room temperature for 24 h and then acidified using glacial acetic acid (720 mg, 1.5 ml) and concentrated by evaporation under reduced pressure at a max. bath temperature of 40'C,and the residue was

partitioned between water (500 ml) and ethyl acetate (400 ml). The aqueous phase

25    was extracted twice w~h ethyl acetate (300 ml each time). The combined organic phases were shaken with a solution of NaHC03 (10 g) in water (500 ml) and then once again with pure water. The organic phase was dried over Na2S04 and concentrated by evaporation under reduced pressure at a max. bath temperature of 40'C.39.8 g of (RS)-N-{2-[3-oxo-2-(2,2,2-trifluoroethyl}-6-trifluoromethyl-2,3-<lihydro-


1H-isoindol-1-yl]acetyl}guanidine, content 87% by NMR, inclusion compound wtth 0.5 equivalent of ethyl acetate, were obtained as pale yellow crystals, yield: 87%. Mp: 164-166'Con gentle heating, escape of ethyl acetate from approximately 100'C.

5    Enantiomeric ratio 49:51 by HPLC (Chiracel OD/21, 250 x 4.6 mm, 4:1 n-heptane/2-propanol, 1 mUmin, 30'C).

Example 7

(S )-N-{2-[3-0xo-2-(2 ,2 ,2-trifluoroethyl)-6-trifluoromethyl-2,3-dihydro-1 H-isoindol-1-yl]-

10   acetyl}guanidine hydrogenfumarate hydrate           
0NH2           
F F~Nj__NH2        o   
F~~/\ ...,J    X  HO~OH   
~ rF    XHO    0   
    2       
0   F           

(S)-N-{2-[3-0xo-2-(2,2,2-trifluoroethyl)-6-trifluoromethyl-2,3-dihydro-1 H-isoindol-1-yl]-acetyl}guanidine [inclusion compound with 0.5 equivalent of2-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 solution formed was concentrated by evaporation under reduced pressure. The residue was taken up with dichloromethane (2 I) and the mixture concentrated again by evaporation under reduced pressure. The residue was suspended in water (1.5 1), filtered wtth suction at room temperature and dried under air at room temperature

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 were obtained as colorless crystals, mp. 21 O'C.

Determination of the NHE inhibttion

25

The inhibitory concentration IC50 for NHE-1 inhibition was determined as follows:

The NHE-1 inhibition IC5o was determined in an FLIPR assay by measurement of the recovery in pH; in transfected cell lines which express human NHE-1.

The assay was carried out in an FLIPR (fluorescent imaging plate reader) with black-walled 96-well microliter plates having clear bases. The transfected cell lines expressing the different NHE subtypes (the parental cell line LAP-1 shows no

5    endogenous NHE activity as a consequence of mutagenesis and subsequent selection) were seeded the preceding day at a density of -25 000 cells/well. The growth medium for the transfected cells (lscove +1 0% fetal calf serum) additionally contained G418 as a selection antibiotic in order to ensure the presence of the
transfected sequences.

10

The actual assay started with the removal of the growth medium and addition of 100 ~I of loading buffer per well (5 ~M BCECF-AM [2',7'-bis(carboxyethyl)-5(and--6-)carboxyfluorescein, acetoxymethyl ester] in 20 mM NH4CI, 115 mM choline chloride, 1 mM MgCiz, 1 mM CaCiz, 5 mM KCI, 20 mM HEPES, 5 mM glucose;

15    pH 7.4 [adjusted with KOH]). The cells were then incubated at 37'Cfor 20 minutes. This incubation led to loading of the cells with the fluorescent dye whose fluorescence intensity depends on pHi, and with NH4CI which makes the cells

slightly alkaline.

The nonfluorescent dye precursor BCECF-AM is, as the ester, membrane-

20    penmeable. Inside the cell, esterases release the actual BCECF dye which is membrane-impenmeable.

After this incubation for 20 minutes, the loading buffer which contained NH4CI and

free BCECF-AM was removed by washing three limes in a cell washer (Tecan

25    Columbus) with in each case 400 ~I of washing buffer (133.8 mM choline chloride, 4.7 mM KCI, 1.25 mM MgCiz, 1.25 mM CaCiz, 0.97 mM KzHP04, 0.23 mM KHzP04,

5 mM HEPES, 5 mM glucose; pH 7.4 [adjusted with KOHl). The residual volume remaining in the wells was 90 ~I (50--125 ~I possible). This washing step removed the free BCECF-AM and led, as a consequence of the removal of the external NH4+

30    ions, to intracellular acidification (-pH; 6.3-6.4).
 

50

Since the equilibrium of intracellular NH4+ with NH3 and H+ was disturbed by the removal of the extracellular NH4+ and by the subsequent 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 eventually
5    lead to cell death if it persists long enough. It was important at this point that the washing buffer was sodium-free (<1 mM), since extracellular sodium ions would lead to instantaneous recovery in the pH; as a result of the activity of the cloned NHE

isoforms.

It was likewise important for all buffers used (loading buffer, washing buffer, recovery

10    buffer) not to contain any HC03- ions, since the presence of bicarbonate would lead to activation of interfering bicarbonate-dependent pHi regulatory systems which are present in the parentallAP-1 cell line.

The microttter plates with the acidified cells were then (up to 20 minutes after the

15    acidification) transferred to the FLIPR. In the FLIPR, the intracellular fluorescent dye was excited by light having a wavelength of 488 nm which was generated by an argon laser, and the measured parameters (laser power, illumination time and aperture of the CCD camera incorporated in the FLIPR) were selected such that the average fluorescence signal per well was between 30 000 and 35 000 relative

20    fluorescence units.

The actual measurement in the FLIPR started with an image being taken by the CCD camera every two seconds under software control. After ten seconds, the recovery of the intracellular pH was initiated by adding 90 ~I of recovery buffer (133.8 mM NaCI,

25    4.7 mM KCI, 1.25 mM MgCI2, 1.25 mM CaCI2, 0.97 mM K2HP04, 0.23 mM KH2P04, 10 mM HE PES, 5 mM glucose; pH 7.4 [adjusted with NaOH]) by means of the 96-well pipettor incorporated in the FLI PR.

The positive controls (100% NHE activity) used were wells to which pure recovery

buffer had been added, while negative controls (0% NHE activity) received washing

30    buffer. Recovery buffer with twice the concentration of test substance was added to
 

51

all other wells. Measurement in the FLIPR ended after 60 measurements (two minutes).

The raw data were exported into the ActivttyBase program. This program firstly

5    calculated the NHE activities for each tested substance concentration and, from these, the ICso values for the substances. Since the course of pH; recovery was not linear over the entire experiment, but rather fell at the end owing to decreasing NHE activity at higher pH; values, it was important to select, for evaluation of the

measurement, the part in which the increase in fluorescence of the positive controls 10 was linear.

Substance    NHE1 inhibition ICso [nM]
       
{S)-N-{2-[3-0xo-2-(2,2,2-trifluoroethyl)-6-       
trifluoromethyl-2,3-dihydro-1 H-isoindol-1-    0.3   
yl]acetyl}guanidine       

52

What is claimed is;

1.    A process for preparing compounds of the fomnula I

NH2

NANH2

R1    R3

R2    0

5    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, 3

or 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

R1~~-R3

R2    0

IV

fR5
R1~N-R3

R2    0

VII

a) fomnylating the amide of the fomnula IV and then cyclizing to the compound of the formula VI,
 

53

b)    reacting the compound of the formula VI wtth an alkoxycarbonylmethylene-triphenylphosphorane, wtth a 1-alkoxy-1-trimethylsiloxyethylene or with a trialkyl phosphonoacetate to give the compound of the formula VII, and

c)    reacting the compound of the formula VII with guanidine to give the compound of 5 the formula I,

where, in the compounds of the formulae IV, VI and VII, R1 to R3 are each as defined in formula I and

R5 is alkoxy having 1, 2, 3 or 4 carbon atoms; and salts thereof.

10

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    is alkyl having 1, 2, 3 or 4 carbon atoms;
    R4    is hydrogen, trifluoromethyl or cycloalkyl having 3, 4, 5, 6 or
20        7 carbon atoms;

and salts thereof;

wherein
 

54

    +    ---Clfj   
R1_,_{        R1    ~N-R3   
               
R2II0    Ill    R2    0   
            IV   
               
               
---CYCH~    R1~:-R3   
R1   ~N-R3       
R2    0    R2    0   
    v        VI   

a) the compound of the formula II is reacted with the amine of the formula Ill to give

the amide of the formula IV,

5    b) the amide of the formula IV is formylated at the ortho-position to the amide function to give the formyl amide of the formula V,
c)    the formyl amide of the formula Vis cyclized to the compound of the formula VI,

d)    the compound of the formula VI is reacted wtth an alkoxycarbonylmethylenetriphenylphosphorane, with a 1-alkoxy-

10    1-trimethylsiloxyethylene or with a trialkyl phosphonoacetate to give the compound of the formula VII and
e) the compound of the formula VII is reacted with guanidine to give the compound of the formula I,
where, in the compounds of the formulae II, Ill, 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 and

X is Cl, Br, OH or alkoxy having 1, 2, 3 or 4 carbon atoms; and salts thereof.

20    3.    The process as claimed in claim 1 and/or 2, in which the process steps are
 

55

each independently conducted continuously or batchwise.

4. The process as claimed in one or more of claims 1, 2 or 3, wherein the compound of the formula I is defined as N-{2-[3-oxo-2-(2,2,2-trifluoroethyl)-

5    6-trifluoromethyl-2,3-dihydro-1H-isoindol-1-yl]acetyl}guanidine, and pharmaceutically acceptable salts thereof.

5. A process for preparing compounds of the formula I

~ ;)::H, I

R1~N-R3

R2    0

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, 3

or 4 carbon atoms;

15    R3    is Alk-R4 or trifluoromethyl;

Alk    is alkyl having 1, 2, 3 or4 carbon atoms;

R4    is hydrogen, trifluoromethyl or cycloalkyl having 3, 4, 5, 6 or 7 carbon atoms;

and salts thereof; 20 which comprises
R1~: _,d~            of>lH,   
            Nd.,_   
    R1        R3NH,   
R2    OH    R2OH    R2    0       
                       
IX        XI    *       
                   
 

56

a)    reacting the amine of the formula IX via a diazonium salt with an alkyl acrylate to give the cinnamic acid derivative of the formula XI,
b)    reacting the compound of the formula XI with the amine of the formula Ill and with guanidine to give the acylguanidine of the formula I,

5    where, in the compounds of the formulae Ill, 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.

10    6. The process for preparing compounds of the formula I as claimed in claim 5 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, 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

-CYNH,
R1 __)0"yo

R2    IX    OH





H,NR3    ,-d~        ~~OH   
Ill        A,B       
        R1~N-R3   
    R2    XII   HN,R3    R2    XIII  O   
        Jo~~""        Js   
        ON~H,    B,C    *OR7   
        NH,    R1        R3   
    R1    R3               
                R2    0   
    R2    0            XIV   
                       

a) the nitro compounds of the formula VIII is converted to the amine of the

formula IX,

5    b) the amine of the formula IX is converted to the diazonium salt of the formula X,

c)    the diazonium salt of the formula X is reacted with an alkyl acrylate to give the cinnamic acid derivative 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 derivative of the formula XIII and subsequenHy by reaction with guanidine with activation to give the acylguanidine of the formula I (alternative A), or, after formation of the isoindolone derivative ofthe formula XIII, in the presence of a

base, from the compound of the fonmula XII, by converting the compound of the formula XIII to the ester of the formula XIV and subsequently by reacting with guanidine to give the acylguanidine of the fonmula I (alternative B), or

by converting the compound of the fonmula XII in the presence of a strong base to

5    the ester of the fonmula XIV and subsequently by reacting with guanidine to the acylguanidine of the fonmula I (alternative C), or
by directly reacting the compound of the formula XII with guanidine in the presence of a base with simultaneously proceeding guanylation and cyclization to give the
isoindolone of the fonmula I (alternative D),

10    where, in the compounds of the fonmulae VIII, IX, X, XI, XII, XIII and XIV, R1 to R3 are each as defined in fonmula I and
R6 and R7 are each independently alkoxy having 1, 2, 3 or 4 carbon atoms; and salts thereof.

15    7. The process as claimed in claim 6, wherein, in process step e), alternative Dis used.


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.

20

9.    The process as claimed in claim 5 and/or 6, in which the process steps are

each independently conducted continuously or batchwise.

10.    The process as claimed in one or more of claims 5 to 9, wherein the compound 25 of the fonmula I is defined as N-{2-[3-<Jxo-2-(2,2,2-trifluoroethyl}-6-trifiuoromethyl-
2,3-dihydro-1 H-isoindol-1-yl]acetyl}guanidine, and phanmaceutically acceptable salts thereof.

11.    A compound of the fonmula XII
 

59

~ ;R6
R1h0
R2  HN,
XII    R3

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, 3 or 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 of the formula XII as claimed in claim 11 for use as a synthetic intermediate.

13.  A process for isolating compounds of the formula Ia and lb

~ ;NJ::H
"Y<-~ :
R2    0

20

where

R1 and R2
 
10
 

60

are each independently hydrogen, F, Cl, trifluoromethoxy, 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    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

R1~N-R3 '

R2    0

Ia    lb

a) converting the compound of the formula I to salts of a 2,3-0-acylated D- or L-tartaric acid and obtaining the two salts of the formulae ~a and ~b separately by crystallization, and

15    b) releasing the free bases of the formulae Ia and lb from the two salts of the formulae ~a and ~b respectively,

where, in the compounds of the formulae I, ~a and ~b. R1 to R3 are each as defined in the formulae Ia and lb


RS is alkyl having 1, 2, 3, 4, 5 or 6 carbon atoms or phenyl which is unsubstituted or substituted by 1, 2 or 3 substituents from the group ofF, Cl, Br, I, alkyl having 1, 2, 3

5    or 4 carbon atoms or alkoxy having 1, 2, 3 or 4 carbon atoms.

14. The process as claimed in claim 13, wherein the undesired enantiomer of the formulae Ia or lb is racemized again.

10    15. The process as claimed in claim 13 and/or 14, wherein the compounds of the formulae Ia and lb are (R}-N-{2-[3-oxo-2-(2,2,2-trifluoroethyl}-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-<lihydro-1 H-isoindol-1-yl]acetyl}guanidine.

15    16.   A compound of the formula >Wa and/or >Wb

0    NH2

(lN~NH2

Rt~N-R3
~    xR*/2

R2    0   
>Wa    XVb   
       

where

R1 and R2

20    are each independently hydrogen, F, Cl, trifluoromethoxy, 2,2,2-trifluoroethoxy, trifluoromethyl, 2,2,2-trifluoroethyl or alkyl having 1, 2, 3
or 4 carbon atoms;

R3    is Alk-R4 or trifluoromelhyl;
 

62

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



or

5

RB is alkyl having 1, 2, 3, 4, 5 or 6 carbon atoms or phenyl which is unsubstituted or substituted by 1, 2 or 3 substituents from the group ofF, Cl, Br, I, alkyl having 1, 2, 3 or 4 carbon atoms or alkoxy having 1, 2, 3 or 4 carbon atoms.

10    17. (S)-N-{2-[3-<>xo-2-(2,2,2-trifluoroethyl)-6-trifluoromethyl-2,3-dihydro-1 H-isoindol-1-yl]acetyl)guanidine hydrogenfumarate hydrate of the formula XVI

\?    NH2
F F    ,~,)N__ NH 2    0
F~N~F XHO~OH0  XVI
X H20
0    F

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 medicament.

19.    19. The use of a compound of the formulae XV a and/or XVb as claimed in claim 16 or of the formula XVI as claimed in claim 17, alone or in combination with other

20    medicaments or active ingredients, for producing a medicament for the treatment or prophylaxis of acute or chronic damage, disorders or indirect sequelae of organs and tissues caused by ischemic or by reperfusion events, for the treatment or prophylaxis of arrhythmias, of life-threatening cardiac ventricular fibrillation, of myocardial

infarction, of angina pectoris, for the treatment or prophylaxis of ischemic states of 25 the heart, of ischemic states of the peripheral and central nervous system or of
 

63

stroke or of ischemic states of peripheral organs and tissues, for the treatment or prophylaxis of states of shock, of diseases in which cellular proliferation represents a primary or secondary cause, of cancer, of metastasis, of prostate hypertrophy and of prostate hyperplasia, of atherosclerosis or of disturbances of lipid metabolism, of

5    high blood pressure, of essential hypertension, of disorders of the central nervous system, of disorders resulting from overexcitability of the CNS, epilepsy or centrally induced convulsions, of disorders of the central nervous system, especially of anxiety states, depressions or psychoses, for the treatment or prophylaxis of non-

insulin-dependent diabetes mellitus {NIDDM) or late damage from diabetes, of

10    thromboses, of disorders resulting from endothelial dysfunction, of intenmittent claudicating, for the treatment or prophylaxis of fibrotic disorders of internal organs, fibrotic disorders of the liver, fibrotic disorders of the kidney, fibrotic disorders of vessels and fibrotic disorders of the heart, for the treatment or prophylaxis of heart failure or of congestive heart failure, of acute or chronic inflammatory disorders, of
15    disorders caused by protozoa, of malaria or of coccidiosis in poultry, and for use for surgical operations and organ transplantations, for preserving and storing transplants for surgical procedures, for use in bypass operations, for use in resuscitation after cardiac arrest, for preventing age-related tissue change, for producing a medicament

directed against aging or for prolonging life, for the treatment and reduction of

20    cardiotoxic effects in thyrotoxicosis or for producing a diagnostic aid.

20. A medicine for human, veterinary and/or phytoprotective use, comprising an effective amount of a compound of the formulae XV a or XVb as claimed in claim 16 together with pharmaceutically acceptable excipients and additives.
25

21. A medicine for human, veterinary or phytoprotective use, comprising an effective amount of a compound of the fonmulae XV a or XVb as claimed in claim 16 together with pharmaceutically acceptable excipients and additives in combination with other pharmacological active ingredients or medicaments.
30

22. A medicine for human, veterinary and/or phytoprotective use, comprising an effective amount of a compound of the fonmula XVI as claimed in claim 17 together with phanmaceutically acceptable excipients and additives.
 

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