EA043636B1 - ANTIBACTERIAL 3,4-DIHYDRO-1H-[1,8]NAPHTHYRIDINONES, SUBSTITUTED WITH CYCLOPENTA[c]pyrrole - Google Patents

Description

The invention relates to new compounds of formula (I) that inhibit the activity of the Fabl enzyme, which are therefore useful for the treatment of bacterial infections. It also relates to pharmaceutical compositions containing these compounds and chemical methods for preparing such compounds.

The compounds of the present invention are antibacterial compounds that inhibit the Fabl protein NADH-dependent enzyme enoyl-acyl transfer protein (ACP) reductase in the fatty acid biosynthetic pathway. Fatty acid synthase (FAS) is involved in the common saturated fatty acid biosynthetic pathway of all organisms, but the structural organization of FAS varies significantly among them. Distinctive characteristics of vertebrate and yeast FAS are that all enzymatic activities are carried on one or two polypeptide chains, and that the acyl transfer protein (ACP) is present as a complex. In contrast, in bacterial FAS, each step of synthesis is catalyzed by a specific monofunctional enzyme, and ACP is a separate protein. For this reason, it is possible to selectively inhibit bacterial FAS by blocking one of the stages of synthesis using an inhibitory agent. NADH-dependent enoyl-ACP reductase (Fabl) is involved in the last step of a four-step reaction involved in each cycle of bacterial fatty acid biosynthesis. Thus, the Fabl enzyme is a biosynthetic enzyme in the general synthesis pathway of bacterial fatty acid biosynthesis.

The Fabl enzyme has been shown to be a critical target in major pathogens such as E. Coli (Heath et al., J. Biol. Chern. 1995, 270, 26538; Bergler et al., Eur. J. Biochem. 2000, 275, 4654). Therefore, Fabl inhibitory compounds may be useful as antibacterial agents.

Compounds having an inhibitory effect on the Fabl enzyme are disclosed in WO-01/26652, WO-01/26654 and WO-01/27103. Substituted naphthyridinone compounds having Fabl inhibitory activity are disclosed in WO-03/088897, WO2007/043835 and WO-2008/098374. International patent application WO 2007/053131 discloses various compounds for putative use as Fabl inhibitors. International patent application WO 2011/061214 also discloses various compounds for putative use as Fabl inhibitors. However, none of these documents disclose a fused bicyclic moiety that is attached directly to a carbonyl moiety located at the a-position of the alkene.

The present invention relates to a compound of formula (I)

where A is -OC- or the ⁷⁷⁷⁷⁷⁷ bond is a single or double bond, X is carbon or nitrogen, and if X is nitrogen, the bond is a single bond;

Zi represents CH or N;

R ¹ is hydrogen, C^alkyl or halogen;

R ² is hydrogen, Smalkyl or halogen;

R ³ is hydrogen, C1. ₆ alkyl, hydroxyl or halogen;

R ⁴ is hydrogen; halogen; C1. ₆ alkyl; With _2.balkenyl ; With _2.bElkynyl ; C1. ₆ alkyloxy; C1.4alkyloxycarbonyl; aminocarbonyl; mono- or di(C1.4alkyl)aminocarbonyl; aryl; aryloxy; arylcarbonyl; arylsulfonyl; heteroaryl; C1. ₆ alkyl substituted with a cyano group; C1. ₆ alkyl substituted with aryl or aryloxy; or C1. ₆ alkyl substituted with heteroaryl; aryl is phenyl; phenyl substituted with one, two or three substituents, each of them individually selected from halogen, hydroxyl, C-alkyl, C1.4alkoxy, polyhalogenC/C-alkyl, polyhalogenC1.4alkoxy, cyano-, nitro- and amino groups; heteroaryl is furanyl, thiophenyl, pyrrolyl, pyrazolyl, imidazolyl, isoxazolyl, thiazolyl, triazolyl, tetrazolyl, isothiazolyl, thiadiazolyl, oxadiazolyl, pyridinyl, pyridazinyl, pyrimidinyl, pyrazinyl, benzo[1,3]dioxolyl, benzofuranyl, benzothia solyl, indolyl, 2, 3-dihydro-1H-indolyl, tetrahydrothiophenyl or quinolinyl, wherein each heteroaryl may be substituted with one or two substituents, each individually selected from halogen, cyano, C1. ₄ alkyl, C1. ₄ alkoxy, C1. ₄ alkylcarbonyl or phenyl;

or a pharmaceutically acceptable acid addition salt thereof.

As used in the preceding definitions, the term halogen is general for fluorine, chlorine, bromine and iodine atoms;

C1. ₄ alkyl denotes straight and branched chain hydrocarbon radicals containing from 1 to 4 carbon atoms, such as, for example, methyl, ethyl, propyl, butyl, 1-methylethyl, 2-methylpropyl and the like;

C1. ₆ alkyl includes C^alkyl and its higher homologues containing 5 or 6 carbon atoms, such as, for example, 2-methylbutyl, pentyl, hexyl, etc.;

- 1043636 _polyhaloC1-4 alkyl is defined as polyhalogen-substituted _C1-4 alkyl (as defined herein and above) substituted with 2-6 halogen atoms, such as difluoromethyl, trifluoromethyl, trifluoroethyl, and the like.

As used herein, whenever the term compound of formula (I) is used, it is intended that it also includes the pharmaceutical addition salts that the compounds of formula (I) are capable of forming and the solvates that the compounds of formula (I) are capable of forming or pharmaceutically acceptable acid addition salts of compounds of formula (I).

The definition of a compound of formula (I) essentially includes all stereoisomers of a compound of formula (I), either as a pure stereoisomer or as a mixture of two or more stereoisomers. Enantiomers are stereoisomers that are incompatible mirror images of each other. A 1:1 mixture of enantiomer pairs is a racemate or racemic mixture. Diastereomers (or diastereoisomers) are stereoisomers that are not enantiomers, that is, they are not related as mirror images. If the compound contains a disubstituted cycloalkyl group, the substituents may be in the cis or trans configuration. Therefore, this invention includes enantiomers, diastereomers, racemates, cis-isomers, trans-isomers and mixtures thereof.

The absolute configuration is determined according to the Cahn-Ingold-Prelog system. The configuration of an asymmetric atom is indicated with either R or S. Selected compounds whose absolute configuration is not known can be indicated with (+) or (-), depending on the direction in which they rotate plane-polarized light. If a specific stereoisomer is specified, this means that said stereoisomer is substantially free from other isomers, that is, associated with less than 50%, preferably less than 20%, more preferably less than 10%, even more preferably less than 5%, in particular less less than 2% and most preferably less than 1% of other isomers. Thus, if a compound of formula (I) is indicated as (R), this means that the compound contains substantially no (S) isomer; if a compound of formula (I) is indicated, for example, as E, this means that the compound contains substantially no Z isomer; if a compound of formula (I) is indicated, for example, as cis, this means that the compound contains substantially no trans isomer.

The terms stereoisomers or stereochemically isomeric forms hereinafter are used interchangeably.

Those skilled in the art can readily determine the absolute stereochemical configuration of the compounds of formula (I) and the intermediates used in their preparation using well known methods such as, for example, x-ray diffraction.

Certain compounds of formula (I) may also exist in their tautomeric form. Such forms, although not expressly indicated in the above formula, are intended to be included within the scope of the present invention.

In addition, some compounds of formula (I) and intermediates used in their preparation may exhibit polymorphism. It should be understood that the present invention covers any polymorphic forms having properties useful in the therapeutic treatment of the conditions described above.

The pharmaceutically acceptable acid addition salts mentioned herein and previously include non-toxic, therapeutically active acid addition salt forms that can be formed by a compound of formula (I). Pharmaceutically acceptable acid addition salts can be readily prepared by treating the base form with such an appropriate acid. Suitable acids include, for example, inorganic acids such as hydrohalic acids, for example hydrochloric or hydrobromic acid, sulfuric, nitric, phosphoric and the like acids; or organic acids such as, for example, acetic, propionic, glycolic, lactic, pyruvic, oxalic (i.e. ethanedioic), malonic, succinic (i.e. butanedioic), maleic, fumaric, malic, tartaric, citric, methanesulfonic, ethanesulfonic, benzenesulfonic , p-toluenesulfonic, cyclamic, salicylic, p-aminosalicylic, pamic and similar acids.

Conversely, said salt forms can be converted by treatment with an appropriate base to the free base form.

The compounds of formula (I) can exist in both unsolvated and solvated forms. The term solvate is used in this context to refer to a molecular association comprising the compounds of this invention and one or more molecules of a pharmaceutically acceptable solvent, such as water or ethanol. The term hydrate is used when the solvent is water.

The term FabI is known in the art and refers to bacterial enzymes that are believed to act as enoyl-acyl transfer protein (ACP) reductase in the last step of the four reactions involved in each cycle of bacterial fatty acid biosynthesis. This enzyme is believed to be widely distributed in bacteria.

Compounds of formula (I) that may be mentioned include those in which:

(i) Z1 represents CH, and therefore the compound of formula I is the following:

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where: (ii) if R ¹ or R ² are halogen, they are preferably F or C1;

(iii) R ¹ is hydrogen or C1_ ₄ alkyl and/or (iv) R ² is hydrogen or C1_ ₄ alkyl.

Preferred compounds of formula (I) include those in which A is a double bond (rather than a triple bond), that is, preferably A is V

Compounds of interest are those compounds of formula (I) where one or more of the following restrictions apply:

a) R ¹ and R ² represent hydrogen; or

b) R ³ represents hydrogen; or

c) R ³ represents hydrogen, halogen or hydroxyl; or

d) R ⁴ represents hydrogen or halogen; or

e) R ⁴ represents aryl; or

f) R ⁴ represents C1_ ₆ alkyl; or

g) R ⁴ is aryloxy or arylsulfonyl; or

h) R ⁴ represents C1_ ₆ alkyl substituted with aryl; or

i) R ⁴ represents heteroaryl; or

j) R ⁴ represents C1_ ₆ alkyl substituted with heteroaryl; or

j) heteroaryl is furanyl, thiophenyl, pyrazolyl, isoxazolyl, thiazolyl, triazolyl, tetrazolyl, thiadiazolyl, pyridinyl or pyrimidinyl; or

1) X represents carbon; or

m) X represents nitrogen and bond ⁷⁷⁷⁷⁷¹ is a single bond.

The first group of compounds are compounds of formula (I)

R ¹ where A is -C=C- or ^r2 = the bond is a single bond or a double bond, X is carbon or nitrogen, and if X is nitrogen, then the ^ ⁷⁷⁷⁷ ^ bond is a single bond;

R ¹ is hydrogen;

R is hydrogen;

R ³ is hydrogen, hydroxyl or halogen;

R ⁴ is hydrogen, halogen; C1_ ₆ alkyl; C1_ ₆ alkyloxy; C1_ ₄ alkyloxycarbonyl; aminocarbonyl; mono- or di(C1_ ₄ alkyl)aminocarbonyl; aryl; aryloxy; arylsulfonyl; heteroaryl; C1_ ₆ alkyl substituted with a cyano group; C1_ ₆ alkyl substituted with aryl; or C1_ ₆ alkyl substituted with heteroaryl; aryl is phenyl; phenyl substituted with one substituent selected from halogen, _{C1_4} alkyl, _{C1_4} alkyloxy and cyano; heteroaryl is furanyl, thiophenyl, pyrazolyl, isoxazolyl, thiazolyl, triazolyl, tetrazolyl, thiadiazolyl, pyridinyl or pyrimidinyl; wherein each heteroaryl may be substituted with one substituent selected from halogen, cyano, _{C1_4} alkyl, _{C1_4} alkyloxy or _{C1_4} alkyl carbonyl;

or a pharmaceutically acceptable acid addition salt thereof.

The second group of compounds of formula (I) are those compounds of formula (I) wherein A is -C^C-.

The third group of compounds of formula (I) are those compounds of formula (I) where A is R ¹ ϊ

Preferred compounds of formula (I) include those in which the X-containing ring is one of the following:

_c j _s single enantiomer (cis) single enantiomer (cis) that is, double rings containing a cis configuration at the junction of the rings (a trans configuration will cause strain in the ring), which may be racemic or single enantiomers. As explained here and below, if the absolute stereochemistry of the single enantiomer is not/was not known, the chiral carbons at the ring junction may be indicated by thick or dashed lines (rather than a wedge).

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More preferred compounds of formula (I) include those wherein the fused bicyclic X-containing ring is one of the following:

where in the above fused double rings the compounds may be racemic or a single enantiomer (if appropriate symmetry is absent and the existence of enantiomers is possible), as described here and above. Preferably in compounds of formula (I):

(i) there is at least one substituent R ³ or R ⁴ that is not hydrogen;

(ii) one of R ³ and R ⁴ (eg R ³ ) is hydrogen, hydroxyl or halogen (eg fluorine) and the other of R ³ and R ⁴ (eg R ⁴ ) is a substituent other than hydrogen ;

(iii) R ³ is hydrogen, hydroxyl or halogen (eg, fluorine) and is more preferably hydrogen (ie, R ³ is substantially not present);

(iv) R ⁴ is a substituent other than hydrogen (ie, an R ⁴ substituent that is not hydrogen is present);

(v) R ⁴ represents a substituent other than hydrogen which is attached to X, in which any of the above may be taken together or in combination.

For example, (iii), (iv) and/or (v) may be taken in combination to provide the particularly preferred compounds of formula (I) below

in which R ⁴ represents a substituent other than hydrogen. Particularly preferred substituents that may be represented by R ⁴ (here and elsewhere in the document) include:

(i) optionally substituted aryl;

(ii) optionally substituted heteroaryl (iii) C _1-6 alkyl substituted with aryl or heteroaryl (the latter of the two groups aryl and heteroaryl are themselves optionally substituted as defined herein);

(iv) aryloxy (in which the aryl moiety is optionally substituted as defined herein);

(v) arylsulfonyl (wherein the aryl moiety is optionally substituted as defined herein);

(vi) C _1-6 alkyl, which is unsubstituted (eg, ethyl, methyl, isopropyl);

(vii) di(C _1-4 alkyl)aminocarbonyl (eg -C(O)N(CH ₃ ) ₂ );

(viii) aminocarbonyl (-C(O)NH2);

(ix) C _1-4 alkyloxycarbonyl (eg -C(O)O-CH ₂ CH ₃ );

(x) halogen (eg fluorine);

(xi) C _2-6 alkynyl (eg -C=C);

(xii) C _1-6 alkoxy (eg -OCH ₃ ).

It is particularly preferred that the R ⁴ group contains an aromatic moiety, and therefore (i), (ii), (iii), (iv) and (v) above are particularly preferred.

When R ⁴ is the above (i), the aryl group is preferably phenyl, which group may be unsubstituted or substituted with one or two (for example, one) substituent selected from C _1-4 alkyloxy, halogen, C _{1 -4} alkyl or cyano groups (for example, OCH3, chlorine, fluorine, methyl or cyano group).

In the case where R ⁴ is the above (ii), the heteroaryl group is a monocyclic 5- or 6-membered ring containing from one to four heteroatoms, for example, thienyl (for example, 2- or 3-thienyl), pyridyl (for example , 4-pyridyl or 3-pyridyl), pyrazolyl (e.g. 5-pyrazolyl, 4-pyrazolyl or 1-pyrazolyl), furanyl (e.g. 2- or 3-furanyl), thiazolyl (e.g. 2-thiazolyl), isoxazolyl ( e.g. 4-isoxazolyl), pyrrolyl (e.g. 1-pyrrolyl), triazolyl (e.g. 1,2,3-triazol-1-yl, 1,2,3-triazol-2-yl or 1,2,4- triazol-2-yl), thiadiazolyl (e.g. 1,3,4-thiadiazol-2-yl), pyrimidinyl (e.g. 5-pyrimidinyl), tetrazolyl (e.g. 1,2,3,4-tetrazol2-yl, 1 ,2,3,4-tetrazol-1-yl), imidazolyl (e.g. 2-imidazolyl). Such heteroaryl groups may be unsubstituted or substituted with one or two (eg two or preferably one) substituent(s) selected from halogen, cyano, C _1-4 alkyl (eg C _1-2 alkyl), C _1-4 alkyloxy (e.g. _C1-2 alkyloxy) and _C1-4 alkylcarbonyl (e.g. _C1-2 alkylcarbonyl), e.g. _-OCH3 , methyl, halogen (e.g. chlorine), cyano and -C(O) _-CH3 .

In the case where R ⁴ is the above (iii), the C _1-6 alkyl group is methyl, that is, -CH3, substituted by aryl (eg, phenyl, such as unsubstituted phenyl) or heteroaryl (eg, 5- or 6- member monocyclic heteroaryl group containing one or

- 4 043636 two (eg one) heteroatom(s), thus forming, for example, a thienyl group such as a 2thienyl group; and such heteroaryl group is preferably unsubstituted).

In the case where R ⁴ is (iv) or (v) above, aryl is preferably unsubstituted phenyl, and therefore the R ⁴ group is -O-phenyl or -S(O) ₂ -phenyl.

More preferably, the R ⁴ group is (i) or (ii) above, ie aryl or heteroaryl. Even more preferably, the R ⁴ group is (i) above, especially unsubstituted phenyl.

Most preferred compounds of formula (I) include those in which the X-containing fused bicyclic moiety is:

r ⁴ - ^{R4 -} wherein R ⁴ is as defined herein. Compounds which contain either an N(R ⁴ ) moiety or a C(R ⁴ ) moiety immediately adjacent to the double bond may be useful. This occurs because the shape of the nitrogen atom (for example, being flatter in nature compared to a CR ⁴ moiety that is not directly adjacent to the double bond), or the presence of a double bond in the X-containing ring can help orient the R ⁴ group ( if present) such that the compound as a whole (eg in terms of the orientation of the R ⁴ substituent) exhibits better/improved binding properties towards the bacterial enzyme FabI. Therefore, these compounds of the present invention may be useful in that the presence of a double bond may result in improved binding to or inhibition of the FabI enzyme. Therefore, the compounds of the present invention may be useful compounds (eg, compared to known compounds) due to these properties, which may thereby lead to better potency, effectiveness, and the like.

Compounds of formula (I) can generally be prepared by reacting an intermediate of formula (II) with an intermediate of formula (III) in at least one inert solvent and optionally in the presence of at least one suitable coupling reagent and/or suitable base, said the method further optionally involves converting the compound of formula (I) to its addition salt and/or obtaining stereochemically isomeric forms thereof.

It may be convenient to activate the carboxylic acid of formula (III) by adding an effective amount of a reaction accelerator. Non-limiting examples of such reaction accelerators include carbonyldiimidazole, N,N'-dicyclohexylcarbodiimide or 1-(3-dimethylaminopropyl)-3ethylcarbodiimide, hydroxybenzotriazole, benzotriazolyl-oxy-tris-(dimethylamino)phosphonium hexafluorophosphate, tetrapyrrolidinephosphonium hexafluorophosphate, bromotripyrrolidinephosphonium hexafluorophosphate, or their functional derivative.

Compounds of formula (I) can also be prepared by reacting an intermediate of formula (II) with an intermediate of formula (IV) wherein Y is hydroxyl or halogen. The reaction can be carried out in an inert solvent, such as, for example, dichloromethane or dimethylformamide, and optionally in the presence of a suitable base, such as, for example, diisopropylethylamine (DIPEA).

? /i=\ /=°

V\Xy ^{NH + NH}

R ^3N (II)(IV)

Compounds of formula (I) in which A is -C(R ² )=C(R ¹ )- can also be prepared by reacting an intermediate of formula (V) with an intermediate of formula (VI),

where X _a1 represents a suitable leaving group, such as a suitable halide group (for example, chloro-, iodo- and especially bromo-), and other designations as defined here and above, under suitable reaction conditions, for example, under coupling reaction conditions in in the presence of a metal catalyst, (for example, a coupling reaction in the presence of a precious metal, where the precious metal is, for example, palladium-based), especially under Heck reaction conditions using preferably a palladium-based catalyst such as palladium acetate, tetrakis(triphenylphosphine)palladium( 0), bis-(triphenylphosphine)palladium(P) dichloride, [1,1'-bis-(diphenylphosphino)ferrocene]palladium(H) dichloride or the like (preferably the catalyst is

- 5 043636 palladium acetate), for example, optionally in the presence of a suitable solvent (for example, acetonitrile or the like), a base (for example, an amine base such as N,Ndiisopropylamine or the like) and a ligand (for example, triphenylphosphine, tri-O -tolylphosphine or the like). The reaction can be carried out in a sealed tube and/or in a microwave oven.

The starting materials and some intermediate compounds are known compounds and can be purchased or prepared according to conventional reaction procedures generally known in the art.

For compounds in which Z ₁ represents CH, intermediates (IV) and (VI) can be prepared as described herein or according to conventional reaction procedures generally known in the art. This may also be applicable in the case of corresponding intermediates in which Z ₁ represents N. However, such compounds can also be prepared according to the following scheme:

CAS 16298-03-6

Conditions:

a) NBS, ACN, reflux, 3 hours, 70%;

b) LiAlH4 1M in THF, THF, 5°C to room temperature, overnight, 20%;

c) PBr ₃ , DCM, room temperature, overnight, 90%;

f) dimethyl malonate, NaOMe in MeOH, MeOH, room temperature, overnight, 25%;

g) NaOH, MeOH, reflux, 4 hours, HCl, reflux, overnight;

h) DIEA, Pd(OAc) ₂ , tri-O-tolylphosphine, ACN, DMF, microwave, 180°C, 25 min.

The compounds of formula (I) obtained in the methods described here and above can be synthesized in the form of racemic mixtures of enantiomers, which can be separated from each other following separation procedures known in the art. These compounds of formula (I), which are obtained in racemic form, can be converted into the corresponding diastereomeric salt forms by reaction with a suitable chiral acid. Said diastereomeric salt forms are then separated, for example by selective or fractional crystallization, and the enantiomers are isolated from there using alkali. Alternatively, liquid chromatography using a chiral stationary phase is used to separate the enantiomeric forms of the compounds of formula (I). Said pure stereochemical isomeric forms can also be prepared from the corresponding pure stereochemical isomeric forms of suitable starting materials, provided that the reaction proceeds stereospecifically. If a specific stereoisomer is desired, it is preferred that said compound be synthesized by stereospecific preparation methods. These methods advantageously use enantiomerically pure starting materials.

The compounds described herein are inhibitors of the FabI enzyme, as illustrated by the examples below (including pharmacological example 1). Given these FabI enzyme inhibitory properties, the compounds described herein are useful in the treatment of bacterial infections.

For example, these compounds are useful in the treatment of bacterial infections, such as, for example, upper respiratory tract infections (eg, otitis media, bacterial tracheitis, acute epiglottitis, thyroiditis), lower respiratory tract infections (eg, empyema, lung abscess), cardiac (eg , infective endocarditis), gastrointestinal (eg, secretory diarrhea, splenic abscess, retroperitoneal abscess), central nervous system (eg, cerebral abscess), eye (eg, blepharitis, conjunctivitis, keratitis, endophthalmitis, preseptal and orbital cellulitis, dacryocystitis), kidney and urinary tract (eg, epididymitis, intrarenal and perirenal abscess, toxic shock syndrome), skin (eg, impetigo, folliculitis, skin abscesses, cellulitis, wound infections, bacterial myositis), and bones and joints (eg, septic arthritis, osteomyelitis ). In addition, the compounds may be useful in combination with known antibiotics.

Thus, the present invention also relates to compounds of formula (I) for use as a medicine, especially for use in the treatment of bacterial infections, in particular bacterial infections caused by bacteria expressing the FabI enzyme. The present compounds may subsequently be used in the manufacture of a medicament for the treatment of bacterial infections, in particular bacterial infections caused by a bacterium expressing the FabI enzyme.

In addition, the present invention provides a method for treating bacterial infections which comprises administering to a subject in need thereof a compound of formula (I) that inhibits the FabI enzyme.

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The subject in need of treatment has a bacterial infection or has been in contact with an infectious bacterium, the symptoms of which can be alleviated by administration of a therapeutically effective amount of the compounds of the present invention. For example, a subject in need of treatment may have an infection against which compounds of formula (I) may be administered as treatment. In another example, the subject in need of treatment may have an open wound or burn, for which compounds of formula (I) may be administered as treatment. Typically the subject is treated against an existing bacterial infection.

The subject may have a bacterial infection caused by

Bacillus anthracis, Citrobacter sp., Escherichia coll,

Francisella tularensis, Haemophilus influenza, Listeria monocytogenes, Moraxella catarrhalis, Mycobacterium tuberculosis, Neisseria meningitidis, Proteus mirabilis, Proteus vulgaris, Salmonella sp., Serratia sp., Shigella sp., Stenotrophomonas maltophilia, Staphylococcus aureus or Staphylococcus epidermi di s.

Preferably, the subject is treated (prophylactic or therapeutic) against a bacterial infection caused by a bacterium that expresses the FabI enzyme.

The term treat and cure as used herein refers to curative, palliative and prophylactic treatment, including reversing, alleviating, slowing the progress of the disease, disorder or condition to which such term applies, or one or more symptoms of such disease, disorder or condition.

A therapeutically effective amount of a compound of this invention is an amount that, when administered to a subject in need of treatment, improves the subject's prognosis, eg, delays the onset and/or reduces the severity of one or more symptoms in the subject associated with a bacterial infection. The amount of the disclosed compound required to administer to a subject will depend on the specific disease, the route of administration, and characteristics of the subject, such as general health, presence of other diseases, age, sex, genotype, body weight, and drug tolerance. One skilled in the art will be able to determine the required dosages depending on these and other factors.

Compounds can be tested in one of several biological studies to determine the concentration of the compound that is required for a given pharmacological effect.

In addition, the present invention provides pharmaceutical compositions containing at least one pharmaceutically acceptable carrier and a therapeutically effective amount of a compound of formula (I).

For the purpose of preparing pharmaceutical compositions in accordance with the present invention, an effective amount of a particular compound, in base form or in acid addition salt form, as the active ingredient is combined in a uniform mixture with at least one pharmaceutically acceptable carrier, which carrier can take a wide variety of forms in depending on the form of the drug desired for administration. These pharmaceutical compositions are preferably in unit dosage form, preferably suitable for oral administration, rectal administration, transdermal administration or parenteral injection.

For example, when preparing the compositions in oral dosage form, any of the usual liquid pharmaceutical carriers, such as, for example, water, glycols, oils, alcohols, etc., can be used in the case of oral liquid preparations such as suspensions, syrups, infusions and solutions; or solid pharmaceutical carriers such as starches, sugars, kaolin, lubricants, binders, disintegrants and the like, in the case of powders, pills, capsules and tablets. Because of their ease of administration, tablets and capsules represent the most convenient oral dosage unit form, in which case solid pharmaceutical carriers are obviously used. In the case of compositions for parenteral injection, the pharmaceutical carrier will primarily comprise sterile water, although other components may be included to improve the solubility of the active ingredient. Injectable solutions can be prepared by using a pharmaceutical carrier containing saline, glucose, or a mixture thereof. Injectable suspensions can be prepared by using suitable liquid carriers, suspending agents, and the like. In compositions suitable for transdermal administration, the pharmaceutical carrier may optionally contain a penetration enhancing agent and/or a suitable wetting agent, optionally combined with suitable additives of any nature in small amounts that do not have a significant deleterious effect on the skin. Said additives may be selected to facilitate delivery of the active ingredient to the skin and/or to facilitate the preparation of the desired compositions. These topical compositions can be administered in a variety of ways, for example, as a transdermal patch, spot application or ointment. The addition salts of the compounds of formula (I), due to their increased solubility in water compared to the corresponding base form, are more suitable for the preparation of aqueous compositions.

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It is particularly convenient to formulate the pharmaceutical compositions of this invention in unit dosage form for ease of administration and uniformity of dosage. Unit dosage form, as used herein, refers to physically discrete units suitable as unitary dosages, each unit containing a predetermined amount of the active ingredient calculated to produce the desired therapeutic effect, together with the required pharmaceutical carrier. Examples of such unit dosage forms are tablets (including divisible or film-coated tablets), capsules, pills, powder packets, wafers, injections or suspensions, heaped teaspoons, heaped tablespoons, and the like. ., as well as their allocated multiples.

When administered orally, the pharmaceutical compositions of this invention may take the form of a solid dosage form, for example, tablets (either chewable or swallowable), capsules, or gel capsules, prepared by conventional methods using pharmaceutically acceptable excipients and carriers such as binders. agents (e.g., pregelatinized corn starch, polyvinylpyrrolidone, hydroxypropyl methylcellulose, etc.), fillers (e.g., lactose, microcrystalline cellulose, calcium phosphate, etc.), lubricants (e.g., magnesium stearate, talc, silica and the like), disintegrating agents (for example, potato starch, sodium starch glycolate, etc.), wetting agents (for example, sodium lauryl sulfate), etc. Such tablets may also be coated by methods known in the art.

Liquid preparations for oral administration may take the form of, for example, solutions, syrups or suspensions, or may be formulated as a dry product for mixing with water and/or other suitable liquid carrier before use. Such liquid preparations may be prepared by conventional methods, optionally with other pharmaceutically acceptable additives such as suspending agents (eg sorbitol syrup, methylcellulose, hydroxypropyl methylcellulose or hydrogenated edible fats), emulsifiers (eg lecithin or gum arabic), non-aqueous vehicles (eg almond oil, fatty esters or ethyl alcohol), sweeteners, flavorings, masking agents and preservatives (for example, methyl or propyl phydroxybenzoates or sorbic acid).

Pharmaceutically acceptable sweeteners suitable for the pharmaceutical compositions of this invention include at least one intense sweetener such as aspartame, acesulfame potassium, sodium cyclamate, alitame, dihydrochalcone sweetener, monellin, stevioside sucralose (4,1',6'-trichloro4,1 ',6'-trideoxygalactosucrose) or preferably saccharin, sodium or calcium saccharin, and optionally at least one bulk sweetener such as sorbitol, mannitol, fructose, sucrose, maltose, isomalt, glucose, hydrogenated glucose syrup, xylitol, caramel or honey . Intense sweeteners are usually used in small quantities. For example, in the case of sodium saccharin, said concentration may vary from about 0.04% to 0.1% (w/v) of the final formulation. The bulk sweetener can be effectively used in higher concentrations, ranging from about 10% to about 35%, preferably from about 10% to 15% (w/v).

Pharmaceutically acceptable flavors that can mask bitter-tasting ingredients in low dosage formulations are preferably fruit flavors such as cherry, raspberry, blackcurrant or strawberry flavors. A combination of two flavors can give very good results. High dosage formulations may require stronger pharmaceutically acceptable flavors such as caramel chocolate, cool mint, fantasy, etc. Each flavoring agent may be present in the final composition at a concentration ranging from about 0.05% to 1% (w/v). Combinations of the mentioned strong flavoring agents are advantageously used. It is preferable to use a flavoring agent that does not undergo any change or reduction in taste and/or color under the conditions of formulation.

Compositions of formula (I) can be formulated for parenteral administration by injection, preferably intravenous, intramuscular or subcutaneous injection, for example, bolus injection or continuous intravenous infusion. Injectable formulations may be formulated as unit dose, for example, in ampoules or multi-dose containers, including an added preservative. They may be formulated as suspensions, solutions or emulsions in oily or aqueous vehicles, and may contain formulation agents such as isotonicating, suspending, stabilizing and/or dispersing agents. Alternatively, the active ingredient may be provided in powder form for mixing with a suitable vehicle, for example, sterile, pyrogen-free water, before use.

The compounds of formula (I) can be formulated into rectal compositions such as suppositories or retention enemas, for example containing traditional suppository bases such as cocoa butter and/or other glycerides.

Those skilled in the treatment of bacterial diseases associated with FabI enzyme inhibition can readily determine a therapeutically effective amount of a compound of formula (I) from the results

- 8 043636 tests presented here and below. In general, it is expected that a therapeutically effective dose will be from about 0.001 mg/kg to about 50 mg/kg body weight, more preferably from about 0.01 mg/kg to about 10 mg/kg body weight of the patient being treated. It may be appropriate to administer the therapeutically effective dose in two or more sub-doses at appropriate intervals throughout the day. Said sub-doses may be formulated in unit dosage forms, each containing from about 0.1 mg to about 1000 mg, in particular from about 1 to about 500 mg of active ingredient per unit dose form.

The exact dosage and frequency of administration will depend on the particular compound of formula (I) used, the particular condition being treated, the severity of the condition being treated, the age, weight and general physical condition of the particular patient, as well as other drug treatments the patient may be taking, such as this is well known to those skilled in the art. In addition, said therapeutically effective amount may be reduced or increased depending on the response of the patient undergoing treatment and/or depending on the assessment of the attending physician prescribing the compounds of the present invention. The effective daily amount ranges given above and below are guidelines only.

Compounds of formula (I) may have the advantage that they may be more potent, less toxic, longer acting, more potent, produce fewer side effects, be more easily absorbed, and/or have an improved pharmacokinetic curve (e.g., have a higher biological availability and/or lower clearance), and/or have beneficial pharmacological, physical or chemical properties compared to compounds known in the prior art, both when used in the above cases and in other cases. Compounds may also exhibit such advantages due to the presence of an NR ⁴ moiety or a CR ⁴ moiety that is immediately adjacent to the double bond in the X-containing ring.

For example, compounds of formula (I) may have the advantage that they have good or improved thermodynamic stability (for example, compared to compounds known in the prior art; and, for example, as determined by a known method and/or the method described herein ). The compounds of formula (I) may also have the advantage that they have a broad spectrum of activity compared to antibacterial agents (for example, a broader spectrum of antibacterial activity compared to compounds known in the prior art; and, for example, as determined by known tests and /or the tests described here). The compounds of formula (I) may also have the advantage of having good or improved in vivo pharmacokinetics and oral bioavailability. They may also have the advantage of having good or improved in vivo efficacy. For example, the compounds of the present invention may be adapted for intravenous formulation/dosing and, therefore, may demonstrate improved in vivo efficacy when administered intravenously. Compounds may also exhibit such advantages due to the presence of an NR ⁴ moiety or a CR ⁴ moiety that is immediately adjacent to the double bond in the X-containing ring.

Experimental part.

Abbreviations.

DMF is defined as N,N-dimethylformamide, DCM or CH2Cl2 is defined as dichloromethane, MeOH is defined as methanol, EtOH is defined as ethanol, MgSO ₄ is defined as magnesium sulfate and THF is defined as tetrahydrofuran, AcOEt or EtOAc is defined as ethyl acetate, DIPEA is defined as diisopropylethylamine, EDCI is defined as N'(ethylcarbonimidoyl)-N,N-dimethyl-1,3-propanediamine monohydrochloride, HOBT is defined as 1hydroxy-1H-benzotriazole, DIPA is defined as diisopropylamine, K2CO3 is defined as potassium carbonate, TFA is defined as trifluoroacetic acid, NH4OH is defined as as ammonium hydroxide, NaHCO ₃ is defined as sodium bicarbonate, Et2O is defined as diethyl ether, Na2SO4 is defined as sodium sulfate, CH3CN is defined as acetonitrile, NaOH is defined as sodium hydroxide, n-BuLi is defined as n-butyllithium, i-PrOH is defined as isopropanol, Pd(OAc)2 is defined as palladium acetate, DMA is defined as dimethylacetamide, Et ₃ N is defined as triethylamine.

Stereochemical representation.

Compounds of formula (I) have at least two asymmetric carbon atoms, as shown below, where the asymmetric carbon atoms are identified by the symbol*:

n

Due to ring stress, only cis forms can be obtained in a system of two ring-shaped five-membered rings, but not trans forms.

- 9 043636

Compounds of formula (I), wherein the system of two annular five-membered rings has a cis configuration

N

Each of the cis compounds described above consists of a racemic mixture of two enantiomers, and bolded or dashed bonds have been used to indicate these respective stereochemical configurations.

When such a cis compound resolves into two separate enantiomers, the stereochemical configuration of one of these enantiomers is designated R* or S*, indicating the relative stereochemistry. Accordingly, one enantiomer, designated (R*,S*), can have either the absolute (R,S) configuration or the (c,R) configuration. If the absolute stereochemistry of a particular chiral carbon atom in one enantiomer is known, the bold and dashed bonds have been replaced by wedge bonds to indicate that the compound is a single enantiomer of known absolute stereochemistry.

A. Synthesis of intermediates.

Example A. 1.

intermediate connection (1)

A solution of 6-bromo-3,4-dihydro-1H-[1,8]naphthyridin-2-one (1.0 g, 4.4 mmol), tert-butyl acrylate (2.56 ml, 17.62 mmol) and N,N-Diisopropylethylamine (1.46 ml, 8.81 mmol) in acetonitrile (20 ml) and DMF (7 ml) was stirred and degassed using nitrogen gas for 10 min. Tri-o-tolylphosphine (0.27 g, 0.88 mmol) and palladium(II) acetate (47% based on Pd) (0.099 g, 0.44 mol) were added and the resulting mixture was microwaved (1600 W , 180°C, 35 min). The reaction mixture was evaporated to dryness, dissolved in DCM/methanol (8/2) (50 ml), filtered through a thin pad of celite and washed with dichloromethane. The organic layer was washed with water, dried (MgSO ₄ ), filtered and evaporated to dryness. The residue was dissolved in cold ethanol (10 ml) and stirred at 5°C for 5 min, the precipitate was filtered off, washed with cold ethanol (3 ml) and dried under vacuum to obtain 950 mg of intermediate (1).

intermediate connection (2)

Intermediate (1) (4.1 g, 14.95 mmol) was dissolved in trifluoroacetic acid (23.2 ml) in DCM (41 ml). The reaction composition was stirred at room temperature for 30 minutes. The reaction mixture was concentrated under reduced pressure. The resulting solid was triturated with diethyl ether, filtered and dried under vacuum to give 3.97 g of intermediate (2).

intermediate connection (3)

Intermediate (2) was triturated overnight in HCl in dioxane (4 M, 48 ml), the solid was filtered, washed with diethyl ether and dried under vacuum to give 3.9 g of intermediate (3).

Example A.2.

a) Receipt

intermediate connection (4)

A solution of allyl-prop-2-ynyl-carbamic acid tert-butyl ester (CAS 147528-20-9, 45 g, 0.23 mol), cobalt carbonyl (17.5 g, 46.1 mmol) and 1.1, 3,3-Tetramethyl-2-thiourea (36.6 g, 0.277 mmol) in toluene (1.8 L) was stirred and heated at 70°C for 5 hours in an autoclave under CO pressure (2-3 bar). The resulting mixture was filtered through a thin pad of celite and evaporated to dryness. The residue was dissolved in DCM and filtered through a fine pad of celite to obtain a clear solution. It was evaporated to dryness to obtain 85.7 g of a crude residue. It was purified by preparative liquid chromatography on (silica gel 20-45 µm, 1000 g, mobile phase (95/5 to 80/20 DCM/AcOEt gradient). The purified fractions were collected and the solvent was evaporated to yield 36.5 g of intermediate ( 4).

intermediate connection (5)

- 10 043636

A mixture of intermediate (4) (37.6 g, 0.168 mol) and 10% palladium on carbon (7.5 g) in ethyl acetate (750 ml) was hydrogenated at room temperature for 30 min at 3 bar in a closed reactor. The resulting mixture was filtered through a fine pad of celite and evaporated to dryness to yield 38.2 g of intermediate (5).

c) Receipt o ° / f ₃ cso—/2% a ^-0-0- \ ( ^cis > ⁰ n intermediate connections (6)

H-BuLi 1.6 M in hexane (64 ml, 0.102 mol) was added dropwise at -20°C, under N ₂ atmosphere, to a solution of diisopropylamine (14.3 ml, 0.102 mol) in dry THF (140 ml), the mixture was then stirred at -20°C for 20 minutes. A solution of intermediate (5) (19.1 g, 84.8 mmol) in dry THF (190 ml) was then added at -78°C and the resulting mixture was stirred for 1 hour at -78°C. A solution of N-phenyl-trifluoromethane sulfonimide (36.4 g, 0.102 mol) in dry THF (110 ml) was added at -78°C, then the mixture was allowed to warm to room temperature and stirred overnight. The mixture was evaporated to dryness. The residue was dissolved in DCM, washed with aqueous NaHCO ₃ solution, dried (MgSO ₄ ) and evaporated to dryness to obtain 27.7 g of intermediate (6).

d) Receipt V o \ /—\ CT A ^-0-0 χ ^- ( ^cis ) n intermediate connection (7)

A solution of intermediate (6) (9.3 g, 26.0 mmol) and phenylboric acid (3.81 g, 31.2 mmol) in a solution of 2 M potassium carbonate (26 ml) and ethylene glycol dimethyl ether (93 ml) was purged N ₂ for 10 min, then tetrakis(triphenylphosphine)palladium (3.0 g, 2.6 mmol) was added. The closed reactor was heated at 80°C using a single CEM Mars multimode chamber microwave oven with power ranging from 0 to 400 W for 30 min. The resulting solution was cooled to room temperature, water and EtOAc were added, the organic layer was separated, washed with water and then brine, dried (MgSO4) and evaporated to dryness. The residue was purified by flash chromatography on silica gel (330 g, 15-40 µm, heptane/EtOAc 100/0 to 80/20). The purified fractions were collected and evaporated to dryness to obtain 4.3 g of intermediate (7).

f) Receipt \ /—xJL' ^NH ( ^cis ) Η intermediate connection (8)

Trifluoroacetic acid (44 ml) was added dropwise to a solution of intermediate (7) (14.5 g, 50.8 mmol) in CH ₂ Cl ₂ (44 ml). The resulting solution was stirred at room temperature for 30 minutes, then the mixture was cooled to 5°C. 3N was slowly added. NaOH until alkaline, the mixture was extracted twice using CH ₂ Cl ₂ . The combined organic layer was washed with 3N. NaOH and then water was dried over MgSO ₄ and evaporated to give 8.8 g of racemic intermediate (8). _________________________________________________

f) Receipt n n intermediate connection (9) And And th intermediate connections (10)

Intermediate (8) was purified and resolved using chiral SFC on (CHIRALPAK AD-H 5 µm 250x20 mm). Mobile phase (0.3% isopropylamine, 73% CO ₂ , 27% iPrOH). The pure fractions were collected and the solvent was removed to yield 3.9 g of intermediate (10) (R*,S*) ([a] _D ²⁰ = -53.19° (589 nm, concentration 0.3365 w/v .%, DMF, 20°C)) and 4 g of intermediate (9) (S*,R*) ([«]d ²⁰ = +38.6° (589 nm, concentration 0.285 wt./vol. %, DMF, 20°C)).

Intermediate connection (9).

¹ H NMR (400 MHz, DMSO-a ₆ ) δ (ppm) 7.43 (d, J=7.6 Hz, 2H), 7.32 (t, J=7.6 Hz, 2H) , 7.20-7.26 (m, 1H), 6.07 (d, J=2.0 Hz, 1H), 3.30-3.39 (m, 1H), 2.77-2.94 (m, 4H), 2.66 (dd, J=3.0, 11.1 Hz, 1H), 2.58 (dd, J=3.0, 11.1 Hz, 1H), 2.46 ( d, J=15.7 Hz, 1H).

Intermediate connection (10).

1H NMR (400 MHz, DMSO-b ₆ ) δ (ppm) 7.43 (d, J=7.6 Hz, 2H), 7.32 (t, J=7.6 Hz, 2H), 7.20-7.26 (m, 1H), 6.07 (d, J=2.0 Hz, 1H), 3.30-3.39 (m, 1H), 2.77-2.94 ( m, 4H), 2.66 (dd, J=3.0, 11.1 Hz, 1H), 2.58 (dd, J=3.0, 11.1 Hz, 1H), 2.46 (d , J=15.7 Hz, 1H).

Example A.3.

a) Receipt /Α-Ά /° Г у—/ Д' N-Д, (cis) ^{s /} о—Z_ Л \ intermediate connections (11)

A solution of intermediate (6) (44.4 g, 111.82 mmol) and 3-thiophenylboronic acid (17.17 g, 134.19 mmol) in a solution of 2 M potassium carbonate (112 ml) and ethylene glycol dimethyl ether

- 11 043636 (444 ml) the open container was purged with N2 for 10 min, then tetrakis(triphenylphosphine) palladium (12.92 g, 223.65 mmol) was added. The solution was heated at 78°C using a CEM Mars single-chamber microwave oven with power ranging from 0 to 400 W for 1 hour. The mixture was cooled to room temperature and water and EtOAc were added. The resulting mixture was filtered through a celite pad. The organic layer was separated, washed with water, then with brine, dried over MgSO ₄ and evaporated to dryness. The residue was purified by preparative liquid chromatography on (silica gel 20-45 μm, 1000 g, mobile phase (80% heptane, 20% AcOEt)). The purified fractions were collected and concentrated to yield 16 g of intermediate (11).

b) Receipt m S «--/ ^NH ( ^cis ) Η intermediate connections (12)

Trifluoroacetic acid (14.37 ml, 186.47 mmol) was added to a solution of intermediate (11) (5.72 g, 18.65 mmol) in CH ₂ Cl ₂ (57 ml). The reaction mixture was stirred at room temperature for 3 hours. K ₂ CO ₃ (10% aqueous solution, 50 ml) and then solid K ₂ CO ₃ was added at 0° C. to make the solution alkaline. The organic layer was separated, washed with water, dried (MgSO ₄ ) and evaporated to dryness. The residue was purified by preparative liquid chromatography on (silica gel 20-45 µm, 1000 g, mobile phase (1% NH4OH, 93% DCM, 7% MeOH)). The purified fractions were collected and concentrated to yield 12 g of intermediate (12).

c) Receipt n intermediate connections (13) And n intermediate connections (14)

Intermediate (12) was purified and resolved using chiral SFC on (CHIRALPAK

AD-H 5 µm 250x20 mm). Mobile phase (0.3% isopropylamine, 80% CO ₂ , 20% methanol). The pure fractions were collected and the solvent was removed to yield 5.8 g of intermediate (14) (R*,S*) ([a] _D ²⁰ = -12.4° (589 nm, 0.5 w/v concentration .%, DCM, 20°C)) and 5.6 g of intermediate (13) (S*,R*) ([a]D ²⁰ =+9.43° (589 nm, concentration 0.35 wt ./vol.%, DCM, 20°C)).

Intermediate connection (13).

1H NMR (500 MHz, DMSO-d ₆ ) δ (ppm) 7.49 (yes, J=2.5, 5.0 Hz, 1H), 7.31 (d, J=5.0 Hz , 1H), 7.29 (d, J=2.5 Hz, 1H), 5.88 (d, J=1.9 Hz, 1H), 3.28-3.33 (br.s, 1H) , 2.75-2.87 (m, 4H), 2.61 (yes, J=2.8, 10.7 Hz, 1H), 2.54 (dd, J=3.3, 10.9 Hz , 1H), 2.40-2.15(m, 2H).

Intermediate connection (14).

1H NMR (500 MHz, DMSO-d6) δ (ppm) 7.49 (dd, J=2.5, 5.0 Hz, 1H), 7.31 (d, J=5.0 Hz, 1H), 7.29 (d, J=2.5 Hz, 1H), 5.88 (d, J=1.9 Hz, 1H), 3.28-3.33 (br.s, 1H), 2.75-2.87 (m, 4H), 2.61 (dd, J=2.8, 10.7 Hz, 1H), 2.54 (dd, J=3.3, 10.9 Hz, 1H), 2.40-2.15 (m, 2H).

Example A.4.

intermediate connection (15)

A solution of intermediate (6) (108 g, 0.302 mol) and pyridine-4-boric acid (49.5 g, 0.363 mol) in 2 M aqueous potassium carbonate (302 ml, 0.604 mol) and ethylene glycol dimethyl ether (1.1 l) purged using N ₂ for 5 min, then tetrakis(triphenylphosphine) palladium (34.9 g, 0.030 mol) was added, the mixture was heated at 78°C using a multi-mode microwave oven (CEM Mars 5) with power in the range from 0 to 800 W for 1 hour, cooled to room temperature, added EtOAc, the organic layer was separated, washed with water and then brine, dried over MgSO ₄ and evaporated to dryness. The residue was purified by preparative liquid chromatography on (silica gel 15-40 µm, 300 g, mobile phase (0.1% NH4OH, 97% DCM, 3% iPrOH). The purified fractions were collected and the solvent was removed to yield 47.6 g of intermediate 15.

Intermediate (16) was purified and isolated by chromatography on Chiralpak AD (20 µm, 2000 g, 110 mm) at 750 ml/min. The mobile phase was 100% methanol. The pure fractions were collected and evaporated to dryness to obtain 18.7 g of intermediate (18) (R*,S*) ([a] _D ²⁰ = +55.75° (589 nm, concentration 0.339 w/v .%, DMF, 20°C)) and 20.7 g of intermediate (17) (S*,R*) ([a] _D ²⁰ = -68.38° (589 nm, concentration 0.253 wt./ vol.%, DMF, 20°C)).

- 12 043636

Intermediate connection (17).

¹ H NMR (500 MHz, DMSO-d ₆ ) δ (ppm) 8.52 (d, J=6.0 Hz, 2H), 7.41 (d, J=6.0 Hz, 2H) , 6.50 (s, 1H), 3.363.61 (m, 4H), 2.81-3.02 (m, 3H), 2.61-2.53 (m, 1H), 1.36 (s , 9H).

Intermediate connection (18).

¹ H NMR (500 MHz, DMSO-d6) δ (ppm) 8.52 (d, J=6.0 Hz, 2H), 7.41 (d, J=6.0 Hz, 2H), 6.50 (s, 1H), 3.363.61 (m, 4H), 2.81-3.02 (m, 3H), 2.61-2.53 (m, 1H), 1.36 (s, 9H).

Example A.5

Receipt - ^ns| R intermediate connections (19)

Intermediate (18) (24.8 g, 86.6 mmol) was added to dioxane HCl (4 M, 108 mL) at 5° C., then the mixture was stirred at room temperature for 90 minutes. The resulting precipitate was filtered, washed with diethyl ether and dried under vacuum at 70°C to obtain 21.1 g of intermediate (19).

Receipt n - SFU ¹⁴¹⁴ ' ^NS1 n intermediate connections (20)

Intermediate (20) was prepared similarly, starting with intermediate (17).

Example A-6.

a) Receipt ABOUT \ intermediate connections (21)

The reaction was carried out in 4 batches of 0.5 g each of 6-bromo-3,4-dihydro-1H-[1,8]naphthyridin-2-one. Solution of 6-bromo-3,4-dihydro-1H-[1,8]naphthyridin-2-one (0.5 g, 2.20 mmol), bis-(pinacolato)diboron (0.67 g, 2.64 mmol) and potassium acetate (0.648 g, 6.61 mmol) in DMF (5 ml), and CH3CN (10 ml) were stirred and degassed using nitrogen for 10 min. 1,1'-Bis(diphenylphosphine)ferrocenedichloropalladium (II) (0.161 g, 0.22 mmol) was added and heated at 120°C using a microwave oven (Biotage initiator 60) with power ranging from 0 to 400 W for 40 min. The mixture was evaporated to dryness, the residue was dissolved in DCM and water, filtered through a thin pad of celite. The organic layer of the filtrate was separated, washed with water, dried (MgSO ₄ ) and evaporated to dryness. The residue was dissolved in EtOH, filtered and dried to give 0.36 g of intermediate (21).

b) Receipt 1° N N Yu N intermediate connections (22)

Intermediate (21) (1.0 g, 3.65 mmol), tert-butylpropionate (0.426 ml, 3.04 mmol), silver(I) oxide (1.06 g, 4.56 mmol) and K2CO3 (0 .84 g, 6.08 mmol) in CH3CN (10 ml) and DMF (5 ml) were purged with N ₂ gas, then palladium(II) acetate (47% Pd) (0.034 g, 0.152 mmol) was added and the mixture was heated at 100 °C using a single-mode microwave oven (Biotage initiator 60) with power ranging from 0 to 400 W for 20 min. Water and EtOAc were added, the mixture was filtered through a fine pad of celite, the organic layer was separated, washed with water and then dried with brine (MgSO ₄ ) and evaporated to dryness. The resulting residue was purified by flash chromatography on silica gel (15-40 µm, 30 g cartridge, CH ₂ Cl ₂ to CH ₂ Cl ₂ /CH ₃ OH/NH ₄ OH: 98.5/1.5/0.1 ). The purified fractions were collected and evaporated to dryness to yield 0.037 g of intermediate (22). _______________________________________________________________

c) Receipt lYI N N O N intermediate connections (23)

Intermediate (22) (0.053 g, 0.195 mmol) was dissolved in TFA/DCM solution (0.37 ml/0.5 ml). The reaction mixture was stirred at room temperature for 30 minutes. The reaction mixture was concentrated under reduced pressure. The resulting solid was triturated with diethyl ether, filtered and dried under vacuum (80°C) to obtain 0.032 g of intermediate (23).

Example A.7 _________________________________________________

a) Receipt intermediate connections (24)

Microwave conditions: Biotage, 90°C, 25 min, slow after 30 s pre-heating

- 13 043636 stirring. A solution of bromobenzene (0.228 ml, 2.64 mmol), cis-2-tert-butyloxy-carbonylhexahydropyrrolo[3.4]pyrrole (0.6 g, 2.82 mmol) and sodium tert-butoxide (0.624 g, 6.5 mmol) in toluene (well dried with molecular sieves) (15 ml) was stirred and degassed using nitrogen for 10 min. Tris-(dibenzylideneacetone)dipalladium(0) (0.198 g, 0.216 mmol) and 2(di-tert-butylphosphino)biphenyl (0.065 g, 0.216 mmol) were added and the resulting mixture was re-irradiated following the microwave conditions described above. Water and ethyl acetate were added and the organic layer was separated and then dried (MgSO ₄ ), filtered and concentrated. The resulting residue was purified by flash chromatography on silica gel (15-40 µm, 40 g, heptane/EtOAc 80/20). The purified fractions were collected and concentrated to give intermediate (24). ______________________________________________________________

b) Receipt n n intermediate connections (25)

TFA (4.54 mL, 58.95 mmol) was added to a solution of intermediate (24) (1.7 g, 5.9 mmol) in DCM (15 mL). The reaction mixture was stirred at room temperature for 2 hours, water and DCM were added, K ₂ CO ₃ (10% aqueous solution) was added to make alkalinized, and the organic layer was separated, washed with water, dried (MgSO4) and evaporated to dryness with obtaining intermediate (25) as an oil.

The following compounds were prepared using the same procedure as in Example A.7, where bromobenzene was replaced by 2-bromothiophene, 2-bromoanisole, 2-bromo-1-methylbenzene, 2-bromo-1chlorobenzene, 3-bromopyridine, 2 -bromothiazole, 4-bromo-1-chlorobenzene or 3-bromo-1-chlorobenzene, respectively.

n \ P ⁿ f V- N Rp NH N 0-<f> ⁿ n intermediate connection (26) intermediate connection (27) intermediate connection (28) n n n rV ^N GK ^NH Η intermediate connection (29) intermediate connection (30) intermediate connection (31) n A- n n intermediate connection (32) intermediate connection (33)

Example A.

a) Receipt °° / F3C-S-0—С NCO-+- (cis) ⁰ n intermediate connections (34)

The reaction was carried out under N2 atmosphere. n-BuLi (1.6 M in hexane) (3.33 ml, 5.33 mmol) was added dropwise at -20°C to a solution of DIPA (0.749 ml, 5.33 mol) in THF (8 ml), then the mixture was stirred at 20°C for 20 minutes. A solution of intermediate (4) (1.0 g, 4.44 mmol) in THF (10 ml) was then added at -78°C and the resulting mixture was stirred for 30 minutes at -78°C. A solution of N-phenyltrifluoromethane-methanesulfonimide (1.4 g, 4.88 mol) in THF (6 ml) was added at -78°C, then the mixture was allowed to warm to room temperature and stirred overnight. The mixture was concentrated and the residue was purified by flash chromatography on silica gel (40 g, 15-40 µm, heptane/EtOAc 70/30). The purified fractions were collected and evaporated to dryness, yielding intermediate (34).

b) Receipt Q-^T^M-C-Q—(Cis) Η intermediate connections (35)

Reaction in a nitrogen atmosphere. Microwave conditions: Biotage initiator 60, 80°C, 20 min. A solution of intermediate (34) (0.42 g, 0.881 mmol) and thiophene-2-boric acid (0.135 g, 1.06 mmol) in K2CO3 (2 M, 0.88 ml) and ethylene glycol dimethyl ether (4 ml) purged with N2 for 10 min, then added tetrakis(triphenylphosphine)palladium(0) (0.102 g, 0.088 mmol). The mixture was irradiated under the microwave conditions described above, cooled to room temperature, water and EtOAc were added, the organic layer was separated, washed with water and then dried with brine (MgSO ₄ ) and evaporated to dryness. The resulting residue was purified by flash chromatography on silica gel (10 g, 15-40 µm, heptane 100 to heptane/EtOAc 80/20). The purified fractions were collected and evaporated to dryness to yield intermediate (35).

- 14 043636

c) Receipt

intermediate connection (36)

A mixture of intermediate (35) (0.226 g, 0.776 mmol) in TFA (0.7 ml) and DCM (4 ml) was stirred at room temperature for 1 hour, then the reaction mixture was poured into K ₂ CO ₃ (10% aqueous solution) and extracted using DCM. The organic layer was separated, washed with water, dried (MgSO ₄ ) and evaporated to dryness to give intermediate (36).

The following compounds were prepared using the same procedure as in Example A.8b/A.8c, where thiophene-2-boric acid was replaced by 2-methoxyphenylboric acid or formic acid, respectively.

intermediate connection (37)

Example A.9.

Microwave conditions: Biotage 60, 120°C, 30 min. A mixture of cis-2-tert-butyloxycarbonyl-hexahydropyrrolo[3,4]pyrrole (0.027 g, 0.13 mmol), 2-bromopropane (0.018 ml, 0.19 mmol) and triethylamine (0.088 ml, 0.64 mol) in DMF ( 0.2 ml) were exposed to radiation under the microwave conditions described above. Water and EtOAc were added, the organic layer was separated, the aqueous layer was extracted twice using EtOAc, the combined organic phase was washed with water and brine, dried (MgSO ₄ ) and evaporated to dryness, yielding intermediate (37).

intermediate connection (38)

TFA (0.62 g, 8.02 mmol) was added to a solution of intermediate (37) (0.204 g, 0.8 mmol) in DCM (2 ml). The reaction mixture was stirred at room temperature for 3 hours, water and DCM were added, 10% K ₂ CO ₃ was added to alkalize, solid NaCl was added to saturation, and the organic layer was separated, washed with water, dried (MgSO ₄ ) and evaporated to dry state to obtain intermediate compound (38) as an oil.

The following compounds were prepared using the same procedure as in Example A.9, where 2-bromopropane was replaced by propargyl bromide, benzylsulfonyl chloride or methanesulfonate, respectively.

pF n ⁿ intermediate intermediate intermediate connection (39) connection (40) connection (41)

Example A.10.

a) Receipt _HO ^Y ? / 7\ 1 \ ( ^cis ) intermediate A- ^{n n} connections (44)

Reaction in N ₂ atmosphere. BuLi (1.6 M in hexane) (4.8 ml, 7.70 mmol) was added dropwise at -78°C to a solution of thiazole (0.5 ml, 7.05 mol) in Et ₂ O (5 ml) , then the mixture was stirred for 30 minutes. A solution of intermediate (5) (1.44 g, 6.41 mmol) in Et ₂ O (7 ml) was added, then the mixture was stirred and allowed to warm to room temperature over 2 hours. Water and EtOAc were added, the organic layer was separated, washed with water, then with brine, dried over MgSO ₄ and evaporated to dryness. The resulting residue was purified by flash chromatography on silica gel (50 g, 15-40 µm, heptane/EtOAc 80/20 to heptane/EtOAc 50/50). The purified fractions were collected and evaporated to dryness to yield intermediate (44).

intermediate connection (45)

A mixture of intermediate (44) (1.05 g, 3.38 mmol) in HCl (37% in H ₂ O) (7 ml) in a sealed tube was heated at 140°C using a microwave oven (Biotage Initiator EXP 60) with power ranging from 0 to 400 W for 1 hour. The reaction mixture was poured into K ₂ CO ₃ (10% aqueous solution), the organic layer was separated, dried (MgSO ₄ ) and evaporated to dryness to obtain 0.2 3 g residue (1). The aqueous layer was evaporated to dryness and the solid was suspended in DCM and stirred for 10 minutes. The suspension was filtered and the filtrate was evaporated to dryness to obtain 0.29 g of residue (2). Residues (1) and (2) were combined for purification, which was carried out with

- 15 043636 using flash chromatography on silica gel (15-40 µm, 30 g, from CH2Cl ₂ to CH ₂ Cl ₂ /CH ₃ OH/NH ₄ OH: 90/10/1). The purified fractions were collected and evaporated to dryness to yield 0.41 g of intermediate (45).

Example A.11.

a) Receipt 'ufn-s-'D- (CIS) intermediate connections (46)

Diethylaminosulfur trifluoride (1.24 ml, 10.12 mmol) was added dropwise to a solution of intermediate (5) (0.570 g, 2.53 mmol) in DCM (6 ml), cooled in an ice bath at 5°C, and the mixture was stirred for 1 hour at 5°C and then overnight at room temperature. The mixture was cooled at 0°C and saturated NaHCO ₃ was added. The organic layer was extracted using CH ₂ Cl ₂ , dried (MgSO ₄ ), filtered and concentrated to give intermediate (46).

b) Receipt y /\ 1 / ⁿ (as) Η intermediate connections (47)

TFA (0.39 g, 5.12 mmol) was added to a solution of intermediate (46) (0.146 g, 0.51 mmol) in DCM (1.2 ml). The reaction mixture was stirred at room temperature for 3 hours, water and DCM were added, K ₂ CO ₃ (10% aqueous solution) was added to make alkalinized, and the organic layer was separated, washed with water, dried (MgSO ₄ ) and evaporated to dryness to obtain intermediate (47) as an oil.

Example A.12.

a) Receipt ¥ o \ /—\ ΖΙΓ /Ν ^- s—o—4— (cis) n intermediate connections (48)

A mixture of intermediate (7) (0.3 g, 1.05 mmol) and 10% dry Pd/C (0.06 g) in MeOH (15 ml) was hydrogenated at room temperature and atmospheric pressure for 2 hours. The resulting mixture was filtered through a fine pad of celite, washed using DCM, and the filtrate evaporated to dryness to yield intermediate (48).

b) Receipt \ /—\ T / ^nh ( ^cis ) Η intermediate connections (49)

A mixture of intermediate (48) (0.286 g, 0.995 mmol) and TFA (0.9 ml) in DCM (6 ml) was stirred at room temperature for 30 min, then the reaction mixture was poured into K ₂ CO ₃ (10% aqueous solution ) and extracted using DCM. The organic layer was separated, washed with water, dried (MgSO ₄ ) and evaporated to dryness to give intermediate (49).

Example A.13.

a) Receipt C| _H __/ θ / \ /—(хУс· ⁰ \ Η intermediate connections (50)

Reaction in N2 atmosphere. Microwave conditions: Biotage initiator 60, 80°C, 20 min. A solution of intermediate (38) (0.45 g, 1.26 mmol) and 2-chlorophenylboronic acid (0.236 g, 1.51 mmol) in K2CO3 (2 M, 1.26 ml) and ethylene glycol dimethyl ether (5 ml ) purged with N2 for 10 min, then tetrakis(triphenylphosphine)palladium(0) (0.146 g, 0.126 mmol) was added. The mixture was irradiated under the treatment conditions described above, cooled to room temperature, water and DCM were added, the organic layer was separated, washed with water, dried (MgSO ₄ ) and evaporated to dryness. The residue was purified by preparative liquid chromatography on (silica gel 15 μm, 150x30.0 mm). Mobile phase (100% DCM). The target fractions were collected and the solvent was evaporated to give intermediate (50).

b) Receipt \ /—Cl X ( ^cis ) Η intermediate connections (51)

A mixture of intermediate (50) (0.3 g, 0.938 mmol) and TFA (0.9 ml) in DCM (6 ml) was stirred at room temperature for 30 min, then the reaction mixture was poured into K ₂ CO ₃ (10% aqueous solution) and extracted using DCM. The organic layer was separated, washed with water, dried (MgSO ₄ ) and evaporated to dryness to give intermediate (51).

The following compounds were prepared using the same procedure as in Example A.13, where 2-chlorophenylboronic acid was replaced by 2-methylphenylboric acid, 1-methyl-1Hpyrazole-5-boron pinacol ester, furan-2-boric acid, 2-fluorophenylboric acid, furan-3boric acid, 2-cyanophenylboric acid, 5-dimethylisoxazole-4-boric acid, pyridine-3boric acid, 1-methyl-4-(4,4,5,5-tetramethyl-1, 3,2-dioxaboran-2-yl)-1H-pyrazole, benzylzinc bromide, 2-chloropyridine-3-boric acid, pyrimidyl-5-boric acid pinacolate, 1-BOC-16 043636 pyrazole-4-boric acid pinacol ester, 5-methylfuran-2-boric acid or 4-methoxy-3-pyridinylboric acid, respectively.

\ /—у ₇ ^nh (cis) ы /—\Т/ ^nh ( ^cis > ^Ν γ Хф/ Г 2-у Т NH (cis) Η \ A n intermediate intermediate intermediate connection (52) connection (53) connection (54) y 2—(cis) 1 \-y f NH (cis) Ρ ^Ν n \ /—\J(/ ^nh (cis) Η Η Η intermediate intermediate intermediate connection (55) connection (56) connection (57) / ΐ №®\ / -W ^NH ( ^cis ) \% (cis) ^/-у 1 _Z ^NH ( ^cis ) \ Η Η Η intermediate intermediate intermediate connection (58) connection (59) connection (60) s u--u f NH (CIS) { \—(Th(cis) ы С /—\I/ ^NH ^s) /\n Ύι Η intermediate intermediate intermediate connection (61) connection (62) connection (63) ы/—у ^(cis) £ ₂ \ J2 /NH ( ^cis ) Η n O- intermediate intermediate intermediate connection (64) connection (65) connection (66)

Example A.14.

a) Receipt _s O , |G (I \-co—T (cis) H intermediate connections (67)

Intermediate (34) (2.798 mmol), palladium(P)acetate (47% Pd) (0.14 mmol), K ₂ CO ₃ (4.198 mmol), trimethylacetic acid (0.84 mmol) and tricyclohexylphosphonium tetrafluoroborate (0.196 mmol ) purged with N ₂ in a sealed tube. Thiazole (4.198 mmol) and DMA (10 ml) were added and the reaction mixture was heated at 100°C overnight. Water and EtOAc were added, the organic layer was separated, washed with water and brine, dried over MgSO ₄ and evaporated to dryness. The resulting residue was purified by silica gel flash chromatography (30 g cartridge, 15-40 µm, heptane/EtOAc 80/20 to heptane/EtOAc 60/40). The purified fractions were collected and evaporated to dryness, yielding intermediate (67).

b) Receipt <04 intermediate connections (68)

A solution of intermediate (67) (0.24 g, 0.821 mmol) in TFA (0.8 ml) and DCM (5 ml) was stirred at room temperature for 30 min, then the reaction mixture was poured into K ₂ CO ₃ (10% aqueous solution) and extracted using DCM. The organic layer was separated, washed with water, dried (MgSO ₄ ) and evaporated to dryness, yielding 0.1 g of intermediate (68).

Example A.15.

a) Receipt n \_ /—( ^cis ) Η intermediate connections (69)

Pd(OAc) ₂ (1/3 mg, 0.0056 mmol) was added to a solution of intermediate (34) (0.1 g, 0.28 mmol), 1,3-bis(diphenylphosphine)propane (4.6 mg, 0.011 mmol) and potassium acetate (0.041 g, 0.42 mmol) in EtOH (0.25 ml) and THF (2 ml) under nitrogen. The mixture was stirred under a carbon monoxide atmosphere at 5 bar for 18 hours in a stainless steel autoclave to give intermediate (69).

b) Receipt n y --y T NH (CIS) 'Ί ^/ N intermediate connection (70)

A solution of intermediate (69) (0.2 g, 0.711 mmol) in HCl (4 M in dioxane) (2 ml) was stirred at room temperature for 30 min, then evaporated to dryness, yielding 0.13 g of intermediate ( 70).

Example A.16.

- 17 043636

a) Receipt o ft / /—\ JL, ^nco— C (cis) but \ n intermediate connections (71)

Pd(OAc)2 (25 mg, 0.112 mmol) was added to a solution of intermediate (34) (2.0 g, 5.6 mmol), 1,3-bis(diphenylphosphine)propane (92 mg, 0.22 mmol ) and potassium acetate (0.82 g, 8.4 mmol) in EtOH (5 ml) and THF (40 ml) under nitrogen, then the mixture was stirred under carbon monoxide at 5 bar at 100°C for 18 h in a stainless steel autoclave. The reaction mixture was poured into water and EtOAc, the organic layer was separated, washed with water and then brine, dried (MgSO ₄ ), filtered and evaporated to dryness. The resulting residue was purified by flash chromatography on silica gel (15-40 µm, 40 g, heptane/EtOAc 90/10 to heptane/EtOAc 70/30). The purified fractions were collected and evaporated to dryness to yield 0.61 g of intermediate (71).

b) Receipt /gftg· ^(cis> intermediate connections (72)

Mixture of intermediate (71) (0.3 g, 1.18 mmol), dimethylamine in THF (2 M, 1.18 mL, 2.37 mmol), EDCI (0.27 g, 1.42 mmol), HOBt (0.19 g, 6.21 mmol) and triethylamine (0.25 ml, 1.78 mmol) in DCM (3 ml) and THF (3 ml) were stirred overnight at room temperature. Water and DCM were added, the organic layer was separated, dried (MgSO ₄ ) and evaporated to dryness, yielding 0.37 g of intermediate (72).

c) Receipt n / /G NH (cis) -d Ag intermediate connections (73)

A solution of intermediate (72) (0.37 g, 1.32 mmol) in HCl (4 M in dioxane) (4 ml) was stirred at room temperature for 30 min, then the reaction mixture was poured into K ₂ CO ₃ (10% aqueous solution) and extracted using DCM. The organic layer was separated, washed with water, dried (MgSO4) and evaporated to dryness to give intermediate (73).

Example A.17.

a) Receipt ° /-^Р\ θ / -\Т/С-О-Л (cis) h ₂ n '-Д '' Η intermediate connections (74)

A mixture of intermediate (71) (0.3 g, 1.18 mmol), 1,1,1,3,3,3-hexamethyldisilazane (0.23 g, 1.42 mmol), EDCI (0.27 g, 1.42 mmol), HOBt (0.19 g, 6.21 mmol) and triethylamine (0.25 ml, 1.78 mmol) in DCM (3 ml) and THF (3 ml) were stirred overnight at room temperature . Water and DCM were added, the organic layer was separated, dried (MgSO ₄ ) and evaporated to dryness. The resulting residue was purified by flash chromatography on silica gel (15-40 µm, 10 g, CH ₂ Cl ₂ to CH ₂ Cl ₂ /CH ₃ OH/NH ₄ OH: 94/6/0.1). The purified fractions were collected and evaporated to dryness to yield 0.16 g of intermediate (74).

b) Receipt /H (cis) h ₂ n Η intermediate connections (75)

A solution of intermediate (74) (0.16 g, 0.634 mmol) in HCl (4 M in dioxane) (2 ml) was stirred at room temperature for 30 min, then evaporated to dryness, yielding 0.1 g of intermediate ( 75).

Example A.18.

a) Receipt ^N-C-O-^- (cis) intermediate connections (76)

A solution of intermediate (34) (0.2 g, 0.56 mmol), bis(pinacolato)diboron (0.171 g, 0.67 mmol) and potassium acetate (0.165 g, 1.68 mmol) in dioxane (2 ml ) were stirred and degassed using N2 for 10 min. 1,1'-Bis(diphenylphosphine)ferrocenedichloropalladium(II) (0.041 g, 0.056 mmol) was added and the reaction mixture was heated at 100°C using a microwave oven (Biotage Initiator EXP 60) with power ranging from 0 to 400 W at for 20 minutes. Water and EtOAc were added, the organic layer was separated, washed with water, then brine, dried over MgSO ₄ and evaporated to dryness. The resulting residue was purified by flash chromatography on silica gel (10 g, 15-40 µm, heptane/EtOAc 85/15 to heptane/EtOAc 70/30). The purified fractions were collected and evaporated to dryness to yield intermediate (76).

b) Receipt _s /DA a / J mTTo-g ( ^cis ) ⁿ 'n mn \ n intermediate connections (77)

- 18 043636

A solution of intermediate (76) (0.45 g, 1.34 mmol) and 2-bromo-5-methyl-1,3,4-thiadiazole (0.288 g, 1.61 mmol) in K ₂ CO ₃ (2 M , 1.34 ml, 2.69 mmol) and ethylene glycol dimethyl ether (5 ml) were stirred and degassed with N ₂ for 10 min. Tetrakis(triphenylphosphine)palladium(0) (0.155 g, 0.134 mmol) was added and the reaction mixture was heated at 150°C using a single-mode microwave oven (Biotage Initiator EXP 60) power ranging from 0 to 400 W for 5 min. Water and EtOAc were added, the organic layer was separated, washed with brine, dried (MgSO ₄ ) and evaporated to dryness. The resulting residue was purified by flash chromatography on silica gel (30 g cartridge, 15-40 µm, DCM to DCM/MeOH/NH4OH: 98/2/0, 1). The purified fractions were collected and evaporated to dryness to yield intermediate (77).

c) Receipt ы УГ 6--/ NH (cis) ⁿ 'n / Η intermediate connections (78)

A solution of intermediate (77) (0.14 g, 0.455 mmol) in HCl (4 M in dioxane) (2 ml) was stirred at room temperature for 30 min, then the reaction mixture was poured into 10% aqueous K2CO3 and extracted, using DCM. The organic layer was separated, washed with water, dried (MgSO ₄ ) and evaporated to dryness to give 81 mg of intermediate (78).

Example A.19.

a) Receipt y _YI^NCO—(CIS) ⁿ intermediate connections (79)

BuLi (1.6 M in hexane) (4.2 ml, 6.66 mmol) was added dropwise to a solution of 1-methylimidazole (0.53 ml, 6.66 mol) in THF (5 ml) under nitrogen at - 78°C, then the resulting mixture was stirred at 0°C for 1 hour. The reaction mixture was cooled to -78°C, and a solution of intermediate (5) (91.0 g, 4.44 mmol) in THF (10 ml) was added. The reaction mixture was stirred for 2 hours at -78°C, then allowed to cool to room temperature and stirred overnight. Water and EtOAc were added, the organic layer was separated, washed with water and brine, dried over MgSO ₄ and evaporated to dryness. The resulting residue was purified by flash chromatography on silica gel (15-40 µm, 30g, CH ₂ Cl ₂ to CH ₂ Cl ₂ /CH ₃ OH/NH ₄ OH: 95/5/0.1). The purified fractions were collected and evaporated to dryness to yield 0.54 g of intermediate (79).

b) Receipt intermediate connections (80)

A mixture of intermediate (79) (0.54 g, 1.76 mmol) in HCl (37% in H ₂ O) (5 ml) in a sealed tube was heated at 140°C using a single-mode microwave oven (Biotage Initiator EXP 60) with power ranging from 0 to 400 W for 1 hour. The reaction mixture was evaporated to dryness to obtain 0.47 g of intermediate (80).

Example A.20.

a) Receipt F ₃ CSQ~^^ / ( ^cis ) n \ intermediate connections (81)

The reaction was carried out under anhydrous conditions under argon, monitoring its progress using thin layer chromatography (silica gel, petroleum ether/ethyl acetate 1/1, UV/phosphomolybdic acid). n-butyllithium, 2.5M in hexane (4.28 ml, 10.7 mmol), was added dropwise (5 min) to a solution of diisopropylamine (1.51 ml, 10.7 mmol) in THF (16 ml) at - 20°C. The mixture was stirred for 15 minutes at -20°C and then cooled to -78°C. A solution of intermediate (95) (2.00 g, 8.88 mmol) in THF (20 ml) was added (5 min) at -78°C. The mixture was stirred at -78°C for 2 hours. A solution of 2-[N,N-bis-(trifluoromethylsulfonyl)amino]pyridine (3.50 g, 9.77 mmol) in THF (12.5 ml) was added (5 min) at -78°C. The mixture was then allowed to return to room temperature and stirred for 17 hours. The mixture was heated at 50°C for 4 hours. The reaction mixture was quenched by adding saturated aqueous ammonium chloride (100 ml) and extracted with ethyl acetate (3x100 ml). The combined organic layers were dried (sodium sulfate), filtered and concentrated. Dichloromethane (50 ml) was added to the resulting residue (6.07 g), the mixture was filtered to obtain 1.30 g of a white solid, the filtrate was concentrated and then purified by silica gel flash column chromatography (eluent: petroleum ether/EtOAc 100 /0 to 60/40). The product fractions were collected and the solvent was evaporated to give 1.02 g of intermediate (81).

b) Receipt o .o X/*—SD-U' / ^(cis) n \ intermediate connections (82)

The reaction was carried out in an argon atmosphere, monitoring its progress using a thin-layer

- 19 043636 chromatography (petroleum ether/ethyl acetate 8/2, UV/phosphomolybdic acid). 5-acetyl-2thienylboronic acid (0.057 g, 0.336 mmol) and 2 M aqueous potassium carbonate (0.280 ml, 0.560 mmol) were added to a solution of intermediate (81) (0.100 g, 0.280 mmol) in 1,2-dimethoxyethane (5 ml ). The mixture was purged with argon and tetrakis(triphenylphosphine)palladium(0) (0.032 g, 0.028 mmol) was added. The mixture was then heated at 80°C overnight. The mixture was cooled to room temperature, water (10 ml) and EtOAc (10 ml) were added. The organic layer was separated, washed with water (10 ml), then with brine (10 ml), dried (sodium sulfate), filtered and evaporated under vacuum to dryness. The residue was purified by column chromatography on silica gel (eluent: petroleum ether/ethyl acetate 8/2). The target fractions were collected and the solvent was evaporated to obtain 0.076 g of intermediate (82).

c) Receipt II n ( ^cis ) Η intermediate connections (83)

The reaction was carried out under anhydrous conditions in an argon atmosphere, monitoring its progress using thin layer chromatography (petroleum ether/ethyl acetate 9/1, UV). HCl, 4 M in dioxane (3.33 ml, 13.3 mmol) was added to a solution of intermediate (82) (0.444 g, 1.33 mmol) in dioxane (9 ml). The reaction mixture was stirred at room temperature for 70 hours and then concentrated to dryness, yielding 0.370 g of intermediate (83).

The following compounds were prepared using the same procedure as in Example A.20b/A.20c, where 5-acetyl-2-thienylboronic acid was replaced by 4-methylthiophene-2-boric acid, 2-chlorothiophene-3-boric acid, 4-methyl-3-thiophene-boric acid, 2-acetyl-3-thiophene-boric acid, 5-cyanothiophene-2-boric acid, 5-chlorothiophen-2-boric acid, pinacol ester 5-methylthiophene-2- boric acid, 3-methylthiophene-2-boric acid pinacol ester or 3-methoxythiophene-2-boric acid pinacol ester, respectively.

Η (cis) f\\JC NH H (cis) L /—\Ji ^Xnh \H (cis) intermediate connection (84) intermediate connection (85) intermediate connection (86) (cis) N<±,s /-J% Lz—\ JL / ^NH H (cis) Cl _s ΐ T У-f TC ,NH H (cis) intermediate connection (87) intermediate connection (88) intermediate connection (89) Η (cis) ы [ί><τ>\H (cis) G f U NH o— n (cis) intermediate intermediate intermediate compound (90) compound (91) compound (92)

Example A.21.

a) Receipt —/( ^cis ) intermediate connections (93)

The reaction was carried out under anhydrous conditions in an argon atmosphere, monitoring its progress using thin layer chromatography (silica gel, eluent: petroleum ether/ethyl acetate 9/1, phosphomolybdic acid). Methyllithium 1.6 M in diethyl ether (3.29 ml, 5.26 mmol) was added to a suspension of copper(I) iodide (0.794 g, 4.17 mmol) in THF (5.0 ml) at 0°C. After 1 h, a solution of intermediate (81) (0.355 g, 0.993 mmol) in THF (2.1 mL) was added via tube at 0° C., washing with THF (2.1 mL). The reaction mixture was stirred at room temperature overnight. The mixture was quenched with saturated NH ₄ Cl solution (14 ml) and dried to dryness. The residue was purified by column chromatography on silica gel (eluent: pentane/ethyl acetate 95/5). The product fractions were collected and the solvent was evaporated to give 0.180 g of intermediate (93).

b) Receipt -f NH (cis) H intermediate connection (94)

The reaction was carried out under anhydrous conditions in an argon atmosphere, monitoring its progress using thin layer chromatography (silica gel, eluent: petroleum ether/ethyl acetate 9/1, phosphomolybdic acid). HCl (4 M in dioxane) (2.02 g, 8.06 mmol) was added to a solution of intermediate (93) (0.180 g, 0.806 mmol) in 1,4-dioxane (4.3 ml), the solution was stirred at room temperature for 65 hours, then concentrated to dryness to give 0.141 g of intermediate (94) (110%).

- 20 043636

Example A.22.

a) Receipt /^-C\ ° / /t/X ^- ( ^cis ) n intermediate connection (95)

The reaction was carried out under anhydrous conditions and monitored using thin layer chromatography (silica gel, eluent: petroleum ether/ethyl acetate 50/50, developer: UV/phosphomolybdic acid. Solution of intermediate (4) (6.93 g, 31.0 mmol ) in THF (180 ml) was hydrogenated at room temperature (atmospheric pressure) with palladium on carbon, 10% (wt.) loading (1.65 g), as catalyst for 15 hours. The catalyst was filtered on clarcel, the precipitate the filter was washed with dichloromethane (50 ml) and the combined filtrates were concentrated under reduced pressure to dryness.The resulting residue (7.26 g) was purified by silica gel column chromatography (eluent: petroleum ether/ethyl acetate 80/20 to 50/50) The product fractions were collected and the solvent was evaporated to give 6.70 g of intermediate (95).

b) Receipt /\ 1 n-c-o—e— (as) n intermediate connection (96)

The reaction was carried out under anhydrous conditions under argon, monitoring its progress using thin layer chromatography (silica gel, eluent: petroleum ether/ethyl acetate 6/4, DCIP). Lanthanum lithium trichloride complex 0.6 M in THF (3.70 mL, 2.22 mmol) was added to a solution of intermediate (95) (0.500 g, 2.22 mmol) in THF (15 mL). The mixture was stirred at room temperature for 1 hour, then cooled to 0°C. A solution of ethylmagnesium bromide, 0.1 M in THF (2.66 mL, 2.66 mmol) was added dropwise and the reaction mixture was allowed to warm to room temperature and stirred for 18 hours. The reaction mixture was quenched by adding saturated aqueous ammonium chloride (50 ml) and extracted with ethyl acetate (3x50 ml). The combined organic layers were dried (Na ₂ SO ₄ ), filtered and concentrated. The resulting residue (0.635 g) was purified by column chromatography on silica gel (eluent: petroleum ether/ethyl acetate 9/1 to 7/3). The product fractions were collected and the solvent was evaporated. The resulting residue (0.410 g) was purified by column chromatography on silica gel (eluent: petroleum ether/ethyl acetate 8/2). The product fractions were collected and the solvent was evaporated to give 0.235 g of intermediate (96).

c) Receipt n /— ^(cis) Η intermediate connection (97)

The reaction was carried out under anhydrous conditions in an argon atmosphere, monitoring its progress using NMR. HCl in dioxane (4M, 2.30 mL, 9.20 mmol) was added to a solution of intermediate (96) (0.235 g, 0.920 mmol) in dioxane (2 mL).

The reaction mixture was stirred at 60°C for 18 hours.

After cooling to room temperature, the precipitate was filtered on a glass frit and washed with diethyl ether (20 ml) to obtain 0.126 g of solid. The filtrate was evaporated to dryness to obtain 0.077 g of residue.

The solid and residue were combined and dissolved in dioxane (2 ml). 4 M HCl in dioxane (2.30 mL, 9.20 mmol) was added and the mixture was stirred at 60°C for 24 hours, then at 100°C for 72 hours. The reaction mixture was concentrated to dryness to give 0.158 g of intermediate connections (97).

Example A.23.

a) Receipt but—C I n-c-o—(cis) n intermediate connection (98)

The reaction was carried out under anhydrous conditions in an argon atmosphere, monitoring its progress using thin layer chromatography (silica gel, eluent: petroleum ether/ethyl acetate 1/1, phosphomolybdic acid). Sodium borohydride (0.893 g, 23.6 mmol) was added portionwise over 30 min to a solution of intermediate (95) (2.66 g, 11.8 mmol) in MeOH (60 mL) at 0°C. The reaction mixture was stirred for 1 hour at 0°C and then concentrated to dryness. The residue was diluted with ethyl acetate (200 ml) and washed with water (100 ml), 1 M aqueous hydrochloric acid (100 ml) and brine (100 ml). The organic layer was dried (Na2SO4), filtered and concentrated to give 2.27 g of intermediate (98).

b) Receipt ° ° / —s—о -/jf/Vc-o “С ( ^cis ) ⁰ UL intermediate connection (99)

The reaction was carried out under anhydrous conditions in an argon atmosphere, monitoring its progress using thin layer chromatography (silica gel, eluent: petroleum ether/ethyl acetate 1/1, phosphomo

- 21 043636 libdic acid). Methanesulfonyl chloride (0.930 ml, 11.9 mmol) was added dropwise to a solution of intermediate (98) (2.27 g, 9.98 mmol) and triethylamine (4.17 ml, 29.9 mmol) in DCM (50 ml) at 0°C. The reaction mixture was stirred for 1 hour at room temperature and then concentrated to dryness. The residue was diluted with ethyl acetate (200 ml) and washed with water (100 ml), brine (100 ml), 1 M aqueous hydrochloric acid (100 ml) and brine again (100 ml). The organic layer was dried (Na ₂ SO ₄ ), filtered and concentrated. The resulting residue (2.52 g) was purified by silica gel column chromatography (eluent: petroleum ether/ethyl acetate 8/2 to 5/5). The product fractions were collected and the solvent was evaporated to give 2.39 g of intermediate (99).

c) Receipt intermediate connections (100)

The reaction was carried out under anhydrous conditions in an argon atmosphere, monitoring its progress using thin layer chromatography (silica gel, eluent: petroleum ether/ethyl acetate 8/2, ninhydrin/phosphomolybdic acid). Intermediate (99) (0.300 g, 0.982 mmol) was dissolved in DMF (3 ml) and the mixture was cooled to 0°C. Pyrrole (0.102 ml, 1.47 mmol) and sodium hydride, 60% dispersion in mineral oil (0.0589 g, 1.47 mmol) were added and the reaction mixture was stirred at room temperature for 18 hours. The reaction mixture was diluted with ethyl acetate (50 ml ) and washed with water (2x50 ml) and then with saline (3x50 ml). The organic layer was dried (Na ₂ SO ₄ ), filtered and concentrated. The resulting residue (0.2-90 g) was purified by column chromatography on silica gel (eluent: petroleum ether/ethyl acetate 98/2 to 95/5 and then to 90/10). The product fractions were collected and the solvent was evaporated to give 0.175 g of intermediate (100).

d) Receipt oA|o« I* intermediate connections (101)

The reaction was carried out under anhydrous conditions in an argon atmosphere, monitoring its progress using thin layer chromatography (silica gel, eluent: petroleum ether/ethyl acetate 6/4, phosphomolybdic acid). 4 M HCl in dioxane (1.58 ml, 6.33 mmol) was added to a solution of intermediate (100) (0.175 g, 0.633 mmol) in dioxane (3 ml). The reaction mixture was stirred at 50°C for 2 hours and concentrated to dryness to give 0.135 g of intermediate (101).

The following compounds were prepared using the same procedure as in Example A.23c/A.23b, where the pyrrole was replaced by tetrazole, pyrazole, 1,2,4-triazole, 1,2,3-triazole or phenol, respectively.

Example A.24.

a) Receipt AA o. —o—6 JT N-c-o—e— (cis) n intermediate connection (109)

The reaction was carried out under anhydrous conditions in an argon atmosphere, monitoring its progress using thin layer chromatography (silica gel, eluent: petroleum ether/ethyl acetate 8/2, ninhydrin/phosphomolybdic acid). A 25% (w/w) solution of sodium methoxide in methanol (0.449 mL, 1.96 mmol) was added to a solution of intermediate (99) (0.300 g, 0.982 mmol) in MeOH (4 mL). The reaction mixture was stirred at reflux for 20 hours. The reaction mixture was concentrated to dryness. The residue was diluted with ethyl acetate (50 ml) and washed with water (50 ml) and then brine (50 ml). The organic layer was dried (Na ₂ SO ₄ ), filtered and concentrated. The resulting residue was purified by column chromatography on silica gel (eluent: petroleum ether/ethyl acetate 100/0 to 97/3 and then to 1/1). The product fractions were collected and the solvent was evaporated to give 0.182 g of intermediate (109).

- 22 043636

intermediate connection (110)

The reaction was carried out under anhydrous conditions in an argon atmosphere, monitoring its progress using thin layer chromatography (silica gel, eluent: petroleum ether/ethyl acetate 8/2, ninhydrin/phosphomolybdic acid). 4 M HCl in dioxane (1.88 ml, 7.54 mmol) was added to a solution of intermediate (109) (0.182 g, 0.754 mmol) in dioxane (4 ml). The reaction mixture was stirred at room temperature for 18 hours and then at 50°C for 2 hours. The reaction mixture was concentrated to dryness to obtain 0.139 g of intermediate (110).

Some of the intermediates used in the preparation of the final compounds can be purchased, e.g.

B. Preparation of final compounds.

Example B.1.

intermediate connection (14)

Mixture of intermediate (3) (9.4 g, 44.3 mmol), intermediate (9) (8.5 g, 44.3 mmol), hydroxybenzotriazole (6.0 g, 4.4.3 mmol), triethylamine (15.4 ml, 0.111 mmol) in CH2Cl2 (160 ml) and THF (160 ml) were stirred overnight at room temperature. Water (175 ml) was added and the precipitate was filtered off and washed with water/EtOH (50 ml). The solid was suspended in EtOH (50 ml) and stirred for 15 minutes. The resulting suspension was filtered and dried under vacuum at 70°C to obtain 7.3 g of compound (14) in the form of a white powder (mp = 266°C), ([a] _D ²⁰ = -105.1° (589 nm, concentration 0.1275 w/v%, CH2CL 20°C).

1H NMR (500 MHz, DMSOC) δ (ppm) 10.64 (d, J=7.6 Hz, 1H), 8.33 (dd, J=1.7, 9.6 Hz, 1H) , 8.04 (d, J=17.3 Hz, 1H), 7.48 (d, J=7.6 Hz, 2H), 7.31-7.42 (m, 3 N), 7.23 -7.28 (m, 1H), 6.99 (t, J=15.0 Hz, 1H), 6.20 (d, J=6.0 Hz, 1H), 3.38-4.04 ( m, 5H), 3.09-3.21 (m, 1H), 2.85-3.04 (m, 3H), 2.55-2.67 (m, 3H).

Example B.2.

Mixture of intermediate (14) (5.8 g, 30.32 mmol), intermediate (3) (7.72 g, 30.32 mmol), 1-hydroxybenzotriazole (4.92 g, 30.38 mmol), EDCI (6.97 g, 30.38 mmol), triethylamine (14.71 ml, 106.12 mmol) in CH ₂ Cl ₂ (100 ml) and THF (100 ml) were stirred overnight at room temperature. The mixture was poured into water. The precipitate was filtered off, washed twice with EtOH, and dried under vacuum at 65°C. This precipitate was recrystallized from EtOH, filtered and dried under vacuum at 62°C to obtain 9.02 g of compound (44) in the form of a white powder (mp=264°C), ([a]D ²⁰ =+170.12° (589 nm, concentration 0.2075 w/v%, C^Clz, 20°C).

1H NMR (400 MHz, DMSO-a ₆ ) δ (ppm) 10.63 (d, J=4.5 Hz, 1H), 8.32 (d, J=5.1 Hz, 1H), 8.03 (d, J=10.6 Hz, 1H), 7.52 (dd, J=2.8, 4.8 Hz, 1H), 7.41 (br.s, 1H), 7.36 (dd, J=4.8, 9.3 Hz, 2H), 6.98 (dd, J=9.1, 15.7 Hz, 1H), 6.01 (br.s, 1H), 3.35- 4.03 (m, 5H), 2.94-3.21 (m, 2H), 2.90 (kV, J=7.9 Hz, 3H), 2.52-2.62 (m, 2H) .

Example B.3. _____________

intermediate connection (40)

Mixture of intermediate (19) (21.1 g, 81.4 mmol), intermediate (3) (17.3 g, 61.8 mmol), 1-hydroxybenzotriazole (11.0 g, 81.4 mmol), EDCI (15.6 g, 81.4 mmol) and triethylamine (47 ml, 0.339 mmol) in CH ₂ Cl ₂ (350 ml) and THF (350 ml) were stirred overnight at room temperature. Water was added to the mixture. The precipitate was filtered, washed with water/EtOH and then EtOH, and dried under vacuum at 70°C to obtain 12.7 g of compound (40) as a white powder (mp=271°C), ([a]D ²⁰ =+116.08° (589 nm, concentration 0.2145 w/v%, C^Ch, 20°C).

¹ H NMR (400 MHz, DMSO-d6) δ (ppm) 10.63 (d, J=5.1 Hz, 1H), 8.52 (d, J=5.6 Hz, 2H), 8.33 (d, J=6.1 Hz, 1H), 8.03 (d, J=13.6 Hz, 1H), 7.41-7.46 (d, J=15.7 Hz, 2H ), 7.38 (d, J=4.0 Hz, 1H), 6.98 (dd, J=11.6, 15.7 Hz, 1H), 6.53 (d, J=8.1 Hz , 1H), 3.37-4.04 (m, 5H), 2.86-3.22 (m, 5H), 2.58-2.70 (m, 2H).

intermediate connection (41)

Compound (41) was prepared similarly by reacting intermediate (20) with intermediate (3) following the same procedure.

- 23 043636

Ή NMR (500 MHz, DMSO-06) δ (ppm) 10.63 (d, J=5.1 Hz, 1H), 8.52 (d, J=5.6 Hz, 2H), 8 .33 (d, J=6.1 Hz, 1H), 8.03 (d, J=13.6 Hz, 1H), 7.41-7.46 (d, J=15.7 Hz, 2H) , 7.38 (d, J=4.0 Hz, 1H), 6.98 (dd, J=11.6, 15.7 Hz, 1H), 6.53 (d, J=8.1 Hz, 1H), 3.37-4.04 (m, 5H), 2.86-3.22 (m, 5H), 2.58-2.70 (m, 2H).

(Md ²⁰ = -115.85° (589 nm, concentration 0.183 wt./vol.%, C^Ch, 20°C)).

Example B.4. ______________________________________________

a) Receipt > ^,JU ca n n intermediate connection (111)

The reaction was carried out in an argon atmosphere, monitoring its progress using thin layer chromatography (silica gel, CH ₂ Cl ₂ /methanol/triethylamine 95/5/0.1, UV/phosphomolybdic acid). 1-(3-Dimethylaminopropyl)-3-ethylcarbodiimide (.HCl) (1.70 g, 8.87 mmol) was added to a mixture of intermediate (3) (2.02 g, 7.39 mmol), crude cis-hexahydro -cyclopenta[c]pyrrol5(III)one (1.85 g, maximum 8.89 mmol), 1-hydroxybenzotriazole monohydrate (1.36 g, 8.87 mmol) and N-ethyldiisopropylamine (6.32 ml, 36, 9 mmol) in DMF (75 ml). The mixture was stirred at room temperature for 18 hours. The mixture was concentrated under reduced pressure, diluted with dichloromethane (150 ml) and washed with saturated aqueous NaHCO ₃ (100 ml). The aqueous layer was again extracted with dichloromethane (2x150 ml). The combined organic layers were washed with brine (400 ml), dried (Na ₂ SO ₄ ), filtered and concentrated under reduced pressure to dryness. The resulting residue was purified by column chromatography on silica gel (eluent: dichloromethane/methanol 100/0 to 94/6). The product fractions were collected and the solvent was evaporated. The alkaline aqueous layers were again extracted with dichloromethane (3x300 ml). The combined organic layers were washed with brine (900 ml), dried (Na ₂ SO ₄ ), filtered and concentrated under reduced pressure to dryness. The resulting residue was purified by column chromatography on silica gel (eluent: dichloromethane/methanol 100/0 to 94/6). The product fractions were collected and the solvent was evaporated. The target residues were combined to yield 1.58 g of intermediate (111).

b) Receipt '-- But intermediate connections (71)

The reaction was carried out under anhydrous conditions in an argon atmosphere, monitoring its progress using thin layer chromatography (petroleum ether/ethyl acetate 95/5, UV/phosphomolybdic acid). Lanthanum trichloride lithium chloride complex 0.6 M in THF (2.38 ml, 1.43 mmol) was added to a suspension of intermediate (111) (0.464 g, 1.43 mmol) in THF (18 ml). The mixture was stirred at room temperature for 1 hour, then cooled to 0°C. A solution of phenylmagnesium bromide, 0.1 M in THF (3.57 mL, 3.57 mmol) was added dropwise. The reaction mixture was stirred and allowed to warm back to room temperature over 3 days. Additionally, a 1.0 M solution of phenylmagnesium bromide in THF (2.85 mL, 2.85 mmol, 2 equivalents) was added dropwise. The mixture was stirred at room temperature for 2 hours. The reaction mixture was quenched by adding saturated aqueous ammonium chloride (30 ml) and extracted with ethyl acetate (3x30 ml). The combined organic layers were dried (Na ₂ SO ₄ ), filtered and concentrated under reduced pressure to dryness. The resulting residue was purified by flash column chromatography on silica gel (eluent: CH ₂ Cl ₂ /MeOH 100/0 to 95/5). The solvent from the collected product fractions was evaporated. The residue was triturated with diethyl ether (2x3 ml) and then dried under vacuum to give 0.077 g of compound (71).

Example B.5

intermediate connection (73)

A mixture of intermediate (23) (0.032 g, 0.097 mmol), intermediate (8) (0.032 g, 0.145 mmol), EDCI (0.022 g, 0.0116 mmol), HOBt (0.016 g, 0.116 mmol) and triethylamine (0.049 ml, 0.349 mmol) in DCM (1 ml) and THF (1 ml) were stirred overnight at room temperature. Water was added, the mixture was extracted using DCM, the organic layer was separated, washed with water, dried (MgSO ₄ ) and evaporated to dryness. The residue was recrystallized from EtOH, the solid was filtered, washed with EtOH and dried (vacuum, 70°C) to obtain 0.015 g of compound (73).

In table F-1 lists compounds that were prepared according to one of the above examples.

- 24 043636

Table F-1

(cis) Conn. No. 1; Note B.l Conn. No. 2; Note IN 1 (cis) Conn. No. 3; Note B.l V-s Conn. No. 4; Note IN 1 X JL/\ N N o ρ η h (cis) Conn. No. 5; Note B.l O S Η Η ό Conn. No. 6; Note B.l NN O [| ^H H (cis) Comp. No. 7; Note Bl a ||V ^H h Conn. No. 8; Note Bl c Conn. No. 9; Note B.l 5¼ (cis) Conn. No. 10; Note B.l Conn. No. 11; Note B.l ^N ·^ (cis) Comp. No. 12; Note Bl Conn. No. 13; Note Bl ( [a] _D ²⁰ =+l04, 17 ° (589 nm, concentration 0.096 wt./vol.%, CH ₂ C1 ₂ , 20 ° C) Conn. No. 14; Note B.l N-\(Cis) Conn. No. 15; Note IN 1 Conn. No. 16; Note B.l

- 25 043636

Conn. No. 17; Note B.l Conn. No. 18; Note B.l сД-^ХХ ^Vo (cis) ^S^xx Conn. No. 19; Note B.l Conn. No. 20; Note B.l o >' ^'W· O-^(cis) Conn. No. 21; Note B.l Conn. No. 22; Note B.l H. JL ^x /X a ^{n n} Conn. No. 23; Note B.l Conn. No. 24; Note B.l H Α^^ΐχ^/ΐχ/χ^\n? ΤΧλ Jx AL NN N ^HH ° \ (cis) YES". (cis) Conn. No. 25; Note B.l Conn. No. 26; Note B.l Ν=^ (cis) (cis) Conn. No. 27; Note B.l Conn. No. 28; Note B.l W Wo (Cis) Conn. No. 29; Note B.l Conn. No. 30; Note B.l

- 26 043636

J/Oca ^Η (cis) ⁿ Conn. No. 31; Note IN 1 ^Vs _(cis) Conn. No. 32; Note IN 1 vAXV··qp N N'^O _F ^z Η Η Conn. No. 33; Note IN 1 (cis) Conn. No. 34; Note IN 1 Conn. No. 35; Note IN 1 Cl Conn. No. 36; Note IN 1 o n X ϋΟ' ΓΧΐ O ^{n n} Conn. No. 37; Note IN 1 Conn. No. 38; Note IN 1 X(cis) rCP". Conn. No. 39; Note IN 1 Conn. No. 40; Note V.Z Hi", n n Conn. No. 41; Note V.Z Conn. No. 42; Note IN 1 1 JL Η Η (cis) rAH." Conn. No. 43; Note IN 1 Conn. No. 44; Note AT 2 °(cis)

- 27 043636

- 28 043636

C. Connection identification.

C1. LCMS.

The following methods were used to characterize the compounds of the present invention using LCMS.

General technique A.

LC measurements were performed using an Acquity UPLC (Ultra Performance Liquid Chromatography) system (Waters) comprising a binary pump with degasser, autosampler, diode array detector (DAD) and column as specified in the respective methods below, the column maintained at 40°C. The flow from the column was directed to the MS detector. The MS detector was equipped with an electrospray ionization source. The capillary needle voltage was 3 kV and the Quattro (Waters Triple Quadrupole Mass Spectrometer) temperature was maintained at 130°C. Nitrogen was used as a nebulizer gas. Data collection and processing were carried out using the Waters-Micromass MassLynx-Openlynx data processing system.

General methodology B.

HPLC measurements were performed using an Alliance HT 2795 system (Waters) comprising a quaternary pump with degasser, autosampler, diode array detector (DAD) and column as specified in the respective methods below, the column maintained at 30°C. The column flow was separated for the MS spectrometer. MS detection was performed with an electrospray ionization source. The capillary needle voltage was 3 kV and the source temperature in the LCT (Waters Time of Flight Zspray™ mass spectrometer) was maintained at 100°C. Nitrogen was used as a nebulizer gas. Data collection and processing were carried out using the WatersMicromass MassLynx-Openlynx data processing system.

Method 1.

In addition to general procedure A, reverse phase UPLC was performed on a Waters Acquity C18 BEN (ethyl siloxane/silica bridged) column (1.7 μm, 2.1 x 100 mm) at a flow rate of 0.35 mL/min. Two mobile phases (mobile phase A: 95% 7 mM ammonium acetate/5% acetonitrile; mobile phase B: 100% acetonitrile) were used to implement a gradient condition from 90% A and 10% B (hold for 0.5 min) to 8% A and 92% B, hold for 2 min, and return to initial conditions after 0.5 min, hold for 1.5 min. Used

- 29 043636 volume of injected sample 2 µl. The cone voltage was 20 V for positive and negative ionization modes. Mass spectra were acquired by scanning from 100 to 1000 in 0.2 s using a delay time of 0.1 s between scans.

Method 2.

In addition to general procedure A, reversed-phase UPLC was performed on a Waters Acquity C18 BEN (ethyl siloxane/silica bridged) column (1.7 μm, 2.1 x 100 mm) at a flow rate of 0.343 mL/min. Two mobile phases (mobile phase A: 95% 7 mM ammonium acetate/5% acetonitrile; mobile phase B: 100% acetonitrile) were used to implement a gradient condition from 84.2% A and 15.8% B (hold for 0. 49 min) to 10.5% A and 89.5% B, holding for 2.18 min and returning to initial conditions after 0.73 min, holding for 0.73 min. An injection volume of 2 μl was used. The cone voltage was 20 V for positive and negative ionization modes. Mass spectra were acquired by scanning from 100 to 1000 in 0.2 s using a delay time of 0.1 s between scans.

Method 3.

In addition to general procedure B, reverse phase HPLC was performed on a Waters Xbridge C18 column (3.5 μm, 4.6 x 100 mm) at a flow rate of 0.8 mL/min. Two mobile phases (mobile phase A: 100% 7 mM ammonium acetate; mobile phase B: 100% acetonitrile) were used to maintain a gradient condition from 80% A to 20% B (hold for 0.5 min) to 90% B over 4.5 min, 90% B for 4 min, and re-equilibrate at baseline conditions for 3 min. An injection volume of 5 μl was used. The cone voltage was 20 V for positive and negative ionization modes. Mass spectra were acquired by scanning from 100 to 1000 in 0.4 s using a delay time of 0.3 s between scans.

Method 4.

In addition to general procedure B: reverse phase HPLC was performed on a Waters Atlantis C18 column (5 μm, 3.9 x 100 mm) at a flow rate of 0.8 ml/min. Three mobile phases (mobile phase A: 100% 7 mM ammonium acetate; mobile phase B: 100% acetonitrile; mobile phase C: 0.2% formic acid + 99.8% ultrapure water) were used to maintain the gradient condition from 50% A and 50% C (hold for 1.5 min) to 10% A, 80% B and 10% C in 4.5 min, hold for 4 min, and re-equilibrate at baseline conditions for 3 min. An injection volume of 5 μl was used. The cone voltage was 20 V for positive and negative ionization modes. Mass spectra were acquired by scanning from 100 to 1000 in 0.4 s using a delay time of 0.3 s between scans.

Method 5.

HPLC measurements were performed using a 1100/1200 HPLC system (Alliance) comprising a quaternary pump with degasser, autosampler, diode array detector (DAD), and column as specified in the respective methods below, the column maintained at room temperature. The MS detector (MS-Agilent simple quadrupole) was equipped with an APCI electrospray ionization source. Nitrogen was used as a nebulizer gas. Data collection and processing were carried out using the Chemstation data processing system.

Reverse-phase HPLC was performed on a Nucleosil C18 column (3 μm, 3x150 mm) with a flow rate of 0.42 ml/min. Two mobile phases (mobile phase A: water/TFA (0.1%); mobile phase B: 100% acetonitrile) were used to maintain a gradient condition from 98% A in 3 min, to 100% B in 12 min, 100% B for 5 min, then return to 98% A for 2 min, and re-equilibrate at baseline conditions with 98% A for 6 min. An injection volume of 2 μl was used. The capillary needle voltage was 3 kV, the corona discharge was held at 1 μA, and the source temperature was maintained at 250 °C. Variable voltage was used for the fragmenter. Mass spectra were obtained using electrospray ionization and CIAD in positive mode, scanning from 100 to 1100 AU. eat.

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Table C.1

LC/MS data

Connection no. Vr. beat MN ⁺ Method Connection no. Vr. beat MN ⁺ Method 2 5, 12 419 3 41 1.98 387 2 3 2.53 392 2 42 2.12 310 2 6 2.84 412 1 43 2.76 400 2 7 2.43 411 2 44 2, 63 392 2 9 2.74 420 2 45 2.58 392 2 10 2.53 392 2 46 2.16 382 2 eleven 2.74 400 2 47 2.24 421 2 12 1.96 387 2 48 5, 4 353 4 13 3.23 392 1 49 1.85 408 2 14 2, 63 386 2 50 1.78 388 2 15 2.92 426 2 51 1.84 376 2 17 2, 64 403 2 52 13.46 324 5 18 2, 65 423 2 53 14.13 338 5 19 2.44 376 2 54 14.38 404 5 21 2, 66 404 2 57 2.59 390 2 22 2.4 376 2 58 11.35 380 5 23 2.22 405 2 59 15.16 426 5 26 1.5 353 2 60 14.79 406 5 thirty 1.78 351 2 61 2.35 417 2 31 8.79 342 5 62 13.58 434 5 32 2.07 393 2 63 15, 03 406 5 33 2.12 453 2 64 16 381 2 34 2.1 348 2 67 14.01 417 5 35 2.72 423 2 68 15, 62 426 5 36 2.73 423 2 69 15, 09 406 5 37 11.2 379 5 71 13, 1 404 5 38 11.4 379 5 72 14.51 422 5 39 12.26 379 5 73 2.78 384 2 40 1.99 387 2

C2. Melting points.

For some compounds, melting points were obtained using a Kofler stage, consisting of a heated plate with a linear temperature gradient, a sliding pointer, and a temperature scale in degrees Celsius.

For a number of compounds, melting points were determined using differential scanning calorimetry (DSC). Melting points were measured with a temperature gradient of 10°C/min starting at 25°C. The maximum temperature was 350°C.

For a number of compounds, melting points were determined using a Buchi B-560 melting point apparatus. The heating medium was a metal block. The melting of the sample was observed visually using a magnifying lens and high light contrast. Melting points were measured with a temperature gradient of either 3 or 10°C/min. The maximum temperature was 300°C.

The remaining melting points were determined using open capillary tubes.

Table C.2

Melting point data Comm. no. Melting point

274.95°C

Method

DSC connection no. Melting point method

249.0-Buchi

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D. Pharmacological examples.

D.1. FabI enzyme inhibition: Staphylococcus aureus FabI enzyme inhibition study.

The FabI enzyme inhibition assay was performed in a half-area, 384-well microtiter plate. Compounds were assessed in 40-μl samples of mixtures containing 100 mM NaADA, pH 6.5 (ADA=N-[2-acetamido]-2-iminodiacetic acid), 250 μM crotonyl-CoA, 625 μM NADH and 50 μg/ml S aureus ATCC 29213 FabI. Inhibitors typically ranged from 50 to 0.39 μM. The reaction mixtures were incubated for 30 min at room temperature and the reaction was stopped by adding 200 mM Tris buffer (pH 9.0) to create a pH shift. The composition of NADH was monitored by measuring changes in absorbance at 340. By comparing the measured values with those of negative (no compound) and positive (no enzyme) controls, the percentage inhibition of enzyme activity of the compounds was determined. The best-fitting curve was determined using the least squares method. From this, the IC ₅₀ value (expressed in μg/ml) was obtained, resulting in 50% inhibition of enzyme activity.

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Table D.1

S values. aureus FabI IC ₅₀ Connection no. Fabl 1C ₅₀ cg/ml Connection no. Fabl 1C ₅₀ cg/ml 1 0.32 35 1.25 2 0.78 36 0.93 3 0.29 37 3.37 4 0.70 38 2.08 5 ~0.6 39 0.56 6 3.73 40 0.39 8 0.50 41 0.44 9 0.75 42 0.83 10 0.53 43 0.60 eleven 0.48 44 0.46 12 0.44 45 0.45 13 0.39 46 0.54 14 0.40 47 0.43 15 0.48 48 2.93 17 0.38 49 0.44 18 0.44 51 0.54 19 ~0.62 52 0.50 20 1.07 53 0.36 21 0.65 54 1.84 22 0.58 57 0.62 23 0.41 59 0.76 24 0.58 60 0.59 25 0.51 61 0.54 26 0.41 62 0.44 27 0.6 63 0.63 29 1.04 64 1, 62 thirty 2, 66 67 0.63 31 1.42 68 0.99 32 0.46 71 2.55 33 3, 06 72 0.43 34 1, 67 73 0.80

D.2. In vitro method for testing the antibacterial effect of compounds against various strains of bacteria.

Preparation of bacterial suspensions for sensitivity determination.

The bacteria used were Staphylococcus aureus ATCC 29213, methicillin-resistant Staphylococcus aureus (MRSA) ATCC 700788, and Escherichia coli ATCC 35218. The bacteria used in this study were grown overnight in flasks containing 100 ml of Mueller-Hinton broth (Difco cat. no. 0757 -17) in sterile deionized water, with shaking, at 37°C. The prepared solution was stored at -70°C until use.

Bacteria were incubated on a trypsin soy agar plate containing 5% sheep blood (Becton Dickinson cat. no. 254053) for 18-24 hours at 35°C under aerobic conditions (first pass). For the second pass, fresh Mueller-Hinton broth was inoculated using 5-10 colonies and grown overnight at 35°C until cloudy (reaching the addition phase) under aerobic conditions. The bacterial suspension is then adjusted to 0.5 McFarland density and further diluted 1:100 in Mueller Hinton broth. This is used as seed.

The results (for STA ATCC 29213) are presented in table. D2 below.

Antibacterial sensitivity test.

Definition.

MIC (Minimum Inhibitory Concentration) studies were performed using a broth microdilution method in a 96-well format (flat bottom microtiter plate) with a final volume of 0.1 mL Mueller Hinton broth containing two-fold dilution series of compounds with a bacterial inoculum of 5 x 105 CFU/mL (standard seed size according to CLSI guidelines). Inhibitors typically range from 63 to 0.49 µM. The final DMSO concentration in the study was 1.25% (maximum permissible DMSO concentration = 6%). In studies testing the effect of human serum on the activity of a compound against S. aureus, human serum was added at a final concentration of 10%. The plates were incubated at 35°C for 16-20 hours. At the end of incubation, bacterial growth was quantified fluorometrically. To do this, resazurin was added to all wells and the plates were re-incubated. Incubi time

- 33 043636 formation depends on the type of bacterium. A color change from blue to pink indicates bacterial growth. Fluorescence was measured in a computer-controlled fluorometer (Fluoroskan Ascent FL, Labsystems) at an excitation wavelength of 540 nm and an emission wavelength of 590 nm. The percentage of growth inhibition achieved by the compounds was calculated according to standard methods. IC ₅₀ (expressed in μg/ml) was determined as the 90% bacterial growth inhibition concentration. A set of reference compounds were tested simultaneously to confirm quality control.

The results are presented in table. D2 (STA+10% HS) is lower.

Cytotoxicity study.

The cytotoxicity of the compounds was assessed using the MTT assay. Human HelaM cells grown in 96-well plates were subjected to serial dilutions of test compounds (final volume of 0.2 ml) and incubated for 72 hours at 37°C and 5% CO ₂ . Inhibitors typically range from 25 to 0.8 µM. The final concentration of DMSO in the study was 0.5%. MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, tetrazole) was added and reduced to violet formazan only in living cells. Solubilization of formazan crystals was achieved by adding 100 μL of 2-propanol. Cell viability was determined by measuring the absorbance of reduced formazan, which produces a violet color, at 540 nm and 690 nm. The absorbance measured at 690 nm was automatically subtracted from the absorbance at 540 nm to eliminate the effect of nonspecific absorption. The percentage of cytotoxicity achieved by the compounds was calculated according to standard methods. Cytotoxicity was expressed as CC ₅₀ , the concentration that causes a 50% reduction in cell viability.

The results are presented in table. D2 (TOX HELAM) below.

Table D2

Representative Example Data

Conn. No. STA (361.159) IC90 dg/ml STA + 10% HS (361.169) IC90 dg/ml TOX HELAM (222.125) CC50 dg/ml 1 0.09 0.17 >3.8547 2 1, 02 1.09 >19.4696 3 0.03 0.06 >3.25636 5 0.64 1.14 7.92 8 1, 15 1.52 >10.5122 9 0.33 0.69 4.77 10 0.08 0.13 >3.915 eleven 0.37 1.76 6, 19 12 0.33 0.53 >9.70744 13 0.31 0.43 >9.68257 14 0.19 0.19 >9.68257 15 0.74 0.72 >9.78279 17 0.37 0.38 >10.1103 18 0.21 0.37 >10.6233 19 0.18 0, 12 >9.43038 21 0.13 0.29 >4.0346 22 0.23 0.25 >9.43038 23 0.67 0.79 >10.3108 24 4, 05 2.44 >3.9549 26 1, 11 1.11 >3.8646

Example E.

E.1. Thermodynamic solubility/solubility in aqueous solution.

The study of the solubility curve versus pH was carried out at ambient temperature over a 4-day period. A saturation solubility study was performed to determine the maximum solubility in a specific buffer solution. The compound was added to the appropriate buffer solution until saturation point was reached. The flask was then shaken for 4 days at ambient temperature. After 4 days, the solutions were filtered and injected into UPLC and the concentration was determined using a conventional HPLC method.

Results. _____________________________________________________

Conn. No. 14 Conn. No. 1 Conn. No. 41 Conn. No. 2 Buffer pH 2 <0.01 <0.002 1.18 <0.01 10% ΗΡ-β-CD 0.076 BUT BUT BUT

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buffer pH 2 20% ΗΡ-β-CD buffer pH 2 0.20 BUT BUT BUT Buffer pH 4 <0.01 <0.002 <0.01 <0.01 10% ΗΡ-β-CD buffer pH 4 0.069 0.177 1.1 0, 11 20% ΗΡ-β-CD buffer pH 4 0.18 0.308 >1.15 0.28 Buffer pH 7.4 <0.01 <0.002 0.13 <0.01 10% ΗΡ-β-CD buffer pH 7.4 0.089 0, 100 0.49 0.14 20% ΗΡ-β-CD buffer pH 7.4 0.20 0.417 0.56 0.33

BUT means not determined.

E.2. Antimicrobial spectrum of action.

Minimum inhibitory concentrations (MICs) were determined according to the Clinical and Laboratory Standards Institute (CLSI) methodology compared to aerobic bacteria (CLSI M07-A8) (see Clinical and Laboratory Standards Institute. 2009. Methods for dilution antimicrobial susceptibility tests for bacteria that grow aerobically. CLSI document M07-A8, Vol. 29, No. 2.) by microdilution with cationically modified Mueller-Hinton medium broth (CA-MMB) for most organisms, with the exception of Haemophilus influenza, where Haemophilis test medium broth was used ( HTM). Descriptions of individual organisms can be found in the table. Where possible, standard ATCC strains were tested.

The inoculum density for susceptibility testing was standardized to produce a final inoculum of approximately 5 x 10 ⁵ CFU/ml. The broth MIC was defined as the lowest drug concentration that prevents visible growth after 16-24 hours (depending on species) of incubation at 35°C-37°C.

Description of individual study organisms

Organism Characteristics MIC of the test medium Staphylococcus aureus ATCC 29213; reference strain MS SA EOM Staphylococcus aureus ATCC 43300; reference strain MRSA EOM Staphylococcus aureus NRS119; LZD-R; SCCmec IV; origin: USA EOM Staphylococcus aureus NRS120; LZD-R; SCCmec IV; origin: USA EOM Staphylococcus aureus NRS121; LZD-R; SCCmec IV; origin: USA EOM Escherichia coll ATCC 25922; reference strain EOM Escherichia coli Tol C mutant EOM Haemophilus influenzae ATCC 49247; reference strain NTM broth Moraxella catarrhalis ATCC 8176; b-lactamase negative EOM

Stock solutions of compounds were prepared in DMSO at a concentration of 1 mg/ml. Linezolid was prepared in DMSO at a concentration of 2 mg/ml. Stock solutions of all compounds were diluted in CA-MNV to obtain a range of twofold dilutions, depending on the sensitivity of the organism being tested.

Results (if available). __________________________________________

Organism Compound No. and MIC90 (tsg/ml) 14 1 44 2 41 10 22 12 S. aureus ATCC 29213 0.03 0.016 0.03 0.25 0.03 0.015 0.06 0.125 S. aureus ATCC 43300 0.03 0.016 0.03 0.5 0.03 0.03 0.125 0.125

- 35 043636

S. aureus NRS119 0.03 0.03 0.03 0.06 S. aureus NRS120 0.03 0.016 0.03 0.06 S. aureus NRS121 0.03 0.016 0.06 0.06 Mutant E. coll tolC 0.25 <0.03 >8 0.25 1 0.125 1 0.25 E.col ATCC 25922 4 >32 >8 >8 8 >8 >8 >8 H. influenza ATCC 49247 0.25 >8 >8 0.5 >8 4 1 M. catarrhalis ATCC 8176 0.015 0.25 0, 12

E.3. In vivo pharmacokinetics and oral bioavailability.

The in vivo pharmacokinetics and oral bioavailability of the example compounds were/are being studied in male Swiss (fed) mice following a single intravenous (i.v.) bolus and oral (p.o.) administration. For solutions of compositions i.v. and p.o. the compound was/is dissolved in a 20% ΗΡ-β-CD solution. The pH of the composition was/is approximately pH 4. All i.v. the formulations were isotonic.

Results.

Conn. No. 14 Connection no. 1 Conn. No. 10 Conn. No. 44 Connection no. 12 i.v. Dose (mg/kg) 2.5 2.5 2.5 2.5 2.5 η 3 3 3 3 3 Co (ng/ml) 2929 2921 4154 4524 2333 Plasma clearance Cl (l/h/kg) 0.33 0.35 0.64 0.49 2.2 Vd _z (l/kg) 1.3 1.5 1.2 0.9 3.7 AUCo-inf (ng.h/ml) 7464 7074 3992 5037 1124 Half-life (tl/ ₂ ) (H) 2.7 2.9 1.3 1.3 1.1 p.o. Dose (mg/kg) 10 5 10 10 10 η 3 3 3 3 3 Stakh (NG/ML) 2950 1720 3537 2670 275 Tschakh (I) 20 2.0 10 1.0 1.0 AUCo-inf (ng.h/ml) 21394 12158 12376 14527 914 AUCo-last Half-life (ti/ ₂ ) (h) 3.2 3.1 2.2 2.8 n.d. Oral bioavailability (%) 72 86 81 59 21

E.4. Efficacy In vivo.

The concept of studying the in vivo effect of an antibacterial compound by treating intraperitoneally infected mice was introduced in 1911 when studying the action of optochin against pneumococci (Morgenroth and Levy, 1911). The model's popularity is attributed to its ease of use with short experimental durations, reproducible infections, and simple endpoints.

Method.

Staphylococcus aureus strain ATCC 29213 was used to infect female Swiss albino mice. A bacterial culture of brain heart extract (BHI) broth was inoculated the day before infection, incubated at 37°C overnight, and diluted with fresh BHI broth to the required concentration. Er. (intraperitoneal) injection of ~5x10 ⁸ -5x10 ⁹ colony-forming units (CFU) was performed in any of the lateral lower quadrants of the abdomen. After infection, mice were kept in cages and monitored daily for signs of infection or death. Both the p.o. and iv pathways were used to treat mice, and each mouse was individually treated with

Claims (18)

administration through a gastric tube or by i.v. introduction. In this model, both solutions (p.o. and i.v.) and suspensions (p.o.) were tested. The parameter used to monitor the progression of infection and the effect of treatment was death or survival of animals within 3 days after infection. Because death may occur as a result of toxicity, a control group of 3 mice was included and treated with the highest dose of test compound (in suspension studies). Results. In vivo antibacterial activity in peritonitis models of S. aureus infection (ATCC 29213) following oral intravenous administration using solutions. Compound Path of infection Sowing material (magazine- nary entry 10) Compound Processing path Treatment dose (mg per kg) % survivability 44 IP 8, 9 Solution 20%CD+lHCl RO, QD 1; 5 57; : 100 14 IP 8.7 20%CD+2H2T IV, QD 2.5; 5 75; : 100 Control mice experienced mortality rates of 80 and 100% in each of the respective trials. CLAIM 1. Compound of formula (I)? / ^|= SU° ^(|) R ³ where A represents -C=C- or ^r2 bond represents a single bond or a double bond; X is carbon or nitrogen, and if X is nitrogen, then the bond is a single bond; Z1 represents CH or N; R ¹ represents hydrogen, C _1-4 alkyl or halogen; R ² represents hydrogen, C _1-4 alkyl or halogen; R ³ represents hydrogen, C _1-6 alkyl, hydroxyl or halogen; R ⁴ represents hydrogen; halogen; C _1-6 alkyl; C _2-6 alkenyl; C _2-6 alkynyl; C _1-6 alkyloxy; C _1-4 alkyloxycarbonyl; aminocarbonyl; mono- or di(C _1-4 alkyl)aminocarbonyl; aryl; aryloxy; arylcarbonyl; arylsulfonyl; heteroaryl; C _1-6 alkyl substituted with cyano group; C _1-6 alkyl substituted with aryl or aryloxy; or C _1-6 alkyl substituted with heteroaryl; aryl is phenyl; phenyl substituted with one, two or three substituents, each of them individually selected from halogen, hydroxyl, C _1-4 alkyl, C _1-4 alkoxy, polyhaloC _1-4 alkyl, polyhaloC _1-4 alkoxy, cyano-, nitro- and amino groups; heteroaryl is furanyl, thiophenyl, pyrrolyl, pyrazolyl, imidazolyl, isoxazolyl, thiazolyl, triazolyl, tetrazolyl, isothiazolyl, thiadiazolyl, oxadiazolyl, pyridinyl, pyridazinyl, pyrimidinyl, pyrazinyl, benzo[1,3]dioxolyl, benzofuranyl, benzothiazolyl, indolyl, 2 ,3-dihydro-1Nindolyl, tetrahydrothiophenyl or quinolinyl, wherein each heteroaryl may be substituted with one or two substituents, each individually selected from halogen, cyano, C _1-4 alkyl, C _1-4 alkoxy, C _1-4 alkylcarbonyl or phenyl; or a pharmaceutically acceptable acid addition salt thereof. 2. The compound according to claim 1, where Z ₁ represents CH; R ¹ represents hydrogen or C _1-4 alkyl; R ² represents hydrogen or C _1-4 alkyl. 3. The compound according to claim 1 or 2, where A is -C=C- or the bond is a single bond or a double bond; X represents carbon or nitrogen, and if X represents nitrogen, then the ^ ⁷⁷⁷⁷ ^ bond represents a single bond; R ¹ is hydrogen; R ² is hydrogen; - 37 043636 R ³ is hydrogen, hydroxyl or halogen; R ⁴ is hydrogen; halogen; With _1-6 alkyl; C1 _-6 alkyloxy; C1 _-4 alkyloxycarbonyl; aminocarbonyl; mono- or di(C _1-4 alkyl)aminocarbonyl; aryl; aryloxy; arylsulfonyl; heteroaryl; C _1-6 alkyl substituted with a cyano group; C _1-6 alkyl substituted with aryl or aryloxy; or With _1-6 alkyl substituted with heteroaryl; aryl is phenyl; phenyl substituted with one substituent selected from halogen, C _1-4 alkyl, C _1-4 alkyloxy and cyano groups; heteroaryl is furanyl, thiophenyl, pyrazolyl, isoxazolyl, thiazolyl, triazolyl, tetrazolyl, thiadiazolyl, pyridinyl or pyrimidinyl, wherein each heteroaryl may be substituted with one substituent selected from halogen, cyano, C _1-4 alkyl, C _1-4 alkoxy or C _1-4 alkylcarbonyl; or a pharmaceutically acceptable acid addition salt thereof. 4. A compound according to any one of claims 1 to 3, where R ¹ is hydrogen and R ² is hydrogen. 5. A compound according to any one of claims 1 to 4, where R ³ represents hydrogen. 6. A compound according to any one of claims 1 to 5, where R ⁴ is aryl. 7. A compound according to any one of claims 1 to 5, where R ⁴ is heteroaryl. 8. A compound according to any one of claims 1-5, where R ⁴ is C _1-6 alkyl substituted with aryl. 9. A compound according to any one of claims 1 to 8, wherein X represents nitrogen and the bond is a single bond. 10. Connection according to any one of claims 1-9, where A represents 11. Connection selected from the following: - 38 043636 - 39 043636 - 40 043636 - 41 043636 or a pharmaceutically acceptable salt thereof. 12. A pharmaceutical composition for the treatment of bacterial infections, containing a pharmaceutically acceptable carrier and a therapeutically effective amount of a compound according to any one of paragraphs. 1 11. 13. A method for preparing a pharmaceutical composition according to claim 12, wherein a therapeutically effective amount of a compound according to any one of claims 1 to 11 is thoroughly mixed with a pharmaceutically acceptable carrier. 14. Use of a compound of formula (I) according to any one of claims 1 to 11 as a medicinal agent for the treatment of bacterial infections. 15. Use of a compound of formula (I) according to any one of claims 1 to 11 for the treatment of bacterial infections. - 42 043636 16. Use according to claim 15, wherein the bacterial infection is caused by a bacterium that expresses the enzyme Fabl. 17. A method for preparing a compound of formula (I) according to claim 1 or a pharmaceutically acceptable acid addition salt thereof, comprising reacting an intermediate of formula (II) with an intermediate of formula (III) (II) (III) where A, X, R ³ , R ⁴ and Ζι are as defined in claim 1, and subsequently converting the compound of formula (I) into a pharmaceutically acceptable acid addition salt by reacting the compound of formula (I) with acid. 18. A method for preparing a compound of formula (I) according to claim 1 or a pharmaceutically acceptable acid addition salt thereof, wherein in the compound of formula (I) A is -C(R ² )=C(R ¹ )-, comprising reacting an intermediate of the formula (V) with an intermediate of formula (VI) (I) where X _a i represents a leaving group representing a halide group, and X, Z _b R ¹ , R ² , R ³ and R ⁴ are as defined in claim 1; and then converting the compound of formula (I) into a pharmaceutically acceptable acid addition salt by reacting the compound of formula (I) with an acid. Eurasian Patent Organization, EAPO Russia, 109012, Moscow, Maly Cherkassky lane, 2