(19) TZZ ¥ZZZ_Z_T

(11) EP 2 300 010 B1

(12) EUROPEAN PATENT SPECIFICATION

(45) Date of publication and mention (51) Int Cl.: of the grant of the patent: C07D 403/12 (2006.01) C07D 241/26 (2006.01) 04.03.2015 Bulletin 2015/10 C07D 401/12 (2006.01)

(21) Application number: 09761700.5 (86) International application number: PCT/EP2009/057060 (22) Date of filing: 08.06.2009 (87) International publication number: WO 2009/150137 (17.12.2009 Gazette 2009/51)

(54) PYRAZINE DERIVATIVES AS EPITHELIAL SODIUM CHANNEL BLOCKERS PYRAZINDERIVATE ALS BLOCKIERER DES EPITHELIALEN NATRIUMKANALS DÉRIVÉS DE PYRAZINE EN TANT QUE BLOQUEUR DU CANAL SODIQUE EPITHELIAL

(84) Designated Contracting States: (56) References cited: AT BE BG CH CY CZ DE DK EE ES FI FR GB GR WO-A-2005/025496 WO-A-2006/023573 HR HU IE IS IT LI LT LU LV MC MK MT NL NO PL WO-A-2007/071400 PT RO SE SI SK TR • JH LI ET AL: "Stereoselective blockade of (30) Priority: 10.06.2008 EP 08157980 amphibian epithelial sodium channels by amiloride analogs" JOURNAL OF (43) Date of publication of application: PHARMACOLOGY AND EXPERIMENTAL 30.03.2011 Bulletin 2011/13 THERAPEUTICS, AMERICAN SOCIETY FOR PHARMACOLOGY AND EXPERIMENTAL (73) Proprietor: Novartis AG THERAPEUTICS, US, vol. 267, no. 3, 1 December 4056 Basel (CH) 1993 (1993-12-01), pages 1081-1084, XP008093856 ISSN: 0022-3565 (72) Inventors: • RUSS T ET AL: "Preparation and diuretic • COLLINGWOOD, Stephen Paul propertiesof novel amiloride analogues" ARCHIV Horsham Sussex RH12 5AB (GB) DER PHARMAZIE, VCH • DEVEREUX, Nicholas James VERLAGSGESELLSCHAFT MBH, WEINHEIM, Horsham Sussex RH12 5AB (GB) DE, vol. 325, no. 12, 1 December 1992 • HOWSHAM, Catherine (1992-12-01), pages 761-767, XP002496825 ISSN: Horsham Sussex RH12 5AB (GB) 0365-6233 • HUNT, Peter Horsham Sussex RH12 5AB (GB) Remarks: • HUNT, Thomas Anthony Thefile contains technical information submitted after Horsham Sussex RH12 5AB (GB) the application was filed and not included in this specification (74) Representative: Mueller, Philippe et al Novartis Pharma AG Patent Department 4002 Basel (CH)

Note: Within nine months of the publication of the mention of the grant of the European patent in the European Patent Bulletin, any person may give notice to the European Patent Office of opposition to that patent, in accordance with the Implementing Regulations. Notice of opposition shall not be deemed to have been filed until the opposition fee has been paid. (Art. 99(1) European Patent Convention). EP 2 300 010 B1

Printed by Jouve, 75001 PARIS (FR) EP 2 300 010 B1

Description

[0001] This invention relates to organic compounds, their preparation and use as pharmaceuticals. [0002] The compounds of the invention typically include a suitable, negatively-charged counter ion. This may be any 5 suitable counter ion and in particular may be any suitable pharmaceutically acceptable counter ion. Examples of suitable counter ions include, but are not limited to: fluoride, chloride, bromide, iodide, nitrate, sulfate, phosphate, hexafluoro- phosphate, formate, acetate, trifluoroacetate, propionate, butyrate, lactate, citrate, tartrate, malate, maleate, succinate, benzoate, p-chlorobenzoate, diphenylacetate, triphenylacetate, o-hydroxybenzoate, p-hydroxybenzoate, 1-hydroxy- naphthalene-2-carboxylate, 3-hydroxynaphthalene-2-carboxylate, methanesulfonate and benzenesulfonate. 10 [0003] In one aspect, there is provided a compound according to the Formula Ia:

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or pharmaceutically acceptable salts, solvates or hydrates thereof, wherein 6 6a R is -(C0-C6 alkylene)-R , wherein the alkylene linker is optionally substituted by one or more groups selected from C1-C3 alkyl, halo and OH; 25 6a 11 12 13 14 R is selected from H, a C 3-C10 carbocyclic group, NR R , C(O)NR R , C6-C10 aryl, 5- to 6-membered heteroaryl, 5- to 6-membered heterocyclyl and a group of the formula P-(CH2)m-Q, wherein each of the ring systems is optionally substituted by one or more substituents selected from the List Z; a- a a P is selected from a bond, -O-, -C(O)-, -C(O)O-, -NHC(=N)NH- and -S(O2)-,-S(O2)NR , -NR C(O)-,-NR C(O)O-, a a b -NR S(O2)- and -NR C(O)NR -; 30 a b R and R are each independently selected from H, C 1-C6 alkyl, a C 3-C10 carbocyclic group and -(C 1-C3 alkylene)-C3-C10 carbocyclic; Q is selected from H, C1-C6 alkyl, a C3-C10 carbocyclic group, C6-C10 aryl, 5- to 6-membered heteroaryl and 5- to 6- membered heterocyclyl, wherein the carbocyclic, C6-C10 aryl, 5- to 6-membered heteroaryl and 5- to 6-membered heterocyclyl groups are each optionally substituted by one or more substituents selected from List Z; 35 m is 0, 1, 2 or 3; 7 8 10 R , R and R are each independently selected from C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, a C3-C10 carbocyclic group, -(C1-C3 alkylene)-C3-C10 carbocyclic group, C6-C10 aryl, 5- to 6-membered heteroaryl, 5- to 6-membered het- erocyclyl or a group of the formula (CH 2)d-A-(CH2)b-B, wherein the carbocyclic, C6-C10 aryl, 5- to 6-membered heteroaryl and 5- to 6-membered heterocyclyl groups are each optionally substituted by one or more substituents selected from 40 List Z; and wherein the alkyl and alkylene groups are optionally substituted by one or more halogen atoms, OH groups or phenyl groups; or R7 and R8, together with the nitrogen atom to which they are attached form a 4- to 10-membered nitrogen-containing heterocyclic group which is optionally substituted by one or more substituents selected from the List Z; or R6 and R 8, together with the atoms to which they are attached form a 4- to 10-membered nitrogen-containing heterocyclic 45 group which is optionally substituted by one or more substituents selected from the List Z; c c A is selected from a bond, -O-, -C(O)-, -C(O)NR-, -C(O)O-, -OC(O)-, -NHC(=N)NH-, -S(O2)- -S(O2)NR -, c c c c c c d c -NR C(O)-,-NR C(O)O-, -OC(O)NR -, -NR S(O2)-,-C(O)NR S(O2)-, -NR C(O)NR -, -NR -, -C6-C10 aryl-, -C3-C10 car- bocyclyl-, -5- to 6-membered heteroaryl-, -5- to 6-membered heterocyclyl-, -C6-C10 aryl-O-, -O-C6-C10 aryl-, -O-C 3-C10 carbocyclyl- and -C3-C10 carbocyclyl-O-, wherein the C6-C10 aryl, carbocyclyl, 5- to 6-membered heteroaryl and 5- to 50 6-membered heterocyclyl groups are each optionally substituted by one or more substituents selected from List Z; x y z B is selected from H, C 1-C6 alkyl, a C 3-C10 carbocyclic group, NR R , C(O)OR , C6-C10 aryl, 5- to 6-membered heteroaryl and 5- to 6-membered heterocyclyl, wherein the carbocyclic, C6-C10 aryl, 5- to 6-membered heteroaryl and 5- to 6- membered heterocyclyl groups are each optionally substituted by one or more substituents selected from List Z; d is 1, 2, 3, 4, 5, 6 or 7; 55 b is 0, 1, 2 or 3; c d x y R , R , R and R are each independently selected from H, 1 C-C6 alkyl, a C3-C10 carbocyclic group and -(C1-C3 alkylene)-C3-C10 carbocyclic; z R is H or C1-C6 alkyl;

2 EP 2 300 010 B1

9 R is H or C1-C6 alkyl; or 6 9 R and R , together with the carbon atom to which they are attached, form a C3-C10 carbocyclic group or a 4- to 10- membered 5- to 6-membered heterocyclyl group, wherein each of the ring systems is optionally substituted by one or more substituents selected from the List Z; 5 11 R is selected from H, C1-C6 alkyl, a C3-C10 carbocyclic group and -(C1-C3 alkylene)-C3-C10 carbocyclic; 12 R is selected from H, C 1-C6 alkyl, a C 3-C10 carbocyclic group, -(C 1-C3 alkylene)-C3-C10 carbocyclic, -C(O)C 1-C6 alkyl, -C(O)C3-C10 carbocyclic, -C(O)(C1-C3 alkylene)-C3-C10 carbocyclic, -C(O)(CH2)x C6-C10 aryl, -C(O)(CH2)x 5- to 6- membered heteroaryl, -C(O)(CH2)x 5- to 6-membered heterocyclyl, -C(O)Oalkyl, C(O)OC6-C10 aryl, (CH2)x C6-C10 aryl, (CH2)x 5- to 6-membered heteroaryl and - (CH 2)x 5- to 6-membered heterocyclyl, wherein each of the ring systems 10 is optionally substituted by one or more substituents selected from the List Z; 13 R is selected from H, C1-C6 alkyl, a C3-C10 carbocyclic group and -(C1-C3 alkylene)-C3-C10 carbocyclic; 14 R is selected from H, C 1-C6 alkyl, a C3-C10 carbocyclic group, -(C 1-C3 alkylene)-C3-C10 carbocyclic, - (CH2)z C6-C10 aryl, (CH2)z 5- to 6-membered heteroaryl and (CH 2)z 5- to 6-membered heterocyclyl, wherein each of the ring systems is optionally substituted by one or more substituents selected from the List Z; 15 x is 0, 1, 2 or 3; z is 0, 1, 2 or 3; each Z is independently selected from OH, C6-C10 aryl, 5- to 6-membered heteroaryl, 5- to 6-membered heterocyclyl,

benzyl, C1-C6 alkyl optionally substituted by one or more halogen atoms, CN or OH groups, 1C-C6 alkoxy optionally 15 17 substituted by one or more halogen atoms, CN or OH groups, -OC6-C10 aryl, -Obenzyl, -O(CH 2)aC(O)E, NR (SO2)R , 20 15 16 18 15 17 15 17 15 16 17 15 17 15 17 15 (SO2)NR R , (SO2)R , NR C(O)R , C(O)NR R , NR C(O)NR R , NR C(O)OR , NR R , C(O)OR , 15 15 17 15 OC(O)R , OC(O)NR , C(O)R , SR , CN, NO2, and halogen; and when there are two or more Z substitutents, two Z substituents together with the atoms to which they are attached optionally form a 5- to 7-membered carbocyclic or a 4- to 7-membered heterocyclic substituent fused to the ring system;

a is 0, 1, 2, 3 or 4, where the alkylene group is optionally substituted by OH or NH 2 when a is 1, 2, 3 or 4; 25 E is NR15R17 or OR17; 15 16 each R and R is independently selected from H, C 1-C6 alkyl, a C 3-C10 carbocyclic group and -(C 1-C3 alkylene)-C3-C10 carbocyclic; 17 each R is selected from H, C1-C6 alkyl, a C3-C10 carbocyclic group, -(C1-C3 alkylene)-C3-C10 carbocyclic, C6-C10 aryl, 5- to 6-membered heteroaryl and 5- to 6-membered heterocyclyl, where each of the ring systems is optionally 30 substituted by OH, halo, C1-C3 alkyl and C1-C3 alkoxy; and 18 R is selected from H, C1-C6 alkyl, a C3-C10 carbocyclic group, -(C1-C3 alkylene)-C3-C10 carbocyclic, C6-C10 aryl, 5- to 6-membered heteroaryl, 5- to 6-membered heterocyclyl and NHC(=NH)NH2, where each of the ring systems is op- tionally substituted by OH, halo, C1-C3 alkyl and C1-C3 alkoxy. [0004] In a still further embodiment of the invention as defined anywhere above, R9 is H. Suitably, R9 is H and R6 is 35 9 6 selected from H and C1-C6 alkyl. Optionally, R and R are both H. [0005] In a yet further embodiment of the invention as defined anywhere above, there is provided a compound according to Formula I selected from:

Structure 40

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55 and pharmaceutically acceptable salts, solvates and hydrates thereof.

18 EP 2 300 010 B1

Definitions

[0006] Terms used in the specification have the following meanings:

5 "Optionally substituted" means the group referred to can be substituted at one or more positions by any one or any combination of the radicals listed thereafter. "optionally substitutedby one or moreZ groups" denotesthat the relevantgroup may include one or more substituents, each independently selected from the groups included within the definition of Z. Thus, where there are two or more Z group substituents, these may be the same or different. 10 "Halo" or "halogen", as used herein, may be fluorine, chlorine, bromine or iodine.

"C1-C6 alkyl", as used herein, denotes straight chain or branched alkyl group having 1-6 carbon atoms. If a different number of carbon atoms is specified, such as C 4 or C3, then the definition is to be amended accordingly. "C2-C6 alkenyl", as used herein, denotes straight chain or branched alkenyl group having 2-6 carbon atoms and one or more carbon-carbon double bonds. If a different number of carbon atoms is specified, such as C 4 or C3, then 15 the definition is to be amended accordingly. "C2-C6 alkynyl", as used herein, denotes straight chain or branched alkynyl group having 2-6 carbon atoms and one or more carbon-carbon triple bonds. If a different number of carbon atoms is specified, such as C4 or C3, then the definition is to be amended accordingly.

"C1-C6-alkoxy", as used herein, denotes straight chain or branched alkoxy having 1-6 carbon atoms. If a different 20 number of carbon atoms is specified, such as C 4 or C3, then the definition is to be amended accordingly.

[0007] The term "alkylene" denotes a straight chain or branched saturated hydrocarbon linking group containing the relevant number of carbon atoms.

[0008] The terms "-(C1-C6 alkylene)-" or "-(C1-C4 alkylene)-" denote a hydrocarbon linking group having the relevant 25 number of carbon atoms. If a different number of carbon atoms is specified, such as C4 or C3, then the definition is to be amended accordingly. [0009] "a C3-C10 carbocyclic group", "C3-C10 carbocyclic" or "C3-C10 carbocyclyl", as used herein, denotes a non- aromatic carbocyclic group having 3- to 10-ring carbon atoms that is saturated or partially unsaturated. This term includes

both C3-C10 cycloalkyl and C5-C10 cycloalkenyl groups, as well as bicyclic and spirocyclic groups. Where the group is 30 bicyclic, it may comprise an aromatic or non-aromatic monocyclic ring fused to a non-aromatic monocyclic ring. Optionally,

the ring system contains 3-6 carbon atoms, i.e. a 3C-C6 carbocyclic group. Examples of C3-C10-carbocyclic groups include but are not limited to cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl or cyclooctyl or a bicyclic group, such as bicyclooctyl, bicyclononyl including indanyl and bicyclodecyl. If a different number of carbon atoms is specified, such as C6, then the definition is to be amended accordingly. 35 [0010] "C3-C10 cycloalkyl", as used herein denotes a saturated carbocyclic ring system having 3- to 10-ring carbon atoms. Optionally, the ring system contains 5 or 6 carbon atoms, i.e. C 5-C6 cycloalkyl. [0011] "C5-C10 cycloalkenyl", as used herein, denotes a non-aromatic carbocyclic group having 5- to 10-ring carbon atoms that contains one or more carbon-carbon double bonds. Optionally, the ring system contains 5 or 6 carbon atoms, i.e. C5-C6 cycloalkenyl. 40 [0012] The terms "aryl" and "C6-C15-Aromatic carbocyclic group", as used herein, denote an aromatic group having 6- to 15-ring carbon atoms. Examples of C6-C15-aromatic carbocyclic groups include, but are not limited to, phenyl, phenylene, benzenetriyl, naphthyl, naphthylene, naphthalenetriyl or anthrylene. If a different number of carbon atoms

is specified, such as C10 (e.g. a C6-C10 aryl group), then the definition is to be amended accordingly. In certain embod- iments, aryl is phenyl or naphthylenyl. In further embodiments, aryl is phenyl. 45 [0013] The terms "heterocyclic group" and "4- to 10-Membered heterocyclic group" refer to 4- to 10-membered hete- rocyclic rings containing at least one ring heteroatom selected from the group consisting of nitrogen, oxygen and sulphur, which may be saturated or partially saturated. The group includes benzofused heterocyclic ring systems. Examples of such heterocyclic groups include, but are not limited to, pyrrolidine, piperidine, piperazine, pyrrolidinone, morpholine, tetrahyrofuran, tetrahydrothiophene, tetrahydrothiopyran, tetrahydropyran, 1,4-dioxane and 1,4-oxathiane. Suitably, the 50 heterocyclic group may be a 5- to 6-membered group. [0014] The terms "heteroaryl group" and "heteroaromatic group" denote a 5- to 10-membered aromatic heterocyclic ring containing at least one ring heteroatom selected from the group consisting of nitrogen, oxygen and sulphur. Examples of such heteroaryl groups include, but are not limited to, pyrrole, pyrazole, imidazole, triazole, tetrazole, pyridine, pyri- dazine, pyrimidine, pyrazine, triazine, quinoline, isoquinoline, indole, isoindole, indolizine, indazole, benzimidazole, pu- 55 rine, quinolizine, cinnoline, quinazoline, quinoxaline, naphthyridine, phthalazine, pteridine, acridine, phenazine and phen- anthroline. Suitably, the heteroaryl group may be a 5- to 6-membered group. [0015] It is to be understood that any and all embodiments of the present invention may be taken in conjunction with any other embodiment to describe additional embodiments of the present invention. Furthermore, any elements of an

19 EP 2 300 010 B1

embodiment are intended to be combinable with any and all other elements from any of the embodiments to describe additional embodiments. It is understood by those skilled in the art that combinations of substituents where not possible are not an aspect of the present invention. [0016] A second aspect of the present invention provides for the use of a compound of formula (I) in any of the 5 aforementioned embodiments for the manufacture of a medicament for the treatment of an inflammatory or allergic condition, particularly an inflammatory or obstructive airways disease, or mucosal hydration. [0017] An embodiment of the present invention provides for the use of a compound of formula (I) in any of the afore- mentioned embodiments for the manufacture of a medicament for the treatment of an inflammatory or allergic condition selected from cystic fibrosis, primary ciliary dyskinesia, chronic bronchitis, chronic obstructive pulmonary disease, asth- 10 ma, respiratory tract infections, lung carcinoma, xerostomia and keratoconjunctivitis sire. [0018] Throughout this specification and in the claims that follow, unless the context requires otherwise, the word "comprise", or variations, such as "comprises" or "comprising", will be understood to imply the inclusion of a stated integer or step or group of integers or steps but not the exclusion of any other integer or step or group of integers or steps. [0019] Especially suitable compounds of formula (I) are those described hereinafter in the Examples. 15 [0020] As mentioned above, the skilled person will appreciate that the compounds of Formula (I) will typically include a counter ion. Thus, the compounds of the invention may be considered as follows:

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wherein R1, R2, R3, R4, R5, R6, R7, R8, R9 and R10 are as defined above and [Counter ion]- is a suitable counter ion, optionally selected from fluoride, chloride, bromide, iodide, nitrate, sulfate, phosphate, hexafluorophosphate, formate, 30 acetate, trifluoroacetate, propionate, butyrate, lactate, citrate, tartrate, malate, maleate, succinate, benzoate, p-chlo- robenzoate, diphenylacetate, triphenylacetate, o-hydroxybenzoate, p-hydroxybenzoate, 1-hydroxynaphthalene-2-car- boxylate, 3-hydroxynaphthalene-2-carboxylate, methanesulfonate and benzenesulfonate. [0021] The skilled person will also appreciate that the counter ion may exist in different stoichiometries. Thus, the two ionic components of the compound may not exist in a 1:1 ratio. For example, certain counter ions may exist in a "2-" 35 form such that the ratio of the cationic component to counter ion may be 2:1. Other stoichiometries are, of course, possible and all such combinations are considered to be within the scope of the invention. [0022] Pharmaceutically acceptable solvates in accordance with the invention include those wherein the solvent of

crystallisation may be isotopically substituted e.g. D 2O, d6-acetone or d6-DMSO. [0023] Some compounds of the invention contain at least one asymmetric carbon atom and thus they exist in individual 40 optically active isomeric forms or as mixtures thereof, e.g. as racemic mixtures. In cases where additional asymmetric centres exist the present invention also embraces both individual optically active isomers as well as mixtures, e.g. diastereomeric mixtures, thereof. [0024] The invention includes all such forms, in particular the pure isomeric forms. The different isomeric forms may be separated or resolved one from the other by conventional methods, or any given isomer may be obtained by con- 45 ventional synthetic methods or; by stereospecific or asymmetric syntheses. Since the compounds of the invention are intended for use in pharmaceutical compositions it will readily be understood that they are each preferably provided in substantially pure form, for example at least 60% pure, more suitably at least 75% pure and preferably at least 85%, especially at least 98% pure (% are on a weight for weight basis). Impure preparations of the compounds may be used for preparing the more pure forms used in the pharmaceutical compositions; these less pure preparations of the com- 50 poundsshould contain atleast 1 %, moresuitably atleast 5% and preferablyfrom 10 to 59% of a compoundof the invention. [0025] The invention includes all pharmaceutically acceptable isotopically-labelled compounds of formula I wherein one or more atoms are replaced by atoms having the same atomic number, but an atomic mass or mass number different from the atomic mass or mass number usually found in nature. Examples of isotopes suitable for inclusion in the com- pounds of the invention include isotopes of hydrogen e.g. 2H and 3H, carbon e.g. 11C, 13C and 14C, chlorine e.g. 36Cl, 55 fluorine e.g. 18F, iodine e.g. 123I and 125I, nitrogen e.g. 13N and 15N, oxygen e.g. 15O, 17O and 18O, and sulfur e.g. 35S. [0026] Certain isotopically-labelled compounds of formula I, for example those incorporating a radioactive isotope, are useful in drug and/or substrate tissue distribution studies. The radioactive isotopes tritium ( 3H) and carbon-14 (14C) are particularly useful for this purpose in view of their ease of incorporation and ready means of detection. Substitution

20 EP 2 300 010 B1

with heavier isotopes such as deuterium ( 2H) may afford certain therapeutic advantages that result from greater metabolic stability, for example increased in vivo half-life or reduced dosage requirements, and hence may be preferred in some circumstances. Substitution with positron emitting isotopes, such as 11C, 18F, 15O, and 13N can be useful in Positron Emission Topography (PET) studies for examining substrate receptor occupancy. 5 [0027] Isotopically-labelled compounds of formula I can generally be prepared by conventional techniques known to those skilled in the art or by processes analogous to those described in the accompanying examples using an appropriate isotopically-labelled reagent in place of the non-labelled reagent previously used. [0028] Some of the compounds of Formula I may exist in different tautomeric forms. Tautomerism is well known to those skilled in the art and the skilled person will readily appreciate which groups are able to tautomerise to form the 10 different tautomeric forms. The invention includes all tautomeric forms of the compounds of Formula I.

Synthesis

[0029] Generally, compounds according to Formula I can be synthesized by the routes described in Scheme 1 and 15 the Examples. [0030] For instance, intermediate 1 can be reacted with intermediate 2 in the presence of a suitable coupling agent and organic base in organic solvent to provide a compound of Formula 3 as the free base. Reaction of this intermediate with a suitable alkylating agent, R10-X, wherein X is a suitable leaving group, provides a compound of Formula I. The leaving group X is well known in the art and may be any suitable group that facilitates alkylation. Examples of the leaving 20 group X include chloride, bromide, iodide, mesylate and tosylate groups. [0031] Intermediates can be prepared from methods known by those skilled in the art or are commercially available.

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45 [0032] The compounds of Formula (I) can be obtained in the form of hydrates or solvates containing a solvent used for crystallisation. Compounds of formula (I) can be recovered from reaction mixtures and purified in a conventional manner. Isomers, such as stereoisomers, may be obtained in a conventional manner, e.g., by fractional crystallisation or asymmetric synthesis from correspondingly asymmetrically substituted, e.g., optically active, starting materials. 50 [0033] The compounds of formula (I) can be prepared, e.g., using the reactions and techniques described below and in the Examples. The reactions may be performed in a solvent appropriate to the reagents and materials employed and suitable for the transformations being effected. It will be understood by those sk illed in the art of organic synthesis that the functionality present on the molecule should be consistent with the transformations proposed. This will sometimes require a judgment to modify the order of the synthetic steps or to select one particular process scheme over another 55 in order to obtain a desired compound of the invention. [0034] The various substituents on the synthetic intermediates and final products shown in the following reaction schemes can be present in their fully elaborated forms, with suitable protecting groups where required as understood

21 EP 2 300 010 B1

by one skilled in the art, or in precursor forms which can later be elaborated into their final forms by methods familiar to one skilled in the art. The substituents can also be added at various stages throughout the synthetic sequence or after completion of the synthetic sequence. In many cases, commonly used functional group manipulations can be used to transform one intermediate into another intermediate, or one compound of formula (I) into another compound of formula 5 (I). Examples of such manipulations are conversion of an ester or a ketone to an alcohol; conversion of an ester to a ketone; interconversions of esters, acids and amides; alkylation, acylation and sulfonylation of alcohols and amines; and many others. Substituents can also be added using common reactions, such as alkylation, acylation, halogenation or oxidation. Such manipulations are well-known in the art, and many reference works summarize procedures and methods for such manipulations. Some reference works which gives examples and references to the primary literature 10 of organic synthesis for many functional group manipulations, as well as other transformations commonly used in the art of organic synthesis are March’s Organic Chemistry, 5th Edition, Wiley and Chichester, Eds. (2001); Comprehensive Organic Transformations, Larock, Ed., VCH (1989); Comprehensive Organic Functional Group Transformations, Kat- ritzky et al. (series editors), Pergamon (1995); and Comprehensive Organic Synthesis, Trost and Fleming (series editors), Pergamon (1991). It will also be recognized that another major consideration in the planning of any synthetic route in 15 this field is the judicious choice of the protecting group used for protection of the reactive functional groups present in the compounds described in this invention. Multiple protecting groups within the same molecule can be chosen such that each of these protecting groups can either be removed without removal of other protecting groups in the same molecule, or several protecting groups can be removed using the same reaction step, depending upon the outcome desired. An authoritative account describing many alternatives to the trained practitioner is Greene and Wuts, Protective 20 Groups in Organic Synthesis, Wiley and Sons (1999).

Pharmacological activity

[0035] Having regard to their blockade of the epithelial sodium channel (ENaC), compounds of formula (I), hereinafter 25 alternately referred to as "agents of the invention", are useful in the treatment of conditions which respond to the blockade of the epithelial sodium channel, particularly conditions benefiting from mucosal hydration. [0036] Diseases treatable by blockade of the epithelial sodium channel, include diseases associated with the regulation of fluid volumes across epithelial membranes. For example, the volume of airway surface liquid is a key regulator of mucociliary clearance and the maintenance of lung health. The blockade of the epithelial sodium channel will promote 30 fluid accumulation on the mucosal side of the airway epithelium thereby promoting mucus clearance and preventing the accumulation of mucus and sputum in respiratory tissues (including lung airways). Such diseases include respiratory diseases, such as cystic fibrosis, primary ciliary dyskinesia, chronic bronchitis, chronic obstructive pulmonary disease (COPD), asthma, respiratory tract infections (acute and chronic; viral and bacterial) and lung carcinoma. Diseases treatable by blockade of the epithelial sodium channel also include diseases other than respiratory diseases that are 35 associated with abnormal fluid regulation across an epithelium, perhaps involving abnormal physiology of the protective surface liquids on their surface, e.g., xerostomia (dry mouth) or keratoconjunctivitis sire (dry eye). Furthermore, blockade of the epithelial sodium channel in the kidney could be used to promote diuresis and thereby induce a hypotensive effect. [0037] Treatment in accordance with the invention may be symptomatic or prophylactic. [0038] Asthma includes both intrinsic (non-allergic) asthma and extrinsic (allergic) asthma, mild asthma, moderate 40 asthma, severe asthma, bronchitic asthma, exercise-induced asthma, occupational asthma and asthma induced following bacterial infection. Treatment of asthma is also to be understood as embracing treatment of subjects, e.g., of less than 4 or 5 years of age, exhibiting wheezing symptoms and diagnosed or diagnosable as "wheezy infants", an established patient category of major medical concern and now often identified as incipient or early-phase asthmatics. (For conven- ience this particular asthmatic condition is referred to as "wheezy-infant syndrome".) 45 [0039] Prophylactic efficacy in the treatment of asthma will be evidenced by reduced frequency or severity of symp- tomatic attack, e.g., of acute asthmatic or bronchoconstrictor attack, improvement in lung function or improved airways hyperreactivity. It may further be evidenced by reduced requirement for other, symptomatic therapy, i.e., therapy for or intended to restrict or abort symptomatic attack when it occurs, e.g., anti-inflammatory (e.g., cortico-steroid) or bron- chodilatory.Prophylactic benefit inasthma may, inparticular, be apparentin subjects proneto "morning dipping". "Morning 50 dipping" is a recognized asthmatic syndrome, common to a substantial percentage of asthmatics and characterized by asthma attack, e.g., between the hours of about 4-6 am, i.e., at a time normally substantially distant from any previously administered symptomatic asthma therapy. Chronic obstructive pulmonary disease includes chronic bronchitis or dys- pnea associated therewith, emphysema, as well as exacerbation of airways hyperreactivity consequent to other drug therapy, in particular, other inhaled drug therapy. The invention is also applicable to the treatment of bronchitis of whatever 55 type or genesis including, e.g., acute, arachidic, catarrhal, croupus, chronic or phthinoid bronchitis. [0040] The suitability of epithelial sodium channel blocker as a treatment of a disease benefiting from mucosal hydration, may be tested by determining the inhibitory effect of the epithelial sodium channel blocker on: the ion channel/ion transport function in suitable isolated cells or confluent epithelia using the methods described in Hirsh et al., J Pharm Exp Ther

22 EP 2 300 010 B1

(2004). [0041] Epithelial sodium channel blockers, including the compounds of formula (I), are also useful as co-therapeutic agents for use in combination with other drug substances, such as anti-inflammatory, bronchodilatory, antihistamine or anti-tussive drug substances, particularly in the treatment of cystic fibrosis or obstructive or inflammatory airways diseases 5 such as those mentioned hereinbefore, e.g., as potentiators of therapeutic activity of such drugs or as a means of reducing required dosaging or potential side effects of such drugs. [0042] The epithelial sodium channel blocker may be mixed with the other drug substance in a fixed pharmaceutical composition or it may be administered separately, before, simultaneously with or after the other drug substance. [0043] Accordingly, the invention includes as a further aspect a combination of epithelial sodium channel blocker with 10 osmotic agents (hypertonic saline, dextran, mannitol, Xylitol) + modifiers of CFTR function, both wild-type and mutant (correctors + potentiators), e.g., those described in WO 2007/021982, WO 2006/099256, WO 2006/127588, WO 2004/080972, WO 2005/026137, WO 2005/035514, WO 2005/075435, WO 2004/111014, WO 2006/101740, WO 2004/110352, WO 2005/120497 and US 2005/0176761, an anti-inflammatory, bronchodilatory, antihistamine, anti-tus- sive, antibiotic or DNase drug substance, said epithelial sodium channel blocker and said drug substance being in the 15 same or different pharmaceutical composition. [0044] Suitable antibiotics include macrolide antibiotics, e.g., tobramycin (TOBI™). [0045] Suitable DNase drug substances include dornase alfa (Pulmozyme™), a highly-purified solution of recombinant human deoxyribonuclease I (rhDNase), which selectively cleaves DNA. Dornase alfa is used to treat cystic fibrosis. [0046] Other useful combinations of epithelial sodium channel blockers with anti-inflammatory drugs are those with 20 antagonists of chemokine receptors, e.g., CCR-1, CCR-2, CCR-3, CCR-4, CCR-5, CCR-6, CCR-7, CCR-8, CCR-9 and CCR10, CXCR1, CXCR2, CXCR3, CXCR4, CXCR5, particularly CCR-5 antagonists, such as Schering-Plough antag- onists SC-351125, SCH-55700 and SCH-D; Takeda antagonists, such as N-[[4-[[[6,7-dihydro-2-(4-methyl-phenyl)-5H- benzo-cyclohepten-8-yl]carbonyl]amino]phenyl]-methyl]tetrahydro-N,N-dimethyl-2H-pyran-4-amin-ium chloride (TAK- 770); and CCR-5 antagonists described in USP 6,166,037 (particularly claims 18 and 19), WO 00/66558 (particularly 25 claim 8), WO 00/66559 (particularly claim 9), WO 04/018425 and WO 04/026873. [0047] Suitable anti-inflammatory drugs include steroids, in particular, glucocorticosteroids, such as budesonide, bec- lamethasone dipropionate, fluticasone propionate, ciclesonide or mometasone furoate, or steroids described in WO 02/88167, WO 02/12266, WO 02/100879, WO 02/00679 (especially those of Examples 3, 11, 14, 17, 19, 26, 34, 37, 39, 51, 60, 67, 72, 73, 90, 99 and 101), WO 03/35668, WO 03/48181, WO 03/62259, WO 03/64445, WO 03/72592, WO 30 04/39827 and WO 04/66920; non-steroidal glucocorticoid receptor agonists, such as those described in DE 10261874, WO 00/00531, WO 02/10143, WO 03/82280, WO 03/82787, WO 03/86294, WO 03/104195, WO 03/101932, WO 04/05229, WO 04/18429, WO 04/19935 and WO 04/26248; LTD4 antagonists, such as montelukast and zafirlukast; PDE4 inhibitors, such as cilomilast (Ariflo ® GlaxoSmithKline), Roflumilast (Byk Gulden),V-11294A (Napp), BAY19-8004 (Bayer), SCH-351591 (Schering-Plough), Arofylline (Almirall Prodesfarma), PD189659/PD168787 (Parke-Davis), AWD- 35 12-281 (Asta Medica), CDC-801 (Celgene), SelCID(TM) CC-10004 (Celgene), VM554/UM565 (Vernalis), T-440 (Tan- abe), KW-4490(Kyowa Hakko Kogyo), andthose disclosed inWO 92/19594, WO93/19749, WO 93/19750, WO93/19751, WO 98/18796, WO 99/16766, WO 01/13953, WO 03/104204, WO 03/104205, WO 03/39544, WO 04/000814, WO 04/000839, WO 04/005258, WO 04/018450, WO 04/018451, WO 04/018457, WO 04/018465, WO 04/018431, WO 04/018449, WO 04/018450, WO 04/018451, WO 04/018457, WO 04/018465, WO 04/019944, WO 04/019945, WO 40 04/045607 and WO 04/037805; adenosine A2B receptor antagonists such as those described in WO 02/42298; and beta-2 adrenoceptor agonists, such as albuterol (salbutamol), metaproterenol, terbutaline, salmeterol fenoterol, pro- caterol, and especially, formoterol, carmoterol and pharmaceutically acceptable salts thereof, and compounds (in free or salt or solvate form) of formula (I) of WO 0075114, which document is incorporated herein by reference, preferably compounds of the Examples thereof, especially a compound of formula: 45

50

55

corresponding to indacaterol and pharmaceutically acceptable salts thereof, as well as compounds (in free or salt or solvate form) of formula (I) of WO 04/16601, and also compounds of EP 1440966, JP 05025045, WO 93/18007, WO

23 EP 2 300 010 B1

99/64035, USP 2002/0055651, WO 01/42193, WO 01/83462, WO 02/66422, WO 02/70490, WO 02/76933, WO 03/24439, WO 03/42160, WO 03/42164, WO 03/72539, WO 03/91204, WO 03/99764, WO 04/16578, WO 04/22547, WO 04/32921, WO 04/33412, WO 04/37768, WO 04/37773, WO 04/37807, WO 04/39762, WO 04/39766, WO 04/45618, WO 04/46083, WO 04/80964, WO 04/108765 and WO 04/108676. 5 [0048] Suitable bronchodilatory drugs include anticholinergic or antimuscarinic agents, in particular, ipratropium bro- mide, oxitropium bromide, tiotropium salts and CHF 4226 (Chiesi), and glycopyrrolate, but also those described in EP 424021, USP 3,714,357, USP 5,171,744, WO 01/04118, WO 02/00652, WO 02/51841, WO 02/53564, WO 03/00840, WO 03/33495, WO 03/53966, WO 03/87094, WO 04/018422 and WO 04/05285. [0049] Suitable dual anti-inflammatory and bronchodilatory drugs include dual beta-2 adrenoceptor agonist/muscarinic 10 antagonists such as those disclosed in USP 2004/0167167, WO 04/74246 and WO 04/74812. [0050] Suitable antihistamine drug substances include cetirizine hydrochloride, acetaminophen, clemastine fumarate, promethazine, loratidine, desloratidine, diphenhydramine and fexofenadine hydrochloride, activastine, astemizole, aze- lastine, ebastine, epinastine, mizolastine and tefenadine, as well as those disclosed in JP 2004107299, WO 03/099807 and WO 04/026841. 15 In accordance with the foregoing, the invention also provides as a further aspect compounds of formula (Ia) for use in treatment of a condition responsive to blockade of the epithelial sodium channel, e.g., diseases associated with the regulation of fluid volumes across epithelial membranes, particularly an obstructive airways disease, which comprises administering to a subject, particularly a human subject, in need thereof a compound of formula (I). In another aspect the invention provides a compound of formula (I) for use in the manufacture of a medicament for the 20 treatment of a condition responsive to blockade of the epithelial sodium channel, particularly an obstructive airways disease, e.g., cystic fibrosis and COPD. The agents of the invention may be administered by any appropriate route, e.g. orally, e.g., in the form of a tablet or capsule; parenterally, e.g., intravenously; by inhalation, e.g., in the treatment of an obstructive airways disease; intra- nasally, e.g., in the treatment of allergic rhinitis; topically to the skin; or rectally. In a further aspect, the invention also 25 provides a pharmaceutical composition comprising a compound of formula (I) optionally together with a pharmaceutically acceptable diluent or carrier therefor. The composition may contain a co-therapeutic agent, such as an anti-inflammatory, broncho-dilatory, antihistamine or anti-tussive drug as hereinbefore described. Such compositions may be prepared using conventional diluents or excipients and techniques known in the galenic art. Thus oral dosage forms may include tablets and capsules. Formulations for topical administration may take the form of creams, ointments, gels or transdermal 30 delivery systems, e.g., patches. Compositions for inhalation may comprise aerosol or other atomizable formulations or dry powder formulations. When the composition comprises an aerosol formulation, it preferably contains, e.g., a hydro- fluoro-alkane (HFA) propellant, such as HFA134a or HFA227 or a mixture of these, and may contain one or more co- solvents known in the art, such as ethanol (up to 20% by weight), and/or one or more surfactants, such as oleic acid or sorbitan trioleate, and/or one or more bulking agents, such as lactose. 35 [0051] When the composition comprises a dry powder formulation, it preferably contains, e.g., the compound of formula (I) having a particle diameter up to 10 microns, optionally together with a diluent or carrier, such as lactose, of the desired particle size distribution and a compound that helps to protect against product performance deterioration due to moisture, e.g., magnesium stearate. When the composition comprises a nebulised formulation, it preferably contains, e.g., the compound of formula (I) either dissolved, or suspended, in a vehicle containing water, a co-solvent, such as ethanol or 40 propylene glycol and a stabilizer, which may be a surfactant. [0052] Further aspects of the invention include:

(a) a compound of formula (I) in inhalable form, e.g., in an aerosol or other atomisable composition or in inhalable particulate, e.g., micronised form; 45 (b) an inhalable medicament comprising a compound of formula (I) in inhalable form;

(c) a pharmaceutical product comprising a compound of formula (I) in inhalable form in association with an inhalation device; and 50 (d) an inhalation device containing a compound of formula I in inhalable form.

[0053] Dosages of compounds of formula (I) employed in practising the present invention will of course vary depending, e.g., on the particular condition to be treated, the effect desired and the mode of administration. In general, suitable daily 55 dosages for administration by inhalation are of the order of 0.005-10 mg, while for oral administration suitable daily doses are of the order of 0.05-100 mg.

24 EP 2 300 010 B1

Pharmaceutical Use and Assay

[0054] Compounds of formula (I), hereinafter referred to alternatively as "agents of the invention", are useful as phar- maceuticals. In particular, the compounds have good ENaC blocker activity and may be tested in the following assays. 5 Cell culture

[0055] Human Bronchial Epithelial cells (HBECs) (Cambrex) were cultured under air-liquid interface conditions to provide a well differentiated mucociliary phenotype. 10 [0056] HBECs were cultured using a modification of the method described by Gray and colleagues (Gray et al., 1996). Cells were seeded in plastic T-162 flasks and were grown in bronchial epithelial cell growth medium (BEGM; Cambrex) supplemented with bovine pituitary extract (52m g/mL), hydrocortisone (0.5 mg/mL), human recombinant epidermal growth factor (0.5 ng/mL), epinephrine (0.5 mg/mL), transferrin (10 mg/mL), insulin (5 mg/mL), retinoic acid (0.1 mg/mL), triiodothyronine (6.5 mg/mL), gentamycin (50 mg/mL) and amphotericin B (50 ng/mL). Medium was changed every 48 15 hours until cells were 90% confluent. Cells were then passaged and seeded (8.25 x 10 5 cells/insert) on polycarbonate Snapwell inserts (Costar) in differentiation media containing 50% DMEM in BEGM with the same supplements as above but without triiodothyronine and a final retinoic acid concentration of 50 nM (all-trans retinoic acid). Cells were maintained submerged for the first 7 days in culture, after which time they were exposed to an apical air interface for the remainder of the culture period. At this time, media was changed to DMEM:F12 media containing 2% v/v Ultroser G for the remainder 20 of culture. Amphotericin B was removed from all media 3 feeds prior to use in the Ussing Chambers. Cells were used between days 7 and 21 after establishment of the apical-air interface. At all stages of culture, cells were maintained at 37°C in 5% CO2 in an air incubator.

Short circuit current (ISC) measurements 25 [0057] Snapwell inserts were mounted in Vertical Diffusion Chambers (Costar) and were bathed with continuously gassed Ringer solution (5% CO2 in O2; pH 7.4) maintained at 37°C containing (in mM): 120 NaCl, 25 NaHCO3, 3.3 KH2PO4, 0.8 K2HPO4, 1.2 CaCl2, 1.2 MgCl2, and 10 glucose. The solution osmolarity was between 280 and 300 mOs- mol/kg H2O for all physiological salt solutions used. Cells were voltage clamped to 0 mV (model EVC4000; WPI). RT 30 was measured by applying a 1- or 2-mV pulse at 30-s intervals and calculating RT by Ohm’s law. Data were recorded using a PowerLab workstation (ADInstruments). [0058] Test compounds were prepared as a 10 mM stock solution in DMSO (95%). Serial 3-fold dilutions were freshly prepared in an appropriate vehicle (distilled H 2O or Ringers solution). The initial concentration was added to the apical chamber as a 1000x concentrate in 5 mL, resulting in a final 1x concentration the 5 mL volume of the Ussing chamber. 35 Subsequent additions of compound were added in a 3.3 mL volume of the 1000x serially diluted stock solution. At the completion of the concentration-response experiment, amiloride (10 mM) was added into the apical chamber to enable

the total amiloride-sensitive current to be measured. An amiloride control 50IC was established at the start of each experiment. [0059] Results are expressed as the mean % inhibition of the amiloride-sensitive ISC. Concentration-response curves 40 were plotted and IC50 values generated using GraphPad Prism 3.02. Cell inserts were typically run in duplicate and the IC50 calculated on the mean % inhibition data. [0060] Compounds of the Examples, herein below, generally have IC50 values in the data measurements described above below 10 mM. For example, the compounds detailed below have the IC50 values shown.

45 Example IC50 (mM) Example IC50 (mM) 10.25380.81 50.02500.24 93.46510.42 50 11 5.32 53 0.60 13 0.17 63 0.043 14 0.046 65 0.15 55 15 0.052 78 0.062 16 0.64 94 0.25

25 EP 2 300 010 B1

(continued)

Example IC50 (mM) Example IC50 (mM) 17 0.0075 100 0.29 5 18 0.037 108 0.54 19 0.020 120 0.004 20 0.002 121 0.01 10 21 5.69 122 0.13 22 0.051 124 0.025 23 0.034 125 0.24 26 0.012 126 2.27 15 27 0.22 129 1.14 29 0.025 130 5.81 30 0.14 132 1.49 20 33 0.15 133 6.52 35 0.41

[0061] The invention is illustrated by the following Examples. 25 EXAMPLES

[0062] Suitable compounds of the present invention include compounds of formula Ia which as shown in Table 1 below. The method of preparation being described hereinafter. 30 TABLE 1 Ex. Structure M+

35 1 315

40

2 301

45

50 3 301

55

26 EP 2 300 010 B1

(continued)

Ex. Structure M+

5

4 329

10

5 315

15

20 6 315

25

7 313

30

35 8 313

40

9 327

45

50 10 299

55

27 EP 2 300 010 B1

(continued)

Ex. Structure M+

5

11 299

10

15 12 287

20

13 363

25

30 14 582

35 15 568

40 16 403

45

17 567 50

55

28 EP 2 300 010 B1

(continued)

Ex. Structure M+

5

10 18 675

15

20

19 626

25

30

20 541

35

40

21 0.011

45

50

22 584

55

29 EP 2 300 010 B1

(continued)

Ex. Structure M+

5

23 617 10

15

24 599 20

25

30 25 555

35

26 661

40

45 27 407

50

55

30 EP 2 300 010 B1

(continued)

Ex. Structure M+

5

28 493 10

15

29 601

20

30 434

25

31 459

30

32 526

35

33 468

40

34 540

45

35 504

50

36 478

55

31 EP 2 300 010 B1

(continued)

Ex. Structure M+

5 37 514

10 38 452

15 39 454

20 40 464

25 41 478

30 42 420

35 43 434

40 44 440

45 45 406

50 46 420

55 47 494

32 EP 2 300 010 B1

(continued)

Ex. Structure M+

5 48 512

10

49 661

15

50 540

20

51 468

25

52 454

30

53 434

35

54 448

40

55 448

45

56 434

50

57 420

55

33 EP 2 300 010 B1

(continued)

Ex. Structure M+

5 58 452

10 59 473

15 60 464

61 482 20

62 462 25

63 554 30

64 568 35

65 496

40

66 476

45

67 482

50

68 463

55

34 EP 2 300 010 B1

(continued)

Ex. Structure M+

5 69 462

10

70 464

15

20 71 487

25 72 492

30 73 468

35 74 448

40

75 448

45

76 478

50

77 434

55

35 EP 2 300 010 B1

(continued)

Ex. Structure M+

5 78 568

10 79 582

15 80 510

81 482 20

82 520 25

83 490

30

84 506

35

85 496

40 86 476

45 87 462

50 88 476

89 462 55

36 EP 2 300 010 B1

(continued)

Ex. Structure M+

5 90 522

10 91 448

15 92 522

20 93 536

25 94 578

95 533 30

96 512

35

97 498

40

98 498

45

99 484

50

100 504

55

37 EP 2 300 010 B1

(continued)

Ex. Structure M+

5 101 484

10 102 518

15 103 484

20 104 498

25 105 470

30

106 530

35

40

107 539 45

50

108 490

55

38 EP 2 300 010 B1

(continued)

Ex. Structure M+

5 109 470

10 110 456

15 111 476

20

112 456

25

113 442

30

114 484

35 115 504

40 116 490

45 117 470

50 118 470

55

39 EP 2 300 010 B1

(continued)

Ex. Structure M+

5 119 456

10

120 449

15

121 435

20

122 513 25

30

123 393

35

124 509

40

125 661 45

50

126 467

55

40 EP 2 300 010 B1

(continued)

Ex. Structure M+

5

127 483

10

15 128 585

20

129 363

25

130 273 30

35 131 400

40

132 465

45

133 451 50

55 [0063] Referring to the examples that follow, compounds of the preferred embodiments are synthesized using the methods described herein, or other methods, which are known in the art. [0064] It should be understood that the organic compounds according to the preferred embodiments may exhibit the phenomenon of tautomerism. As the chemical structures within this specification can only represent one of the possible

41 EP 2 300 010 B1

tautomeric forms, it should be understood that the preferred embodiments encompasses any tautomeric form of the drawn structure. [0065] It is understood that the invention is not limited to the embodiments set forth herein for illustration, but embraces all such forms thereof as come within the scope of the above disclosure. 5 General Conditions:

[0066] Mass spectra are run on LCMS systems using electrospray ionization. These are either Agilent 1100 HPLC/Mi- cromass Platform Mass Spectrometer combinations or Waters Acquity UPLC with SQD Mass Spectrometer. [M+H]+ 10 and M+ refers to mono-isotopic molecular weights. [0067] The various starting materials, intermediates, and compounds of the preferred embodiments may be isolated and purified, where appropriate, using conventional techniques such as precipitation, filtration, crystallization, evapora- tion, distillation, and chromatography. Unless otherwise stated, all starting materials are obtained from commercial suppliersand used withoutfurther purification. Saltsmay be preparedfrom compounds by known salt-forming procedures. 15 [0068] In addition various trade reagents and materials available have been utilized. Such reagents and materials include: [Isolute™ (available from Biotage) and Celite® (available from Aldrich)] and can be readily obtained from the suppliers indicated. [0069] For the examples below as well as throughout the application, the following abbreviations have the following meanings. If not defined, the terms have their generally accepted meanings. 20 Abbreviations:

[0070]

25 RT room temperature DMF dimethyl-formamide THF tetrahydrofuran MeOH methanol DCM dichloromethane 30 EtOAc ethyl acetate EtOH ethanol LCMS liquid chromatographic mass spectroscopy TEA triethylamine TFA trifluoroacetic acid 35 HPLC high performance liquid chromatography DMSO dimethyl sulfoxide

Et3N triethylamine HPLC high performance liquid chromatography HATU N-[(dimethylamino)-1H-1,2,3-triazolo[4,5-B]pyridin-1-ylmethylene]-N-methylmethanaminiumhexafluoro- 40 phosphate CDI 1,1’-carbonyl-diimidazole DEAD diethylazodicarboxylate PS polymer-supported 9-BBN 9-borabicyclo[3.3.1]nonane 45 dppf (diphenylphosphino)ferrocene

Preparation of final compounds

Example 1 50 ((R)-1-{[(3,5-Diamino-6-chloro-pyrazine-2-carbonyl)-amino]-methyl}-butyl)-trimethylammonium iodide

Step 1: [(R)-1-(1,3-Dioxo-1,3-dihydro-isoindol-2-ylmethyl)-butyl]-carbamic acid tert-butyl ester

55 [0071] To a mixture of phthalimide (1.43 g, 9.72 mmol), ((R)-1-hydroxymethyl-butyl)-carbamic acid tert-butyl ester (prepared according to the procedure described in Tetrahedron Letters (1987), 28(48), 6069-72) (1.97 g, 9.69 mmol) and triphenylphosphine (2.55 g, 9.72 mmol) in DCM (25 mL) at 0 °C is added DEAD (1.6 mL, 10.2 mmol) dropwise. The reaction mixture is stirred at RT overnight. The reaction mixture is adsorbed onto silica gel and purification by column

42 EP 2 300 010 B1

chromatography (SiO2, EtOAc/iso-hexane, gradient of 0-15% EtOAc) affords the title compound as white solid. [M+H]+ 233. [0072] Step 2: ((R)-1-Aminomethyl-butyl-carbamic acid tert-butyl ester A mixture of [(R)-1-(1,3-dioxo-1,3-dihydro-iso- indol-2-ylmethyl)-butyl]-carbamic acid tert-butyl ester (1.2 g, 3.61 mmol) and hydrazine monohydrate (1.0 mL, 20.6 mmol) 5 in EtOH (25 mL) and DCM (75 mL) is stirred at RT for 48 h. The precipitated solid is removed by filtration and washed with DCM. The filtrate is concentrated in vacuo to afford the title compound as white solid that is used without further purification.

Step 3: ((R)-1-{[(3,5-Diamino-6-chloro-pyrazine-2-carbonyl)-amino]-methyl}-butyl)-carbamic acid tert-butyl ester 10 [0073] A mixture of ((R)-1-aminomethyl-butyl)-carbamic acid tert-butyl ester (0.70 g, 3.46 mmol), 3,5-diamino-6-chloro- pyrazine-2-carboxylic acid (0.65 g, 3.45 mmol), N-methylmorpholine (1.5 mL, 13.6 mmol) and HATU (1.32 g, 3.47 mmol) in anhydrous DMF (50 mL) is stirred at RT for 16 h. The reaction mixture is concentratedin vacuo and the resulting residue is purified by column chromatography (basic alumina, 0-1% methanol in DCM) to afford the title compound as 15 pale yellow solid. [M+H]+ 373.

Step 4: 3,5-Diamino-6-chloro-pyrazine-2-carboxylic acid ((R)-2-amino-pentyl)-amide hydrochloride

[0074] To a solution of ((R)-1-{[(3,5-diamino-6-chloro-pyrazine-2-carbonyl)-amino]-methyl}-butyl)-carbamic acid tert- 20 butyl ester (0.65 g, 1.74 mmol) in 1,4-dioxane (15 mL) is added HCl (30 mL of a 4 M solution in 1,4-dioxane, 120 mmol) and the reaction mixture is stirred at RT for 16h. The reaction mixture is concentratedin vacuo and the yellow solid obtained is triturated with diethyl ether; the ether layer is decanted and the product is dissolved in minimal MeOH and is precipitated by the addition of diethyl ether. The solvent is decanted and the resulting solid is dried under vacuum to afford the title compound. [M+H]+ 273. 25 Step 5: ((R)-1-{[(3,5-Diamino-6-chloro-pyrazine-2-carbonyl)-amino]-methyl}-butyl)-trimethyl-ammonium iodide

[0075] To a mixture of 3,5-Diamino-6-chloro-pyrazine-2-carboxylic acid ((R)-2-amino-pentyl)-amide hydrochloride (0.08 g, 0.26 mmol) and potassium carbonate (0.20 g, 1.45 mmol) in acetonitrile (15 mL) is added iodomethane (0.10 30 mL, 1.60 mmol). The reaction mixture is stirred at RT for 16 h after which time the reaction mixture is filtered through Celite® (filter material). The filtrate is concentrated and the resulting solid is dissolved in acetonitrile and filtered throu gh Celite®. The filtrate is concentrated and then triturated with diethyl ether and the solvent is decanted. The resulting solid is dried under vacuum to afford the title compound as a yellow solid. M+ 315. 1H NMR (400 MHz, DMSO) δ 8.30 (1H, t), 7.36 (2H, br), 7.10 (2H, br), 3.70-3.64 (1H, m), 3.51-3.43 (2H, m), 3.02 (9H, s), 1.85-1.79 (1H, m), 1.61-1.41 (3H, m), 35 0.92 (3H, t).

Example 2

((S)-1-{[(3,5-Diamino-6-chloro-pyrazine-2-carbonyl)-amino]-methyl}-propyl)-trimethyl-ammonium iodide 40 [0076] The title compound is prepared by an analogous procedure to Example 1 by replacing ((R)-1-aminomethyl- butyl)-carbamic acid tert-butyl ester in Step 3 with ((S)-1-aminomethyl-propyl)-carbamic acid tert-butyl ester. M + 301.

Example 3 45 ((R)-1-{[(3,5-Diamino-6-chloro-pyrazine-2-carbonyl)-amino]-methyl}-propyl)-trimethyl-ammonium iodide

[0077] The title compound is prepared by an analogous procedure to Example 1 by replacing ((R)-1-aminomethyl- butyl)-carbamic acid tert-butyl ester in Step 3 with ((R)-1-aminomethyl-propyl)-carbamic acid tert-butyl ester (prepared 50 according to the procedure described in WO2007125331, page 217-218). M+301.

Example 4

((S)-1-{[(3,5-Diamino-6-chloro-pyrazine-2-carbonyl)-amino]-methyl}-pentyl)-trimethyl-ammonium iodide 55 [0078] The title compound is prepared by an analogous procedure to Example 1 by replacing ((R)-1-hydroxymethyl- butyl)-carbamic acid tert-butyl ester in Step 1 with ((S)-1-hydroxymethyl-pentyl)-carbamic acid tert-butyl ester (prepared according to the procedure described in Tetrahedron Letters 1987, 28(48), 6069-72). M+ 329.

43 EP 2 300 010 B1

Example 5

((S)-1-{[(3,5-Diamino-6-chloro-pyrazine-2-carbonyl)-amino]-methyl}-butyl)-trimethyl-ammonium trifluoroacetate

5 [0079] The title compound is prepared by an analogous procedure to Example 1 by replacing ((R)-1-hydroxymethyl- butyl)-carbamic acid tert-butyl ester in Step 1 with ((S)-1-hydroxymethyl-butyl)-carbamic acid tert-butyl ester (prepared + 1 according to the procedure described in US 2007/0032433 page 232). M 315. H NMR (400 MHz, DMSO-d6) δ 8.30 (1H, t), 7.20 (2H, br), 7.09 (2H, br), 3.73-3.62 (1H, m), 3.55-3.48 (2H, m), 3.08 (9H, s), 1.90-1.78 (1H, m), 1.65-1.47 (3H, m), 0.93 (3H, t). 10 Example 6

((S)-1-{[(3,5-Diamino-6-chloro-pyrazine-2-carbonyl)-amino]-methyl}-2-methyl-propyl)-trimethyl-ammonium iodide

15 [0080] The title compound is prepared by an analogous procedure to Example 1 by replacing ((R)-1-hydroxymethyl- butyl)-carbamic acid tert-butyl ester in Step 1 with N-(tert-butoxycarbonyl)-L-valinol. M+ 315.

Example 7

20 (R)-2-{[(3,5-Diamino-6-chloro-pyrazine-2-carbonyl)-amino]-methyl}-1,1-dimethyl-piperidinium iodide

[0081] The title compound is prepared by an analogous procedure to Example 1 by replacing ((R)-1-aminomethyl- butyl)-carbamic acid tert-butyl ester in Step 3 with (R)-2-aminomethyl-1-N-Boc-piperidine. M + 313.

25 Example 8

(S)-2-{[(3,5-Diamino-6-chloro-pyrazine-2-carbonyl)-amino]-methyl}-1,1-dimethyl-piperidinium iodide

[0082] The title compound is prepared by an analogous procedure to Example 1 by replacing ((R)-1-aminomethyl- 30 butyl)-carbamic acid tert-butyl ester in Step 3 with (S)-2-aminomethyl-1-N-Boc-piperidine. M + 313.

Example 9

(1-{[(3,5-Diamino-6-chloro-pyrazine-2-carbonyl)-amino]-methyl}-cyclopentyl)-trimethyl-ammonium iodide 35 [0083] The title compound is prepared by an analogous procedure to Example 1 by replacing ((R)-1-aminomethyl- butyl)-carbamic acid tert-butyl ester in Step 3 with (1-aminomethyl-cyclopentyl)-carbamic acid tert-butyl ester. M+ 327. 1 H NMR (400 MHz, DMSO-d6) δ 8.27 (1H, t), 7.50 (2H, br), 7.12 (2H, br), 3.69 (2H, d), 3.09 (9H, s), 2.19-2.09 (2H, m), 1.95-1.88 (2H, m), 1.67-1.53 (4H, m). 40 Example 10

(S)-2-{[(3,5-Diamino-6-chloro-pyrazine-2-carbonyl)-amino]-methyl}-1,1-dimethyl-pyrrolidinium iodide

45 [0084] The title compound is prepared by an analogous procedure to Example 1 by replacing ((R)-1-aminomethyl- butyl)-carbamic acid tert-butyl ester in Step 3 with (S)-2-aminomethyl-1-N-Boc-pyrrolidine. M + 299.

Example 11

50 (R)-2-{[(3,5-Diamino-6-chloro-pyrazine-2-carbonyl)-amino]-methyl}-1,1-dimethyl-pyrrolidinium iodide

[0085] The title compound is prepared by an analogous procedure to Example 1 by replacing ((R)-1-aminomethyl- butyl)-carbamic acid tert-butyl ester in Step 3 with (R)-2-aminomethyl-1-N-Boc-pyrrolidine. M + 299. 1H NMR (400 MHz, DMSO-d6) δ 8.39 (1H, t), 7.60 (2H, br), 7.10 (2H, br), 3.81-3.71 (1H, m), 3.69-3.43 (4H, m), 3.11 (3H, s), 3.00 (3H, s), 55 2.23-2.19 (1H, m), 2.05-1.88 (3H, m).

44 EP 2 300 010 B1

Example 12

{(S)T2-[(3,5-Diamino-6-chloro-pyrazine-2-carbonyl)-amino]-1-methyl-ethyl}-trimethyl-ammonium iodide

5 [0086] The title compound is prepared by an analogous procedure to Example 1 by replacing ((R)-1-hydroxymethyl- butyl)-carbamic acid tert-butyl ester in Step 1 with ((S)-2-hydroxy-1-methyl-ethyl)-carbamic acid tert-butyl ester. M + 287.

Example 13

10 {(S)-1-Benzyl-2-[(3,5-diamino-6-chloro-pyrazine-2-carbonyl)-amino]-ethyl}-trimethylammonium iodide

[0087] The title compound is prepared using an analogous procedure to Example 1 by replacing 3,5-diamino-6-chloro- pyrazine-2-carboxylic acid ((R)-2-amino-pentyl)-amide hydrochloride in Step 4 with 3,5-diamino-6-chloro-pyrazine-2- + 1 carboxylic acid ((S)-2-amino-3-phenyl-propyl)-amide (Intermediate A). M 363. H NMR (400 MHz, CD 3OD) δ 7.54-7.00 15 (5H, m), 3.85-3.56 (2H, m), 3.57 (1H, dd), 3.41-3.33 (m, 2H), 3.22 (9H, s).

Example 14

((S)-5-[3-(4-Benzyloxy-phenyl)-propionylamino]-1-{[(3,5-diamino-6-chloro-pyrazine-2-carbonyl)-amino]-me- 20 thyl}-pentyl)-trimethyl-ammonium iodide

Step 1: ((S)-5-[3-(4-Benzyloxy-phenyl)-propionylamino]-1-{[(3,5-diamino-6-chloro-pyrazine-2-carbonyl)-amino]-me- thyl}-pentyl)-carbamic acid tert-butyl ester

25 [0088] To a solution of 3-(4-benzyloxyphenyl)propionic acid (0.19 g, 0.74 mmol) and HATU (0.29 g, 0.75 mmol) in DMF (10 mL) is added N-methylmorpholine (0.32 mL, 2.91 mmol) followed by ((S)-5-amino-1-{[(3,5-diamino-6-chloro- pyrazine-2-carbonyl)-amino]-methyl}-pentyl)-carbamic acid tert-butyl ester (Intermediate B) (0.30 g, 0.75 mmol). The resulting mixture is stirred at RT for 16 h. The solvent is removedin vacuo and the residue is purified by column chromatography (basic alumina, 2% MeOH in DCM) to afford the title compound as yellow solid. [M-Boc+H] + 540. 30 Step 2: 3,5-Diamino-6-chloro-pyrazine-2-carboxylic acid {(S)-2-amino-6-[3-(4-benzyloxy-phenyl)-propionylami- no]-hexyl}-amide hydrochloride

[0089] A solution of ((S)-5-[3-(4-benzyloxy-phenyl)-propionylamino]-1-{[(3,5-diamino-6-chloro-pyrazine-2-carbon- 35 yl)-amino]-methyl}-pentyl)-carbamic acid tert-butyl ester (0.29 g, 0.45 mmol) in 1,4-dioxane (3 mL) and HCl (5 mL of a 4 M solution in 1,4-dioxane, 20 mmol) is stirred at RT for 2 h. The solvent is removed in vacuo and the resulting residue is triturated with diethyl ether and then the diethyl ether is decanted. The remaining solid is dissolved in a minimal amount of MeOH then diethyl ether is added to cause precipitation. The supernatant solvents are decanted and the remaining solid is washed with diethyl ether then dried under vacuum to afford the title compound as yellow solid. [M+H]+ 540. 40 Step 3: ((S)-5-[3-(4-Benzyloxy-phenyl)-propionylamino]-1-{[(3,5-diamino-6-chloro-pyrazine-2-carbonyl)-amino]-me- thyl}-pentyl)-trimethyl-ammonium iodide

[0090] The title compound is prepared using an analogous procedure to Example 1 by replacing 3,5-diamino-6-chloro- 45 pyrazine-2-carboxylic acid ((R)-2-amino-pentyl)-amide hydrochloride in Step 5 with 3,5-diamino-6-chloro-pyrazine-2- carboxylic acid {(S)-2-amino-6-[3-(4-benzyloxy-phenyl)-propionylamino]-hexyl}-amide hydrochloride M + 582.

Example 15

50 ((S)-5-[2-(4-Benzyloxy-phenyl)-acetylamino]-1-{[(3,5-diamino-6-chloro-pyrazine-2-carbonyl)-amino]-methyl}-pentyl)-tri- methyl-ammonium iodide

[0091] The title compound is prepared using an analogous procedure to Example 14 by replacing 3-(4-benzyloxyphe- + 1 nyl)propionic acid in Step 1 with (4-benzyloxy-phenyl)-acetic acid. M 568. H NMR (400 MHz, DMSO-d6) δ 8.31 (1H, 55 t), 7.97 (1H, t), 7.42-7.31 (4H, m), 7.14 (2H, d), 7.13-7.09 (1H, m), 7.05 (4H, br), 6.90 (2H, d), 5.04 (2H, s), 3.50-3.40 (4H, m), 3.30 (2H, s), 3.05 (9H, s), 1.49-1.32 (6H, m).

45 EP 2 300 010 B1

Example 16

4-Benzyl-1-{2-[(3,5-diamino-6-chloro-pyrazine-2-carbonyl)-amino]-ethyl}-1-methyl-piperidinium iodide

5 Step 1: 3,5-Diamino-6-chloro-pyrazine-2-carboxylic acid [2-(4-benzyl-piperidin-1-yl)-ethyl] amide

[0092] A solution of 3,5-diamino-6-chloro-pyrazine-2-carboxylic acid (0.10 g, 0.53 mmol) and CDI (0.172 g, 1.06 mmol) in DMF (2 mL) is stirred at RT for 1 h. To this reaction mixture is added 2-(4-benzylpiperidinol)-1-ethanamine (0.139 g, 0.63 mmol) and stirring is continued at RT for a further 48 h. The reaction mixture is diluted with water (20 mL) and 10 extracted with EtOAc (3 x 20 mL). The combined organic portions are dried (MgSO 4) and concentrated in vacuo to afford a yellow solid which is recrystallised from acetonitrile to yield the title compound as off-white crystals. [M+H]+ 389.

Step 2: 4-Benzyl-1-{2-[(3,5-diamino-6-chloro-pyrazine-2-carbonyl)-amino]-ethyl}-1-methyl-piperidinium iodide

15 [0093] A solution of 3,5-diamino-6-chloro-pyrazine-2-carboxylic acid [2-(4-benzyl-piperidin-1-yl)-ethyl]-amide (39 mg, 0.10 mmol) and methyl iodide (0.031 mL, 0.50 mmol) in acetone (2 mL) is heated at reflux for 3 h. After cooling to RT + 1 the solvent is removed in vacuo to afford the title compound as yellow solid. M 403. H NMR (400 MHz, DMSO-d6) δ 8.30 (1H, t), 7.62 (2H, br), 7.39-7.36 (2H, m), 7.29-7.23 (3H, m), 7.16 (2H, br), 3.73-3.47 (6H, m), 3.35-3.28 (2H, m), 3.14 (3H, s), 2.67 (2H, d), 1.92-1.81 (1H, m), 1.74-1.68 (4H, m). 20 Example 17

Allyl-{2-[(3,5-diamino-6-chloro-pyrazine-2-carbonyl)-amino]-ethyl}-bis-[3-(4-methoxyphenyl)-propyl]-ammonium trifluor- oacetate 25 [0094] To a solution of 3,5-diamino-6-chloro-pyrazine-2-carboxylic acid (2-{bis-[3-(4-methoxy-phenyl)-propyl]-ami- no}-ethyl)-amide trifluoroacetate (Intermediate D) (0.20 g, 0.31 mmol) in acetone (5 mL) is added sodium carbonate (99 mg, 0.94 mmol) followed by allyl bromide (0.082 mL, 0.94 mmol). The reaction mixture is heated at reflux for 72 h, cooled to RT then filtered to remove inorganic salts. The filtrate is concentrated and purification by reverse phase column 30 chromatography (Isolute™ C18, 0-100% acetonitrile in water with 0.1% TFA) affords the title compound. M+ 567. 1H NMR (400 MHz, DMSO-d6) δ 8.32 (1H, t), 7.76 (2H, br), 7.16 (2H, br), 7.11 (4H, d), 6.85 (4H, d), 5.95 (1H, ddt) 5.62 (1H, d), 5.57 (1H, d), 3.99 (2H, d), 3.72 (6H, s), 3.58-3.54 (2H, m), 3.30 (2H, t), 3.24-3.20 (4H, m), 2.50 (4H, t), 1.97-1.78 (4H, m).

35 Example 18

{2-[(3,5-Diamino-6-chloro-pyrazine-2-carbonyl)-amino]-ethyl}-tris-[3-(4-methoxyphenyl)-propyl]-ammonium trifluoroac- etate

40 [0095] The title compound is prepared using an analogous procedure to Example 17 by replacing allyl bromide with + 1 1-(3-bromo-propyl)-4-methoxy-benzene. M 675. H NMR (400 MHz, CD3OD) 7.07 (6H, d), 6.83 (6H, d), 3.88 (9H, s), 3.59 (2H, t), 3.39 (2H, t), 3.26-3.21 (6H, m), 2.54 (6H, t), 1.80 (6H, m).

Example 19 45 {2-[(3,5-Diamino-6-chloro-pyrazine-2-carbonyl)-amino]-ethyl}-(2-diethylamino-ethyl)-bis-[3-(4-methoxy-phenyl)-pro- pyl]-ammonium trifluoroacetate

[0096] The title compound is prepared using an analogous procedure to Example 17 by replacing allyl bromide with 50 diethylaminoethyl bromide. M+ 626.

Example 20

{2-[(3,5-Diamino-6-chloro-pyrazine-2-carbonyl)-amino]-ethyl}-bis-[3-(4-methoxy-phenyl)-propyl]-methyl-ammonium trif- 55 luoroacetate

[0097] The title compound is prepared using an analogous procedure to Example 17 by replacing allyl bromide with + 1 methyl iodide. M 541. H NMR (400 MHz, DMSO- d6) δ 8.32 (1H, t), 7.75 (2H, br), 7.14 (4H, d), 6.97 (2H, br), 6.83 (4H,

46 EP 2 300 010 B1

d), 3.73 (6H, s), 3.57 (2H, dt), 3.38 (2H, t), 3.34-3.41 (4H, m), 3.03 (3H, s), 2.49 (4H, t), 1.94-1.87 (4H, m).

Example 21

5 {2-[(3,5-Diamino-6-chloro-pyrazine-2-carbonyl)-amino]-ethyl}-bis-[3-(4-methoxy-phenyl)-propyl]-propyl-ammonium trif- luoroacetate

[0098] The title compound is prepared using an analogous procedure to Example 17 by replacing allyl bromide with propyl iodide. M+ 569. 10 Example 22

Carbamoylmethyl-{2-[(3,5-diamino-6-chloro-pyrazine-2-carbonyl)-amino]-ethyl}-bis-[3-(4-methoxy-phenyl)-propyl]-am- monium trifluoroacetate 15 [0099] The title compound is prepared using an analogous procedure to Example 17 by replacing allyl bromide with 2-bromoacetamide. M+ 584.

Example 23 20 Benzyl-{2-[(3,5-diamino-6-chloro-pyrazine-2-carbonyl)-amino]-ethyl}-bis-[3-(4-methoxy-phenyl)-propyl]-ammonium trif- luoroacetate

[0100] The title compound is prepared using an analogous procedure to Example 17 by replacing allyl bromide with 25 benzyl bromide. M+ 617.

Example 24

{2-[(3,5-Diamino-6-chloro-pyrazine-2-carbonyl)-amino]-ethyl}-methoxycarbonylmethyl-bis-[3-(4-methoxy-phenyl)-pro- 30 pyl]-ammonium trifluoroacetate

[0101] The title compound is prepared using an analogous procedure to Example 17 by replacing allyl bromide with methyl bromoacetate. M+ 599.

35 Example 25

{2-[(3,5-Diamino-6-chloro-pyrazine-2-carbonyl)-amino]-ethyl}-ethyl-bis-[3-(4-methoxy-phenyl)-propyl]-ammonium trif- luoroacetate

40 [0102] The title compound is prepared using an analogous procedure to Example 17 by replacing allyl bromide with ethyl iodide. M+ 555.

Example 26

45 {4-[3-(3-Chloro-4-ethoxy-phenyl)-ureido]-butyl}-{2-[(3,5-diamino-6-chloro-pyrazine-2-carbonyl)-amino]-ethyl)-[3-(4- methoxy-phenyl)-propyl]-methyl-ammonium bromide

Step 1: 3,5-Diamino-6-chloro-pyrazine-2-carboxylic acid [2-({4-[3-(3-chloro-4-ethoxy-phenyl)-ureido]-butyl}-methyl-ami- no)-ethyl]-amide trifluoroacetate 50 [0103] A mixture comprising 3,5-diamino-6-chloro-pyrazine-2-carboxylic acid {2-[(4-amino-butyl)-methyl-ami- no]-ethyl}-amide (Intermediate F),(0.20 g, 0.63 mmol) and 1-ethoxy-2-chloro-4-isocyanato-benzene (Intermediate G) (0.91 g, 0.95 mmol) in DMF (3 mL) is stirred under an inert atmosphere of argon at 50 °C for 48 h. Purification by reverse phase column chromatography (Isolute™ C18, 0-100% acetonitrile in water with 0.1%TFA) affords the title compound. 55 [M+H]+ 513.

47 EP 2 300 010 B1

Step 2: {4-[3-(3-Chloro-4-ethoxy-phenyl)-ureido]-butyl}-{2-[(3,5-diamino-6-chloro-pyrazine-2-carbonyl)-ami- no]-ethyl)-[3-(4-methoxy-phenyl)-propyl]-methyl-ammonium bromide

[0104] A mixture comprising 3,5-diamino-6-chloro-pyrazine-2-carboxylic acid [2-({4-[3-(3-chloro-4-ethoxy-phenyl)-ure- 5 ido]-butyl}-methyl-amino)-ethyl]-amide trifluoroacetate (435 mg, 0.85 mmol), 1-(3-bromo-propyl)-4-methoxy-benzene (1.83 g, 7.99 mmol) and sodium carbonate (270 mg, 2.54 mmol) in acetone (8 mL) is heated at reflux for 11 days. After cooling to RT, the mixture is filtered and concentrated in vacuo. DCM is added and the precipitated solid is collected by + 1 filtration. The solid is crystallised from ethanol to afford the title compound. M 661. H NMR (400 MHz, DMSO-d6) δ 8.50 (1H, s), 8.29 (1H, t), 7.63 (2H, br), 7.62 (1H, d), 7.15 (1H, dd), 7.10 (2H, d), 6.99 (1H, d), 6.93 (2H, br), 6.81 (2H, 10 d), 6.23 (1H, t), 4.01 (2H, q), 3.71 (3H, s), 3.61-3.56 (2H, m), 3.39-3.29 (4H, m), 3.08 (2H, q), 3.03 (3H, s), 2.49 (2H, t), 2.00-1.91 (2H, m), 1.72-1.64 (2H, m), 1.42 (2H, quint), 1.32 (3H, t).

Example 27

15 {2-[(3,5-Diamino-6-chloro-pyrazine-2-carbonyl)-amino]-ethyl}-[3-(4-methoxy-phenyl)-propyl]-dimethyl-ammonium bro- mide

[0105] A mixture comprising 3,5-diamino-6-chloro-pyrazine-2-carboxylic acid (2-dimethylamino-ethyl)-amide (Inter- mediate H) (1.8 g, 7.0 mmol) and 1-(3-bromo-propyl)-4-methoxy-benzene (3.2 g, 14.0 mmol) in acetone (50 mL) is 20 heated at reflux for 72 h. After cooling to RT, the solvent is removedin vacuo and the resulting solid is recrystallised + 1 from ethanol to afford the title compound. M 407. H NMR (400 MHz, DMSO- d6) δ 8.37 (1H, t), 7.76 (2H, br), 7.19 (2H, br), 7.17 (2H, d), 6.90 (2H, d), 3.79 (3H, s), 3.67 (2H, q), 3.49 (2H, t), 3.15 (6H, s), 2.55 (2H, t), 2.02 (2H, quintet).

Example 28 25 {2-[(3,5-Diamino-6-chloro-pyrazine-2-carbonyl)-amino]-ethyl}-{2-[3-(4-isobutoxy-phenyl)-ureido]-ethyl}-dimethyl-am- monium chloride

[0106] The title compound is prepared using an analogous procedure to Example 27 by replacing 1-(3-bromo-propyl)- 30 4-methoxy-benzene with 1-(2-chloro-ethyl)-3-(4-isobutoxy-phenyl)-urea (Intermediate I). M+ 493.

Example 29

{2-[(3,5-Diamino-6-chloro-pyrazine-2-carbonyl)-amino]-ethyl)-[3-(4-{3-[4-((S)-2,3-dihydroxy-propoxy)-phenyl]-pro- 35 poxy}-phenyl)-propyl]-dimethyl-ammonium bromide

[0107] 3,5-Diamino-6-chloro-pyrazine-2-carboxylic acid (2-dimethylamino-ethyl)-amide (Intermediate H) (0.65 g, 2.52 mmol) and (S)-3-(4-{3-[4-(3-bromo-propyl)-phenoxy]-propyl}-phenoxy)-propane-1,2-diol (Intermediate J) (1.60 g, 3.78 mmol) in butan-2-one (25 mL) is heated at reflux overnight during which time precipitation of white solid is observed. 40 The reaction mixture is filtered to collect the product, washed with warm acetone and dried under vacuum to afford the + 1 title compound. M 601. H NMR (400 MHz, DMSO-d6) δ 8.29 (1H, t), 7.30 (1H, br), 7.12 (2H, d), 7.11 (2H, br), 7.07 (2H, d), 6.95 (1H, br), 6.84 (2H, d), 6.82 (2H, d), 4.89 (1H, d), 4.63 (1H, t), 3.94 (1H, dd), 3.89 (2H, t), 3.81 (1H, dd), 3.7 7 (1H, m), 3.59 (2H, m), 3.43 (2H, m), 3.42 (2H, m), 3.34 (2H, m), 3.08 (6H, s), 2.66 (2H, m), 2.47 (2H, m), 1.96 (2H, m), 1.93 (2H, m). 45 Example 30

{2-[(3.5-Diamino-6-chloro-pyrazine-2-carbonyl)-aminol-ethyl}-dimethyl-[2-(3-phenyl-propionylamino)-ethyl]-ammonium hexafluorophosphate 50 [0108] A mixture of (2-amino-ethyl)-{2-[(3,5-diamino-6-chloro-pyrazine-2-carbonyl)-amino]-ethyl}-dimethyl-ammoni- um bromide (Intermediate K) (0.50 g, 1.31 mmol), 3-phenylpropionic acid (0.20 g, 1.33 mmol), N-methylmorpholine (0.60 mL, 5.46 mmol) and HATU (0.50 g, 1.33 mmol) in anhydrous DMF (10 mL) is stirred under a nitrogen atmosphere at RT for 16 h. The reaction mixture is concentrated in vacuo and is purified by column chromatography (basic alumina, 55 + 1 0-3% MeOH in DCM) to afford the title compound. M 434. H NMR (400 MHz, DMSO-d6) δ 8.26 (1H, t), 8.16 (1H, t), 7.40 (2H, br), 7.28-7.23 (2H, m), 7.20-7.17 (3H, m), 7.15 (2H, br), 3.62-3.59 (2H, m), 3.49-3.29 (6H, m), 3.07 (6H, s), 2.81 (2H, t), 2.41 (2H, t).

48 EP 2 300 010 B1

Example 31

{2-[(3,5-Diamino-6-chloro-pyrazine-2-carbonyl)-amino]-ethyl}-[2-(2-1H-indol-3-yl-acetylamino)-ethyl]-dimethyl-ammo- nium hexafluorophosphate 5 [0109] The title compound is prepared using an analogous procedure to Example 30 by replacing 3-phenylpropionic acid with (1H-indol-3-yl)-acetic acid. M + 459.

Example 32 10 {2-[2-(4-Benzyloxy-phenyl)-acetylamino]-ethyl}-{2-[(3,5-diamino-6-chloro-pyrazine-2-carbonyl)-amino]-ethyl}-dimethyl- ammonium hexafluorophosphate

[0110] The title compound is prepared using an analogous procedure to Example 30 by replacing 3-phenylpropionic 15 acid with (4-benzyloxy-phenyl)-acetic acid. M+ 526.

Example 33

{2-[3-(4-Chloro-phenyl)-propionylamino]-ethyl}-{2-[(3,5-diamino-6-chloro-pyrazine-2-carbonyl)-amino]-ethyl}-dimethyl- 20 ammonium hexafluorophosphate

[0111] The title compound is prepared using an analogous procedure to Example 30 by replacing 3-phenylpropionic acid with 3-(4-chloro-phenyl)-propionic acid. M+ 468.

25 Example 34

{2-[3-(4-Benzyloxy-phenyl)-propionylamino]-ethyl}-{2-[(3,5-diamino-6-chloropyrazine-2-carbonyl)-amino]-ethyl}-dime- thyl-ammonium hexafluorophosphate

30 [0112] The title compound is prepared using an analogous procedure to Example 30 by replacing 3-phenylpropionic acid with 3-(4-benzyloxy-phenyl)-propionic acid. M+ 540.

Example 35

35 {2-[(3,5-Diamino-6-chloro-pyrazine-2-carbonyl)-amino]-ethyl}-{2-[2-(6-methoxy-1,2,3,4-tetrahydro-naphthalen-1-yl- acetylamino]-ethyl}-dimethyl-ammonium hexafluorophosphate

[0113] The title compound is prepared using an analogous procedure to Example 30 by replacing 3-phenylpropionic acid with (6-methoxy-1,2,3,4-tetrahydro-naphthalen-1-yl)-acetic acid. M+ 504. 40 Example 36

[2-(3-Benzo[1,3]dioxol-5-yl-propionylamino)-ethyl]-{2-[(3,5-diamino-6-chloropyrazine-2-carbonyl)-amino]-ethyl}-dime- thyl-ammonium hexafluorophosphate 45 [0114] The title compound is prepared using an analogous procedure to Example 30 by replacing 3-phenylpropionic acid with 3-Benzo[1,3]dioxol-5-yl-propionic acid. M + 478.

Example 37 50 {2-[(3,5-Diamino-6-chloro-pyrazine-2-carbonyl)-amino]-ethyl}-{2-[3-(4-methoxy-naphthalen-1-yl)-propionylami- no]-ethyl}-dimethyl-ammonium hexafluorophosphate

[0115] The title compound is prepared using an analogous procedure to Example 30 by replacing 3-phenylpropionic 55 acid with 3-(4-methoxy-naphthalen-1-yl)-propionic acid. M+ 514.

49 EP 2 300 010 B1

Example 38

{2-[(3,5-Diamino-6-chloro-pyrazine-2-carbonyl)-amino]-ethyl-{[3-(4-fluoro-phenyl)-propionylamino]-ethyl}-dimethyl-am- monium hexafluorophosphate 5 [0116] The title compound is prepared using an analogous procedure to Example 30 by replacing 3-phenylpropionic acid with 3-(4-fluoro-phenyl)-propionic acid. M+ 452.

Example 39 10 {2-[2-(4-Chloro-phenyl)-acetylamino]-ethyl}-{2-[(3,5-diamino-6-chloro-pyrazine-2-carbonyl)-amino]-ethyl}-dimethyl-am- monium hexafluorophosphate

[0117] The title compound is prepared using an analogous procedure to Example 30 by replacing 3-phenylpropionic 15 acid with (4-chloro-phenyl)-acetic acid. M+ 454.

Example 40

{2-[(3,5-Diamino-6-chloro-pyrazine-2-carbonyl)-amino]-ethyl}-{2-[3-(4-methoxy-phenyl)-propionylamino]-ethyl}-dime- 20 thyl-ammonium hexafluorophosphate

[0118] The title compound is prepared using an analogous procedure to Example 30 by replacing 3-phenylpropionic acid with 3-(4-methoxy-phenyl)-propionic acid. M + 464.

25 Example 41

{2-[(3,5-Diamino-6-chloro-pyrazine-2-carbonyl)-amino]-ethyl}-{2-[4-(4-methoxy-phenyl)-butyrylamino]-ethyl}-dimethyl- ammonium hexafluorophosphate

30 [0119] The title compound is prepared using an analogous procedure to Example 30 by replacing 3-phenylpropionic acid with 4-(4-methoxy-phenyl)-butyric acid. M+ 478.

Example 42

35 {2-[(3,5-Diamino-6-chloro-pyrazine-2-carbonyl)-amino]-ethyl}-dimethyl-(2-phenylacetylamino-ethyl)-ammonium hex- afluorophosphate

[0120] The title compound is prepared using an analogous procedure to Example 30 by replacing 3-phenylpropionic acid with phenyl-acetic acid. M+ 420. 40 Example 43

{2-[(3,5-Diamino-6-chloro-pyrazine-2-carbonyl)-amino]-ethyl}-dimethyl-[2-(2-p-tolyl-acetylamino)-ethyl]-ammonium hexafluorophosphate 45 [0121] The title compound is prepared using an analogous procedure to Example 30 by replacing 3-phenylpropionic acid with p-tolyl-acetic acid. M+ 434.

Example 44 50 [2-(4-Chloro-benzoylamino)-ethyl]-{2-[(3.5-diamino-6-chloro-pyrazine-2-carbonyl)-amino]-ethyl}-dimethyl-ammonium hexafluorophosphate

[0122] The title compound is prepared using an analogous procedure to Example 30 by replacing 3-phenylpropionic 55 acid with 4-chloro-benzoic acid. M+ 440.

50 EP 2 300 010 B1

Example 45

(2-Benzoylamino-ethyl)-{2-[(3,5-diamino-6-chloro-pyrazine-2-carbonyl)-amino]-ethyl}-dimethyl-ammonium hexafluoro- phosphate 5 [0123] The title compound is prepared using an analogous procedure to Example 30 by 3-phenylpropionic acid with benzoic acid. M+ 406.

Example 46 10 {2-[(3,5-Diamino-6-chloro-pyrazine-2-carbonyl)-amino]-ethyl}-dimethyl-[2-(4-methyl-benzoylamino)-ethyl]-ammonium hexafluorophosphate

[0124] The title compound is prepared using an analogous procedure to Example 30 by replacing 3-phenylpropionic 15 acid with 4-methyl-benzoic acid. M+ 420.

Example 47

{2-[(3,5-Diamino-6-chloro-pyrazine-2-carbonyl)-amino]-ethyl}-{2-[3-(3,4-dimethoxy-phenyl)-propionylami- 20 no]-ethyl}-dimethyl-ammonium hexafluorophosphate

[0125] The title compound is prepared using an analogous procedure to Example 30 by replacing 3-phenylpropionic acid with 3-(3,4-dimethoxy-phenyl)-propionic acid. M+ 494.

25 Example 48

{2-[(3,5-Diamino-6-chloro-pyrazine-2-carbonyl)-amino]-ethyl}-{2-[3-(4-methanesulfonyl-phenyl)-propionylami- no]-ethyl}-dimethyl-ammonium hexafluorophosphate

30 [0126] The title compound is prepared using an analogous procedure to Example 30 by replacing 3-phenylpropionic acid with 3-(4-methanesulfonyl-phenyl)-propionic acid. M + 512.

Example 49

35 {2-[(3,5-Diamino-6-chloro-pyrazine-2-carbonyl)-amino]-ethyl}-(2-{3-[3-(diaminomethylene-sulfamoyl)-4-methoxy-phe- nyl]-3-phenyl-propionylamino}-ethyl)-dimethyl-ammonium hexafluorophosphate

[0127] The title compound is prepared using an analogous procedure to Example 30 by replacing 3-phenylpropionic acid with 3-[3-(Diaminomethylene-sulfamoyl)-4-methoxyphenyl]-3-phenyl-propionic acid (Intermediate L). M+ 661. 40 Example 50

{3-[2-(4-Benzyloxy-phenyl)-acetylamino]-propyl}-{2-[(3,5-diamino-6-chloro-pyrazine-2-carbonyl)-aminol-ethyl}-dime- thyl-ammonium bromide 45 [0128] A mixture of (3-amino-propyl)-{2-[(3,5-diamino-6-chloro-pyrazine-2-carbonyl)-amino]-ethyl}-dimethyl-ammoni- um bromide (Intermediate M) (1.0 g, 2.52 mmol), 4-benzyloxyphenylacetic acid (0.61 g, 2.52 mmol), N-methylmorpholine (1.0 mL, 9.10 mmol), N,N’-dicyclohexylcarbodiimide (0.52 g, 2.52 mmol) and 1-hydroxybenzotriazole (0.34 g, 2.52 mmol) in anhydrous DMF (20 mL) is stirred under a nitrogen atmosphere at RT for 16 h. The reaction mixture is concentrated 50 under vacuum and is purified by column chromatography (basic alumina, 0-3% MeOH in DCM) to afford the title compound + 1 as a white solid. M 540. H NMR (400 MHz, DMSO-d6) δ 8.23 (1H, t), 8.07 (1H, t), 7.52 (2H, br), 7.41-7.28 (5H, m), 7.16 (2H, d), 7.09 (2H, br), 6.91 (2H, d), 5.05 (2H, s), 3.58-3.3.51 (2H, m), 3.40-3.25 (6H, m), 3.11-3.03 (2H, m), 3.02 (6H, s), 1.87-1.75 (2H, m).

55

51 EP 2 300 010 B1

Example 51

{3-[2-(4-Chloro-phenyl)-acetylamino]-propyl}-{2-[(3,5-diamino-6-chloro-pyrazine-2-carbonyl)-amino]-ethyl}-dimethyl- ammonium hexafluorophosphate 5 [0129] A mixture of (3-amino-propyl)-{2-[(3,5-diamino-6-chloro-pyrazine-2-carbonyl)-amino]-ethyl}-dimethyl-ammoni- um bromide (Intermediate M) (0.25 g, 0.63 mmol), (4-chloro-phenyl)-acetic acid (0.11 g, 0.64 mmol), N-methylmorpholine (0.25 mL, 2.27 mmol) and HATU (0.24 g, 0.63 mmol) in anhydrous DMF (8 mL) is stirred under a nitrogen atmosphere at RT for 16 h. The reaction mixture is concentrated in vacuo and is purified by column chromatography (basic alumina, 10 + 1 0-4% MeOH in DCM) to afford the title compound. M 468. H NMR (400 MHz, DMSO-d6) δ 8.22 (1H, t), 8.14 (1H, t), 7.45 (2H, br), 7.32 (2H, d), 7.24 (2H, d), 7.08 (2H, br), 3.57-3.51 (2H, m), 3.43 (2H, s), 3.42-3.25 (4H, m), 3.12-3.03 (2H, m), 3.02 (6H, s), 1.86-1.78 (2H, m).

Example 52 15 [3-(4-Chloro-benzoylamino)-propyl]-{2-[(3,5-diamino-6-chloro-pyrazine-2-carbonyl)-amino]-ethyl}-dimethyl-ammonium hexafluorophosphate

[0130] The title compound is prepared using an analogous procedure to Example 51 by replacing (4-chloro-phe- 20 nyl)-acetic acid with 4-chloro-benzoic acid. M+ 454.

Example 53

{2-[(3,5-Diamino-6-chloro-pyrazine-2-carbonyl)-amino]-ethyl}-dimethyl-(3-phenylacetylamino-propyl)-ammonium hex- 25 afluorophosphate

[0131] The title compound is prepared using an analogous procedure to Example 51 by replacing (4-chloro-phe- nyl)-acetic acid with phenyl acetic acid. M + 434.

30 Example 54

{2-[(3,5-Diamino-6-chloro-pyrazine-2-carbonyl)-amino]-ethyl}-dimethyl-[3-(2-p-tolyl-acetylamino)-propyl]-ammonium hexafluorophosphate

35 [0132] The title compound is prepared using an analogous procedure to Example 51 by replacing (4-chloro-phe- nyl)-acetic acid with p-tolyl-acetic acid. M+ 448.

Example 55

40 {2-[(3,5-Diamino-6-chloro-pyrazine-2-carbonyl)-amino]-ethyl}-dimethyl-[3-(3-phenyl-propionylamino)-propyl]-ammoni- um hexafluorophosphate

[0133] The title compound is prepared using an analogous procedure to Example 51 by replacing (4-chloro-phe- nyl)-acetic acid with 3-phenyl-propionic acid. M+ 448. 45 Example 56

{2-[(3,5-Diamino-6-chloro-pyrazine-2-carbonyl)-amino]-ethyl}-dimethyl-[3-(4-methyl-benzoylamino)-propyl]-ammonium hexafluorophosphate 50 [0134] The title compound is prepared using an analogous procedure to Example 51 by replacing (4-chloro-phe- nyl)-acetic acid with 4-methyl-benzoic acid. M+ 434.

55

52 EP 2 300 010 B1

Example 57

(3-Benzoylamino-propyl)-{2-[(3,5-diamino-6-chloro-pyrazine-2-carbonyl)-amino]-ethyl}-dimethyl-ammonium hexafluor- ophosphate 5 [0135] The title compound is prepared using an analogous procedure to Example 51 by replacing (4-chloro-phe- nyl)-acetic acid with benzoic acid. M+ 420

Example 58 10 {2-[(3,5-Diamino-6-chloro-pyrazine-2-carbonyl)-amino]-ethyl}-{3-[2-(4-fluoro-phenyl)-acetylamino]-propyl}-dimethyl- ammonium hexafluorophosphate

[0136] The title compound is prepared using an analogous procedure to Example 51 by replacing (4-chloro-phe- 15 nyl)-acetic acid with (4-fluoro-phenyl)-acetic acid. M+ 452.

Example 59

{2-[(3,5-Diamino-6-chloro-pyrazine-2-carbonyl)-amino]-ethyl}-[3-(2-1H-indol-3-yl-acetylamino)-propyl]-dimethyl-ammo- 20 nium hexafluorophosphate

[0137] The title compound is prepared using an analogous procedure to Example 51 by replacing (4-chloro-phe- nyl)-acetic acid with (1H-indol-3-yl)-acetic acid. M+ 473.

25 Example 60

{2-[(3,5-Diamino-6-chloro-pyrazine-2-carbonyl)-amino]-ethyl}-{3-[2-(4-methoxyphenyl)-acetylamino]-propyl}-dimethyl- ammonium hexafluorophosphate

30 [0138] The title compound is prepared using an analogous procedure to Example 51 by replacing (4-chloro-phe- nyl)-acetic acid with (4-methoxy-phenyl)-acetic acid. M+ 464.

Example 61

35 {3-[3-(4-Chloro-phenyl)-propionylamino]-propyl}-{2-[(3,5-diamino-6-chloro-pyrazine-2-carbonyl)-amino]-ethyl}-dime- thyl-ammonium hexafluorohosphate

[0139] The title compound is prepared using an analogous procedure to Example 51 by replacing (4-chloro-phe- nyl)-acetic acid with 3-(4-chloro-phenyl)-propionic acid. M+ 482. 40 Example 62

{2-[(3,5-Diamino-6-chloro-pyrazine-2-carbonyl)-amino]-ethyl}-dimethyl-[3-(3-p-tolyl-propionylamino)-propyl]-ammoni- um hexafluorophosphate 45 [0140] The title compound is prepared using an analogous procedure to Example 51 by replacing (4-chloro-phe- nyl)-acetic acid with 3-p-tolyl-propionic acid. M+ 462.

Example 63 50 {4-[2-(4-Benzyloxy-phenyl)-acetylamino]-butyl}-{2-[(3,5-diamino-6-chloro-pyrazine-2-carbonyl)-amino]-ethyl}-dimethyl- ammonium bromide

[0141] To a solution of 4-Benzyloxyphenyl acetic acid (0.013 g, 0.30 mmol) in DMF (1.5 mL) is added HATU (0.12 g, 55 0.3 mmol), N-methylmorpholine (0.13 mL, 1.2 mmol) and (4-amino-butyl)-{2-[(3,5-diamino-6-chloro-pyrazine-2-carbon- yl)-amino]-ethyl}-dimethyl-ammonium bromide (Intermediate N) (0.1 g, 0.30 mmol). The solution is stirred at RT for 2 h. The solvent is removed in vacuo and the residue is purified by reverse phase chromatography (Isolute™ C18, 0-100% + 1 acetonitrile in water with 0.1% TFA) to yield the title compound. M 535. H NMR (400 MHz, DMSO-d6) δ 8.25 (1H, t),

53 EP 2 300 010 B1

8.05 (1H, t), 7.25 (2H, br), 7.42-7.32 (5H, m), 7.17 (2H, d), 7.09 (2H, br), 6.92 (2H, d), 5.06 (2H, s), 3.61-3.53 (2H, m), 3.39-3.33 (4H, m), 3.32 (2H, s), 3.10-3.03 (2H, m), 3.02 (6H, s), 1.61-1.49 (2H, m), 1.44-1.33 (2H, m).

Example 64 5 {4-[3-(4-Benzyloxy-phenyl)-propionylamino]-butyl}-{2-[(3,5-diamino-6-chloropyrazine-2-carbonyl)-amino]-ethyl}-dime- thyl-ammonium hexafluorophosphate

[0142] The title compound is prepared using an analogous procedure to Example 63 by replacing 4-benzyloxypheny- 10 lacetic acid with 3-(4-benzyloxy-phenyl)-propionic acid. M+ 568.

Example 65

{4-[3-(4-Chloro-phenyl)-propionylamino]-butyl}-{2-[(3,5-diamino-6-chloro-pyrazine-2-carbonyl)-amino]-ethyl}-dimethyl- 15 ammonium hexafluorophosphate

[0143] The title compound is prepared using an analogous procedure to Example 63 by replacing 4-benzyloxypheny- lacetic acid with 3-(4-chloro-phenyl)-propionic acid. M+ 496.

20 Example 66

{2-[(3,5-Diamino-6-chloro-pyrazine-2-carbonyl)-amino]-ethyl}-dimethyl-[4-(3-p-tolyl-propionylamino)-butyl]-ammonium hexafluorophosphate

25 [0144] The title compound is prepared using an analogous procedure to Example 63 by replacing 4-benzyloxypheny- lacetic acid with 3-para-tolyl-propionic acid. M+ 476.

Example 67

30 {4-[2-(4-Chloro-phenyl)-acetylamino]-butyl}-{2-[(3,5-diamino-6-chloro-pyrazine-2-carbonyl)-amino]-ethyl}-dimethyl-am- monium hexafluorophosphate

[0145] The title compound is prepared using an analogous procedure to Example 63 by replacing 4-benzyloxypheny- lacetic acid with (4-chloro-phenyl)-acetic acid. M+ 482. 35 Example 68

{2-[(3,5-Diamino-6-chloro-pyrazine-2-carbonyl)-amino]-ethyl}-dimethyl-[4-(2-p-tolyl-acetylamino)-butyl]-ammonium hexafluorophosphate 40 [0146] The title compound is prepared using an analogous procedure to Example 63 by replacing 4-benzyloxypheny- lacetic acid with para-tolyl-acetic acid. M+ 463.

Example 69 45 {2-[(3,5-Diamino-6-chloro-pyrazine-2-carbonyl)-amino]-ethyl}-dimethyl-[4-(3-phenyl-propionylamino)-butyl]-ammonium hexafluorophosphate

[0147] The title compound is prepared using an analogous procedure to Example 63 by replacing 4-benzyloxypheny- 50 lacetic acid with 3-phenyl-propionic acid. M+ 462

Example 70

{2-[(3,5-Diamino-6-chloro-pyrazine-2-carbonyl)-amino]-ethyl}-{4-[2-(4-hydroxyphenyl)-acetylamino]-butyl}-dimethyl- 55 ammonium trifluoroacetate

[0148] The title compound is prepared using an analogous procedure to Example 63 by replacing 4-benzyloxypheny- lacetic acid with (4-hydroxy-phenyl)-acetic acid. M+ 464.

54 EP 2 300 010 B1

Example 71

{2-[(3,5-Diamino-6-chloro-pyrazine-2-carbonyl)-amino]-ethyl}-[4-(2-1H-indol-3-yl-acetylamino)-butyl]-dimethyl-ammo- nium hexafluorophosphate 5 [0149] The title compound is prepared using an analogous procedure to Example 63 by replacing 4-benzyloxypheny- lacetic acid with (1H-indol-3-yl)-acetic acid. M+ 487.

Example 72 10 {2-[(3,5-Diamino-6-chloro-pyrazine-2-carbonyl)-amino]-ethyl}-{4-[3-(4-methoxy-phenyl)-propionylamino]-butyl}-dime- thyl-ammonium hexafluorophosphate

[0150] The title compound is prepared using an analogous procedure to Example 63 by replacing 4-benzyloxypheny- 15 lacetic acid with 3-(4-methoxy-phenyl)-propionic acid. M+ 492.

Example 73

[4-(4-Chloro-benzoylamino)-butyl]-{2-[(3,5-diamino-6-chloro-pyrazine-2-carbonyl)-amino]-ethyl}-dimethyl-ammonium 20 hexafluorophosphate

[0151] The title compound is prepared using an analogous procedure to Example 63 by replacing 4-benzyloxypheny- lacetic acid with 4-chloro-benzoic acid. M+ 468.

25 Example 74

{2-[(3,5-Diamino-6-chloro-pyrazine-2-carbonyl)-amino]-ethyl}-dimethyl-(4-phenylacetylamino-butyl)-ammonium hex- afluorophosphate

30 [0152] The title compound is prepared using an analogous procedure to Example 63 by replacing 4-benzyloxypheny- lacetic acid with phenyl-acetic acid. M+ 448.

Example 75

35 {2-[(3,5-Diamino-6-chloro-pyrazine-2-carbonyl)-amino]-ethyl}-dimethyl-[4-(4-methyl-benzoylamino)-butyl]-ammonium hexafluorophosphate

[0153] The title compound is prepared using an analogous procedure to Example 63 by replacing 4-benzyloxypheny- lacetic acid with 4-methyl-benzoic acid. M+ 448. 40 Example 76

{2-[(3,5-Diamino-6-chloro-pyrazine-2-carbonyl)-amino]-ethyl}-{4-[2-(4-methoxy-phenyl)-acetylamino]-butyl}-dimethyl- ammonium hexafluorophosphate 45 [0154] The title compound is prepared using an analogous procedure to Example 63 by replacing 4-benzyloxypheny- lacetic acid with (4-methoxy-phenyl)-acetic acid. M+ 478.

Example 77 50 (4-Benzoylamino-butyl)-{2-[(3,5-diamino-6-chloro-pyrazine-2-carbonyl)-amino]-ethyl}-dimethyl-ammonium hexafluoro- phosphate

[0155] The title compound is prepared using an analogous procedure to Example 63 by replacing 4-benzyloxypheny- 55 lacetic acid with benzoic acid. M+ 434.

55 EP 2 300 010 B1

Example 78

{5-[2-(4-Benzyloxy-phenyl)-acetylamino]-pentyl}-{2-[(3,5-diamino-6-chloro-pyrazine-2-carbonyl)-amino]-ethyl}-dime- thyl-ammonium bromide 5 [0156] A solution of (5-amino-pentyl)-{2-[(3,5-diamino-6-chloro-pyrazine-2-carbonyl)-amino]-ethyl}-dimethyl-ammoni- um bromide (Intermediate O) (1.0 g, 2.35 mmol), 4-benzyloxy phenyl acetic acid (0.57 g, 2.35 mmol), N-methylmorpholine (1.0 mL, 9.09 mmol), DCC (0.49 g, 2.35 mmol) and HOBt (0.32 g, 2.36 mmol) in DMF (20 mL) is stirred at RT for 1 h. The reaction mixture is concentrated in vacuo, and the residue is purified by column chromatography (basic alumina, 10 0-4% methanol in DCM) to afford a brown solid which is crystallised from methanol and EtOAc to afford the title compound. M+ 568.

Example 79

15 {5-[3-(4-Benzyloxy-phenyl)-propionylamino]-pentyl}-{2-[(3,5-diamino-6-chloro-pyrazine-2-carbonyl)-ami- no]-ethyl}-dimethyl-ammonium hexafluorophosphate

[0157] The title compound is prepared using an analogous procedure to Example 78 by replacing 4-benzyloxypheny- lacetic acid with 3-(4-benzyloxy-phenyl)-propionic acid. M+ 582. 20 Example 80

{5-[3-(4-Chloro-phenyl)-propionylamino]-pentyl}-{2-[(3,5-diamino-6-chloro-pyrazine-2-carbonyl)-amino]-ethyl}-dime- thyl-ammonium hexafluorophosphate 25 [0158] The title compound is prepared using an analogous procedure to Example 78 by replacing 4-benzyloxypheny- lacetic acid with 3-(4-chloro-phenyl)-propionic acid. M+ 510.

Example 81 30 [5-(4-Chloro-benzoylamino)-pentyl]-{2-[(3,5-diamino-6-chloro-pyrazine-2-carbonyl)-amino]-ethyl}-dimethyl-ammonium hexafluorophosphate

[0159] The title compound is prepared using an analogous procedure to Example 78 by replacing 4-benzyloxypheny- 35 lacetic acid with 4-chloro-benzoic acid. M+ 482.

Example 82

{2-[(3,5-Diamino-6-chloro-pyrazine-2-carbonyl)-amino]-ethyl}-{5-[4-(4-methoxy-phenyl)-butyrylamino]-pentyl}-dime- 40 thyl-ammonium hexafluorophosphate

[0160] The title compound is prepared using an analogous procedure to Example 78 by replacing 4-benzyloxypheny- lacetic acid with 4-(4-methoxy-phenyl)-butyric acid. M+ 520.

45 Example 83

{2-[(3,5-Diamino-6-chloro-pyrazine-2-carbonyl)-amino]-ethyl}-dimethyl-[5-(3-p-tolyl-propionylamino)-pentyl]-ammoni- um hexafluorophosphate

50 [0161] The title compound is prepared using an analogous procedure to Example 78 by replacing 4-benzyloxypheny- lacetic acid with 3-p-tolyl-propionic acid. M+ 490.

Example 84

55 {2-[(3,5-Diamino-6-chloro-pyrazine-2-carbonyl)-amino]-ethyl}-{5-[3-(4-methoxy-phenyl)-propionylamino]-pentyl}-dime- thyl-ammonium hexafluorophosphate

[0162] The title compound is prepared using an analogous procedure to Example 78 by replacing 4-benzyloxypheny-

56 EP 2 300 010 B1

lacetic acid with 3-(4-methoxy-phenyl)-propionic acid. M+ 506.

Example 85

5 {5-[2-(4-Chloro-phenyl)-acetylamino]-pentyl}-{2-[(3,5-diamino-6-chloro-pyrazine-2-carbonyl)-amino]-ethyl}-dimethyl- ammonium hexafluorophosphate

[0163] The title compound is prepared using an analogous procedure to Example 78 by replacing 4-benzyloxypheny- lacetic acid with (4-chloro-phenyl)-acetic acid. M+ 496. 10 Example 86

{2-[(3,5-Diamino-6-chloro-pyrazine-2-carbonyl)-amino]-ethyl}-dimethyl-[5-(2-p-tolyl-acetylamino)-pentyl]-ammonium hexafluorophosphate 15 [0164] The title compound is prepared using an analogous procedure to Example 78 by replacing 4-benzyloxypheny- lacetic acid with p-tolyl-acetic acid. M+ 476.

Example 87 20 {2-[(3,5-Diamino-6-chloro-pyrazine-2-carbonyl)-amino]-ethyl}-dimethyl-[5-(4-methyl-benzoylamino)-pentyl]-ammonium hexafluorophosphate

[0165] The title compound is prepared using an analogous procedure to Example 78 by replacing 4-benzyloxypheny- 25 lacetic acid with 4-methyl-benzoic acid. M+ 462.

Example 88

{2-[(3,5-Diamino-6-chloro-pyrazine-2-carbonyl)-amino]-ethyl}-dimethyl-[5-(3-phenyl-propionylamino)-pentyl]-ammoni- 30 um hexafluorophosphate

[0166] The title compound is prepared using an analogous procedure to Example 78 by replacing 4-benzyloxypheny- lacetic acid with 3-phenyl-propionic acid. M+ 476.

35 Example 89

{2-[(3,5-Diamino-6-chloro-pyrazine-2-carbonyl)-amino]-ethyl}-dimethyl-(5-phenylacetylamino-pentyl)-ammonium hex- afluorophosphate

40 [0167] The title compound is prepared using an analogous procedure to Example 78 by replacing 4-benzyloxypheny- lacetic acid with phenyl-acetic acid. M+ 462.

Example 90

45 {2-[(3,5-Diamino-6-chloro-pyrazine-2-carbonyl)-amino]-ethyl}-{5-[2-(2,4-dimethoxy-phenyl)-acetylamino]-pentyl}-dime- thyl-ammonium hexafluorophosphate

[0168] The title compound is prepared using an analogous procedure to Example 78 by replacing 4-benzyloxypheny- lacetic acid with (2,4-dimethoxy-phenyl)-acetic acid. M+ 522. 50 Example 91

(5-Benzoylamino-pentyl)-{2-[(3,5-diamino-6-chloro-pyrazine-2-carbonyl)-amino]-ethyl}-dimethyl-ammonium hexafluor- ophosphate 55 [0169] The title compound is prepared using an analogous procedure to Example 78 by replacing 4-benzyloxypheny- lacetic acid with benzoic acid. M+ 448.

57 EP 2 300 010 B1

Example 92

{2-[(3,5-Diamino-6-chloro-pyrazine-2-carbonyl)-amino]-ethyl}-{5-[2-(3,4-dimethoxyphenyl)-acetylamino]-pentyl}-dime- thyl-ammonium hexafluorophosphate 5 [0170] The title compound is prepared using an analogous procedure to Example 78 by replacing 4-benzyloxypheny- lacetic acid with (3,4-dimethoxy-phenyl)-acetic acid. M+ 522.

Example 93 10 {2-[(3,5-Diamino-6-chloro-pyrazine-2-carbonyl)-amino]-ethyl}-{5-[3-(3,4-dimethoxyphenyl)-propionylami- no]-pentyl}-dimethyl-ammonium hexafluorophosphate

[0171] The title compound is prepared using an analogous procedure to Example 78 by replacing 4-benzyloxypheny- 15 lacetic acid with 3-(3,4-dimethoxy-phenyl)-propionic acid. M+ 536.

Example 94

[5-(4-Chloro-benzenesulfonylamino)-pentyl]-{2-[(3,5-diamino-6-chloro-pyrazine-2-carbonyl)-amino]-ethyl}-dimethyl- 20 ammonium bromide

[0172] A mixture of (5-amino-pentyl)-{2-[(3,5-diamino-6-chloro-pyrazine-2-carbonyl)-amino]-ethyl}-dimethyl-ammoni- um bromide (Intermediate O) (0.25 g, 0.59 mmol), 4-chlorobenzenesulfonyl chloride (0.13 g, 0.59 mmol) and N-meth- ylmorpholine (0.30 mL, 2.73 mmol) in DMF (5 mL) is stirred at RT for 16 h. The reaction mixture is concentrated in vacuo 25 and is purified by column chromatography (basic alumina, 0-10% MeOH in DCM). The solid that is obtained is dissolved in a minimum amount of MeOH and DCM then precipitated by the addition of diethyl ether. The supernatant solvent mixture is decanted and the product is washed again with diethyl ether then dried under vacuum to give the title compound. + 1 M 578. H NMR (400 MHz, DMSO-d6) δ 8.25 (1H, t), 7.78 (2H, d), 7.74 (1H, t), 7.57 (2H, br), 7.67 (2H, d), 7.11 (2H, br), 3.56 (2H, q), 3.36 (2H, t), 3.28-3.22 (2H, m), 3.04 (6H, s), 2.95 (2H, q), 1.65-1.58 (2H, m), 1.37 (2H, quintet), 1.24-1.1 6 30 (2H, m).

Example 95

[5-(4-Chloro-phenylmethanesulfonylamino)-pentyl]-{2-[(3,5-diamino-6-chloropyrazine-2-carbonyl)-amino]-ethyl}-dime- 35 thyl-ammonium bromide

[0173] The title compound is prepared using an analogous procedure to Example 94 by replacing 4-chlorobenzenesul- fonyl chloride with (4-chloro-phenyl)-methanesulfonyl chloride. M+ 533.

40 Example 96

{2-[(3,5-Diamino-6-chloro-pyrazine-2-carbonyl)-amino]-ethyl}-dimethyl-(5-p-tolylmethanesulfonylamino-pentyl)-ammo- nium bromide

45 [0174] The title compound is prepared using an analogous procedure to Example 94 by replacing 4-chlorobenzenesul- fonyl chloride with p-tolyl-methanesulfonyl chloride. M + 512.

Example 97

50 {2-[(3,5-Diamino-6-chloro-pyrazine-2-carbonyl)-amino]-ethyl}-dimethyl-[5-(toluene-4-sulfonylamino)-pentyl]-ammoni- um bromide

[0175] The title compound is prepared using an analogous procedure to Example 94 by replacing 4-chlorobenzenesul- fonyl chloride with 4-methyl-benzenesulfonyl chloride. M+ 498. 55

58 EP 2 300 010 B1

Example 98

{2-[(3,5-Diamino-6-chloro-pyrazine-2-carbonyl)-amino]-ethyl}-dimethyl-(5-phenylmethanesulfonylamino-pentyl)-am- monium bromide 5 [0176] The title compound is prepared using an analogous procedure to Example 94 by replacing 4-chlorobenzenesul- fonyl chloride with phenyl-methanesulfonyl chloride. M+ 498.

Example 99 10 (5-Benzenesulfonylamino-pentyl)-{2-[(3,5-diamino-6-chloro-pyrazine-2-carbonyl)-amino]-ethyl}-dimethyl-ammonium bromide

[0177] The title compound is prepared using an analogous procedure to Example 94 by replacing 4-chlorobenzenesul- 15 fonyl chloride with benzenesulfonyl chloride. M+ 484.

Example 100

[4-(4-Chloro-benzenesulfonylamino)-butyl]-{2-[(3,5-diamino-6-chloro-pyrazine-2-carbonyl)-amino]-ethyl}-dimethyl-am- 20 monium bromide

[0178] A solution of (4-amino-butyl)-{2-[(3,5-diamino-6-chloro-pyrazine-2-carbonyl)-amino]-ethyl}-dimethyl-ammoni- um bromide (Intermediate N) (0.25 g, 0.60 mmol), 4-chlorobenzenesulfonyl chloride (0.125 g, 0.59 mmol) and N-meth- ylmorpholine (0.3 mL, 2.73 mmol) in DMF (5 mL) is shaken at RT for 16 h. The reaction mixture is concentrated in vacuo, 25 and the residue is purified by chromatography (basic alumina, 0-15% methanol in DCM) to obtain the title compound. M+ 504.

Example 101

30 {2-[(3,5-Diamino-6-chloro-pyrazine-2-carbonyl)-amino]-ethyl}-dimethyl-[4-(toluene-4-sulfonylamino)-butyl]-ammonium bromide

[0179] The title compound is prepared using an analogous procedure to Example 100 by replacing 4-chlorobenze- nesulfonyl chloride with 4-methyl-benzenesulfonyl chloride. M+ 484 35 Example 102

[4-(4-Chloro-phenylmethanesulfonylamino)-butyl]-{2-[(3,5-diamino-6-chloro-pyrazine-2-carbonyl)-amino]-ethyl}-dime- thyl-ammonium bromide 40 [0180] The title compound is prepared using an analogous procedure to Example 100 by replacing 4-chlorobenze- nesulfonyl chloride with (4-chloro-phenyl)-methanesulfonyl chloride. M+ 518.

Example 103 45 {2-[(3,5-Diamino-6-chloro-pyrazine-2-carbonyl)-amino]-ethyl}-dimethyl-(4-phenylmethanesulfonylamino-butyl)-ammo- nium bromide

[0181] The title compound is prepared using an analogous procedure to Example 100 by replacing 4-chlorobenze- 50 nesulfonyl chloride with phenyl-methanesulfonyl chloride. M + 484.

Example 104

{2-[(3,5-Diamino-6-chloro-pyrazine-2-carbonyl)-amino]-ethyl}-dimethyl-(4-p-tolylmethanesulfonylamino-butyl)-ammoni- 55 um bromide

[0182] The title compound is prepared using an analogous procedure to Example 100 by replacing 4-chlorobenze- nesulfonyl chloride with p-tolyl-methanesulfonyl chloride. M+ 498.

59 EP 2 300 010 B1

Example 105

(4-Benzenesulfonylamino-butyl)-{2-[(3,5-diamino-6-chloro-pyrazine-2-carbonyl)-amino]-ethyl}-dimethyl-ammonium bromide 5 [0183] The title compound is prepared using an analogous procedure to Example 100 by replacing 4-chlorobenze- nesulfonyl chloride with benzenesulfonyl chloride. M+ 470.

Example 106 10 {2-[(3,5-Diamino-6-chloro-pyrazine-2-carbonyl)-amino]-ethyl}-[4-(2,4-dimethoxy-benzenesulfonylamino)-butyl]-dime- thyl-ammonium trifluoroacetate

[0184] The title compound is prepared using an analogous procedure to Example 100 by replacing 4-chlorobenze- 15 nesulfonyl chloride with 2,4-dimethoxy-benzenesulfonyl chloride. M+ 530.

Example 107

{4-[4-(2-Cyano-ethoxy)-benzenesulfonylamino]-butyl}-{2-[(3,5-diamino-6-chloro-pyrazine-2-carbonyl)-ami- 20 no]-ethyl}-dimethyl-ammonium trifluoroacetate

[0185] The title compound is prepared using an analogous procedure to Example 100 by replacing 4-chlorobenze- nesulfonyl chloride with 4-(2-cyano-ethoxy)-benzenesulfonyl chloride. M + 539.

25 Example 108

[2-(4-Chloro-phenylmethanesulfonylamino)-ethyl]-{2-[(3,5-diamino-6-chloro-pyrazine-2-carbonyl)-amino]-ethyl}-dime- thyl-ammonium bromide

30 [0186] A solution of (2-amino-ethyl)-{2-[(3,5-diamino-6-chloro-pyrazine-2-carbonyl)-amino]-ethyl}-dimethyl-ammoni- um bromide (Intermediate K) (0.3 g, 0.78 mmol), (4-chlorophenyl)-methanesulfonyl chloride (0.18 g, 0.78 mmol) and N- methylmorpholine (0.35 mL, 3.18 mmol) in DMF (5 mL) is shaken at RT for 16 h. After this time the solvent is removed in vacuo and the residue is subjected to chromatography (basic alumina, 0-15% methanol in DCM) to yield the title compound. M+ 490. 35 Example 109

{2-[(3,5-Diamino-6-chloro-pyrazine-2-carbonyl)-amino]-ethyl}-dimethyl-(2-p-tolylmethanesulfonylamino-ethyl)-ammoni- um bromide 40 [0187] The title compound is prepared using an analogous procedure to Example 108 by replacing (4-chloro-phe- nyl)-methanesulfonyl chloride with p-tolyl-methanesulfonyl chloride. M+ 470.

Example 110 45 {2-[(3,5-Diamino-6-chloro-pyrazine-2-carbonyl)-amino]-ethyl}-dimethyl-[2-(toluene-4-sulfonylamino)-ethyl]-ammonium bromide

[0188] The title compound is prepared using an analogous procedure to Example 108 by replacing (4-chloro-phe- 50 nyl)-methanesulfonyl chloride with 4-methyl-benzenesulfonyl chloride. M + 456.

Example 111

[2-(4-Chloro-benzenesulfonylamino)-ethyl]-{2-[(3,5-diamino-6-chloro-pyrazine-2-carbonyl)-amino]-ethyl}-dimethyl-am- 55 monium bromide

[0189] The title compound is prepared using an analogous procedure to Example 108 by replacing (4-chloro-phe- nyl)-methanesulfonyl chloride with 4-chloro-benzenesulfonyl chloride. M + 476.

60 EP 2 300 010 B1

Example 112

{2-[(3,5-Diamino-6-chloro-pyrazine-2-carbonyl)-amino]-ethyl}-dimethyl-(2-phenylmethanesulfonylamino-ethyl)-ammo- nium bromide 5 [0190] The title compound is prepared using an analogous procedure to Example 108 by replacing (4-chloro-phe- nyl)-methanesulfonyl chloride with phenyl-methanesulfonyl chloride. M + 456.

Example 113 10 (2-Benzenesulfonylamino-ethyl)-{2-[(3,5-diamino-6-chloro-pyrazine-2-carbonyl)-amino]-ethyl}-dimethyl-ammonium bromide

[0191] The title compound is prepared using an analogous procedure to Example 108 by replacing (4-chloro-phe- 15 nyl)-methanesulfonyl chloride with benzenesulfonyl chloride. M + 442.

Example 114

{2-[(3,5-Diamino-6-chloro-pyrazine-2-carbonyl)-amino]-ethyl}-dimethyl-(3-p-tolylmethanesulfonylamino-propyl)-ammo- 20 nium bromide

[0192] A solution of (3-amino-propyl)-{2-[(3,5-diamino-6-chloro-pyrazine-2-carbonyl)-amino]-ethyl}-dimethyl-ammoni- um bromide (Intermediate M) (0.30 g, 0.756 mmol), p-tolyl-methanesulfonyl chloride (0.16 g, 0.76 mmol) and N-meth- ylmorpholine (0.4 mL, 3.64 mmol) in DMF (5 mL) is shaken at RT for 16 h. The solvent is removedin vacuo and the 25 residue is subjected to chromatography (basic alumina; 0-15% methanol in DCM) to yield the title compound. M + 484.

Example 115

[3-(4-Chloro-phenylmethanesulfonylamino)-propyl]-{2-[(3,5-diamino-6-chloropyrazine-2-carbonyl)-amino]-ethyl}-dime- 30 thyl-ammonium bromide

[0193] The title compound is prepared using an analogous procedure to Example 114 replacing p-tolyl-methanesulfonyl chloride with (4-chloro-phenyl)-methanesulfonyl chloride. M+ 504.

35 Example 116

[3-(4-Chloro-benzenesulfonylamino)-propyl]-{2-[(3,5-diamino-6-chloro-pyrazine-2-carbonyl)-amino]-ethyl}-dimethyl- ammonium bromide

40 [0194] The title compound is prepared using an analogous procedure to Example 114 replacing p-tolyl-methanesulfonyl chloride with 4-chloro-benzenesulfonyl chloride. M+ 490.

Example 117

45 {2-[(3,5-Diamino-6-chloro-pyrazine-2-carbonyl)-amino]-ethyl}-dimethyl-(3-phenylmethanesulfonylamino-propyl)-am- monium bromide

[0195] The title compound is prepared using an analogous procedure to Example 114 replacing p-tolyl-methanesulfonyl chloride with phenyl-methanesulfonyl chloride. M+ 470. 50 Example 118

{2-[(3,5-Diamino-6-chloro-pyrazine-2-carbonyl)-amino]-ethyl}-dimethyl-[3-(toluene-4-sulfonylamino)-propyl]-ammoni- um bromide 55 [0196] The title compound is prepared using an analogous procedure to Example 114 replacing p-tolyl-methanesulfonyl chloride with 4-methyl-benzenesulfonyl chloride. M+ 470.

61 EP 2 300 010 B1

Example 119

(3-Benzenesulfonylamino-propyl)-{2-[(3,5-diamino-6-chloro-pyrazine-2-carbonyl)-amino]-ethyl}-dimethyl-ammonium bromide 5 [0197] The title compound is prepared using an analogous procedure to Example 114 replacing p-tolyl-methanesulfonyl chloride with benzenesulfonyl chloride. M+ 456.

Example 120 10 ((S)-1-{[(3,5-Diamino-6-chloro-pyrazine-2-carbonyl)-amino]-methyl}-butyl)-[3-(4-methoxy-phenyl)-propyl]-dimethyl-am- monium trifluoroacetate

Step 1: [(S)-1-(1,3-Dioxo-1,3-dihydro-isoindol-2-ylmethyl)-butyl]-carbamic acid tert-butyl ester 15 [0198] To a mixture of phthalimide (1.43 g, 9.72 mmol), ((S)-1-hydroxymethyl-butyl)-carbamic acid tert-butyl ester (prepared according to the procedure described in US 2007/0032433 page 232) (1.97 g, 9.69 mmol) and triphenylphos- phine (2.55 g, 9.72 mmol) in DCM (25 mL) at 0 °C is added DEAD (1.6 mL, 10.2 mmol) dropwise. The reaction mixture

is stirred at RT overnight. The reaction mixture is adsorbed onto silica gel and purification by chromatograghy (SiO2, 20 0-15% EtOAc in iso-hexane) affords the title compound as white solid. [M+H]+ 233.

Step 2: ((S)-1-Aminomethyl-carbamic acid tert-butyl ester

[0199] A mixture of [(S)-1-(1,3-dioxo-1,3-dihydro-isoindol-2-ylmethyl)-butyl]-carbamic acid tert-butyl ester (2.80 g, 8.42 25 mmol) and hydrazine monohydrate (3.0 mL, 60.0 mmol) in EtOH (50 mL) and DCM (75 mL) is stirred at RT for 48 h. The precipitated solid is collected by filtration and washed with DCM to afford the crude product as a white solid that is used without further purification.

Step 3: ((S)-1-{[(3,5-Diamino-6-chloro-pyrazine-2-carbonyl)-amino]-methyl}-butyl)-carbamic acid tert-butyl ester 30 [0200] A mixture of ((S)-1-aminomethyl-butyl)-carbamic acid tert-butyl ester (1.70 g, 8.40 mmol), 3,5-diamino-6-chloro- pyrazine-2-carboxylic acid (1.59 g, 8.43 mmol), N-methylmorpholine (3.8 mL, 34.5 mmol) and HATU (3.2 g, 8.42 mmol) in anhydrous DMF (50 mL) is stirred at RT for 16 h. The reaction mixture is concentratedin vacuo and the resulting residue is purified by column chromatography (basic alumina, MeOH:DCM) to afford the title compound as yellow solid. 35 [M+H]+ 333.

Step 4: 3,5-Diamino-6-chloro-pyrazine-2-carboxylic acid ((S)-2-amino-pentyl)-amide hydrochloride

[0201] To a solution of ((S)-1-{[(3,5-diamino-6-chloro-pyrazine-2-carbonyl)-amino]-methyl}-butyl)-carbamic acid tert- 40 butyl ester (1.30 g, 3.49 mmol) in 1,4-dioxane (20 mL) is added HCl (50 mL of a 4 M solution in 1,4-dioxane, 200 mmol) and the reaction mixture is stirred at RT for 16 h. The reaction mixture is concentratedin vacuo and the yellow solid obtained is triturated with diethyl ether; the diethyl ether layer is decanted and the product is dissolved in a minimal volume of MeOH and is precipitated by the addition of diethyl ether. The solvent is decanted and the resulting solid is dried under vacuum to afford the title compound. 45 Step 5: 3,5-Diamino-6-chloro-pyrazine-2-carboxylic acid {(S)-2-[3-(4-methoxy-phenyl)-propylamino]-pentyl}-amide hy- drochloride

[0202] 3,5-Diamino-6-chloro-pyrazine-2-carboxylic acid ((S)-2-amino-pentyl)-amide hydrochloride (400 mg, 1.467 50 mmol), 3-(4-methoxyphenyl)propionaldehyde (181 mg, 1.103 mmol) and sodium triacetoxyborohydride (467 mg, 2.205 mmol) are dissolved in DCM (150 mL). The reaction mixture is stirred at reflux overnight. To the reaction mixture is added 1 M NaOH solution (50 mL) and DCM (50 mL). The organic phase is separated and the aqueous phase is extracted

once with DCM. The combined organic phases are washed with brine, dried (MgSO 4), filtered and concentrated in vacuo to afford a yellow solid. Purification by flash column chromatography (SiO 2, MeOH:DCM, gradient 2-10% + 0.1% TEA) 55 affords the title compound. [M+H]+ 421.

62 EP 2 300 010 B1

Step 6 : ((S)-1-{[(3,5-Diamino-6-chloro-pyrazine-2-carbonyl)-amino]-methyl}-butyl)-[3-(4-methoxy-phenyl)-pro- pyl]-dimethyl-ammonium trifluoroacetate

[0203] 3,5-Diamino-6-chloro-pyrazine-2-carboxylic acid {(S)-2-[3-(4-methoxy-phenyl)-propylamino]-pentyl}-amide hy- 5 drochloride (500 mg, 0.935 mmol) is dissolved in butan-2-one (50 mL). Potassium carbonate (646 mg, 4.67 mmol) and MeI (0.234 ml, 3.74 mmol) are added and the reaction mixture is stirred at 80 °C overnight. Further MeI (0.234 ml, 3.74 mmol) is added and reaction is heated at 80 °C for 24 h. The reaction mixture iss filtered through Celite™ washing with MeOH and the filterate concentrated. Purification by reverse phase chromatography (Isolute™ C18, 0-100% acetonitrile + 1 in water with 0.1% TFA) yields the title compound. M 449. H NMR (400 MHz, DMSO-d6) δ 8.39 (1H, t), 7.11 (2H, br), 10 7.01 (2H, d), 6.79 (2H, d), 5.76 (2H, br), 3.70 (3H, s), 3.59-3.41 (2H, m), 3.37-3.29 (1H, m), 3.04 (3H, s), 3.01 (3H, s), 2.55-2.34 (4H, m), 1.97-1.72 (3H, m), 1.61-1.36 (3H, m), 0.91 (3H, t).

Example 121

15 ((S)-1-{[(3,5-Diamino-6-chloro-pyrazine-2-carbonyl)-amino]-methyl}-butyl)-[3-(4-hydroxy-phenyl)-propyl]-dimethyl-am- monium trifluoroacetate

[0204] To a solution of ((S)-1-{[(3,5-diamino-6-chloro-pyrazine-2-carbonyl)-amino]-methyl}-butyl)-[3-(4-methoxy-phe-

nyl)-propyl]-dimethyl-ammonium trifluoroacetate (Example 120) (313 mg, 0.556 mmol) in DCM (100 mL) is added BBr 3 20 (3.34 mL of a 1M solution in heptane, 3.34 mmol). The reaction mixture is stirred at RT overnight. The reaction mixture is quenched cautiously by the addition of water then the organic solvents are removed in vacuo. Purification by reverse phase column chromatography (Isolute™ C18, 0-50% acetonitrile in water with 0.1% TFA) affords the title compound. + 1 M 435. H NMR (400 MHz, DMSO-d6) δ 8.40 (1H, t), 7.13 (2H, br), 6.89 (2H, d), 6.64 (2H, d), 5.29 (3H, br), 3.79-3.68 (1H, m), 3.57-3.43 (3H, m), 3.37-3.25 (1H, m), 3.04 (3H, s), 3.01 (3H, s), 2.41-2.30 (2H, m), 1.95-1.73 (3H, m), 1.64-1.35 25 (3H, m), 0.92 (3H, t).

Example 122

{2-[(3,5-Diamino-6-chloro-pyrazine-2-carbonyl)-amino]-ethyl}-bis-[3-(4-hydroxy-phenyl)-propyl]-methyl-ammonium trif- 30 luoroacetate

[0205] The title compound is prepared using an analogous procedure to Example 121 by replacing ((S)-1-{[(3,5- diamino-6-chloro-pyrazine-2-carbonyl)-amino]-methyl}-butyl)-[3-(4-methoxy-phenyl)-propyl]-dimethyl-ammonium trif- luoroacetate (Example 120) with {2-[(3,5-diamino-6-chloro-pyrazine-2-carbonyl)-amino]-ethyl}-bis-[3-(4-methoxy-phe- 35 nyl)-propyl]-methyl-ammonium trifluoroacetate (Example 20). M+ 513.

Example 123

{2-[(3,5-Diamino-6-chloro-pyrazine-2-carbonyl)-amino]-ethyl}-[3-(4-hydroxy-phenyl)-propyl]-dimethyl-ammonium bro- 40 mide

[0206] The title compound is prepared using an analogous procedure to Example 121 by replacing ((S)-1-{[(3,5- diamino-6-chloro-pyrazine-2-carbonyl)-amino]-methyl}-butyl)-[3-(4-methoxy-phenyl)-propyl]-dimethyl-ammonium trif- luoroacetate (Example 120) with {2-[(3,5-diamino-6-chloro-pyrazine-2-carbonyl)-amino]-ethyl}-[3-(4-methoxy-phe- 45 nyl)-propyl]-dimethyl-ammonium bromide (Example 27). M + 393.

Example 124

((S)-1-{[(3,5-Diamino-6-chloro-pyrazine-2-carbonyl)-amino]-methyl}-butyl)-{3-[4-((S)-2.3-dihydroxy-propoxy)-phe- 50 nyl]-propyl}-dimethyl-ammonium trifluoroacetate

[0207] Triethylamine (63.5 mL, 0.46 mmol) is added to a solution of ((S)-1-{[(3,5-Diamino-6-chloro-pyrazine-2-carbo- nyl)-amino]-methyl}-butyl)-[3-(4-hydroxy-phenyl)-propyl]-dimethyl-ammonium trifluoroacetate (Example 121)(50 mg, 0.091 mmol) in EtOH (5 mL) and the solution is heated to reflux. Once at reflux (S)-glycidol (6.04m L, 0.091 mmol) is 55 added and the reaction is heated at reflux overnight. Two further portions of (S)-glycidol (2 x 6.04 ml, 2 x 0.091 mmol) are added and the reaction is heated at reflux for 24 h. The reaction mixture is concentrated in vacuo and purification by reverse phase chromatography (Isolute™ C18, 0-30% acetonitrile in water with 0.1% TFA) affords the title compound. + 1 M 509. H NMR (400 MHz, DMSO-d6) δ 8.39 (1H, t), 8.20-7.18 (2H, br), 7.24-6.98 (2H, br), 6.99 (2H, d), 6.79 (2H, d),

63 EP 2 300 010 B1

5.70-3.98 (2H, br), 3.93-3.89 (1H, m), 3.85-3.67 (3H, m), 3.60-3.43 (3H, m), 3.42 (2H, d), 3.39-3.28 (1H, m), 3.04 (3H, s), 3.01 (3H, s), 2.48-2.32 (2H, m), 2.00-1.71 (3H, m), 1.61-1.37 (3H, m), 0.91 (3H, t).

Example 125 5 {2-[(3,5-Diamino-6-chloro-pyrazine-2-carbonyl)-amino]-ethyl}-bis-[3-[4-(2,3-dihydroxy-propoxy)-phenyl]-propyl}-me- thyl-ammonium trifluoroacetate

[0208] The title compound is prepared using an analogous procedure to Example 124 by replacing ((S)-1-{[(3,5- 10 diamino-6-chloro-pyrazine-2-carbonyl)-amino]-methyl}-butyl)-[3-(4-hydroxy-phenyl)-propyl]-dimethyl-ammonium trif- luoroacetate (Example 121) with {2-[(3,5-diamino-6-chloro-pyrazine-2-carbonyl)-amino]-ethyl}-bis-[3-(4-hydroxy-phe- nyl)-propyl]-methyl-ammonium trifluoroacetate (Example 122) and by replacing (S)-glycidol with ( 6)-glycidol. M+ 661.

Example 126 15 {2-[(3,5-Diamino-6-chloro-pyrazine-2-carbonyl)-amino]-ethyl}-{3-[4-(2,3-dihydroxy-propoxy)-phenyl]-propyl-dimethyl- ammonium trifluoroacetate

[0209] The title compound is prepared using an analogous procedure to Example 124 by replacing ((S)-1-{[(3,5- 20 Diamino-6-chloro-pyrazine-2-carbonyl)-amino]-methyl}-butyl)-[3-(4-hydroxy-phenyl)-propyl]-dimethyl-ammonium trif- luoroacetate (Example 121) with {2-[(3,5-diamino-6-chloro-pyrazine-2-carbonyl)-amino]-ethyl}-[3-(4-hydroxy-phe- nyl)-propyl]-dimethyl-ammonium bromide (Example 123), and by replacing (S)-glycidol with ( 6)-glycidol. M+ 467.

Example 127 25 [3-(4-Benzyloxy-phenyl)-propyl]-{2-[(3,5-diamino-6-chloro-pyrazine-2-carbonyl)-amino]-ethyl}-dimethyl-ammonium trif- luoroacetate

[0210] The title compound is prepared using an analogous procedure to Example 126 by replacing (6)-glycidol with 30 benzyl bromide. M+ 483.

Example 128

Carboxymethyl-{2-[(3,5-diamino-6-chloro-pyrazine-2-carbonyl)-amino]-ethyl}-bis-[3-(4-methoxy-phenyl)-propyl]-am- 35 monium chloride

[0211] Lithium hydroxide (12 mg, 0.28 mmol) is added to a stirred suspension of {2-[(3,5-Diamino-6-chloro-pyrazine- 2-carbonyl)-amino]-ethyl}-methoxycarbonylmethyl-bis-[3-(4-methoxy-phenyl)-propyl]-ammonium trifluoroacetate (Ex- ample 24) (113 mg, 0.16 mmol) in methanol (9 mL), and the resulting solution is stirred at RT for 3 days. After this time 40 the pH of the solution is adjusted to 1 using 1 N HCl solution, and the solvent then removedin vacuo. The residue is washed with water (10 mL), and dried in vacuo to yield the title compound as an orange solid. M+ 585.

Example 129

45 {2-[(3,5-Diamino-6-chloro-pyrazine-2-carbonyl)-amino]-ethyl}-dimethyl-phenethyl-ammonium trifluoroacetate

[0212] A suspension of 3,5-Diamino-6-chloro-pyrazine-2-carboxylic acid (2-dimethylaminoethyl)-amide (Intermediate H) (100 mg, 0.39 mmol) and phenylethyl iodide (280 ml, 1.9 mmol) in acetonitrile (4 mL) is heated in the microwave at 150 °C for 1 h. The solvent is removed in vacuo to afford a brown oil which is purified by reverse phase chromatography 50 (Isolute™ C18, 0-100% acetonitrile in water with 0.1% TFA) to yield the title compound. M+ 363. 1H NMR (400 MHz, DMSO-d6) δ 8.36 (1H, t), 7.34-7.24 (5H, m), 7.13 (2H, br), 3.66 (2H, t), 3.58-3.50 (4H, m), 3.06-3.02 (2H, m), 3.17 (6H, s).

Example 130

55 {2-[(3,5-Diamino-6-chloro-pyrazine-2-carbonyl)-amino]-ethyl}-trimethyl-ammonium iodide

[0213] Methyl iodide (120 mL, 1.9 mmol) is added to a suspension of 3,5-Diamino-6-chloropyrazine-2-carboxylic acid (2-dimethylamino-ethyl)-amide (Intermediate H) (100 mg, 0.39 mmol) in DCM (4 mL), and the resultant solution is stirred

64 EP 2 300 010 B1

at RT for 2 h. The white solid which has formed is collected by filtration and washed with DCM (5 mL) to afford the title + 1 compound. M 273. H NMR (400 MHz, DMSO-d 6) δ 8.27 (1H, t), 7.09 (2H, br s), 3.61 (2H, m), 3.45 (2H, t), 3.11 (9H, s).

Example 131 5 (7-Carbamoyl-heptyl)-{2-[(3,5-diamino-6-chloro-pyrazine-2-carbonyl)-amino]-ethyl}-dimethyl-ammonium bromide

[0214] A solution of 3,5-Diamino-6-chloro-pyrazine-2-carboxylic acid (2-dimethylaminoethyl)-amide (Intermediate H) (100 mg, 0.39 mmol) and 8-Bromo-octanoic acid amide (Intermediate P) (130 mg, 0.6 mmol) in DMF (3 mL) is heated 10 at 70 °C for 3 days. After this time, the solvent is removed in vacuo to yield an orange oil, which is triturated with acetone (2 x 10 mL) to afford the title compound. M+ 400.

Example 132

15 {2-[(3,5-Diamino-6-chloro-pyrazine-2-carbonyl)-amino]-ethyl}-[3-(4methoxycarbonylmethoxy-phenyl)-propyl]-dimethyl- ammonium trifluoroacetate and Example 133

[3-(4-Carboxymethoxy-phenyl)-propyl]-{2-[(3,5-diamino-6-chloro-pyrazine-2-carbonyl)-amino]-ethyl}-dimethyl-ammoni- um trifluoroacetate 20 Step 1 : {4-[3-({2-[(3,5-Diamino-6-chloro-pyrazine-2-carbonyl)-amino]-ethyl}-methyl-amino)-propyl}-phenoxyl-acetic ac- id methyl ester

[0215] To a solution of 3,5-Diamino-6-chloro-pyrazine-2-carboxylic acid (2-methylaminoethyl)-amide (Intermediate E) 25 (0.97 g, 2.7 mmol) and [4-(3-Bromo-propyl)-phenoxy]-acetic acid methyl ester (Intermediate Q) (2.33 g, 8.12 mmol) in acetone (30 mL) is added sodium carbonate (0.86 g, 8.12 mmol), and the reaction is heated at reflux for 4 days. After this time, the reaction is allowed to cool to RT, filtered, and concentratedin vacuo. The residue is purified by reverse phase chromatography (Isolute™ C18, 0-100% acetonitrile in water with 0.1% TFA) to yield {4-[3-({2-[(3,5-Diamino-6- chloro-pyrazine-2-carbonyl)-amino]-ethyl}-methyl-amino)-propyl]-phenoxy}-acetic acid methyl ester. [M+H]+ 451. 30 Step 2: {2-[(3,5-Diamino-6-chloro-pyrazine-2-carbonyl)-amino]-ethyl}-[3-(4methoxycarbonylmethoxy-phenyl)-pro- pyl]-dimethyl-ammonium trifluoroacetate and

[3-(4-Carboxymethoxy-phenyl)-propyl]-{2-[(3,5-diamino-6-chloro-pyrazine-2-carbonyl)-amino]-ethyl}-dimethyl-ammoni- 35 um trifluoroacetate

[0216] To a solution of {4-[3-({2-[(3,5-Diamino-6-chloro-pyrazine-2-carbonyl)-amino]-ethyl}-methyl-amino)-pro- pyl]-phenoxy}-acetic acid methyl ester (0.45 g, 0.79 mmol) in acetone (10 mL) is added sodium carbonate (0.42 g, 3.98 mmol) and methyl iodide (0.24 ml, 3.98 mmol). The reaction is heated at reflux for 3 h, then allowed to cool to RT. The 40 reaction mixture is filtered, and the solid is washed with methanol. The methanol is removedin vacuo and seperation by reverse phase column chromatography (Isolute™ C18, 0-100% acetonitrile in water with 0.1% TFA) yields the title compounds. {2-[(3,5-Diamino-6-chloro-pyrazine-2-carbonyl)-amino]-ethyl}-[3-(4methoxycarbonylmethoxy-phenyl)-propyl]-dimethyl- + 1 ammonium trifluoroacetate (Example 133) M 465. H NMR (400 MHz, DMSO-d6) δ 8.30 (1H, t), 7.95 (2H, br s), 7.10 45 (2H, d), 6.82 (2H, d), 4.75 (2H, s), 3.60 (2H, t), 3.41 (2H, t), 3.34 (2H, m), 3.07 (6H, s), 2.98 (2H, m), 2.54 (3H, s), 1.94 (2H, m). [3-(4-Carboxymethoxy-phenyl)-propyl]-{2-[(3,5-diamino-6-chloro-pyrazine-2-carbonyl)-amino]-ethyl}-dimethyl-ammoni- + 1 um trifluoroacetate (Example 134) M 451. H NMR (400 MHz, CD3OD) δ 7.12 (2H, d), 6.85 (2H, d), 4.64 (2H, s), 3.74 (2H, t), 3.51 (2H, t), 3.41-3.38 (2H, m), 3.16 (6H, s), 2.59 (2H, t), 2.10 (2H, m). 50 Preparation of Intermediate Compounds

Intermediate A

55 3,5-Diamino-6-chloro-pyrazine-2-carboxylic acid ((S)-2-amino-3-phenyl-propyl)-amide

[0217] To a solution of 3,5-diamino-6-chloro-pyrazine-2-carboxylic acid (0.225 g, 1.2 mmol) and HATU (0.46 g, 1.2 mmol) in DMF (10 mL) is added N-methylmorpholine (0.53 mL, 4.8 mmol) and ((S)-t-aminomethyl-2-phenyl-ethyl)-car-

65 EP 2 300 010 B1

bamic acid tert-butyl ester (0.3 g, 1.2 mmol) and the resulting yellow solution is stirred at RT for 18 h. The solvent is removed in vacuo and the residue is diluted with water (10 mL). The precipitate which forms is collected by filtration and dissolved in DCM (5 mL). TFA (1 mL) is added and the solution stirred at RT for 1 h. The solvent is removed in vacuo, and the residue is purified by reverse phase chromatography (Isolute™ C18, 0-100% acetonitrile in water with 0.1% 5 TFA) to yield the title compound. [M+H]+ 321

Intermediate B

((S)-5-Amino-1-{[(3,5-diamino-6-chloro-pyrazine-2-carbonyl)-amino]-methyl}-pentyl)-carbamic acid tert-butyl ester: 10 Step 1: ((S)-5-Benzyloxycarbonylamino-1-{[(3,5-diamino-6-chloro-pyrazine-2-carbonyl)-amino]-methyl}-pentyl)-car- bamic acid tert-butyl ester:

[0218] To a solution of 3,5-diamino-6-chloro-pyrazine-2-carboxylic acid (0.39 g, 2.05 mmol) and HATU (0.78 g, 2.05 15 mmol) in DMF (10 mL) is added N-methylmorpholine (0.83 g, 8.22 mmol) followed by ((S)-1-aminomethyl-5-benzyloxy- carbonylamino-pentyl)-carbamic acid tert-butyl ester (prepared according to the procedure described in WO1997/01204 page 20) (750 mg, 2.05 mmol). The resulting solution is stirred at RT for 18 h and then concentrated in vacuo to afford an orange oil. The oil is dissolved in MeOH (10 mL) and allowed to stand at RT after which time a cream precipitate forms that is collected by filtration and dried under vacuum to yield the title compound. [M+H] + 536. 20 Step 2: ((S)-5-Amino-1-{[(3,5-diamino-6-chloro-pyrazine-2-carbonyl)-amino]-methyl}-pentyl)-carbamic acid tert-butyl es- ter:

[0219] A suspension of ((S)-5-benzyloxycarbonylamino-1-{[(3,5-diamino-6-chloro-pyrazine-2-carbonyl)-amino]-me- 25 thyl}-pentyl)-carbamic acid tert-butyl ester (0.68 g, 1.27 mmol) in EtOH (20 mL) under an inert atmosphere of2 Nis treated with activated palladium on charcoal (10%). The reaction mixture is then placed under a positive pressure of hydrogen and stirred at RT. After 3 h, the catalyst is removed by filtration through Celite® (filter material). The filtrate is concentrated in vacuo and the resulting colourless oil is dissolved in MeOH (10 mL) and allowed to stand at RT overnight. A cream precipitate forms which is removed by filtration and the solution is concentrated in vacuo. Purification by reverse 30 phase column chromatography (Isolute™ C18, 0-100% acetonitrile in water with 0.1% NH 3) affords the title compound. [M+H]+ 402.

Intermediate C

35 3,5-Diamino-6-chloro-pyrazine-2-carboxylic acid (2-amino-ethyl)-amide

[0220] 3,5-Diamino-6-chloro-pyrazine-2-carboxylic acid methyl ester (2.0 g, 9.88 mmol) in neat ethylene diamine (18 mL) is heated using microwave irradiation in a Personal Chemistry Emrys™ Optimizer microwave reactor at 130 °C for 1 h at 4 bar. The solvent is removed in vacuo, water (10 mL) is added to the orange solid and the suspension is then 40 cooled to 0 °C. The off-white solid is collected by filtration and dried under vacuum at 50 °C to afford the title compound. [M+H]+ 231.

Intermediate D

45 3,5-Diamino-6-chloro-pyrazine-2-carboxylic acid (2-{bis-[3-(4-methoxy-phenyl)-propyl]-amino}-ethyl)-amide trifluoroac- etate

[0221] To a solution of 3,5-diamino-6-chloro-pyrazine-2-carboxylic acid (2-amino-ethyl)-amide (Intermediate C) (0.90 g, 3.91 mmol) in acetone (20 mL) is added 1-(3-bromo-propyl)-4-methoxy-benzene (2.30 g, 10.0 mmol) and sodium 50 carbonate (1.04 g, 10.0 mmol). The reaction mixture is heated at 45 °C for 48 h. After cooling to RT the mixture is filtered to remove the inorganic salts and the solvent removed in vacuo. Purification by reverse phase column chromatography (Isolute™ C18, 0-100% acetonitrile in water with 0.1% TFA) affords the title compound. [M+H]+ 527.

55

66 EP 2 300 010 B1

Intermediate E

3,5-Diamino-6-chloro-pyrazine-2-carboxylic acid (2-methylamino-ethyl)-amide trifluoroacetate

5 Step 1: {2-[(3,5-Diamino-6-chloro-pyrazine-2-carbonyl)-amino]-ethyl}-methyl-carbamic acid tert-butyl ester:

[0222] A solution of 3,5-diamino-6-chloro-pyrazine-2-carboxylic acid (3.85 g, 20.5 mmol) in DMF (90 mL) is treated with CDI (6.64 g, 41 mmol) and stirred at RT for 1 h. To this mixture is added N-(2-aminoethyl)-N-methyl carbamic acid tert-butyl ester (5.0 g, 28.6 mmol) and stirring continues at RT for 72 h. The solvent is removed in vacuo and the residue 10 is partitioned between EtOAc (100 mL) and water (60 mL). The organic portion is separated, dried (MgSO4) and con- centrated in vacuo. The resulting yellow residue is washed with diethyl ether to afford the title compound which is used in the next step without further purification. [M+H] + 345.

Step 2: 3,5-Diamino-6-chloro-pyrazine-2-carboxylic acid (2-methylamino-ethyl)-amide trifluoroacetate 15 [0223] A mixture comprising {2-[(3,5-diamino-6-chloro-pyrazine-2-carbonyl)-amino]-ethyl}-methyl-carbamic acid tert- butyl ester in DCM:TFA (60 mL of a 5:1 DCM/TFA solution) is stirred at RT for 3 h. The solvent is removed in vacuo and the resulting yellow oil is dissolved in water with heating. Any undissolved material is removed by filtration and the filtrate is concentrated in vacuo to afford the title compound. [M+H]+ 245. 20 Intermediate F

3,5-Diamino-6-chloro-pyrazine-2-carboxylic acid {2-[(4-amino-butyl)-methyl-amino]-ethyl}-amide trifluoroacetate

25 Step 1:

[4-({2-[(3,5-Diamino-6-chloro-pyrazine-2-carbonyl)-amino]-ethyl}-methyl-amino)-butyl]-carbamic acid tert-butyl ester:

[0224] To a mixture comprising 3,5-diamino-6-chloro-pyrazine-2-carboxylic acid (2-methylamino-ethyl)-amide (Inter- 30 mediate E) (0.5 g, 2.0 mmol) and 4-(Boc-amino)butyl bromide (0.8 g, 3.1 mmol) in DMF (5 mL) is added sodium carbonate (0.4 g, 4.0 mmol). The resulting mixture is heated at reflux for 3 h and then allowed to cool to RT, filtered and concentrated in vacuo. The crude product is dissolved in DCM and filtered to remove any undissolved impurities. The filtrate is concentrated in vacuo to yield the title compound which is used in the next step without further purification.

35 Step 2: 3,5-Diamino-6-chloro-pyrazine-2-carboxylic acid {2-[(4-amino-butyl)-methyl-amino]-ethyl}-amide:

[0225] [4-({2-[(3,5-Diamino-6-chloro-pyrazine-2-carbonyl)-amino]-ethyl}-methyl-amino)-butyl]-carbamic acid tert-butyl ester (0.54 g, 2.2 mmol) in DCM (10 mL) and TFA (2 mL) is stirred at RT for 1 h. The solvent is removed in vacuo and the resulting oil is dissolved in saturated aqueous sodium carbonate solution. The mixture is concentrated in vacuo and 40 the residue is dissolved in MeOH and filtered to remove the sodium carbonate. The filtrate is concentrated in vacuo to afford the title compound. [M+H]+ 316

Intermediate G

45 1-Ethoxy-2-chloro-4-isocyanato-benzene

[0226] 1-Ethoxy-2-chloro-4-isocyanato-benzene is prepared by addition of triethylamine to a solution of 4-ethoxy-3- chlorophenylamine in EtOAc at 0-5 °C. Triphosgene is added and the reaction mixture is slowly heated to reflux over 2 h. The solvent is removed in vacuo and the crude product is filtered through silica to afford the title compound. 50 Intermediate H

3,5-Diamino-6-chloro-pyrazine-2-carboxylic acid (2-dimethylamino-ethyl)-amide

55 [0227] 3,5-Diamino-6-chloro-pyrazine-2-carboxylic acid methyl ester (25.0 g, 0.12 mol) and N,N-dimethylethylenedi- amine (100 mL) are heated at 100 °C in a miniclave for 20 h. The reaction mixture is concentrated in vacuo and the pale yellow crystals that form are collected by filtration, washed with MeOH and dried under vacuum to afford the title com- pound. [M+H]+ 259.

67 EP 2 300 010 B1

Intermediate I

1-(2-Chloro-ethyl)-3-(4-isobutoxy-phenyl)-urea

5 [0228] A solution of 2-chloroethyl isocyanate (15 g, 142 mmols) is added dropwise to a solution of 4-isobutoxy phe- nylamine (20 g, 121 mmols) in diethyl ether (100 mL) and stirred at RT for 1 h. The solid formed is collected by filtration and recrystallisation from ethanol affords the title compound.

Intermediate J 10 (S)-3-(4-{3-[4-(3-Bromo-propyl)-phenoxy]-propyl}-phenoxy)-propane-1.2-diol

Step 1: (S)-3-[4-(3-Hydroxy-propyl)-phenoxy]-propane-1.2-diol

15 [0229] To a solution of 3-(4-hydroxyphenyl)-1-propanol (15.0 g, 99 mmol) in acetone (300 mL) is added S-(-)-glycidol (10.0 g, 105 mmol) and potassium carbonate (20.0 g, 145 mmol) and the reaction mixture is heated at reflux for 24 h. The resulting mixture is allowed to cool to RT, filtered and the filtrate is concentrated in vacuo. Purification by column + chromatography (SiO2, 0-4% MeOH in DCM) affords the product as a white solid. [M+H] 227. [0230] Step 2: 3-[4-((R)-2,2-Dimethyl-[1,3]dioxolan-4-ylmethoxy)-phenyl]-propan-1-ol To a solution of (S)-3-[4-(3-hy- 20 droxy-propyl)-phenoxy]-propane-1,2-diol (6.5 g, 28.7 mmol) in anhydrous DMF (100 mL) is added 2,2-dimethoxypropane (15.0 mL, 122.0 mmol) and pyridinium p-toluenesulfonate (0.75 g, 2.98 mmol). The reaction mixture is stirred at RT for 16 h and then concentrated in vacuo. The residue is dissolved in EtOAc and the organic portion is washed with 10%

aqueous NaHCO3 solution, water, brine and dried over Na 2SO4. The solvent is removed in vacuo and the crude product is purified by column chromatography (SiO2, EtOAc in petroleum ether, gradient 0-30% EtOAc) to obtain the title com- 25 pound as a colourless oil. [M+H]+ 267.

Step 3: (R)-4-(4-{3-[4-(3-Bromo-propyl)-phenoxy]-propyl}-phenoxymethyl)-2,2-dimethyl-[1,3]dioxolane

[0231] To a mixture of 3-[4-((R)-2,2-dimethyl-[1,3]dioxolan-4-ylmethoxy)-phenyl]-propan-1-ol (6.2 g, 23.3 mmol), 4-(3- 30 bromo-propyl)-phenol (prepared according to the procedure described in WO2006/097744, page 30) (5.0 g, 23.3 mmol) and triphenylphosphine (6.1 g, 23.3 mmol) in DCM (75 mL) at 0 °C is added DEAD (3.7 mL, 23.5 mmol) dropwise and the resulting mixture is stirred at RT overnight. The solvent is removed in vacuo and the residue is triturated with diethyl ether. The precipitated solid is removed by filtration and the filtrate is concentratedin vacuo. Purification of the crude residue by column chromatography (SiO2, EtOAc in petroleum ether, gradient 0-4% EtOAc) affords the title compound 35 as white solid.

Step 4: (S)-3-(4-{3-[4-(3-Bromo-propyl)-phenoxy]-propyl}-phenoxy)-propane-1,2-diol

[0232] To a solution of (R)-4-(4-{3-[4-(3-bromo-propyl)-phenoxy]-propyl}-phenoxymethyl)-2,2-dimethyl-[1,3]dioxolane 40 (5.0 g, 10.8 mmol) in DCM (50 mL) is added TFA (2.0 mL, 26.9 mmol) dropwise followed by water (2.0 mL) and the reaction mixture is stirred at RT overnight. The solvent is removedin vacuo and the residue is treated with aqueous sodium hydrogen carbonate solution. The mixture is extracted with EtOAc (3 x 200 mL) and the combined organic

portions are washed with water, brine, dried over Na 2SO4 and concentrated in vacuo. Recrystallizisation of the residue from diethyl ether-hexane affords the title compound as white solid. 45 Intermediate K

(2-Amino-ethyl)-{2-[(3,5-diamino-6-chloro-pyrazine-2-carbonyl)-amino]-ethyl}-dimethyl-ammonium bromide

50 Step 1: {2-[(3,5-Diamino-6-chloro-pyrazine-2-carbonyl)-amino]-ethyl}-[2-(1,3-dioxo-1,3-dihydro-isoindol-2- yl)-ethyl]-dimethyl-ammonium bromide

[0233] The titlecompound isprepared analogously toExample 27 by replacing 1-(3-bromo-propyl)-4-methoxy-benzene with N-(4-bromoethyl)-phthalimide. 55 Step 2: (2-Amino-ethyl)-{2-[(3,5-diamino-6-chloro-pyrazine-2-carbonyl)-amino]-ethyl}-dimethyl-ammonium bromide

[0234] A mixture of {2-[(3,5-diamino-6-chloro-pyrazine-2-carbonyl)-amino]-ethyl}-[2-(1,3-dioxo-1,3-dihydro-isoindol-2-

68 EP 2 300 010 B1

yl)-ethyl]-dimethyl-ammonium bromide (47 g, 92 mmol) and hydrazine monohydrate (20 mL, 412 mmol) in EtOH (500 mL) is heated at 40 °C for 4 h. After cooling to RT the resulting solid is removed by filtration and washed with diethyl ether. The filtrate is concentrated in vacuo to afford the title product. [M+H]+ 302.

5 Intermediate L

3-[3-(Diaminomethylene-sulfamoyl)-4-methoxy-phenyl]-3-phenyl-propionic acid

Step 1: 3-(4-Methoxy-3-sulfo-phenyl)-3-phenyl-propionic acid methyl ester 10 [0235] To a solution of 3-(4-methoxy-phenyl)-3-phenyl-propionic acid methyl ester (prepared according to the proce- dure reported in Tetrahedron (2006), 62(41), 9610-9621) (2.50 g, 9.25 mmol) in DCM (250 mL) at 0 °C is added chlo- rosulfonic acid (30.7 ml, 462.5 mmol). The reaction mixture is allowed to warm to RT and is stirred at this temperature for 2 h. Ice water is added to cause precipitation of the product. The solvents are decanted and the residual solid is 15 washed with ice water. The solid is then dried under vacuum to afford the title compound that is used without further purification.

Step 2: 3-(3-Chlorosulfonyl-4-methoxy-phenyl)-3-phenyl-propionic acid methyl ester

20 [0236] To a solution of 3-(4-methoxy-3-sulfo-phenyl)-3-phenyl-propionic acid methyl ester (850 mg, 2.43 mmol) in 1,3- dimethyl-2-imidazolidinone (25 mL) is added POCl 3 (0.57 ml, 6.06 mmol) and pyridine (0.33 mL, 4.13 mmol). The reaction mixture is stirred at RT for 1 h. After this time, water (50 mL) is added and the reaction mixture is extracted with DCM

(3 x 50mL). Thecombined organic phases are washed with brine(30 mL)then dried over MgSO 4,filtered andconcentrated in vacuo to afford the title compound as an oil. 25 Step 3: 3-[3-(Diaminomethylene-sulfamoyl)-4-methoxy-phenyl]-3-phenyl-propionic acid methyl ester

[0237] To a solution of 3-(3-chlorosulfonyl-4-methoxy-phenyl)-3-phenyl-propionic acid methyl ester (500 mg, 1.36 mmol) in DCM (50 mL) is added triethylamine (0.57 ml, 4.08mmol) and guanidine hydrochloride (0.14 g, 1.50 mmol). 30 The reaction mixture is stirred at RT overnight, then diluted with DCM (100 mL). The reaction mixture is washed with

an aqueous saturated solution of NaHCO 3 (30 mL), water (30 mL) then brine (30 mL). The organic phase is then dried over MgSO4, filtered and concentrated in vacuo. Purification of the residue by reverse phase chromatography (Isolute™ C18, 0-100% acetonitrile in water with 0.1% TFA) affords the title compound.

35 Step 4: 3-[3-(Diaminomethylene-sulfamoyl)-4-methoxy-phenyl]-3-phenyl-propionic acid

[0238] To a solution of 3-[3-(diaminomethylene-sulfamoyl)-4-methoxy-phenyl]-3-phenyl-propionic acid methyl ester (400 mg, 1.02 mmol) in a mixture of MeOH and water (30 mL of 1:1 solution) is added LiOH (73 mg, 3.06 mmol). The reaction mixture is stirred at RT overnight. 1 M aqueous HCl is added to neutralise the reaction mixture. The resulting 40 solid is collected by filtration and dried under vacuum to afford the title compound that is used without further purification.

Intermediate M

(3-Amino-propyl)-{2-[(3,5-diamino-6-chloro-pyrazine-2-carbonyl)-amino]-ethyl}-dimethyl-ammonium bromide 45 [0239] The title compound is prepared by an analogous procedure to (2-amino-ethyl)-{2-[(3,5-diamino-6-chloro-pyra- zine-2-carbonyl)-amino]-ethyl}-dimethyl-ammonium bromide (Intermediate K) by replacing N-(4-bromoethyl)-phthalim- ide with N-(4-bromopropyl)-phthalimide. M+ 316.

50 Intermediate N

(4-Amino-butyl)-{2-[(3,5-diamino-6-chloro-pyrazine-2-carbonyl)-amino]-ethyl}-dimethyl-ammonium bromide

[0240] The title compound is prepared by an analogous procedure to (2-amino-ethyl)-{2-[(3,5-diamino-6-chloro-pyra- 55 zine-2-carbonyl)-amino]-ethyl}-dimethyl-ammonium bromide (Intermediate K) by replacing N-(4-bromoethyl)-phthalim- ide with N-(4-bromobutyl)-phthalimide. M+ 330.

69 EP 2 300 010 B1

Intermediate O

(5-Amino-pentyl)-{2-[(3,5-diamino-6-chloro-pyrazine-2-carbonyl)-amino]-ethyl}-dimethyl-ammonium bromide

5 [0241] The title compound is prepared by an analogous procedure to (2-amino-ethyl)-{2-[(3,5-diamino-6-chloro-pyra- zine-2-carbonyl)-amino]-ethyl}-dimethyl-ammonium bromide (Intermediate K) by replacing N-(4-bromoethyl)-phthalim- ide with N-(4-bromopentyl)-phthalimide.

Intermediate P 10 8-Bromo octanamide

[0242] The title compound is prepared as described in ’Ortho esters with 2,4,10-Trioxa admanatane Structure as Carboxyl Protecting Groups; Applications in the Synthesis of Substituted Carboxylic Acids by Means of Grignard Rea- 15 gents’, G. Voss and H. Gerlach, Helvetica Chimica Acta, 1983, 66, 2294-2307, p2300

Intermediate Q

[4-(3-Bromo-propyl)-phenoxy]-acetic acid methyl ester 20 Step 1 : [4-(3-Hydroxy-propyl)-phenoxy]-acetic acid methyl ester

[0243] To a solution of 3-(4-hydroxyphenyl)-1-propanol (3 g, 19.7 mmol) in acetone (30 mL) is added potassium carbonate (4.08 g, 29.5 mmol) and methyl bromoacetate (2.8 mL, 29.3 mmol) and the reaction is heated at reflux for 15 25 h. The reaction mixture is allowed to cool to RT, filtered, and the solvent removed in vacuo. The residue is purified by 1 chromatography (SiO2, EtOAc: iso-hexane, 2:3) to afford the title compound. H NMR (400 MHz, CDCl 3) δ 7.14 (2H, d), 6.85 (2H, d), 4.64 (2H, s), 3.82 (3H, s), 3.69 (2H, t), 2.68 (2H, t), 1.82 (2H, m).

Step 2: [4-(3-Bromo-propyl)-phenoxy]-acetic acid methyl ester 30 [0244] To a solution of [4-(3-Hydroxy-propyl)-phenoxy]-acetic acid methyl ester (2.45 g, 10.9 mmol) in DCM (60 mL) is added polymer-supported triphenylphosphine (7.26 g, 21.8 mmol) and carbon tetrabromide (3.47 g, 11.99 mmol) and the resulting reaction mixture is stirred at RT for 15 h. The reaction mixture is filtered to remove the polymer-supported reagent, and then concentrated in vacuo. The residue is purified by chromatography (SiO 2, EtOAc: iso-hexane, 1:5) to 35 1 yield the title compound. H NMR (400 MHz, CDCl3) δ 7.14 (2H, d), 6.86 (2H, d), 4.69 (2H, s), 3.83 (3H, s), 3.40 (2H, t), 2.74 (2H, t), 2.15 (2H, m).

Claims 40 1. A compound having the formula Ia

45

50

or pharmaceutically acceptable salts, solvates or hydrates thereof, wherein 55 6 6a R is -(C0-C6 alkylene)-R , wherein the alkylene linker is optionally substituted by one or more groups selected from C1-C3 alkyl, halo and OH; 6a 11 12 13 14 R is selected from H, a C3-C10 carbocyclic group, NR R , C(O)NR R , C6-C10 aryl, 5- to 6-membered het- eroaryl, 5- to 6-membered heterocyclyl and a group of the formula P-(CH2)m-Q, wherein each of the ring systems

70 EP 2 300 010 B1

is optionally substituted by one or more substituents selected from the List Z; a a a P is selected from a bond, -O-, -C(O)-, -C(O)O-, -NHC(=N)NH- and -S(O 2)-, - S(O2)NR -, -NR C(O)-,-NR C(O)O-, a b -NR S(O2)- and -NRaC(O)NR -; a b R and R are each independently selected from H,1 -C C6 alkyl, a 3 C-C10 carbocyclic group and -(C1-C3 5 alkylene)-C3-C10 carbocyclic; Q is selected from H, C 1-C6 alkyl, a C3-C10 carbocyclic group, C6-C10 aryl, 5- to 6-membered heteroaryl and 5- to 6-membered heterocyclyl, wherein the carbocyclic, C6-C10 aryl, 5- to 6-membered heteroaryl and 5- to 6-membered heterocyclyl groups are each optionally substituted by one or more substituents selected from List Z; m is 0, 1, 2 or 3; 10 7 8 10 R , R and R are each independently selected from C 1-C6 alkyl, C 2-C6 alkenyl, C 2-C6 alkynyl, a C 3-C10 carbocyclic group, -(C1-C3 alkylene)-C3-C10 carbocyclic group, C6-C10 aryl, 5- to 6-membered heteroaryl, 5- to 6-membered heterocyclyl or a group of the formula (CH 2)d-A-(CH2)b-B, wherein the carbocyclic, C6-C10 aryl, 5- to 6-membered heteroaryl and 5- to 6-membered heterocyclyl groups are each optionally substituted by one or more substituents selected from List Z; and wherein the alkyl and alkylene groups are optionally substituted by one or more halogen 15 atoms, OH groups or phenyl groups; or R7 and R 8, together with the nitrogen atom to which they are attached form a 4- to 10-membered nitrogen-containing heterocyclic group which is optionally substituted by one or more substituents selected from the List Z; or R6 and R8, together with the atoms to which they are attached form a 4- to 10-membered nitrogen-containing heterocyclic group which is optionally substituted by one or more substituents selected from the List Z; 20 c c A is selected from a bond, -O-, -C(O)-, -C(O)NR-, -C(O)O-, -OC(O)-, -NHC(=N)NH-, -S(O2)-, -S(O2)NR -, c c c c c c d c -NR C(O)-,-NR C(O)O-, -OC(O)NR -, -NR S(O2)-, - C(O)NR S(O2)-, -NR C(O)NR -, -NR -, -C6-C10 aryl -, -C 3-C10 carbocyclyl-, -5- to 6-membered heteroaryl-, -5- to 6-membered heterocyclyl-, -C6-C10 aryl-O-, -O-aryl-, - O-C 3-C10 carbocyclyl- and -C3-C10 carbocyclyl-O-, wherein the C 6-C10 aryl, carbocyclyl, 5- to 6-membered heteroaryl and 5- to 6-membered heterocyclyl groups are each optionally substituted by one or more substituents selected from List Z; 25 x y z B is selected from H, C 1-C6 alkyl, a C3-C10 carbocyclic group, NR R , C(O)OR , aryl, 5- to 6-membered heteroaryl and 5- to 6-membered heterocyclyl, wherein the carbocyclic, C 6-C10 aryl, 5- to 6-membered heteroaryl and 5- to 6- membered heterocyclyl groups are each optionally substituted by one or more substituents selected from List Z; d is 1, 2, 3, 4, 5, 6 or 7; b is 0, 1, 2 or 3; 30 c d x y R , R , R and R are each independently selected from H, C1-C6 alkyl, a C3-C10 carbocyclic group and -(C1-C3 alkylene)-C3-C10 carbocyclic; z R is H or C1-C6 alkyl; 9 R is H or C1-C6 alkyl; or 6 9 R and R , together with the carbon atom to which they are attached, form a C3-C10 carbocyclic group or a 4- to 35 10-membered 5- to 6-membered heterocyclyl group, wherein each of the ring systems is optionally substituted by one or more substituents selected from the List Z; 11 R is selected from H, C1-C6 alkyl, a C3-C10 carbocyclic group and -(C1-C3 alkylene)-C3-C10 carbocyclic; 12 R is selected from H, C1-C6 alkyl, a C3-C10 carbocyclic group, -(C1-C3 alkylene)-C3-C10 carbocyclic, -C(O)C1-C6 alkyl, -C(O)C3-C10 carbocyclic, -C(O)(C1-C3 alkylene)-C3-C10 carbocyclic, -C(O)(CH2)x C6-C10 aryl, -C(O)(CH2)x 40 5- to 6-membered heteroaryl, -C(O)(CH 2)x 5- to 6-membered heterocyclyl, -C(O)Oalkyl, C(O)OC6-C10 aryl, (CH 2)x C6-C10 aryl, (CH2)x 5- to 6-membered heteroaryl and - (CH 2)x 5- to 6-membered heterocyclyl, wherein each of the ring systems is optionally substituted by one or more substituents selected from the List Z; 13 R is selected from H, C1-C6 alkyl, a C3-C10 carbocyclic group and -(C1-C3 alkylene)-C3-C10 carbocyclic; 14 R is selected from H, C1-C6 alkyl, a C 3-C10 carbocyclic group, -(C 1-C3 alkylene)-C3-C10 carbocyclic, - (CH2)z C6- 45 C10 aryl, (CH2)z 5- to 6-membered heteroaryl and (CH 2)z 5- to 6-membered heterocyclyl, wherein each of the ring systems is optionally substituted by one or more substituents selected from the List Z; x is 0, 1, 2 or 3; z is 0, 1, 2 or 3; each Z is independently selected from OH, C6-C10 aryl, 5- to 6-membered heteroaryl, 5- to 6-membered heterocyclyl, 50 benzyl, C1-C6 alkyl optionally substituted by one or more halogen atoms, CN or OH groups, C 1-C6 alkoxy optionally substituted by one or more halogen atoms, CN or OH groups, -OC6-C10 aryl, -Obenzyl,2 )a -O(CHC(O)E, 15 17 15 16 18 15 17 15 17 15 16 17 15 17 NR (SO2)R , (SO2)NR R , (SO2)R , NR C(O)R , C(O)NR R , NR C(O)NR R , NR C(O)OR , 15 17 15 15 15 17 15 NR R , C(O)OR , OC(O)R , OC(O)NR , C(O)R , SR , CN, NO2, and halogen; and when there are two or more Z substitutents, two Z substituents together with the atoms to which they are attached optionally form a 5- to 55 7-membered carbocyclic or a 4- to 7-membered heterocyclic substituent fused to the ring system; a is 0, 1, 2, 3 or 4, where the alkylene group is optionally substituted by OH or NH 2 when a is 1, 2, 3 or 4; E is NR15R17 or OR17 ; 15 16 each R and R is independently selected from H, 1-C C6 alkyl, a 3 C-C10 carbocyclic group and -(C1-C3

71 EP 2 300 010 B1

alkylene)-C3-C10 carbocyclic; 17 each R is selected from H, C1-C6 alkyl, a C3-C10 carbocyclic group, -(C1-C3 alkylene)-C3-C10 carbocyclic, C6- C10 aryl, 5- to 6-membered heteroaryl and 5- to 6-membered heterocyclyl, where each of the ring systems is optionally substituted by OH, halo, C1-C3 alkyl and C1-C3 alkoxy; and 5 18 R is selected from H, C1-C6 alkyl, a C 3-C10 carbocyclic group, -(C 1-C3 alkylene)-C3-C10 carbocyclic, C6-C10 aryl, 5- to 6-membered heteroaryl, 5- to 6-membered heterocyclyl and NHC(=NH)NH2, where each of the ring systems is optionally substituted by OH, halo, C 1-C3 alkyl and C1-C3 alkoxy.

2. A compound according to Claim 1, wherein R 9 is H. 10 9 6 3. A compound according to any preceding claim, wherein R is H and R is selected from H and C 1-C6 alkyl.

4. A compound according to Claim 1 which is selected from:

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72 EP 2 300 010 B1

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73 EP 2 300 010 B1

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74 EP 2 300 010 B1

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75 EP 2 300 010 B1

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76 EP 2 300 010 B1

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77 EP 2 300 010 B1

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78 EP 2 300 010 B1

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79 EP 2 300 010 B1

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80 EP 2 300 010 B1

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81 EP 2 300 010 B1

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82 EP 2 300 010 B1

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83 EP 2 300 010 B1

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84 EP 2 300 010 B1

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85 EP 2 300 010 B1

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86 EP 2 300 010 B1

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20 and pharmaceutically acceptable salts, solvates and hydrates thereof.

5. A compound according to any of Claims 1 to 4 for use as a pharmaceutical.

6. A pharmaceutical composition comprising a compound according to any of Claims 1 to 4, together with one or more 25 pharmaceutical excipients.

7. The use of a compound according to any of Claims 1 to 4, in the manufacture of a medicament for treatment of an inflammatory or allergic condition.

30 8. The use of a compound according to any of Claims 1 to 4, in the manufacture of a medicament for the treatment of an inflammatory or allergic condition selected from cystic fibrosis, primary ciliary dyskinesia, chronic bronchitis, chronic obstructive pulmonary disease, asthma, respiratory tract infections, lung carcinoma, xerostomia, and kera- toconjunctivitis sire.

35 9. A pharmaceutical composition comprising a compound according to any of Claims 1 to 4 and an anti-inflammatory, bronchodilatory, antihistamine or anti-tussive drug substance.

10. A process for the preparation of compounds of formula (Ia) according to claim 1:

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50 comprising reacting a compound of Formula 3:

55

87 EP 2 300 010 B1

5

10 with an alkylating agent R10X, wherein X is a suitable leaving group and wherein R6, R7, R8, R9 and R10 are as defined in Claim 1.

15 Patentansprüche

1. Verbindung mit der Formel Ia

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oder pharmazeutisch verträgliche Salze, Solvate oder Hydrate davon, wobei 6 6a R -(C0-C6-Alkylen)-R ist, wobei der Alkylenlinker optional durch eine oder mehrere Gruppe(n) substituiert ist, die 35 aus C1-C3-Alkyl, Halo und OH ausgewählt ist/sind; 6a 11 12 13 14 R aus H, einer carbocyclischen C3-C10-Gruppe, NR R , C(O)NR R , C6-C10-Aryl, 5-bis 6-gliedrigem Hete- roaryl, 5- bis 6-gliedrigem Heterocyclyl und einer Gruppe der Formel P-(CH2)m-Q ausgewählt ist, wobei jedes der Ringsysteme optional durch einen oder mehrere Substituenten substituiert ist, der/die aus der Liste Z ausgewählt sind; 40 a a a P aus einer Bindung, -O-, -C(O)-, -C(O)O-, NHC(=N)NH- und 2 -S(O)-, -S(O2)NR -,-NR C(O)-, -NR C(O)O-, a a b -NR S(O2)- und -NR C(O)NR - ausgewählt ist; a b R und R jeweils unabhängig aus H, 1-C C 6-Alkyl, einer carbocyclischen C3-C10-Gruppe und -(C1-C3-Alky- len)-C3-C10-carbocyclus ausgewählt sind; Q aus H, C 1-C6-Alkyl, einer carbocyclischen C 3-C10-Gruppe, C6-C10-Aryl, 5- bis 6-gliedrigem Heteroaryl und 5- bis 45 6-gliedrigem Heterocyclyl ausgewählt ist, wobei die carbocyclischen, C6-C10-Aryl-, 5- bis 6-gliedrigen Heteroaryl- und 5- bis 6-gliedrigen Heterocyclylgruppen jeweils optional durch einen oder mehrere Substituenten substituiert sind, der/die aus der Liste Z ausgewählt ist/sind; m 0, 1, 2 oder 3 ist; 7 8 10 R ,R und R jeweils unabhängig aus 1-C C6-Alkyl, C2-C6-Alkenyl, C2-C6-Alkynyl, einer carbocyclischen 50 C3-C10-Gruppe, carbocyclischen -(C1-C3-Alkylen)-C3-C10-Gruppe, C6-C10-Aryl, 5- bis 6-gliedrigem Heteroaryl, 5- bis 6-gliedrigem Heterocyclyl oder einer Gruppe der Formel (CH 2)d-A-(CH2)b-B ausgewählt sind, wobei die carbo- cyclischen, C6-C10-Aryl-, 5- bis 6-gliedrigen Heteroaryl- und 5- bis 6-gliedrigen Heterocyclylgruppen jeweils optional durch einen oder mehrere Substituenten substituiert sind, der/die aus der Liste Z ausgewählt ist/sind; und wobei die Alkyl- und Alkylengruppen optional durch ein(e) oder mehrere Halogenatom(e), OH-Gruppe(n) oder Phenyl- 55 gruppe(n) substituiert sind; oder R7 und R8 gemeinsam mit dem Stickstoffatom, an dem sie angehaftet sind, eine 4- bis 10-gliedrige stickstoffhaltige heterocyclische Gruppe bilden, die optional durch einen oder mehrere Substituenten substituiert ist, der/die aus der Liste Z ausgewählt ist/sind; oder

88 EP 2 300 010 B1

R6 und R8 gemeinsam mit den Atomen, an denen sie angehaftet sind, eine 4- bis 10-gliedrige stickstoffhaltige heterocyclische Gruppe bilden, die optional durch einen oder mehrere Substituenten substituiert ist, der/die aus der Liste Z ausgewählt ist sind; c c c A aus einer Bindung, -O-, -C(O)-, -C(O)NR -, -C(O)O-, -OC(O)-, -NHC(=N)NH-, -S(O2)-, -S(O2)NR -, -NR C(O)-, 5 c c c c c d c -NR C(O)O-, -OC(O)NR -, -NR S(O2)-, -C(O)NR S(O2)-, - NR C(O)NR -, -NR -, -C6-C10-Aryl-, -C3-C10-Carbocy- clyl-, -5- bis 6-gliedrigem Heteroaryl-, -5- bis 6-gliedrigem Heterocyclyl-, -C 6-C10-Aryl-O-, -O-Aryl-, -O-C3-C10-Car- bocyclyl- und -C3-C10-Carbocyclyl-O- ausgewählt ist, wobei die C6-C10-Aryl-, Carbocyclyl-, 5- bis 6-gliedrigen He- teroaryl- und 5- bis 6-gliedrigen Heterocyclylgruppen jeweils optional durch einen oder mehrere Substituenten sub- stituiert sind, der/die aus der Liste Z ausgewählt ist/sind; 10 x y z B aus H, C1-C6-Alkyl, einer carbocyclischen C3-C10-Gruppe, NR R , C(O)OR , Aryl, 5-bis 6-gliedrigem Heteroaryl und 5- bis 6-gliedrigem Heterocyclyl ausgewählt ist, wobei die carbocyclischen, C6-C10-Aryl-, 5- bis 6-gliedrigen Heteroaryl- und 5- bis 6-gliedrigen Heterocyclylgruppen jeweils optional durch einen oder mehrere Substituenten substituiert sind, der/die aus der Liste Z ausgewählt ist/sind; d 1, 2, 3, 4, 5, 6 oder 7 ist; 15 b 0, 1, 2 oder 3 ist; c d x y R , R , R und R jeweils unabhängig aus H, C1-C6-Alkyl, einer carbocyclischen C3-C10-Gruppe und -(C1-C3-Alky- len-)-C3-C10-carbocyclus ausgewählt sind; z R H oder C1-C6-Alkyl ist; 9 R H oder C1-C6-Alkyl ist; oder 20 6 9 R und R gemeinsam mit dem Kohlenstoffatom, an dem sie angehaftet sind, eine carbocyclische C 3-C10-Gruppe oder eine 4- bis 10-gliedrige 5- bis 6-gliedrige Heterocyclylgruppe bilden, wobei jedes dieser Ringsysteme optional durch einen oder mehrere Substituenten substituiert ist, der/die aus der Liste Z ausgewählt ist/sind; 11 R aus H, C 1-C6-Alkyl, einer carbocyclischen C 3-C10-Gruppe und -(C 1-C3-Alkylen)-C3-C10-carbocyclus ausgewählt ist; 25 12 R aus H, 1-C6 C-Alkyl, einer carbocyclischen 3-C10 C-Gruppe, -(C1-C3-Alkylen)-C3-C10-carbocyclus, -C(O)C1-C6-Alkyl, -C(O)C3-C10-Carbocyclus, -C(O)(C1-C3-Alkylen)-C3-C10-carbocyclus, -C(O)(CH2)x-C6-C10-Aryl, 5- bis 6-gliedrigem C(O)(CH2)x-Heteroaryl, 5- bis 6-gliedrigem -C(O)(CH2)x-Heterocyclyl, -C(O)O-Alkyl, C(O)O- C6-C10-Aryl, (CH2)x-C6-C10-Aryl, 5- bis 6-gliedrigem (CH 2)x-Heteroaryl und 5- bis 6-gliedrigem -(CH 2)x-Heterocyclyl ausgewählt ist, wobei jedes dieser Ringsysteme optional durch einen oder mehrere Substituenten substituiert ist, 30 der/die aus der Liste Z ausgewählt ist/sind; 13 R aus H, C 1-C6-Alkyl, einer carbocyclischen C 3-C10-Gruppe und -(C 1-C3-Alkylen)-C3-C10-carbocyclus ausgewählt ist; 14 R aus H, 1-C6 C-Alkyl, einer carbocyclischen 3-C10 C-Gruppe, -(C1-C3-Alkylen)-C3-C10-carbocyclus, -(CH2)z-C6-C10-Aryl, 5- bis 6-gliedrigem (CH 2)z-Heteroaryl und 5- bis 6-gliedrigem (CH 2)z-Heterocyclyl ausgewählt 35 ist, wobei jedes der Ringsysteme optional durch einen oder mehrere Substituenten substituiert ist, der/die aus der Liste Z ausgewählt ist/sind; x 0, 1, 2 oder 3 ist; z 0, 1, 2 oder 3 ist; Z jeweils unabhängig aus OH, C 6-C10-Aryl, 5- bis 6-gliedrigem Heteroaryl, 5- bis 6-gliedrigem Heterocyclyl, Benzyl, 40 C1-C6-Alkyl, optional substituiert durch ein oder mehrere Halogenatom(e), CN- oder OH-Gruppen, C1-C6-Alkoxy, optional substituiert durch ein oder mehrere Halogenatom(e), CN- oder OH-Gruppen, -OC6-C10-Aryl, -OBenzyl, 15 17 15 16 18 15 17 15 17 15 16 17 -O(CH2)aC(O)E, NR (SO2)R , (SO2)NR R , (SO2)R , NR C(O)R , C(O)NR R , NR C(O)NR R , 15 17 15 17 15 15 15 17 15 NR C(O)OR , NR R , C(O)OR , OC(O)R , OC(O)NR , C(O)R , SR , CN, NO2 und Halogen ausgewählt ist; und wenn zwei oder mehrere Z-Substituenten vorhanden sind, zwei Z-Substituenten gemeinsam mit den Atomen, 45 an denen sie angehaftet sind, optional einen 5- bis 7-gliedrigen carbocyclischen oder einen 4- bis 7-gliedrigen heterocyclischen Substituenten bilden, der an das Ringsystem fusioniert ist;

a 0, 1, 2, 3 oder 4 ist, wobei die Alkylengruppe optional durch OH oder NH 2 substituiert ist, wenn a 1, 2, 3 oder 4 ist; E NR15R17 oder OR17 ist; 15 16 R und R jeweils unabhängig aus H, 1 C-C6-Alkyl, einer carbocyclischen C3-C10-Gruppe und -(C1-C3-Alky- 50 len)-C3-C10-carbocyclus ausgewählt sind; 17 R jeweils aus H, 1-C C 6-Alkyl, einer carbocyclischen 3 C-C10-Gruppe, -(C1-C3-Alkylen)-C3-C10-carbocyclus, C6-C10-Aryl, 5- bis 6-gliedrigem Heteroaryl und 5- bis 6-gliedrigem Heterocyclyl ausgewählt ist, wobei jedes der Ringsysteme optional durch OH, Halo, C1-C3-Alkyl und C1-C3-Alkoxy substituiert ist; und 18 R aus H, C1-C6-Alkyl, einer carbocyclischen C3-C10-Gruppe, -(C1-C3-Alkylen)-C3-C10-carbocyclus, C6-C10-Aryl, 55 5- bis 6-gliedrigem Heteroaryl, 5- bis 6-gliedrigem Heterocyclyl und NHC(=NH)NH 2 ausgewählt ist, wobei jedes der Ringsysteme optional durch OH, Halo, C1-C3-Alkyl und C1-C3-Alkoxy substituiert ist.

2. Verbindung nach Anspruch 1, wobei R9 H ist.

89 EP 2 300 010 B1

9 6 3. Verbindung nach einem vorhergehenden Anspruch, wobei R H ist und R aus H und C1-C6-Alkyl ausgewählt ist.

4. Verbindung nach Anspruch 1, die ausgewählt ist aus:

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90 EP 2 300 010 B1

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92 EP 2 300 010 B1

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93 EP 2 300 010 B1

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94 EP 2 300 010 B1

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95 EP 2 300 010 B1

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35 und pharmazeutisch verträglichen Salzen, Solvaten und Hydraten davon.

5. Verbindung nach einem der Ansprüche 1 bis 4 zur Verwendung als Pharmazeutikum.

6. Pharmazeutische Zusammensetzung, die eine Verbindung nach einem der Ansprüche 1 bis 4 gemeinsam mit einem

40 oder mehreren pharmazeutischen Hilfsstoffen umfasst.

7. Verwendung einer Verbindung nach einem der Ansprüche 1 bis 4 bei der Herstellung eines Medikaments zur Behandlung eines entzündlichen oder allergischen Zustands.

45 8. Verwendung einer Verbindung nach einem der Ansprüche 1 bis 4 bei der Herstellung eines Medikaments zur Behandlung eines entzündlichen oder allergischen Zustands, der aus zystischer Fibrose, primärer Ziliendyskinesie, chronischer Bronchitis, chronisch-obstruktiver Lungenkrankheit, Asthma, Atemwegsinfektionen, Lungenkarzinom, Xerostomie und Keratokonjunktivitis sicca ausgewählt ist.

50 9. Pharmazeutische Zusammensetzung, die eine Verbindung nach einem der Ansprüche 1 bis 4 und eine entzün- dungshemmende, bronchodilatatorische, antihistaminische oder hustenhemmende Arzneimittelsubstanz umfasst.

10. Verfahren zur Herstellung von Verbindungen der Formel (Ia) nach Anspruch 1:

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103 EP 2 300 010 B1

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das das Reagieren einer Verbindung der Formel 3:

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mit einem Alkylierungsmittel R 10X umfasst, wobei X eine geeignete Abgangsgruppe ist und wobei R 6,R7,R8, R9 und 25 R10 wie in Anspruch 1 definiert sind.

Revendications

30 1. Composé ayant la formule Ia :

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ou sels, solvates ou hydrates pharmaceutiquement acceptables de celui-ci, où :

6 6a - R représente - (alkylène en C 0-C6) -R , le bras de liaison alkylène étant facultativement substitué par un ou 45 plusieurs groupes choisis parmi alkyle en C1-C3, halo et OH ; 6a 11 12 13 14 - R est choisi parmi H, un groupe carbocyclique en C3-C10, NR R , C(O)NR R , aryle en C6-C10, hété- roaryle à 5 à 6 chaînons, hétérocyclyle à 5 à 6 chaînons et un groupe de la formule P-(CH2)m-Q, chacun des systèmes de noyau étant facultativement substitué par un ou plusieurs substituants choisis dans la Liste Z ; a a - P est choisi parmi une liaison, -O-, -C(O) -, -C(O)O-, -NHC(=N)NH- et -S(O2)-, -S(O2)NR -, -NR C(O)-, 50 3 a a b -NR C(O)O-, -NR S(O2) - et -NR C(O)NR -; a b - R et R sont chacun indépendamment choisis parmi H, alkyle en C 1-C6, un groupe carbocyclique en C 3-C10 et - (alkylène en C1-C3)-carbocyclique en C3-C10 ; - Q est choisi parmi H, alkyle en C1-C6, un groupe carbocyclique en C3-C10, aryle en C6-C10, hétéroaryle à 5 à 6 chaînons et hétérocyclyle à 5 à 6 chaînons, les groupes carbocyclique, aryle en C 6-C10, hétéroaryle à 5 à 55 6 chaînons et hétérocyclyle à 5 à 6 chaînons étant chacun facultativement substitués par un ou plusieurs substituants choisis dans la Liste Z ; - m est 0, 1, 2 ou 3 ; 7 8 10 - R , R et R sont chacun indépendamment choisis parmi alkyle en C1-C6, alcényle en C2-C6, alcynyle en

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C2-C6, un groupe carbocyclique en C3-C10, - (alkylène en C1-C3)-groupe carbocyclique en C3-C10, aryle en C6-C10, hétéroaryle à 5 à 6 chaînons, hétérocyclyle à 5 à 6 chaînons ou un groupe de la formule (CH2)d-A-(CH2)b-B, les groupes carbocyclique, aryle en C6-C10, hétéroaryle à 5 à 6 chaînons et hétérocyclyle à 5 à 6 chaînons étant chacun facultativement substitués par un ou plusieurs substituants choisis dans la Liste 5 Z ; et les groupes alkyle et alkylène étant facultativement substitués par un ou plusieurs atomes d’halogène, groupes OH ou groupes phényle ; ou - R7 et R8, conjointement avec l’atome d’azote auquel ils sont attachés forment un groupe hétérocyclique contenant de l’azote, à 4 à 10 chaînons, qui est facultativement substitué par un ou plusieurs substituants choisis dans la Liste Z ; ou 10 - R 6 et R 8, conjointement avec les atomes auxquels ils sont attachés forment un groupe hétérocyclique contenant de l’azote, à 4 à 10 chaînons, qui est facultativement substitué par un ou plusieurs substituants choisis dans la Liste Z ; c c - A est choisi parmi une liaison, -O-, -C(O)-, -C(O)NR -, -C(O)O-, -OC(O)-, -NHC(=N)NH- , -S(O 2)-, -S(O2)NR -, c c c c c c d c -NR C(O)-, -NR C(O)O-, -OC(O)NR -, -NR S(O2) -, -C(O)NR S(O2) -, -NR C(O)NR -, -NR -, -aryl en C6-C10-, 15 -carbocyclyl en C3-C10-, -hétéroaryl à 5 à 6 chaînons-, hétérocyclyl à 5 à 6 chaînons-, -aryl en C6-C10-O-, -O- aryl-, -O-carbocyclyl en C 3-C10- et -carbocyclyl en C 3-C10-O-, les groupes aryle en C 6-C10, carbocyclyle, hété- roaryle à 5 à 6 chaînons et hétérocyclyle à 5 à 6 chaînons étant chacun facultativement substitués par un ou plusieurs substituants choisis dans la Liste Z ; x y z - B est choisi parmi H, alkyle en C 1-C6, un groupe carbocyclique en C 3-C10, NR R , C(O)OR , aryle, hétéroaryle 20 à 5 à 6 chaînons et hétérocyclyle à 5 à 6 chaînons, les groupes carbocyclique, aryle en C 6-C10, hétéroaryle à 5 à 6 chaînons et hétérocyclyle à 5 à 6 chaînons étant chacun facultativement substitués par un ou plusieurs substituants choisis dans la Liste Z ; - d est 1, 2, 3, 4, 5, 6 ou 7 ; - b est 0, 1, 2 ou 3 ; 25 c d x y - R , R , R et R sont chacun indépendamment choisis parmi H, alkyle en C 1-C6, un groupe carbocyclique en C3-C10 et - (alkylène en C1-C3)-carbocyclique en C3-C10 ; z - R représente H ou alkyle en C1-C6 ; 9 - R représente H ou alkyle en C 1-C6 ; ou - R6 et R9, conjointement avec l’atome de carbone auquel ils sont attachés, forment un groupe carbocyclique 30 en C3-C10 ou un groupe hétérocyclyle à 4 à 10 chaînons à 5 à 6 chaînons, chacun des systèmes de noyau étant facultativement substitué par un ou plusieurs substituants choisis dans la Liste Z ; 11 - R est choisi parmi H, alkyle en C1-C6, un groupe carbocyclique en C3-C10 et - (alkylène en C1-C3)-carbo- cyclique en C3-C10 ; 12 - R est choisi parmi H, alkyle en C 1-C6, un groupe carbocyclique en C 3-C10, -(alkylène en C 1-C3)-carbocyclique 35 en C3-C10, -C(O)-alkyle en C1-C6, -C(O)-carbocyclique en C3-C10, -C(O)(alkylène en C1-C3)-carbocyclique en C3-C10, -C(0) (CH2)x-aryle en C6-C10, -C(0) (CH2)x-hétéroaryle à 5 à 6 chaînons, -C(O) (CH2)x-hétérocyclyle à 5 à 6 chaînons, -C(O)O-alkyle, C(O)O-aryle en C6-C10, (CH2)x-aryle en C6-C10, (CH2)X-hétéroaryle à 5 à 6 chaînons et - (CH2)X-hétérocyclyle à 5 à 6 chaînons, chacun des systèmes de noyau étant facultativement substitué par un ou plusieurs substituants choisis dans la Liste Z ; 40 13 - R est choisi parmi H, alkyle en C 1-C6, un groupe carbocyclique en C3-C10 et -(alkylène en C 1-C3)-carbocy- clique en C3-C10; 14 - R est choisi parmi H, alkyle en C 1-C6, un groupe carbocyclique en C 3-C10, -(alkylène en C 1-C3)-carbocyclique en C 1-C10,-(CH 2)z-aryleen C 6-C10, (CH 2)z-hétéroaryle à 5 à 6 chaînons et (CH 2)z-hétérocyclyle à 5 à 6 chaînons, chacun des systèmes de noyau étant facultativement substitué par un ou plusieurs substituants choisis dans 45 la Liste Z ; - x est 0, 1, 2 ou 3 ; - z est 0, 1, 2 ou 3 ;

- chaque Z est indépendamment choisi parmi OH, aryle en C 6-C10, hétéroaryle à 5 à 6 chaînons, hétérocyclyle à 5 à 6 chaînons, benzyle, alkyle en C1-C6 facultativement substitué par un ou plusieurs atomes d’halogène, 50 groupes CN ou OH, alcoxy en C 1-C6 facultativement substitué par un ou plusieurs atomes d’halogène, groupes 15 17 15 16 18 CN ou OH, -O-aryle en 6-C C 10, -O-benzyle, -O(CH2)aC(0) E, NR (SO2)R , (SO2)NR R , (SO2)R , NR15C(O)R17, C(O)NR15R17, NR15C(O)NR16R17, NR15C(O)OR17, NR15R17, C(O)OR15, OC(O)R15, 15 17 15 OC(O)NR , C(O)R , SR , CN, NO2 et halogène; et lorsqu’il y a au moins deux substituants Z, deux substi- tuants Z conjointement avec les atomes auxquels ils attachés forment facultativement un substituant carbocy- 55 clique à 5 à 7 chaînons ou un substituant hétérocyclique à 4 à 7 chaînons fusionné au système de noyau ; - a est 0, 1, 2, 3 ou 4, où le groupe alkylène est facultativement substitué par OH ou NH 2 lorsque a est 1, 2, 3 ou 4 ; - E représente NR15R17 ou OR17; 15 16 - chaque R et R est indépendamment choisi parmi H, alkyle en C 1-C6, un groupe carbocyclique en C1-C10

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et -(alkylène en C1-C3)-carbocyclique en C3-C10; 17 - chaque R est choisi parmi H, alkyle en C 1-C6, un groupe carbocyclique en C 3-C10, -(alkylène en C1-C3)-car- bocyclique en C3-C10, aryle en C6-C10, hétéroaryle à 5 à 6 chaînons et hétérocyclyle à 5 à 6 chaînons, où chacun des systèmes de noyau est facultativement substitué par OH, halo, alkyle en C 1-C3 et alcoxy en C 1-C3 ; et 5 18 - R est choisi parmi H, alkyle en C 1-C6, un groupe carbocyclique en C 3-C10, -(alkylène en C 1-C3)-carbocyclique en C3-C10, aryle en C6-C10, hétéroaryle à 5 à 6 chaînons, hétérocyclyle à 5 à 6 chaînons et NHC(=NH)NH2, où chacun des systèmes de noyau est facultativement substitué par OH, halo, alkyle en C 1-C3 et alcoxy en C 1-C3.

2. Composé selon la revendication 1, dans lequel R 9 représente H. 10 3. Composé selon l’une quelconque des revendications précédentes, dans lequel R9 représente H et R6 est choisi parmi H et alkyle en C1-C6.

4. Composé selon la revendication 1 qui est choisi parmi: 15 Structure

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20 et des sels, solvates et hydrates pharmaceutiquement acceptables de ceux-ci.

5. Composé selon l’une quelconque des revendications 1 à 4 en vue d’une utilisation comme produit pharmaceutique.

6. Composition pharmaceutique comprenant un composé selon l’une quelconque des revendications 1 à 4, conjoin- 25 tement avec un ou plusieurs excipients pharmaceutiques.

7. Utilisation d’un composé selon l’une quelconque des revendications 1 à 4, dans la fabrication d’un médicament pour le traitement d’un état inflammatoire ou allergique.

30 8. Utilisation d’un composé selon l’une quelconque des revendications 1 à 4, dans la fabrication d’un médicament pour le traitement d’un état inflammatoire ou allergique choisi parmi la fibrose kystique, la dyskinésie ciliaire primitive, la bronchite chronique, la maladie pulmonaire obstructive chronique, l’asthme, les infections du tractus respiratoire, le carcinome du poumon, la xérostomie et la kératoconjonctivite sèche.

35 9. Composition pharmaceutique comprenant un composé selon l’une quelconque des revendications 1 à 4 et une substance médicamenteuse anti-inflammatoire, bronchodilatatrice, antihistaminique ou antitussive.

10. Procédé de préparation de composés de formule (Ia) selon la revendication 1 :

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REFERENCES CITED IN THE DESCRIPTION

This list of references cited by the applicant is for the reader’s convenience only. It does not form part of the European patent document. Even though great care has been taken in compiling the references, errors or omissions cannot be excluded and the EPO disclaims all liability in this regard.

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