BG66437B1 - Use of quinoline derivatives for the treatment of latent tuberculosis - Google Patents

Abstract

The invention relates to the use of a compound for the preparation of a medicament for the treatment of latent tuberculosis. The compound has the formula or It can be a pharmaceutically acceptable acid or base adjunctive salt, N-oxide, their tautomeric or stereochemical isomeric form. In the formulae R? is hydrogen, halogen, haloalkyl, cyano, hydroxy, Ar, Het, alkyl, alkoxy, alkylthio, alkyloxyalkyl, alkylthioalkyl, Ar-alkyl or di(Ar)alkyl; p has the value of 1, 2, 3 or 4; R? is hydrogen, hydroxy, mercapto, alkyloxy, alkyloxyalkyloxy, alkylthio, mono or di(alkyl)amino, or radical with the formula R? is alkyl, Ar-alkyl, Het or Het-alkyl; q has the value of 0, 1, 2, 3 or 4; each R4 and R5 is independently hydrogen, alkyl or benzyl; or R4 and R5 can be taken together, including the nitrogen to which they are attached; R6 is hydrogen, halogen, haloalkyl, hydroxy, Ar, alkyl, alkoxy, alkylthio, alkyloxyalkyl, alkylthioalkyl, Ar-alkyl, or di(Ar)alkyl; or two adjacent R6 radicals can be taken together in order to form a two-valent radical -CH=CH-CH=CH- r has the value of 1, 2, 3,4 or 5; R7 is hydrogen, alkyl, Ar or Het; R8 is hydrogen or alkyl; R9 is oxo; R8 and R9 together form the radical =N-CH=CH-.

Description

(54) USE OF QUINOLINE COMPOUNDS FOR THE TREATMENT OF LATENT TUBERCULOSIS

Field of technology

The invention relates to the use of a compound of formula (Ia) or (1b) for the treatment of latent tuberculosis.

BACKGROUND OF THE INVENTION

Mycobacterium tuberculosis is the cause of death of more than two million people a year and is the leading cause of death in people living with HIV ¹ . Despite decades of tuberculosis control programs, about 2 million are infected with M. tuberculosis asymptomatically. About 10% of these individuals are at risk of developing tuberculosis during their lifetime ² . The global tuberculosis epidemic is fueled by HIV patients infected with tuberculosis and the increased drug resistance of tuberculosis strains (MDR-TB). Reactivation of latent tuberculosis is a high risk factor for the development of the disease and is the cause of 32% of mortality in HIV-infected patients ¹ . To control the tuberculosis epidemic, it is necessary to discover new drugs that can kill latent or latent bacilli. Latent tuberculosis can be reactivated and cause disease by various factors such as suppression of the host immune system by using immunosuppressants such as antibodies against tumor necrosis factor alpha or interferon-gamma. In the case of HIV-positive patients, the only prophylactic treatment suitable for latent tuberculosis is a two- to three-month course of rifampicin, pyrazinamide ³⁴ . The effectiveness of the course of treatment is not yet clear, and in addition, the duration of treatment is an important limitation due to limited equipment. Therefore, there is a drastic need to find new drugs that can act as chemoprophylactic agents in patients carrying a latent tuberculosis bacillus.

The tubercle bacillus infects healthy patients by inhalation; they are phagocytosed by the alveolar microphages of the lungs. This results in a possible immune response and the formation of granulomas, which consist of macrophages infected with M. tuberculosis surrounded by T cells. After a period of 6-8 weeks, the host immune response causes the death of infected cells through necrosis and accumulation of caseous material with extracellular bacilli surrounded by macrophages, epithelial cells and layers of lymphoid tissue at the periphery ⁵ . In the case of healthy individuals, most of the mycobacteria die in this environment, but a small proportion of the bacilli come to life and are thought to exist in a non-replicating, hypometabolic state and may be resistant to anti-TB drugs such as isoniazid ⁶ . These bacilli can remain in the altered physiological environment even throughout the patient's life without showing any symptoms of disease. However, in 10% of cases, these latent bacilli can reactivate and cause disease. One of the hypotheses for the development of these resistant bacteria is the pathophysiological environment in patients, namely the reduced oxygen pressure, impaired nutrition and acidic pH ⁷ . These factors have been postulated to present these bacteria as phenotypically resistant to essential drugs.

WO 2004/011436 describes substituted quinoline derivatives suitable for the treatment of mycobacterial diseases. This document discloses the antimycobacterial properties of substituted quinoline derivatives against susceptible, resistant mycobacterial strains, but does not disclose their activity against latent, latent, persistent mycobacteria.

It has been found that the compounds of WO 2004/011436, and in particular the compounds of formula 1 (a) and 1 (b) as described below, have sterilizing properties; that they are effective in killing latent mycobacteria, in particular Mycobacterium tuberculosis., and can therefore be used to treat latent tuberculosis. They therefore significantly increase the arsenal to fight tuberculosis.

Explanation of the attached figures

Figure 1. Effect of different drugs on latent Mycobacterium bovis as assessed by luciferase measurement (RLU: relative

66437 B1 luminescent units) (bacteria were suspended in drug-free medium for 5 days after 7 days of anaerobiosis).

Figure 2A. Influence of various latent drugs Mycobacterium bovis, (CFU: colony forming units) (CFU reported 2 days after anaerobiosis).

Figure 2B. Influence of various latent drugs Mycobacterium bovis, (CFU: colony forming units) (CFU reported 5 days after anaerobiosis).

Figure 3. Influence of various latent drugs Mycobacterium tuberculosis, (Wayne model).

Technical essence of the invention

The present invention relates to the use of compound 1 (a) and 1 (b) for the manufacture of a medicament for the treatment of latent tuberculosis, wherein compound 1 (a) or 1 (b) is

(1a) a pharmaceutically acceptable acid or base addition salt thereof, an N-oxide thereof, a tautomeric form thereof or a stereochemically isomeric form thereof in which

R ¹ is hydrogen, halogen, haloalkyl, cyano, hydroxy, Ar, Het, alkyl, alkyloxy, alkylthio, alkyloxy alkyl, alkylthioalkyl, Ar-alkyl or di (Ar) alkyl;

p is a factor equal to 1,2, 3 or 4;

R ² is hydrogen, hydroxy, mercapto, alkyloxy, alkyloxyalkyloxy, alkylthio, mono or di (alkyl) amino or a radical of formula

wherein Y is CH ₂ , O, S, NH or N-alkyl;

R ³ is alkyl, Ar, Ar-alkyl, Het or Het 1Q alkyl;

q is a factor equal to 1,2, 3 or 4;

R ⁴ and R ^{5 are} each independently hydrogen, alkyl or benzyl; or

R ⁴ and R ⁵ together, including N, to which। 5 are linked may form a radical selected from the group consisting of pyrrolidinyl, 2-pyrrolinyl, 3-pyrrolinyl, pyrrolyl, imidazolidinyl, pyrazolidinyl, 2-imidazolinyl, 2-pyrazolinyl, imidazolyl, pyrazolyl, triazolyl, piperidinyl, pyridinyl, 2-pyridinyl , pyrazinyl, triazinyl, morpholinyl and thiomorpholinyl optionally substituted with alkyl, halogen, haloalkyl, hydroxy, alkyloxy, amino, mono- or dialkylamino, alkylthio, alkyloxyalkyl, alkylthioalkyl and pyrimidinyl;

R ⁶ is hydrogen, halogen, haloalkyl, hydroxy, Ar, alkyl, alkyloxy, alkylthio, alkyloxyalkyl, alkylthioalkyl, Ar-alkyl or di (Ar) alkyl; or two adjacent R ⁶ radicals may be taken together to form a divalent radical -CH = CH-CH = CH 2 is a coefficient equal to 1, 2, 3, 4 or 5;

R ⁷ is hydrogen, alkyl, Ar or Het;

R ⁸ is hydrogen or alkyl;

R ⁹ is oxo; or

R ⁸ and R ⁹ together form a radical = HCH = CH-;

Alkyl is a linear or branched hydrocarbon radical having 1 to 6 carbon 4Q atoms; or is a cyclic saturated hydrocarbon radical having 3 to 6 carbon atoms; or is a cyclic saturated hydrocarbon radical having 3 to 6 carbon atoms attached to a linear or branched hydrocarbon radical having 1 to 6 carbon atoms; wherein each carbon atom may be optionally substituted with halogen, hydroxy, alkoxy or oxo;

Ar is a homocycle selected from the group consisting of 5Q phenyl, naphthyl, acenaphthyl, tetrahydronaphthyl,

66437 B1 each homocycle is optionally substituted with 1, 2 or 3 substituents, each substituent is independently selected from the group of hydroxy, halo, cyano, nitro, amino, mono- or dialkylamino, alkyl, haloalkyl, alkyloxy, haloalkyloxy, 5 carboxyl, alkyloxycarbonyl, aminocarbonyl, morpholinyl and mono- or dialkylaminocarbonyl;

Het is a monocyclic heterocycle selected from the group consisting of N-phenoxypiperidinyl, piperidinyl, pyrrolyl, pyrazolyl, imidazolyl, 10 furanyl, thienyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, pyridinyl, pyrimidinyl, pyridazine; or bicyclic heterocycles selected from the group consisting of quinolinyl, quinoxalinyl, indolyl, benzimidazolyl, benzoxazolyl, benzisoxazolyl, benzothiazolyl, benzisothiazolyl, benzofuranyl, benzothienyl, 2,3-dihydrobenzo [1,3] dihydrobenzo [or 1,4] a monocyclic or bicyclic ring may be optionally substituted with 20, 1, 2 or 3 substituents selected from the group consisting of halogen, hydroxy, alkyl, alkyloxy or Arcarbonyl;

halogen is a substituent selected from the group consisting of fluorine, chlorine, bromine and iodine; and 25 haloalkyl is a linear or branched saturated hydrocarbon radical having from 1 to 6 carbon atoms or a cyclic saturated hydrocarbon radical having from 3 to 6 carbon atoms in which one or more carbon atoms are replaced by one or more halogen atoms. .

The present invention also relates to a method of treating patients, including people with latent tuberculosis, which comprises administering to patients a therapeutically effective amount of a compound of the invention.

The compounds according to formula 1 (a) and 1 (b) are interrelated that a compound of formula 1 (b) with R ⁹ equal to oxo is a tautomeric equivalent of a compound of formula 1 (a) with R ² equal to of hydroxy (ketoenol autometry).

Within the scope of this application, alkyl is a linear or branched saturated hydrocarbon radical 45 having 1 to 6 carbon atoms; or is a cyclic saturated hydrocarbon radical having 3 to 6 carbon atoms; or is a cyclic saturated hydrocarbon radical having 3 to 6 carbon atoms attached to a linear or 50 branched saturated hydrocarbon radical having 1 to 6 carbon atoms; wherein each carbon atom may be optionally substituted with halogen, hydroxy, alkyloxy or oxo. Preferably alkyl is methyl, ethyl or cyclohexylmethyl.

Within this invention Ar is a homocycle selected from the group of phenyl, naphthyl, acenaphthyl, tetrahydronaphthyl, each homocycle is optionally substituted with 1, 2 or 3 substituents, each substituent is independently selected from the group of hydroxy, halo, cyano, nitro , amino, mono- or dialkylamino, alkyl, haloalkyl, alkyloxy, haloalkyloxy, carboxyl, alkyloxycarbonyl, aminocarbonyl, morpholinyl and mono- or dialkylaminocarbonyl. Preferably Ar is naphthyl or phenyl, each optionally substituted with 1 or 2 halogen substituents.

Within this application Het is a monocyclic heterocycle selected from the group consisting of N-phenoxypiperidinyl, piperidinyl, pyrrolyl, pyrazolyl, imidazolyl, furanyl, thienyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, pyridinyl, pyridyl, pyrimidines; or bicyclic heterocycles selected from the group consisting of quinolinyl, quinoxalinyl, indolyl, benzimidazolyl, benzoxazolyl, benzisoxazolyl, benzothiazolyl, benzisothiazolyl, benzofuranyl, benzothienyl, 2,3-dihydrobenzo [or 1,4] dioxo; each monocyclic or bicyclic ring may be optionally substituted with 1, 2 or 3 substituents selected from the group consisting of halogen, hydroxy, alkyl, alkyloxy or Ar-carbonyl. Preferably Het is thienyl.

In this application, halogen is a substituent selected from the group consisting of fluorine, chlorine, bromine and iodine and haloalkyl is a linear or branched saturated hydrocarbon radical having 1 to 6 carbon atoms or a cyclic saturated hydrocarbon radical having from 3 to 6 carbon atoms. in which one or more carbon atoms are replaced by one or more halogen atoms. Preferred halogens are bromine, fluorine or chlorine, and the preferred haloalkyl is polyhaloC _1-6 alkyl, which is defined as mono- or polyhalogen substituted C alkyl, for example methyl with one or more fluorine atoms, for example difluoromethyl or trifluoromethyl, 1,1-difluoroethyl others. In case of

66437 B1 more than one halogen atom is attached to the alkyd group to form polyhaloC _1-6 alkyl, they may be the same or different.

In the determination of Het, or when R ⁴ and R ⁵ are taken together, this means that all possible isomeric forms of the heterocycles are included, for example pyrrolyl includes 1H-pyrrolyl and 2H-pyrrolyl.

Ar or Het specified in the definition of the substituents of the compounds of formula (Ia) and (1b) (see for example R ³ ), as noted above or below, may be linked to the remainder of the molecule of formula (Ia) or (1b), by any ring carbon or heteroatom as appropriate, unless otherwise noted. For example, when Het is imidazolyl, it may be 1-imidazolyl, 2-imidazolyl, 4-imidazolyl and the like.

The lines drawn by the substituent in the ring system indicate that the bond may be attached to any suitable ring atom.

When two adjacent R ⁶ radicals are taken together to form a divalent radical of formula -CH = CH-CH = CH-, this means that the two adjacent R ⁶ radicals form together with the phenyl ring to which a naphthyl is attached.

Salts of formula (Ia) and (1b) for therapeutic use are those in which the response is pharmaceutically acceptable. Therefore, salts of acids and bases which are non-pharmaceutically acceptable may also find use, for example in the preparation and purification of pharmaceutically acceptable compounds. All salts, whether pharmaceutically acceptable or not, are included within the scope of the present invention.

Pharmaceutically acceptable addition salts as noted above or below mean that they include therapeutically active, non-toxic acid addition salt forms which the compounds of formula (Ia) and (1b) may form. The latter can be conveniently obtained by treating the base form with suitable acids, e.g. inorganic acids - halogen acids such as hydrochloric, hydrobromic acid, etc. similar; sulfuric acid, nitric acid, phosphoric acid and the like; or organic acids such as acetic, propane, hydroxyacetic, 2-hydroxypropanoic, 2-oxopropanoic, oxalic, malonic, succinic, maleic, fumaric, malic, tartaric, 2-hydroxy-1,2,3-propanetricarboxylic, methanesulfonic, ethanesulfonic, benzenesulfonic , 4-methyl-benzenesulfonic, cyclohexanesulfonic, 2-hydroxybenzoic, 4-amino-2-hydroxybenzoic and the like. The reversible salt form can be recovered by treatment with alkali in free base form.

The compounds of formula (Ia) and (1b) containing acidic protons may be converted into their therapeutically active non-toxic metal or amine addition salt forms by treatment with a suitable organic or inorganic base. Suitable base salt forms include, for example, ammonium salts, alkali or alkaline earth metal salts such as lithium, sodium, potassium, magnesium, calcium salts and the like, salts with organic bases such as primary, secondary or tertiary aliphatic or aromatic amines such as methylamine, ethylamine , isopropylamine, the four butylamine isomers, dimethylamine, diethylamine, diethanolamine, dipropylamine, diisopropylamine, di-n-butylamine, pyrrolidone, piperidine, morpholine, trimethylamine, triethylamine, tripropylamine, quinuclidine, pyridine, quinoline and isoquinoline , 2-amino-2- (hydroxymethyl) 1,3-propanediol, hydrabamine salts and salts of amino acids such as arginine, lysine and the like. The reversible salt form can be returned by treating the acid to the free acid form.

The term addition salt also includes the hydrates and solvent addition forms which the compounds of formula (Ia) or (1b) are able to form. Examples of such forms are, for example, hydrates, alcoholates and the like.

The compounds of any of formulas (1a) and (1b) and some of the intermediates always have at least two stereo centers in their structure, which can lead to at least 4 stereochemically different structures.

The term "stereochemically isomeric forms" as used above or below deijjULWMiwaiwWH

66437 B1 finishes all possible stereoisomeric forms which the compounds of formula (Ia) and (1b), their N-oxides, addition salts or physiological functional derivatives may have. Unless otherwise indicated, the chemical designation of substances denotes the mixture of all possible stereochemically isomeric forms, and said mixtures contain all diastereomers and enantiomers of the basic molecular structure. In particular, stereo centers may have an R- or S-configuration; the substituents in the divalent cyclic (partial) saturated radicals may have a cis- or transconfiguration. Compounds having double bonds may have E (entgegen) or Z (zusammenen) stereochemistry at said double bond. The terms cis-, trans-, R, S, E and Z are well known to those skilled in the art.

Stereochemically isomeric forms of the compounds of formula (Ia) and (1b) are obviously within the scope of the invention.

In accordance with the CAS nomenclature conventions, when there are two stereo centers of a known absolute configuration in the molecule, an index R or S is assigned to the chiral center with the lowest number, the corresponding center (according to the Cahn-lngold- sequence rule). Prelog). The configuration of the second stereo center is denoted using relative indices [R *, R * J or [R *, S *], where R * is always defined as the corresponding center and [R *, R *] means the centers with the same chirality , a [R *, S *] means centers with different chirality. For example, the chiral center at the lowest number in the molecule has the S configuration, and the second center is R, the stereo index is denoted as S- [R *, S *]. If the "alpha" and "beta" positions of the substituent with the highest priority over the asymmetric carbon atom in the ring having the lowest number in the ring are used, it is always in the "alpha" place in the plane defined by the ring. The position of the substituent with the highest priority over another asymmetric carbon atom in the ring, other than the position of the substituent with the highest priority over the reference atom, is marked with "alpha" if it is on the same side of the plane defined by the ring or "Beta" if it is on the other side of the plane defined by the ring.

When specific stereoisomeric forms are indicated, this means that said form is substantially free, for example containing less than 50%, preferably less than 20%, more preferably less than 5%, even more preferably less than less than 5%, further preferably less than 2% and most preferably less than 1% of the other isomer (s). Thus, when a compound of formula (I) is designated as (aS, PR), this means that the compound is substantially free of the (aR, PS) isomer.

The compounds of formula (Ia) and (1b) may be synthesized in the form of a racemic mixture of enantiomers which can be separated from one another following procedures known in the art. The racemic compounds of formula (Ia) or (1b) may be converted into the corresponding diastereosalt forms by reaction with a suitable chiral acid. Said diastereomeric salt forms are obtained sequentially, for example, by selective or fractional crystallization, and the enantiomers are released by alkalization. An alternative way to separate the enantiomeric forms from the compounds of formula (Ia) or (1b) involves liquid chromatography using a chiral stationary phase. Said stereochemically isomeric forms may also be prepared from suitable starting compounds which ensure that the reaction proceeds stereospecifically. If it is necessary to prepare specific stereoisomers, it is preferred that said compounds be synthesized by stereospecific methods of preparation. Enantiomerically pure starting compounds are preferably used in these methods.

Tautomeric forms of the compounds of formula (Ia) or (1b) mean that they include those compounds of formula (Ia) or (1b) in which, for example, an enol group is converted to a keto group (keto-enol tautomerism).

The N-oxide forms of the compounds of formula (Ia) or (1b) mean that they include those compounds of formula (Ia) or (1b) in which one or more tertiary nitrogen atoms are oxidized to so-called N-oxides.

The compounds of formula (Ia) and (1b) may be converted into the corresponding N-oxide forms following art known in the art.

66437 B1 procedures for converting trivalent nitrogen to the oxide form. Said N-oxidation reaction may generally be carried out by reacting the starting compounds of formula (Ia) and (1b) with a suitable organic or inorganic peroxide. Suitable inorganic peroxides include, for example, hydrogen peroxide, alkali or alkaline earth metal peroxides, such as sodium peroxide, potassium peroxide; suitable organic peroxides 10 include peroxy acids such as benzenecarboperoxoic acid, such as 3-chlorobenzenecarboperoxoic acid, peroxoalkanoic acids, such as peroxoacetic acid, alkyd hydroperoxides, such as t-butyl hydroperoxide. Suitable solvents are water, lower alcohols such as ethanol and the like, hydrocarbons such as toluene, ketones such as 2-butanone, halogenated hydrocarbons such as dichloromethane, and mixtures of such solvents. 20

The invention also includes derivatives (commonly referred to as "prodrugs") of the pharmacologically active compounds of the invention that degrade in vivo to the compounds of the invention. Prodrugs 25 usually (but not always) have lower activity at the target receptor than the compounds to which they are degraded. Prodrugs are suitable in cases where the desired compound has chemical or physical properties that make its administration difficult or ineffective. For example, if the desired compound may be poorly soluble, it may be difficult to transport through the mucosal epithelium, or it may have an undesirably short plasma half-life. 35 Further discussions on prodrugs can be found in Stella, V. J. et al., “Prodrugs”, Drug Delivery System, 1985, pp. 112176, and Drugs, 1985, 29, pp. 455-473.

The prodrug forms of the pharmacologically active compounds of the compounds are generally compounds of formula (Ia) and (1b), their pharmaceutically acceptable acid or base addition salts, their stereochemically isomeric forms, their tautomeric forms and their N-oxide forms which have an acid group which is esterified or amidated. Included in such esterified acid groups are groups of formula -COOR ^X , wherein R ^x is C ₁₆ alkyl, phenyl, benzyl or one of the following groups:

Amidated groups include groups of formula -CONR ^y R ^z wherein R ^y is H, C16 alkyl, phenyl or benzyl and R ^z is -OH, H, C16 alkyl, phenyl or benzyl.

The compounds of the invention having an amino group can be modified with a ketone or aldehyde as formaldehyde to give Mannich bases. These bases are hydrolyzed in an aqueous medium with first order kinetics.

When the term "compounds of formula (Ia) or (1b)" is used, this means that their pharmaceutically acceptable acid or base addition salts, their N-oxide forms, their tautomeric forms or their stereochemically isomeric forms are also included. Of particular interest are those compounds of formula (Ia) or (Ib) which are stereochemically pure.

A first embodiment of the present invention relates to the use of the compounds of formula (1a) or (1b) defined above, wherein:

R 'is hydrogen, halogen, haloalkyl, cyano, Ar, Het, alkyl, alkyloxy;

p is a factor equal to 1, 2, 3 or 4; specifically 1 or 2;

R ² is hydrogen, hydroxy, alkyloxy, alkyloxyalkyloxy, alkylthio, or a radical of formula

R ³ is alkyl, Ar, Ar-alkyl, or Het;

q is a factor equal to 1, 2 or 3;

R ⁴ and R ^{5 are} each independently hydrogen, alkyl or benzyl; or

R ⁴ and R ⁵ together, including N to which they are attached may form a radical selected

66437 B1 from the group of pyrrolidinyl, imidazolidinyl, triazolyl, piperazinyl, pyrazinyl, morpholinyl and thiomorpholinyl, each cyclic system optionally substituted with alkyl or pyrimidinyl;

R ⁶ is hydrogen, halogen or alkyl; or two adjacent R ⁶ radicals may be taken together to form a bivalent radical -CH = CH-CHCH- r is a factor equal to 1;

R ⁷ is hydrogen;

R ⁸ is hydrogen or alkyl;

R ⁹ is oxo; or

R ⁸ and R ⁹ together form a radical = HCH = CH-;

alkyl is a linear or branched hydrocarbon radical having 1 to 6 carbon atoms; or is a cyclic saturated hydrocarbon radical having 3 to 6 carbon atoms; or is a cyclic saturated hydrocarbon radical having 3 to 6 carbon atoms attached to a linear or branched hydrocarbon radical having 1 to 6 carbon atoms; wherein each carbon atom may be optionally substituted with halogen or hydroxy;

Ar is a homocycle selected from the group of phenyl, naphthyl, acenaphthyl, tetrahydronaphthyl, each homocycle is optionally substituted with 1, 2 or 3 substituents, each substituent is independently selected from the group of halogen, haloalkyl, cyano and morpholinyl;

Het is a monocyclic heterocycle selected from the group consisting of HaN-phenoxypiperidinyl, piperidinyl, furanyl, thienyl, pyridinyl, pyrimidinyl; or bicyclic heterocycles selected from the group of benzothienyl, 2,3-dihydrobenzo [1,4] dioxinyl or benzo [1,3] dioxolyl, each monocyclic or bicyclic ring may be optionally substituted with 1, 2 or 3 alkyl or Ar -carbonyl substituents;

halogen is a substituent selected from the group consisting of fluorine, chlorine and bromine.

In a second embodiment, the compounds of formula (Ia) or (Ib) or any subgroup thereof, as noted above as an embodiment of the invention, are those compounds according to any of formulas (1a) and (1b). wherein R ¹ is hydrogen, halogen, Ar, alkyl or alkyloxy; preferably R ¹ is halogen; more preferably when R ¹ is bromine.

In a third embodiment, the compounds of formula (Ia) or (Ib) or any subgroup thereof, as noted above as exemplary embodiments of the invention, are those compounds according to any of formulas (Ia) and (Ib) ) in which p is equal to 1 and R ¹ is different from hydrogen.

In a fourth embodiment, the compounds of formula (Ia) or (1b) or any subgroup thereof, as noted above as exemplary embodiments of the invention, are those compounds according to any of formulas (Ia) and (Ib) wherein R ² is hydrogen, alkyloxy or alkylthio; preferably R ² is alkyloxy, in particular C ₄ alkyloxy; more preferably when R ² is methyloxy.

C _1-4 alkyl is a linear or branched saturated hydrocarbon radical having from 1 to 4 carbon atoms, for example methyl, ethyl, propyl, 2-methyl-ethyl and the like.

In a fifth embodiment, the compounds of formula (Ia) or (1b) or any subgroup thereof, as noted above as exemplary embodiments of the invention, are those compounds according to any of formulas (Ia) and (Ib) , wherein R ³ is naphthyl, phenyl or thienyl, each optionally substituted with 1 or 2 substituents, which are preferably halogen or haloalkyl, more preferably halogen; preferably R ³ is naphthyl or phenyl, each substituted with halogen, preferably 3-fluoro; more preferably when R ³ is naphthyl or phenyl; most preferably when R ³ is naphthyl.

In a sixth embodiment, the compounds of formula (Ia) or (1b) or any subgroup thereof, as noted above as exemplary embodiments of the invention, are those compounds according to any of formulas (Ia) and (Ib) , wherein q is zero, 1 or 2; preferably q is equal to 1.

In a seventh embodiment, the compounds of formula (Ia) or (1b) or any subgroup thereof, as noted above as exemplary embodiments of the invention, are those compounds according to any of formulas (Ia) and (Ib) in which R ⁴ and R ⁵ each independently is hydrogen or alkyl, in particular hydrogen or C _{| 4} alkyl, in particular C _1-4 alkyl; preferably hydrogen, methyl or ethyl; best methyl. C _1-4 alkyl is a linear or branched carbon> 437 B1 according to formula (Ia) only, a pharmaceutically acceptable acid or base addition salt thereof, a tautomeric form thereof or a stereochemically isomeric form thereof.

An interesting group of compounds are the compounds of formula (Ia), their pharmaceutically acceptable acid or base addition salts, their stereochemically isomeric forms, their tautomeric forms or their 10 N-oxide forms, in which R ¹ is hydrogen, halogen, Ar, alkyl or alkyloxy; p = 1, R ² is hydrogen, alkyloxy or alkylthio; R ³ is naphthyl, phenyl or thienyl, each optionally substituted with 1 or 2 substituents selected from the group consisting of halogen or 15 haloalkyl; q = 0, 1, 2 or 3; R ⁴ and R ^{5 are} each independently hydrogen or alkyl or R ⁴ and R ⁵ together and including N to which they are attached form a radical selected from the group consisting of imidazolyl, triazolyl, piperidinyl, piperazinyl and thiomorpholinyl; R ⁶ is hydrogen, alkyl or halogen; r is 1 and R ⁷ is hydrogen.

Preferably for the compounds of formula (Ia) and (1b) or any subgroup thereof, as noted above as an exemplary embodiment of the invention, the term "alkyl" represents C _1-6 alkyl, which C _1-6 alkyl is linear or a branched hydrocarbon radical having from 1 to 6 carbon atoms such as methyl, ethyl, propyl, 2-methyl-ethyl, pentyl, 30 hexyl and the like.

Preferably, the compounds are selected from:

• 1- (6-bromo-2-methoxy-quinolin-3-yl) -2 (3,5-difluoro-phenyl) -4-dimethylamino-1-phenyl-butan-2-ol;

• 1- (6-Bromo-2-methoxy-quinolin-3-yl) -4-dimethylamino-2-naphthalen-1-yl-1-phenyl-butan-2-ol corresponding to 6-bromo-alpha- [2- (dimethylamino) ethyl] -2-methoxy-alpha-1-naphthalenyl40 beta-phenyl-3-quinolinethanol;

• 1- (6-bromo-2-methoxy-quinolin-3-yl) -2 (2,5-difluoro-phenyl) -4-dimethylamino-1-phenyl-butan-2-ol;

• 1- (6-bromo-2-methoxy-quinolin-3-yl) -245 (2,3-difluoro-phenyl) -4-dimethylamino-1-phenyl-butan-2-ol;

• 1- (6-bromo-2-methoxy-quinolin-3-yl) -4-dimethylamino-2- (2-fluoro-phenyl) -1-phenyl-butan-2-ol;

· 1- (6-bromo-2-methoxy-quinolin-3-yl) -41 η hydrogen radical having from 1 to 4 carbon atoms, such as methyl, ethyl, propyl, 2-methyl-ethyl and the like.

In an eighth embodiment, the compounds of formula (1a) or (1b) or any subgroup thereof, as noted above as an exemplary embodiment of the invention, are those compounds according to any one of formula (1a) and (1b) wherein R ⁴ and R ⁵ together with N to which they are attached form a radical selected from the group consisting of imidazolyl, triazolyl and thiomorpholinyl optionally substituted with alkyl, halogen, haloalkyl, hydroxy, alkyloxy, alkylthio, alkyloxyalkyl or alkylthioalkyl preferably substituted with alkyl, most preferably substituted with methyl or ethyl.

In a ninth embodiment, the compounds of formula (Ia) or (Ib) or any subgroup thereof, as noted above as an embodiment of the invention, are those compounds according to any formula (Ia) and (Ib) wherein R ⁶ is hydrogen, alkyl or halogen, preferably R ⁶ is hydrogen.

In a tenth embodiment, the compounds of formula (Ia) or (1b) or any subgroup thereof, as noted above as exemplary embodiments of the invention, are those compounds according to any of formulas (1a) and (1b). ) in which d is 1 or 2.

In an eleventh embodiment, the compounds of formula (Ia) or (1b) or any subgroup thereof, as noted above as exemplary embodiments of the invention, are those compounds according to any of formulas (Ia) and (Ib) , R ⁷ is hydrogen or methyl, preferably R ⁷ is hydrogen.

In the twelfth embodiment, the compounds of formula (Ia) or (1b) or any subgroup thereof, as noted above as exemplary embodiments of the invention, are those compounds according to any of formulas (1a) and (1b). , wherein for compounds of formula (1b) only, R ⁸ is alkyl, preferably methyl, and R ⁹ is oxygen.

In a thirteenth embodiment, the compounds of formula (Ia) or (1b) or any subgroup thereof, as noted above as exemplary embodiments of the invention, are those compounds according to any of formulas (1a) and (1b). wherein the compound is a compound

66437 B1 Dimethylamino-2-naphthalen-1-yl-1-p-tolyl-butan2-ol;

• 1- (6-bromo-2-methoxy-quinolin-3-yl) -4-dimethylamino-2-naphthalen-1-yl-1-phenyl-butan-2-ol; 5 • 1- (6-bromo-2-methoxy-quinolin-3-yl) -4-dimethylamino-2- (3-fluoro-phenyl) -1-phenyl-butan-2-ol; and • 1- (6-bromo) -2-Methoxy-quinolin-3-yl) -4-dimethylamino-2-phenyl-1-phenyl-butan-2-ol. 10

Preferred compounds are:

• 1- (6-bromo-2-methoxy-quinolin-3-yl) -2 (2,3-difluoro-phenyl) -4-dimethylamino-1-phenylbutan-2-ol;

• 1- (6-bromo-2-methoxy-quinolin-3-yl) -4-15-dimethylamino-2-naphthalen-1-yl-1-phenyl-butan-2-ol corresponding to 6-bromo-alpha- [2 - (dimethylamino) ethyl] -2-methoxy-alpha-1-naphthalenyl-betaphenyl-3-quinolinethanol; or a pharmaceutically acceptable acid or base attachment thereof, a salt thereof, an oxide thereof, a tautomeric form thereof or a stereochemically isomeric form thereof.

Another interesting group of compounds are the following: compounds 12, 71, 174, 75, 172 and 79, 25 as described in Tables 1 to 6 and in particular compounds 12,71,174 and 75, in particular 12, 71 and 174; a pharmaceutically acceptable acid or base addition salt thereof, an N-oxide thereof, a tautomeric form thereof or a stereochemically isomeric form thereof.

Alternative chemical name of 1- (6-bromo-2-methoxy-quinolin-3-yl) -4-dimethylamino-2-naphthalen-1-yl-1-phenyl-butan-2-ole 6-bromo-alpha [2 - (dimethylamino) ethyl] -2-methoxy-35 alpha-1-naphthalenyl-beta-phenyl-3-quinolinethanol. Said compound can be represented as follows:

Most preferably, one of the following compounds:

6-bromo-alpha [2- (dimethylamino) ethyl] -2-methoxy-alpha-1-naphthalenyl-beta-phenyl-3-quinolineethanol, a pharmaceutically acceptable acid or base addition salt thereof, their N-oxide, their tautomeric form or their stereochemically isomeric form; or

6-bromo-alpha [2- (dimethylamino) ethyl] -2-methoxy-alpha-1-naphthalenyl-beta-phenyl-3-quinolineethanol, or a pharmaceutically acceptable acid or base addition salt thereof; or

6-bromo-alpha [2 (dimethylamino) ethyl] -2-methoxy-alpha-1-naphthalenyl-beta-phenyl-3-quinolineethanol, or a stereochemically isomeric form thereof; or

6-bromo-alpha [2 (dimethylamino) ethyl] -2-methoxy-alpha-1-naphthalenyl-beta-phenyl-3-quinolinetanol, or their N-oxide; or a mixture, in particular a racemic mixture of (aS, pK) -6-bromo-alpha- [2- (dimethylamino) ethyl] -2-methoxy-alpha-1-naphthalenyl-beta-phenyl3-quinolinethanol and (aR, p8 ) -6-bromo-alpha- [2 (dimethylamino) ethyl] -2-methoxy-alpha-1-naphthalenyl-beta-phenyl-3-quinolinethanol, or a pharmaceutically acceptable acid or base addition salt thereof, or a stereochemically isomeric form thereof ; for example compound 14 (diastereoisomer A); or (aS, 3R) -6-6Pomo-alpha- [2- (dimethylamino) ethyl] -2-methoxy-alpha-1-naphthalenyl-beta-phenyl-3-quinolinethanol, for example compound 12, or a pharmaceutically acceptable acid addition or a salt thereof; or (aS, βR) -6-bromo-alpha- [2- (dimethylamino) ethyl] -2-methoxy-alpha-1-naphthalenyl-beta-phenyl-3-quinolineethanol, for example compound 12.

The most preferred compound is (aS, βR) -6-bromo-alpha- [2- (dimethylamino) ethyl] -2-methoxy-alpha-1-naphthalenyl-beta-phenyl-3-quinolineethanol, which corresponds to (1R, 2S) 1- (6-Bromo-2-methoxy-quinolin-3-yl) -4-dimethylamino-2-naphthalen-1-yl-1-phenyl-butan-2-ol. Said compound can be represented as follows:

1

66437 Bl

to be prepared by the methods described in WO 2004/011436, which are incorporated herein by reference.

In general, the compounds of the invention in the sequence of steps 5 are each known to those skilled in the art.

In particular, the compounds of formula (Ia) can be prepared by reacting an intermediate of formula (II) with 10 intermediates of formula (111) according to the following reaction scheme (1):

The compounds of formula (Ia) and (1b) may

Scheme 1

Using BuLi in a mixture of diisopropyl and tetrahydrofuran, all variables are defined as in formula (Ia). Stirring may increase the reaction rate. The reaction is usually carried out in the temperature range between -20 and -70 ° C.

The same reaction procedure can be used in the synthesis of the compounds of formula (1b).

The starting compounds and intermediates of formula (II) and (III) are compounds which are either commercially available or can be prepared by standard reaction procedures known in the art. For example, intermediate (II-a) can be prepared according to the following reaction scheme (2):

66437 B1 where all variables are defined as in formula (Ia). Reaction scheme (2) comprises step (a) in which a suitably substituted aniline is reacted with a suitable acyl chloride such as 3-phenylpropionyl chloride, 3-fluorobenzene-5-propionyl chloride or p-chlorobenzenepropionyl chloride, in the presence of a suitable base such as triethylamino and a suitable reaction a solvent such as metal chloride or ethylene dichloride. The reaction can be carried out at a temperature range from room temperature to reflux temperature. In the next step (b), the adduct obtained in step (a) is reacted with phosphoryl chloride (POCl ₃ ) in the presence of N, N-dimethylformamide (15 Vilsmeier-Haack composition followed by cyclization). The reaction usually takes place in the temperature range from room temperature to reflux temperature. In the next step (c), the specific group R ² is introduced, where the radical R ^{2 is,} for example, C _1-6 alkyloxy or C _1-6 alkylthio. The introduction is carried out by reacting the intermediate obtained in step (b) with a compound H-X-C 1-6 alkyl in which X is S or O.

The intermediates of formula (IIb) can be prepared according to reaction scheme (3), wherein in the first step (a) the substituted indole-2,3-dione is reacted with substituted 3-phenylpropionaldehyde in the presence of a suitable base such as sodium hydroxide (reaction of Pfitzinger), after which the carboxylic acid compound in the next step (b) is decarboxylated at high temperature in the presence of a suitable reaction-inert solvent such as diphenyl ether.

(P-b)

It will be appreciated that in the preceding and subsequent reactions, the reaction products may be isolated from the reaction medium and, if necessary, further purified according to methods well known in the art, such as extraction, crystallization and chromatography. It is also obvious that the reaction products, which exist in more than one enantiometric form, can be isolated from their mixtures by known techniques, in particular by preparative chromatography such as preparative HPLC. Generally, the compounds of formula (I) may be separated into their isomeric forms.

Intermediates of formula (III) are compounds which are either commercially available or can be prepared by standard methods well known in the art. For example, the intermediate of formula (IIIa), wherein R ³ is Ar substituted with R ¹⁰ substituents, wherein each R ¹⁰ is independently selected from the group consisting of hydroxy, halogen, cyano, nitro, amino, mono- or Di (C _1-6). alkyl) amino, ₆ alkyl, polyhaloC ,, alkyl, C ,, alkyloxy, polyhaloC "alkyloxy, carboxyl, C _1-6 alkyloxycarbonyl, aminocarbonyl, morpholinyl and mono- or di (C _{| -C6} alkyl) aminocarbonyl and s is an integer equal to zero , 1, 2 or 3, can be obtained by following reaction scheme (4):

чимо.жда.отмм-л

66437 Bl

Scheme 4

Ο (R'V

Ar ^{C1 V1}

(W-a)

The reaction scheme (4) comprises step (a) in which a suitably substituted Ar, and in particular a suitably substituted phenyl, is reacted by a Friedel-Craft reaction with a suitable acyl chloride such as 3-chloropropyl chloride or 4-chlorobutyryl chloride in the presence of a suitable Lewis acid. such as A1S1 _p FeCl _3, SnCl ₄ or ZnCl ₂ and a suitable reaction-inert solvent such as methylene chloride or ethylene dichloride. The reaction can usually be carried out in the temperature range between room temperature and reflux temperature. In the next step (b), an amino group (-NR ⁴ R ⁵ ) is introduced by reacting the intermediate obtained in step (a) with a primary or secondary amine.

As regards the interpretation of the present invention, the terms "latent tuberculosis", "latent tuberculosis" or "persistent tuberculosis" have the same meaning.

As noted above, the compounds of formula (Ia) and (1b) may be used to treat latent tuberculosis. The exact dosage and mode of administration of the present compounds depends on the particular compound of formula (Ia) and (1b) used, the particular case being treated, the severity of the case being treated, age, weight, sex, diet, the time of administration, the general physical condition of the particular patient, the route of administration, and other medications that the patient may be taking, as is well known to those skilled in the art. It will be appreciated that the effective daily dose may be reduced or increased depending on the response of the subject and / or the judgment of the prescribing physician.

The compounds of the present invention may be administered in a pharmaceutically acceptable form, optionally with a pharmaceutically acceptable carrier.

The pharmaceutical compositions may take different pharmaceutical forms depending on the route of administration. As suitable compositions, all compositions commonly used as systemically administered drugs can be cited. For the preparation of a pharmaceutical composition, an effective amount of the particular compound is combined, optionally in the form of an addition salt form as active ingredient in a homogeneous mixture with a pharmaceutically acceptable carrier, which carrier may be different depending on the desired route of administration. It is desirable for these pharmaceutical compositions to be in unit dosage form, for example, for oral administration or parenteral injection. For example, in preparing a composition for oral dosage form, any pharmaceutical medium may be used, such as water, glycols, oils, alcohols and the like in the case of oral liquid preparations such as suspensions, syrups, elixirs, emulsions and solutions; or solid carriers such as starch, sugar, kaolin, diluents, lubricants, binders, disintegrants and the like in the case of powders, pills, capsules50 / 1

66437 Bl li or tablets. Due to the ease of administration, tablets and capsules represent the most suitable dosage forms for oral administration, in which case a solid pharmaceutical carrier is usually employed. In the case of parenteral compositions, they contain sterile water, at least in large part, as is the use of other components, for example solubilizers. In the preparation of injectable solutions, the carrier comprises a co-solution, a glucose solution or a mixture of saline and glucose solution. Injectable suspensions may also be prepared in which case a suitable liquid carrier, suspending agent and the like may be employed. Also included are solid dosage forms that can be converted to liquid dosage forms immediately prior to use.

Depending on the route of administration, the pharmaceutical compositions comprise from 0.05 to 99% by weight, preferably from 0.1 to 70% by weight. % of the active ingredient and from 1 to 99.5 wt. %, preferably from 30 to 99.9 wt. % pharmaceutically acceptable carrier, all based on the total composition. 25

The pharmaceutical compositions may additionally contain other ingredients known in the art, such as lubricants, stabilizers, buffers, emulsifiers, viscosity regulators, surfactants, preservatives, flavors or colorants.

It is especially important to prepare the above dosage forms in dosage forms for easy administration and uniformity of 35 dosages. The dosage form as used herein refers to discrete physical units suitable for unit doses, each unit containing a predetermined amount of the active ingredient calculated to give the desired therapeutic effect together with the required pharmaceutical carrier. Examples of such dosage forms are tablets (including dashed or coated tablets), capsules, pills, sachets, patches, suppositories, injectable solutions or suspensions, and the like, and packaging. The daily dose of the compound according to the invention will, of course, depend on the compound used, the route of administration, the treatment required and the microbacterial parameters. In general, satisfactory results are obtained when the compound according to the present invention is administered in a daily dose not exceeding 1 or 2 g, for example from 10 to 50 mg / kg body weight.

Experimental part

As emphasized above, the compounds of formula (Ia) and (1b) and their preparation are described in WO 2004/011436 and are incorporated by reference.

For some compounds, the absolute stereochemical configuration of the stereogenic carbon atom has not been determined experimentally. In these cases, the stereochemically isomeric form which is isolated is first designated as "A" and the second as "B", without further reference to the actual stereochemical configuration. Therefore, said "A" and "B" isomeric forms can be unambiguously characterized by one skilled in the art using methods known in the art such as X-ray diffraction (X-ray diffraction).

When "A" and "B" are stereoisomeric mixtures, they may be further separated, the first isolated fraction being marked as "A 1" and "B 1" respectively, and the second as "A2" and "B2" respectively, without further notation. of the current stereochemical configuration. Therefore, the "Al, A2" and "B1, B2" isomeric forms can be unambiguously characterized by one skilled in the art using methods known in the art such as X-ray diffraction.

The present compounds (Tables 1 to 6) are numbered according to the compounds in WO 2004/011436 and can be prepared according to the methods described in WO 2004/011436. The example number in the Tables below corresponds to the example numbers in WO 2004/011436, indicating the procedures by which the compounds can be prepared.

The preparation of compounds 12,13,12a, 13a, 14 and 15 is described in detail below.

Preparation of intermediates

Example A1

Preparation of intermediate 1

66437 Bl

Benzenepropanoyl chloride (0.488 ml) was added dropwise at room temperature to a solution of 4-bromobenzenamine (0.407 mol) in triethanolamine EtN (70 ml) and dichloromethane CH ₂ Cl ₂ (700 ml) and the mixture was stirred at room temperature for one day and night. The mixture was poured into water and concentrated NH ₄ OH and extracted with CH ₂ Cl ₂ . The organic layer was dried (MgSO ₄ ), filtered and the solvent was evaporated.

The residue was crystallized from diethyl ether. The residue (119.67 g) was placed in CH ₂ Cl ₂ and washed with HCl 1N. The organic layer was dried (MgSO ₄ ), filtered and the solvent was evaporated. Yield 107.67 g of intermediate 1.

Example 2

Preparation of intermediate 2

The reaction is carried out twice. POCl ₃ 30 (1.225 mol) was added dropwise and at 10 ° C to N, N-dimethylformamide (DMF) (0.525 mol). Intermediate 1 (prepared according to Al) (0.175 mol) was then added at room temperature. The mixture was stirred overnight at 80 ° C, poured onto ice and extracted with CH ₂ Cl ₂ . The organic layer was dried (MgSO ₄ ), filtered and the solvent was evaporated. The product is used without further purification. Yield 77.62 g of intermediate 2 (67%).

Example AZ

Preparation of intermediate 3

A mixture of intermediate 2 (prepared according to A2) (0.233 mol) in CH ₃ ONa (30%) in methanol (222.32 ml) and methanol (776 ml) was stirred at reflux for one day, then pour on ice and extract with CH ₂ Cl ₂ . The organic layer was separated, dried (MgSO ₄ ), filtered and the solvent was evaporated. The residue was purified by column chromatography over silica gel (eluent: CH ₂ Cl ₂ / cyclohexane 20/80 and then 100/0; 20-45 microm). The pure fraction was collected and the solvent was evaporated. Yield: 25 g of intermediate 3 (Yield = 33%; m.p. 84 ° C) as a white powder.

Preparation of the final compounds 12, 13, 12a, 13a, 14 and 15

Preparation of the final compounds 12, 13, 12a, 13a, 14 and 15

A

66437 Bl

Compound 14 (A)

Compound 15 (B)

Compound 12 (Al)

[a] _D »= -166.98 (c = 0.505g / 100ml in DMF)

Compound 12a (Bl)

_{[(X] d} 20 = -42 56 (c = 0.336g / 100ml in DMF)

Compound 13 (A2)

[a] _D »= + 167.60 (c = 0.472g / 100ml in DMF)

Compound 13a (B2)

[α] _D 20 = + 43.55 (c = 0.349g / 100ml in DMF) nBuLi 1.6M (0.005 mol) was added slowly at -20 ° C under nitrogen purge to a solution of N- (1-methylethyl) -2- propanamine (0.05 mol) in tetrahydrofuran (THF) (80 ml). The mixture was stirred at -20 ° C for 15 minutes and then cooled to -70 ° C. A solution of intermediate 3 (prepared according to Example A3 described above) (0.046 mol) in THF (150 ml) was added slowly. The mixture was stirred at -70 ° C for 30 minutes. A solution of 0.055 mol of 3- (dimethylamino) -1- (1-naphthyl) -140 propanone in THF (120 ml) was added slowly. The mixture was stirred at -70 ° C for 3 h, hydrolyzed at -30 ° C with ice water and extracted with EtOAc ethyl acetate. The organic layer was separated, dried (MgSO ₄ ), filtered and the solvent was evaporated. The residue (29 g) was purified by column chromatography on silica gel (eluent: CH ₂ Cl ₂ / CH ₃ OH / NH ₄ OH; 99.5 / 0.5 / 0.1; 1535 microm). The two fractions were combined and the solvent was evaporated, yielding 3 g of fraction 1 and 4.4 g of fraction 2. Fractions 1 and 2

66437 B1 are crystallized separately from DIPE diisopropyl ether. The precipitate was filtered off and dried, yielding 2.2 g of diastereomer A final compound 14 (Yield 9%, melting point 210 ° C) as a white solid and 4 g of diastereomer B final compound 15 (Yield 16%, temperature of mp 244 ° C) as a white solid. To give the corresponding enantiomers, diastereomer A (final compound 14) was purified by chiral chromatography on silica gel (chiralpack AD) (eluent: hexane / ethanol; 99.95 / 0.05). The two fractions were combined and the solvent was evaporated. Yield: 0.233 g of enantiomer A1 (final compound 12) (mp 118 ° C, [alpha] ₀ ^{2 ()} = -166.98 ° (c = 0.505 g / 100 ml in DMF)) as a white solid and 0.287 g of enantiomer A2 (final compound 13) (mp 120 ° C, [alpha] ^{20 =:} + 167.60 ° (c = 0.472 g / 100 ml in DMF)) as a white solid. Enantiomer A1 was crystallized from ethanol to give a white solid: mp 184 ° C, [alpha] ²⁰ = -188.71 ° (c = 0.621 g / 100ml in DMF). Crystallization of enantiomer A2 from ethanol gave a white solid with a melting point of 175 ° C.

0.2 g of diastereoisomer B (final compound 15) was purified by chiral chromatography on silica gel (chiralpack AD) (eluent ethanol / isopropyl alcohol / N-ethylethanamine; 50/50 / 0.1). The two fractions were combined and the solvent was evaporated. Yield: 78.2 mg of enantiomer B1 and 78.8 mg of enantiomer B2.

Enantiomer B1 was purified by column chromatography over silica gel (eluent CH ₂ Cl ₂ / CH ₃ OH / NH ₄ OH; 99/1 / 0.1; 15-40 microns). One fraction was collected and the solvent was evaporated. Yield: 57 mg of enantiomer B1 (final compound 12a) [alpha] _at ²⁰ = -42.56 ° (c = 0.336 g / 100ml in DMF)). Enantiomer B2 was purified by column chromatography over silica gel (eluent CH ₂ Cl ₂ / CH ₃ OH / NH ₄ OH; 99/1 / 0.1; 15-40 microns). One fraction was collected and the solvent was evaporated. Yield 5 3 mg of enantiomer B2 (final compound 1 To) ([alpha] _D ²⁰ = + 43.55 ° (a = 0.349 g / 100 ml in DMF)).

Tables 1 to 6 describe the compounds of formula (Ia) and (1b).

about

66437 Bl

Table 1

R6

Eat. № Example № R ’ R ² R ³ R ⁶ Stereochemistry and melting points 1 Bl Br OCH, Phenyl N (A1); 194 ° C 2 Bl Br AND ₃ Phenyl N (A2); 191 ° C 3 Bl Br OCH, Phenyl h (A); 200 ° C 4 Bl Br OCH, Phenyl N (IN); 190 ° C 16 Bl Br AND ₃ 4-chlorophenyl N (A); 200 ° C 17 Bl Br OCH, 4-chlorophenyl N (IN); 190 ° C 20 Bl Br OCH, 2-thienyl N (A); 96 ° C 21 Bl Br AND ₃ 2-thienyl N (IN); 176 ° C 22 Bl CH ₃ OCH, phenyl N (A); 148 ° C 23 Bl CH, OCH, phenyl N (IN); 165 ° C 24 Bl Br OCH, 3-thienyl N (A); 162 ° C 25 Bl Br OCH, 3-thienyl N (IN); 160 ° C 26 Bl phenyl OCH, phenyl N (A); 174 ° C

66437 Bl

27 Bl phenyl AND ₃ phenyl N (IN); 192 ° C 28 Bl F AND ₃ phenyl N (A); 190 ° C 29 Bl F AND ₃ phenyl N (IN); 166 ° C 30 Bl Cl OCH, phenyl N (A); 174 ° C 31 Bl Cl AND ₃ phenyl N (IN); 18 GS 32 Bl Br SCH ₃ phenyl N (A); 208 ° C 33 Bl Br SCH ₃ phenyl N (IN); 196 ° C 34 Bl AND ₃ AND ₃ phenyl n (A); 165 ° C 35 Bl AND ₃ AND ₃ phenyl n (IN); 165 ° C 36 Bl Br AND ₃ phenyl C1 (A); 197 ° C 37 Bl Br AND ₃ phenyl C1 (IN); 221 ° C 38 B9 Br AND ₃ 3-fluorophenyl n (A); 198 ° C 39 B9 Br AND ₃ 3-fluorophenyl n (B): 207 ° C 108 B9 Br AND ₃ 3-fluorophenyl n (A1): 160 ° C 109 B9 Br AND ₃ 3-fluorophenyl n (A2); 156 ° C 40 Bl H AND ₃ phenyl n (A): 152 ° C 41 Bl H AND ₃ phenyl n (IN); 160 ° C 42 Bl H AND ₃ CH ₃ n (A); 140 ° C 43 Bl H AND ₃ CH ₃ n (B): 120 ° C 59 B) Br OH phenyl n (A); > 260 ° C 60 Bl Br OH phenyl n (IN); 215 ° C 5 B2 Br OCH _; CH ₃ phenyl n (A); 162 ° C 6 B2 Br AND ₂ CH ₃ phenyl n (IN); Low: 74 ° C 7 B3 Br H phenyl n (A); 98 ° C 8 B3 Br H phenyl n (IN); 180 ° C 12 B7 Br AND ₃ 1-naphthyl n (A1); 118 ° C (foam) a = K b = S; [a] _D ²⁰ = -166.98 (c = 0.505 g / 100mlBDMF) 13 B7 Br AND ₃ 1-naphthyl n (A2); 120 ° C (foam) a = S, b = R; [α] ^ + 167.60 ° (c = 0.472 g / 100mlBDMF) 12a B7 Br OSNz 1-naphthyl n (Bl); [ab ² ° = - 42.56 * 13a B7 Br AND ₃ 1-naphthyl n (B2); [a] _D ^2C '= +43.55 ** 14 B7 Br OCHj 1-naphthyl n (A) 210 ° C 15 B7 Br AND ₃ 1-naphthyl n (B); 244 ° C 45 B7 Br OSNz 2-naphthyl n (A); 262 ° C 46 B7 Br OCH, 2-naphthyl n (B); 162 ° C

he

66437 Bl

67 B8 Vg OSN ₃ 2,5-difluorophenyl N (A); 262 ° C 68 B8 Vg OSN ₃ 2,5-dafluorophenyl N (IN); 208 ° C 110 B8 Vg OSNz 2,5-difluorophenyl N (A1); 167 ° C 111 B8 Vg OSN ₃ 2,5-dafluorophenyl N (A2): oil 69 B1 Vg OSN ₃ 2-fluorophenyl N (A); butter 70 B1 Vg OSN ₃ 2-fluorofnsnyl N (IN); butter 71 B1 Vg OSN ₃ 1-naphthyl СНз (A); 174 ° C 72 B1 Vg OSNz 1-naphthyl СНз (IN); 178 ° C 73 B1 Vg OSNz 1-naphthyl C1 (IN); 174 ° C 74 B1 Vg OSNz 1-naphthyl C1 (A); 110 ° C 75 B1 Vg OSNz N (A); 196 ° C 76 B1 Vg OSNz N (IN); 130 ° C 77 B1 Vg OSNz N (A); 202 ° C 78 B1 Vg OSNz N (IN); 202 ° C 79 B1 Vg --- Ν ^// 'θ 1-naphthyl N (A); > 250 ° C 80 B1 Vg OSNz 4-cyanofsnyl N (A); 224 ° C 81 B1 Vg OSN ₃ 4-cyanophenyl N (IN); 232 ° C 82 B1 CH ₃ AXIS, 1-naphthyl N (A); 202 ° C 83 B1 СНз AXIS, 1-naphthyl n (IN); 198 ° C 84 B1 phenyl OSNz 1-naphthyl n (A); 248 ° C 85 B1 phenyl OSNz 1-naphthyl n (IN); 214 ° C 86 B1 Vg OSNz n (A); 184 ° C 87 B1 Vg OSNz n (IN); 186 ° C 88 B1 Vg SCH ₃ 1-naphthyl n (A); 240 ° C

66437 Bl

89 Bl Br AND ₃ 0 N (A); 236 ° C 90 Bl Br AND ₃ 0 N (IN); 206 ° C 91 Bl H AND ₃ 1-naphthyl N (A); 178 ° C 92 Bl H AND ₃ 1-naphthyl N (IN); 160 ° C 93 Bl H AND ₃ 3-fluorophenyl N (A); 178 ° C 94 Bl H AND ₃ 3-fluorophenyl N (IN); 182 ° C 95 Bl Br AND ₃ 2-phenylethyl N (A); 178 ° C 96 Bl Br AND ₃ 15 2-phenylethyl N (IN); 146 ° C 97 Bl AND ₃ AND ₃ 1-naphthyl N (A); 168 ° C 98 Bl AND ₃ AND ₃ 1-naphthyl N (IN); 154 ° C 113 BI4 Br OCH, 2,3-difluorophenyl N (A); 128 ° C 114 B14 Br AND ₃ 20,3-difluorophenyl N (IN); 213 ° C 115 B15 Br AND ₃ 3,5-difluorophenyl N (A); 192 ° C 116 B15 Br AND ₃ 3,5-difluorophenyl N (IN); 224 ° C 117 B15 Br OCH, 3,5-difluorophenyl N (A1); 161 ° C 118 B15 Br AND ₃ 2 5>. 5-difluorophenyl N (A2); 158 ° C 119 B7 Cl AND ₃ 1-naphthyl N (A); 212 ° C 120 B7 Cl AND ₃ 1-naphthyl N (IN); 236 ° C 122 B7 Br OCH, ³⁰ N (IN); 227 ° C 127 B7 Br AND ₃ 5-bromo-2-naphthyl N (A); 226 ° C 130 B7 Br AND ₃ 3 $ 5-6romo-2-naphthyl N (IN); 220 ° C 131 B7 Br AND ₃ 00 N (A); 206 ° C 134 B9 AND ₃ AND ₃ 4Q 3-fluorophenyl N (A); 172 ° C 135 B9 AND ₃ OCH, 3-fluorophenyl n (IN); 192 ° C 143 B7 Br AND ₃ 3-bromo-1-naphthyl n (A); 234 ° C 150 B7 Br OCH, C-bromo-1-naphthyl n (IN); 212 ° C 159 B8 Br AND ₃ 4 ^. 5-difluorophenyl n (A1); 208 ° C 160 B8 Br OCH., 2,5-difluorophenyl n (A2): 167 ° C 162 B7 Br AND ₃ 6-methoxy-2-naphthyl n (A); 206 ° C 163 B7 Br AND ₃ 6-methoxy-2-naphthyl n (IN); 206 ° C

66437 Bl

164 B9 Vg 0 0 3-fluorophenyl N (A); 118 ° C 165 B9 Vg S 0 0 \ 3-fluorophenyl N (IN); butter 174 B9 and. OSNz 3-fluorophenyl N (A); 159 ° C 175 B9 and. OSN ₃ 3-fluorophenyl N (IN); 196 ° C 176 B7 Vg 0 0 \ 1-naphthyl N (A); butter 179 B9 CN OSN ₃ 3-fluorophenyl N (A); 213 ° C 180 B9 CN OSNz 3-fluorophenyl N (IN); 163 ° C 181 B9 Vg OSNz 4-fluorophenyl N (A); 198 ° C 182 B9 Vg OSNz 4-fluorophenyl N (IN); 238 ° C 183 B1 Vg OSNz 3-trifluoromethylphenyl N (A); 170 ° C 188 B1 Vg OSNz 1,4-pyrimidin-2-yl N (A); 110 ° C 189 B1 Vg OSN ₃ 1,4-pyrimidin-2-yl N (IN); 145 ° C 195 B15 Vg OSN ₃ 3,4-difluorophenyl N (A); 250 ° C 196 B15 Vg OSNz 3,4-difluorophenyl N (IN); 184 ° C 201 B1 Vg AXIS, / --- \ θ N (A); 214 ° C 202 B1 Vg AXIS, N (IN); 246 ° C 203 B9 Op OSNz 3-fluorophenyl n (A); 225 ° C 204 B9 0 ”Ί OSNz 3-fluorophenyl n (IN); 216 ° C 205 B7 Vg OSNz 1-naphthyl F (A); 213 ° C

66437 Bl

205 B7 Br AND ₃ 1-naphthyl F (IN); 213 ° C 207 B15 F OSNz 3,5-difluorophenyl N (A); 232 ° C 208 B15 F OSNz 5 3,5-difluorophenyl N (IN); 188 ° C 212 B7 1K \ JL OSNz 1-naphthyl F (IN); 220 ° C

* ¢ = 0.336 g / 100 mlBDMF ** ¢ = 0.349 g / 100 ml in DMF 10

Table 2:

R ⁶

Eat. № Example № R ¹ R ² 2B ³ R ⁴ R ⁵ Physical data (salt / temp. Top.) And stereochemistry 18 B1 Vg OSNz phenyl CH, CH ₃ CH ₂ CH3 ethanedionate (2: 3); (A); 230 ° C 19 B1 Vg OSNz ф ^ уил CH ₂ CH3 СН.СНз ethanedionate (2: 3); (IN); 150 ° C 44 B4 Vg OSNz phenyl n N (A); 190 ° C 9 B4 Vg OSNz phenyl n N (IN); 204 ° C 141 B7 Vg OSNz 2- ^ phthyl СНз CH ₂ CH ₃ (A); 188 ° C 142 B7 Vg OSNz 2-naphthyl CH ₃ CH ₂ CH ₃ (IN); 202 ° C 230 B12 Vg OSNz 1-naphthyl СНз benzyl /butter 147 B7 Vg OSNz 1-naphthyl CH ₃ CH ₂ CH ₃ (A); 168 ° C 148 B7 Vg OSNz 1-naphthyl 40 СНз CH ₂ CH3 (IN); 212 ° C 56 B13 Vg OSNz 1-naphthyl СНз N (A); 204 ° C 214 B13 Vg OSNz 1-naphthyl CH ₃ N (IN); 225 ° C

1L

66437 Bl

Table 3:

R ⁶

Eat. № Example № R ³ L Stereochemistry and melting points 47 Bl phenyl 1-piperidinyl (A); 190 ° C 48 Bl phenyl 1-piperidinyl (IN); 190 ° C 128 Bl 2-naphthyl 1-piperidinyl (A); 254 ° C 129 Bl 2-naphthyl 1-niperidinyl (IN); 212 ° C 49 Bl phenyl 1-imvdazolyl (A); 216 ° C 50 Bl phenyl 1-imvdazolyl (IN); 230 ° C 51 Bl phenyl 1- (4-methyl) piperazinyl (A); 150 ° C 52 Bl phenyl 1- (4-methyl) piperazinyl (IN); 230 ° C 53 Bl phenyl 1- (1,2,4-triazolyl) (A); 180 ° C 54 Bl phenyl 1 - (1,2,4-triazolyl) (IN); 142 ° C 55 Bl phenyl thiomorpholyl (A); butter 57 B5 phenyl I- (A); 244 ° C 10 B5 phenyl ^N C 1 ' (IN); 198 ° C 58 B6 phenyl ^{+ n} \ O ' (A); 208 ° C 11 B6 phenyl ⁺ <O ' (IN); 208 ° C

66437 Bl

99 YOU 1-naphthyl ^{+ n} C 5 0 ' (A1); 218 ° C 100 B6 1-naphthyl OZ ⁺ / \ (A2): 218 ° C 101 B6 1-naphthyl \ / Z-'O (IN); 175 ° C 102 B5 1-naphthyl 15 ^t Ч ^ 1- (A2): 210 ° C 103 B5 1-naphthyl 20 d (IN); > 210 ° C 121 B5 1-naphthyl 25 I- (A1): 210 ° C 123 B1 phenyl morpholinyl (A); 226 ° C 124 B1 phenyl morpholinyl (IN); 210 ° C 136 B7 2-naphthyl 4-methylpyrazinyl (A); 188 ° C 137 B7 2-naphthyl 3-methylpyrazinyl (IN); 232 ° C 139 B7 2-naphthyl morpholinyl (A); 258 ° C 140 B7 2-naphthyl morpholinyl (IN); 214 ° C 144 B7 2-naphthyl pyrrolidinyl (A); 238 ° C 145 B7 1-naphthyl 2 51 -piperidinyl (A); 212 ° C 146 B7 1-naphthyl 1-piperidinyl (IN); 220 ° C 149 B7 1-naphthyl 4-methylpyrazinyl (IN); 232 ° C 151 B7 3-bromo-1-naphthyl 4-megilirazinyl (A); 178 ° C 152 B7 3-bromo-1-naphthyl methylpyrazinyl (IN); 226 ° C 153 B7 6-bromo-2-naphthyl 4-methylpyrazinyl (A); 208 ° C 154 B7 b-bro.mo-2-naphthyl 4-methylpyrazinyl (IN); 254 ° C 155 B7 6-bromo-2-naphthyl 1-piperidinyl (A); 224 ° C 156 B7 1-naphthyl 4-methylpiperazinyl 45 (A); 200 С 157 B7 6-bromo-2-naphthyl 1-pyrrolidinyl (IN); 220 ° C 158 B7 1-naphthyl morpholinyl (IN); 272 ° C 166 B7 6-bromo-2-naphthyl 1-piperidinyl (IN); 218 ° C

66437 Bl xi.

170 B7 2-naphthyl 1-pyrrolidinyl (A); 238 ° C 171 B7 2-naphthyl 1-pyrrolidinyl (IN); 218 ° C 172 B7 1-naphthyl 1,2,4-triazol-1-yl / 142 ° C 173 B7 1-naphthyl 1,2-imidazol-1-yl (A); 222 ° C 177 B7 6-bromo-2-naphthyl morpholinyl (A); 242 ° C 178 B7 6- € romo-2-naphthyl morpholinyl (IN); 246 ° C 187 B7 1-naphthyl 1,2-imdazol-1-yl (IN); 236 ° C 200 B7 2-naphthyl 0 (A); 254 ° C 209 B7 2-naphthyl / \ ^{N =} \ -N, Ή) \ - / _N _7 (IN); 198 ° C

Table 4;

Eat. № Example № R ³ Q L Stereochemistry and points of melting 61 B1 phenyl 0 N (CH ₃ ) ₂ (A); 220 ° C 62 B1 phenyl 0 N (CH ₃ ) ₂ (IN); 220 ° C 63 B1 phenyl 2 N (CH ₃ ) ₂ (A); 150 ° C 64 B1 phenyl 2 N (CH ₃ ) ₂ (IN); 220 ° C 125 B7 2-naphthyl 2 N (CH ₃ ) _: (A); 229 ° C 126 B7 2-naphthyl 2 N (CH ₃ ) ₂ (IN); 214 ° C 65 B1 phenyl 3 N (CH ₃ ) ₂ (A); 130 ° C 65 B1 phenyl 3 N (CH ₃ ) ₂ (IN); 170 ° C 132 B7 2-naphthyl 2 pyrrolidinyl (A); 227 ° C 133 B7 2-naphthyl 2 pyrrolidinyl (IN); 222 ° C 161 B7 2-naphthyl 2 morpholinyl (IN); 234 ° C 186 B7 1-naphthyl 2 N (CH ₃ ) ₂ (A); 187 ° C

66437 Bl

190 B7 2-naphthyl 3 N (CH ₃ ) ₂ (A); 170 ° C 191 B7 2-naphthyl 3 N (CH ₃ ) ₂ (IN); 145 ° C 192 B7 2-naphthyl 2 N (CH ₂ CH ₃ ) ₂ (A); 90 ° C 193 B7 2-naphthyl 2 N (CH ₂ CH ₃ ) ₂ (IN); 202 ° C 194 B7 1-naphthyl 2 pyrrolidinyl (IN); 206 ° C 197 B7 1-naphthyl 3 N (CH ₃ ) ₂ (A); 160 ° C 198 B7 2-naphthyl 2 morpholinyl (A); 215 ° C 199 B7 1-naphthyl 2 N (CH ₂ CH ₃ ) ₂ (A); 185 ° C 210 B7 1-naphthyl 2 morpholinyl (IN); 222 ° C 211 B7 1-naphthyl 2 morpholinyl (A); 184 ° C

Table 5:

Eat. № Example № R ³ R ⁸ R ⁹ Stereochemistry and melting points 104 B1 phenyl -CH = CH-N = (A); 170 ° C 105 B1 phenyl -CH = CH-N = (IN); 150 ° C 106 B1 phenyl CH ₃ = 0 (A); 224 ° C 107 B1 phenyl CH ₃ = 0 (IN); 180 ° C 138 B7 1-naphthyl N = 0 (A1); > 260 ° C

Table 6:

R6

Ω0

66437 Bl

Eat .№ Note № R ' R ³ R ⁶ Stereochemistry and melting points a l s d 215 B9 N Vg CH ₃ N 3-fluorophenyl N (A); 197 ° C 215 B9 N Vg CH ₃ n 3-fluorophenyl N (IN); 158 ° C 217 B7 N n Vg n 1-naphthyl N (A); 212 ° C 218 B7 N n Vg n 1-naphthyl N (IN); 172 ° C 219 B9 N Vg N СНз 3-fluorophenyl N (A); 220 ° C 220 B9 N Vg N СНз 3-fluorophenyl N (IN); 179 ° C 221 B7 Vg N n n 1-naphthyl N (A); 170 ° C 224 B7 Vg n n n 1-naphthyl N / 205 ° C 222 B7 n Vg n n 1-naphthyl o 3 4 (A); 155 ° C 223 B7 1 n Vg n n 1-naphthyl l 3 4 (IN); 205 ° C 225 B7 n Vg CH ₃ n 1-naphthyl N (A); 238 ° C 226 B7 n Vg СНз n 1-naphthyl n (IN); 208 ° C 227 B15 n Vg CH ₃ n 3,5-difluorophenyl n (A); 195 ° C 228 B15 n Vg СНз n 3,5-difluorophenyl n (IN); 218 ° C 229 B7 n CH ₃ СНз n 1-naphthyl n (A); 238 ° C

A. Investigation of the effect of final compound 12 in the destruction of latent Mycobacterium bovis

Bacterial strains and culture medium

Mycobacterium bovis BCG was obtained from Tibotec Virco (TB0087- (Belgium). M, bovis BCG expressing the luciferase gene in plasmid pSMT1 (kindly provided by Dr. Kris Huygen at the Pasteur Institute, Brassies ⁸ ) was grown in Middlebrook 7H9 medium (Difco, BD271310) c 0 = 05% Tween-80 (Sigma) in the log phase for a period of 3-4 days before the start of the experiment.

To prepare the culture medium, 4.7 g of Middlebrook powder are dissolved in 895 ml of distilled water, and 5 ml of glycerol, 200 microl of Tween 80 are added and boiled in an autoclave at 121 ° C for 15 minutes. Aseptically, 100 ml of Middlebrook OADC enrichment medium was added after cooling to 45 ° C. Stored at

4 ° C for a maximum of one month. The whole medium was incubated for 2 days at 37 ° C to check for contamination. 50 microg / ml hygromycin was added for M. bovis BCG strain expressing luciferase gene (BCG-pSMT1).

I. Study with Mycobacterium bovis BCG Latency study

500 microl of Mycobacterium bovis BCG origin was added to 100 ml of Middlebrook 7H9 nutrient medium with additives in a 250 ml sterile Duran bottle with a magnetic stirrer. Incubation was performed with electromagnetic stirring for 7 days at 37 ° C (500 rpm). Transfer 5 ml aliquots of the culture phase (OD _600nm = 0.5 to 0.8) to tubes with screw caps. Different drugs were added to the individual tubes up to a final concentration of 10 microg / ml. After adding the drugs, all tubes are closed

OP

66437 Bl freely and placed in an anaerobic jar (BBL). Anaerobic gas generator closure is used to obtain an anaerobic medium, and anaerobic strips are used to monitor anaerobic conditions. The addition of individual drugs and the onset of anaerobiosis in the jar is extremely fast, as described earlier ⁹ . The jar was incubated for 7 days at 37 ° C.

CFU study

After 7 days of anaerobiosis, latent cultures were collected by low speed centrifugation (2000 rpm for 10 min). The cells were washed twice with 7H9 medium so that the drugs were separated and resuspended in drug-free medium. CFU of treated and untreated cultures was determined by monitoring swabs on day 0.2 and day 5 to assess bacterial activity.

II. M. bovis BCG assay expressing luciferase gene in plasmid pSMT1 Latency assay

500 ml of Mycobacterium bovis BCG origin is added to 100 ml of Middlebrook 7H9 nutrient medium with additives in a 250 ml sterile Duran bottle with a magnetic stirrer. Incubation was performed with electromagnetic stirring for 7 days at 37 ° C (500 rpm). Transfer 5 ml aliquots of the log culture phase (OD _600nm = 0.5 to 0.8) to tubes with screw caps. Different drugs were added to the individual tubes up to a final concentration of 10 microg / mi. After the addition of the drugs, all tubes were closed freely and placed in an anaerobic jar (BBL) as described previously ⁹ . Anaerobic gas generator closure is used to obtain an anaerobic medium, and anaerobic strips are used to monitor anaerobic conditions. The jar was incubated for 7 days at 37 ° C.

Luciferase assay

After 7 days of anaerobiosis, latent cultures were collected by low speed centrifugation (2000 rpm for 10 min). The cells were washed twice with 7H9 medium so that the drugs were separated and resuspended in drug-free medium. After washing, 250 microl of latent M. Bovis BCG luciferase (pSMT1) was added to 5 different microplates (day 0 to day 5). Each sample was diluted on microplates (5-fold dilution) in medium and re-incubated at 37 ° C for 0 to 5 days. 40 microl of the samples and diluents were added to 140 microl PBS. 20 microl luciferase substrate (1% pdecyl aldehyde in ethanol) was added. To monitor the growth of vital bacteria, the luminescence was measured for 10 s every day from day 0 to day 5. (Use Lumonoskan Ascent Labsystems with injector).

Organization of the experience

66437 Bl

№ sample Strain M. Bovis Sample / compound Microgram / ml 1-2 BCG Control 3 BCG Metronidazole 10 4 BCG Isoniazid 10 5-6 BCG Final compound 12 10 7-8 BCG Final compound 12 1 9-10 BCG Final compound 12 0.1 11-12 BCG / pSMTl Control 13 BCG / pSMTl Final compound 12 10 14 BCG / pSMTl Final compound 12 10 15-16 BCG / pSMTl Final compound 12 10 17-18 BCG / pSMTl Final compound 12 1 19-20 BCG / pSMTl Final compound 12 0.1

Discussion of the results

An in vitro latency model has been developed by Wayne's method to create a latent bacterium by oxygen depletion ^9,10 . In the Wayne model for the mycobacterium located at the bottom of the flask, an oxygen gradient is created, providing anaerobic conditions at the bottom of the flask. This shift to low oxygen concentrations causes the bacterium to shift to latency, leading to irregular expression of various genes, including isocitrate lyase and glycine dehydrogenase ⁷ . These enzymes are responsible for energy production in the absence of oxygen, and the final electron receptors are nitrates, sulfates, and others. as compared to molecular oxygen in the case of aerobic respiration. The energy of the reduced substrates generates an electron chemical gradient.

This experiment used an adaptation of the Wayne model in the proposed experiment, including the use of a Gaspak anaerobic jar in which oxygen was depleted in the chamber by means of a chemical reaction ⁹ . The Gaspak jar is equipped with a 50 lid containing a catalyst. The Gaspak foil bag containing substances that generate hydrogen and CO ₂ is placed in the bacterial culture jar. Open the bag and pipette 10 ml of water. When the jar is closed (the lid is tightened tightly), the separated hydrogen reacts with the oxygen via the catalyst to form water. This gradually depletes the oxygen available in the chamber and also creates an oxygen gradient. Additionally, indicator strips in the jar contain metallic blue, which turns colorless in the absence of oxygen. The change in color of the indicator strips indicates that the required weather conditions have been reached.

For a rapid analysis of the effect of the compound on the latent bacterium M. Bovis, earlier experiments were used as a substitute for apparent latency in general in mycobacteria and in particular in M. tuberculosis' ^1.12 . Luciferase strains are used too often to assess bacterial viability ^13,14 . M. bovis BCG was transformed with the known plasmid pSMT1, which is a vector containing the pro66437 B1 output of a replica of E. coli and mycobacteria ⁸ . The luminescent genes of Vibro harveyi (lux A and B) are under the control of the BCG hsp60 promoter and emit light in the presence of ATP or Flavin mononucleotides (FMNH ₂ ). Dead cells cannot produce these co-factors, which corresponds to a decrease in luminescence.

In this latency study, both the activity of the final compound 12 and the activity of other drugs such as metronidazole and Isoniazid were analyzed. Latent bacteria are not killed by Isoniazid, and in some cases are resistant to rifampicin, but are susceptible to metronidazole, which is known as an antibiotic for anaerobic pathogens ¹⁵ · ¹⁶ . Isoniazid acts as a bactericidal agent and its activity is limited to the destruction of bacillus replication, but does not show a significant sterilizing effect against latent bacilli ¹⁷ .

After 7 days of anaerobiosis, the bacteria were suspended in drug-free medium for 5 days, examining the effect of the various compounds on bacterial vitality by counting luciferase. As shown in Figure 1, Isoniazid had no effect on latent bacteria and they had almost the same developmental characteristics as the control sample, demonstrating latency or non-replicating status of the bacilli tested. In contrast, metronidazole was completely effective in killing latent bacilli over the same time period with a reduction of 2 log ₁₀ compared to the control sample. End compound 12 affects the survival of the latent bacterium in a concentration-dependent manner. At a concentration of the final compound 12 of 10 microg / ml, a decrease of 4-log ₁₀ in bacterial viability was observed compared to the untreated control sample. At concentrations of the compound of 0.1 and 1 microg / ml, the corresponding kill of the latent bacterium is about 0.5 log ₁₀ and 2 log _{10, respectively} .

To find the correlation of the effect of final compound 12 on the destruction of bacteria in relative luminescent units (RLU / ml) with respect to the ability to form colonies (CFU / ml), bacterial counting was performed on 7H10 plates.

A similar relationship was observed between RLU units and CFU units after day 2 and day 4 of 7H10 plate placement. The decrease in CFU numbers compared to those in the untreated control sample showed that the final compound 12 at a concentration of 10 microg / ml, 1 microg / ml and 0.1 microg / ml decreased the vitality by approximately 4, 2.3 and 0.5 log ₁₀ on day 2 and about 6, 4.7 and 1.1 log ₁₀ per day 5. Figure 2 (A and 10 B) shows the CFU data. A close relationship between luminescence and CFU was observed during different stages of the experiment. Interestingly, there is a significant reduction in RLU at present 0 compared to CFU numbers, mainly because the concentration of these cells is very low, which has been shown to be a characteristic of the metabolic state of latent bacilli ⁸ .

The activity of the final compound 12 on the latency (non-replication) of mycobacteria, which is an extremely important effect that will help in the fight against tuberculosis by eliminating the disease in patients who are at risk of developing tuberculosis.

B. Study of the effect of final compound 12 on the eradication of latent Mycobacterium tuberculosis according to a Wayne latency model *

Bacterial strains and culture medium Mycobacterium tuberculosis (H37RV) was cultured in Middlebrook medium with 0.05% Tween.

To prepare the Middlebrook (1X) medium (BD271310) with additives, 4.7 g of Middlebrook powder are dissolved in 895 ml of distilled water and 5 ml of glycerol, 200 ml of Tween 80 are added and autoclaved at 121 ° C for 15 psh. Aseptically, 100 ml of Middlebrook OADC Enrichment BD (211886) was added to the medium after cooling to 45 ° C. Store at 4 ° C for a maximum of one month. The medium is preincubated for 2 days at 37 ° C to check for contamination.

* Wayne L.G. et al .; Infection and Immunity 64 (6), 2062-2069 (1996)

Mycobacterium tuberculosis test (H37RV)

Latency study

1000 microl c origin Mycobacterium

66437 Bl tuberculosis (previous culture) was added to 100 ml of Middlebrook 7H9 nutrient medium with additives in a 250 ml sterile Duran bottle with a magnetic stirrer. Incubation was performed with electromagnetic stirring for 7 days at 37 ° C (500 rpm). 17 ml aliquots of the log culture phase (calculated (OD _600nm = 0.01)) were transferred to 25 ml tubes, the tubes were tightly closed with rubber septa stoppers and incubated on a magnetic stirrer to create anaerobiosis by reducing oxygen. Stirring in tubes was accomplished with 8 mm Teflon stirrers, incubated for 22 days at 37 ° C in an incubator on a 15 magnetic stirrer (120 rpm) until anaerobiosis (metallic blue (1.5 mg / l) After 14 days, different drugs were added to the individual tubes (final concentrations 100,10,1 and 0.1 microg / ml) Metronidazole was added as a control to kill latent bacteria (added at the beginning) Izoniazid was added as a control to show that it has no effect on the development and viability of latent bacteria.

CFU study

After 22 days, the cultures were collected by 10 low speed centrifugation (2000 rpm for 10 min). The cells were washed twice with drug-free medium so that the drugs were separated, resuspended in drug-free medium and incubated. The reduction in CFU compared to untreated control cultures was determined by observing a swab after anaerobiosis to assess bacterial activity.

Organization of the experience

№ sample Sample / compound pg / ml 1-2 control - 3-4 Metronidazole 100 5-6 Isoniazid 10 7-8 Moxifloxacin 10 9-10 1 11-12 Final compound 12 10 13-14 1 15-16 Rifampicin 10 17-18 1

Discussion of the results

The effect of final compound 12 on latent bacteria was demonstrated (Figure 3) using a Wayne latency model. As noted above, this is an in vitro oxygen depletion model that targets the latent response of bacteria ^{18 '23} . In the Wayne model, bacteria were subjected to a gradual decrease in oxygen when incubated45 not in sealed tubes with stirring. By slowly moving aerobically growing bacteria under anaerobic conditions, the bacteria are able to adapt and survive anaerobiosis by transitioning to an anaerobic state. The Wayne model is a well-characterized in vitro latency model.

At a concentration of the final compound 12 of 10 microg / ml, a reduction of

66437 Bl latent bacteria more than 2 log ₁₀ , as seen in the cases of movifloxacin and rifampicin. Isoniazid did not show any effect on the latent bacterium as a control compound, and metromidazole showed good efficacy. 5

Claims (26)

Use of a compound of formula (1a) or (1b) for the manufacture of a medicament for the treatment of latent tuberculosis, wherein the compound of formula (1a) or (1b) is their pharmaceutically acceptable acid or base addition salt, their oxide, their tautomeric form or their stereochemically isomeric form, in which R 1 is hydrogen, halogen, haloalkyl, cyano, hydroxy, Ar, Het, alkyl, alkyloxy, alkylthio, alkyloxyalkyl, alkylthioalkyl, Ar-alkyl or di (Ar) alkyl; p is a factor equal to 1, 2, 3 or 4; R ² is hydrogen, hydroxy, mercapto, alkyloxy, alkyloxyalkyloxy, alkylthio, mono or 40 di (alkyl) amino or a radical of formula wherein Y is CH ₂ , O, S, NH or N-alkyl; R ³ is alkyl, Ar, Ar-alkyl, Het or Het-alkyl; q is a factor equal to 1, 2, 3 or 4; R ⁴ and R ^{5 are} each independently hydrogen, alkyl or benzyl; or R ⁴ and R ⁵ together, including N to which they are attached may form a radical selected from the group consisting of pyrrolidinyl, 2-pyrrolinyl, 3-pyrrolinyl, pyrrolyl, imidazolidinyl, pyrazolidinyl, 2-imidazolinyl, 2-pyrazolinyl, imidazolyl, pyrazolyl, triazols , piperidinyl, pyridinyl, piperazinyl, pyridazinyl, pyrimidinyl, pyrazinyl, triazinyl, morpholinyl and thiomorpholinyl substituted optionally with alkyl, halogen, haloalkyl, hydroxy, alkyloxy, amino, mono- or dialkylamino, alkylthio, alkylthio; R ⁶ is hydrogen, halogen, haloalkyl, hydroxy, Ar, alkyl, alkyloxy, alkylthio, alkyloxyalkyl, alkylthioalkyl, Ar-alkyl or di (Ar) alkyl; or two adjacent R ⁶ radicals may be taken together to form a divalent radical -CH = CH-CH = CH 2 is a coefficient equal to 1, 2, 3, 4 or 5; R ⁷ is hydrogen, alkyl, Ar or Het; R ⁸ is hydrogen or alkyl; R ⁹ is oxo; or R ⁸ and R ⁹ together form a radical = HCH = CH-; Alkyl is a linear or branched hydrocarbon radical having 1 to 6 carbon atoms; or is a cyclic saturated hydrocarbon radical having 3 to 6 carbon atoms; or is a cyclic saturated hydrocarbon radical having 3 to 6 carbon atoms attached to a linear or branched hydrocarbon radical having 1 to 6 carbon atoms; wherein each carbon atom may be optionally substituted with halogen, hydroxy, alkoxy or oxo; Ar is a homocycle selected from the group of phenyl, naphthyl, acenaphthyl, tetrahydronaphthyl, each homocycle is optionally substituted with 1, 2 or 3 substituents, each substituent is independently selected from the group of hydroxy, halo, cyano, nitro, amino, mono- or dialkylamino, alkyl, haloalkyl, alkyloxy, haloalkyloxy, carboxyl, alkyloxycarbonyl, aminocarbonyl, morpholinyl and mono- or dialkylaminocarbonyl; Het is a monocyclic heterocycle selected from the group consisting of HaN-phenoxypiperidinyl, piperidinyl, pyrrolyl, pyrazolyl, imidazolyl, fura66437 B1 nyl, thienyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, pyridinyl, pyrimidinyl; or bicyclic heterocycles selected from the group consisting of quinolinyl, quinoxalinyl, indolyl, benzimidazolyl, benzoxazolyl, benzisoxazolyl, benzothiazolyl, benzisothiazolyl, benzofuranyl, benzothienyl, 2,3-dihydrobenzo [or monoxiol] benzo [1,3] diol or the bicyclic ring may be optionally substituted with 1, 2 or 3 substituents selected from the group consisting of halogen, hydroxy, alkyl, alkyloxy or N-carbonyl; halogen is a substituent selected from the group consisting of fluorine, chlorine, bromine and iodine; and haloalkyl is a linear or branched saturated hydrocarbon radical having from 1 to 6 carbon atoms or a cyclic saturated hydrocarbon radical having from 3 to 6 carbon atoms in which one or more carbon atoms are replaced by one or more halogen atoms. Use according to claim 1, wherein R ¹ is hydrogen, halogen, cyano, Ar, Het, alkyl and alkyloxy; p is a factor equal to 1 or 2; R ² is hydrogen, hydroxy, alkyloxy, alkyloxyalkyloxy, alkylthio, or a radical of formula wherein Y is O; R ³ is alkyl, Ar, Ar-alkyl, or Het; q is a factor equal to zero, 1, 2 or 3; R ⁴ and R ^{5 are} each independently hydrogen, alkyl or benzyl; or R ⁴ and R ⁵ together, including N to which they are attached may form a radical selected from the group consisting of pyrrolidinyl, imidazolyl, triazolyl, piperidinyl, piperazinyl, pyrazinyl, morpholinyl and thiomorpholinyl, each cyclic system being optionally substituted with alkyl or pyrimidinyl; R ⁶ is hydrogen, halogen or alkyl; or two adjacent R ⁶ radicals may be taken together to form a divalent radical of formula-CH = CH-CH = CH 2 O g is a factor of 1; R ⁷ is hydrogen; R ⁸ is hydrogen or alkyl; R ⁹ is oxo; or 5 R ⁸ and R 'together form a radical = HCH = CH-; alkyl is a linear or branched hydrocarbon radical having 1 to 6 carbon atoms; or is a cyclic saturated hydrocarbon radical having 3 to 6 carbon atoms; or is a cyclic saturated hydrocarbon radical having 3 to 6 carbon atoms attached to a linear or branched saturated hydrocarbon radical having 1 to 6 carbon 15 atoms; wherein each carbon atom may be optionally substituted with halogen or hydroxy; Ar is a homocycle selected from the group of phenyl, naphthyl, acenaphthyl, tetrahydronaphthyl, each homocycle is optionally substituted with 1, 2 or 3 substituents, each substituent is independently selected from the group of halogen, haloalkyl, cyano and morpholinyl; Het is a monocyclic heterocycle selected from the group consisting of N-phenoxypiperidinyl, piperidinyl, furanyl, thienyl, pyridinyl, pyrimidinyl; or bicyclic heterocycles selected from the group of benzothienyl, 2,3-dihydrobenzo [1,4] dioxinyl or benzo [1,3] dioxolyl, each monocyclic or bicyclic ring may be optionally substituted with 1, 2 or 3 alkyl or Ar -carbonyl substituents; halogen is a substituent selected from the group consisting of fluorine, chlorine and bromine. Use according to claims 1 or 2, wherein in formula (Ia) or (Ib) R ¹ is hydrogen, halogen, Ar, alkyl or alkyloxy. Use according to claim 3, wherein R ¹ is halogen. Use according to any one of the preceding claims, wherein in formula (1a) or (1b) p is equal to 1. Use according to any one of the preceding claims, wherein in formula (Ia) or (Ib) R ² is hydrogen, alkyloxy or alkylthio. Use according to claim 6, wherein R ² is alkyloxy. Use according to any one of the preceding claims, wherein in formula (Ia) or (Ib) R ³ is naphthyl, phenyl or thienyl, each optionally substituted with 1 or 2 substituents selected from 'ur. j,; ~ -I -i: .. ·. · ...- • -ί ·? · '· ”· ^ -' ·. ^ ··? ·. ·: ^ '^ ·?; * .rr ' ^ll ' ^^ ”'' ^, * '^>: 66437 B1 from the group halogen or haloalkyl. Use according to claim 8, wherein R ³ is naphthyl. Use according to any one of the preceding claims, wherein in formula (Ia) or (Ib) q is equal to 1. Use according to any one of the preceding claims, wherein in formula (Ia) or (Ib) R ⁴ and R ^{5 are} independently hydrogen or alkyl or R ⁴ and R ⁵ together with N to which they are attached form a radical, selected from the group consisting of imidazolyl, triazolyl, piperidinyl, piperazinyl and thiomorpholinyl. Use according to claim 11, wherein in formula (Ia) or (Ib) R ⁴ and R ^{5 are} independently hydrogen or alkyl. Use according to claim 12, wherein in formula (Ia) or (Ib) R ⁴ and R ⁵ are C ₁₄ alkyl. Use according to any one of the preceding claims, wherein in formula (Ia) or (Ib) R ⁶ is hydrogen, alkyl or halogen. Use according to claim 13, wherein R ⁶ is hydrogen. Use according to any one of the preceding claims, wherein in formula (Ia) or (Ib) d is equal to 1. Use according to any one of the preceding claims, wherein in formula (Ia) or (Ib) R ⁷ is hydrogen. Use according to claim 1, wherein in formula (Ia) or (Ib) R ¹ is hydrogen, halogen, Ar, alkyl or alkyloxy; p = 1; R ² is hydrogen, alkyloxy or alkylthio; R ³ is naphthyl, phenyl or thienyl, each optionally substituted with 1 or 2 substituents selected from the group consisting of halogen or haloalkyl; q = 0,1,2 or 3; R ⁴ and R ^{5 are} independently hydrogen or alkyl or R ⁴ and R ⁵ together with N to which they are attached form a radical selected from the group consisting of imidazolyl, triazolyl, piperidinyl, piperazinyl and thiomorpholinyl; R ⁶ is hydrogen, alkyl or halogen; r is 1 and R ⁷ is hydrogen. Use according to claim 1, characterized in that the compound is selected from the group consisting of: 1- (6-bromo-2-methoxy-quinolin-3-yl) -2 (3,5-difluoro-phenyl) -4-dimethylamino-1-phenylbutan-2-ol; • 1- (6-bromo-2-methoxy-quinolin-3-yl) -4-dimethylamino-2-naphthalen-1-yl-1-phenyl-butan-2-ol; • 1- (6-bromo-2-methoxy-quinolin-3-yl) -2 (2,5-difluoro-phenyl) -4-dimethylamino-1-phenyl-butan-2-ol; • 1- (6-bromo-2-methoxy-quinolin-3-yl) -2 (2,3-difluoro-phenyl) -4-dimethylamino-1-phenyl-butan-2-ol; • 1- (6-bromo-2-methoxy-quinolin-3-yl) -410 dimethylamino-2- (2-fluoro-phenyl) -1-phenyl-butan-2-ol; • 1- (6-bromo-2-methoxy-quinolin-3-yl) -4-dimethylamino-2-naphthalen-1-yl-1-p-tolyl-butan-2-ol; 15 · 1- (6-Bromo-2-methoxy-quinolin-3-yl) -4-dimethylamino-2-naphthalen-1-yl-1-phenyl-butan-2-ol; • 1- (6-bromo-2-methoxy-quinolin-3-yl) -4-dimethylamino-2- (3-fluoro-phenyl) -1-phenyl-butan-2-ol, and • 1- (6-bromo- 2-Methoxy-quinolin-3-yl) -4-dimethylamino-2-phenyl-1-phenyl-butan-2-ol; a pharmaceutically acceptable acid or base addition salt thereof, an N-oxide thereof, a tautomeric form thereof or a stereochemically isomeric form thereof. Use according to claim 19, wherein the compound is selected from the group consisting of: • 1- (6-bromo-2-methoxy-quinolin-3-yl) -2 (2,3-difluoro-phenyl) -4-dimethylamino-1-phenyl-butan-2-ol; • 1- (6-Bromo-2-methoxy-quinolin-3-yl) -4-dimethylamino-2-naphthalen-1-yl-1-phenyl-butan-2-ol corresponding to 6-bromo-alpha- [2- (dimethylamino) ethyl] -2-methoxy-alpha-1-naphthalenylbeta-phenyl-3-quinolinethanol; or a pharmaceutically acceptable acid or base addition salt thereof, an N-oxide thereof, a tautomeric form thereof or a stereochemically isomeric form thereof. Use according to claim 1, wherein the compound is 6-bromo-alpha- [2- (dimethylamino) ethyl] -2-methoxy-alpha-1-naphthalenyl-beta-phenyl-3-quinolineethanol, a pharmaceutically acceptable acid addition compound thereof or a base salt, an N-oxide thereof, or a stereochemically isomeric form thereof. Use according to claim 21, wherein the compound is 6-bromo-alpha- [2- (dimethyl- Bl isoniazid versus rifampicin and pyrazinamide for prevention of tuberculosis in HIV-1 infection. Lancer 351; 786-792 (1998). 5. Mitchison, D. A. & Coates, A. R. 5 Predictive in vitro models of the sterilizing activity of antituberculosis drugs. Curr. Pharm. Des. 10, 3285-3295 (2004). 6. Karakousis P. C. et al. Dormancy phenotype displayed by extracellular 10 Mycobacterium Tuberculosis within artificial granulomas in mice. J Exp. Med. 200, 647-657 (2004). 7. Gomez, J. E. & McKinney, J. D. M. tuberculosis persistence, latency and drug 15 tolerance. Tuberculosis. (Edinb.) 84, 29-44 (2004). 8. Snewin, V. A. et al. Assessment of immunity to mycobacterial infection with luciferase reporter constructs. Infect. Immun. 67, 4586-4593 (1999). 20 9. Stover, C. K. et al. A small-molecule nitroimidazopyran drug candidate for the treatment of tuberculosis. Nature 405, 962-966 (2000). 10. Wayne, L. G. Synchronized replication of Mycobacterium tuberculosis. Infect. Immun. 17, 25 528-530 (1977). 11. Boon, C. & Dick, T. Mycobacterium bovis BCG response regulator essential for hypoxic dormancy. J. Bacterio !. 184, 6760-6767 (2002). 12. Hutter, B. & Dick, T. Up-regulation of 30 narX, encoding a purative fused nitrate reductase in anaerobic dormant Mycobacterium bovis BCG. FEMS Microbiol. Lett. 178, 63-69 (1999). 13. Andrew, P. W. & Roberts, I. S. Construction of a bioluminescent mycobacterium 35 and its use for assay of antimycobacterial agents. J. Clin. Microbiol. 31, 2251-2254 (1993). 14. Hickey, M. J. et al. Luciferase in vivo expression technology: use of recombinant mycobacterial reporter strains to evaluate 40 antimycobacterial activity in mice. Antimicrobial. Agents Chemoter. 40, 400-407, (1996). 15. Wayne, L. G. & Sramek, H. A. Metromidazole is bactericidal to dormant cells of mycobacterium tuberculosis. Antimicrobial. Agents Chemoter. 38, 2054-2058 (1994). 16. Wayne, L. G. Dormancy of Mycobacterium tuberculosis and latency of disease. Eur. J Clin. Microbiol. Infect. Dis. 13, 908-914 (1994). lamino) ethyl] -2-methoxy-alpha-1-naphthalenyl-beta-phenyl-3-quinolinetanol, or a pharmaceutically acceptable acid addition salt thereof. Use according to claim 21, wherein the compound is 6-bromo-alpha- [2- (dimethylamino) ethyl] -2-methoxy-alpha-1-naphthalenyl-beta-phenyl-3-quinolineethanol, or a stereochemically isomeric form thereof. Use according to claim 21, wherein the compound is 6-bromo-alpha- [2- (dimethylamino) ethyl] -2-methoxy-alpha-1-naphthalenyl-beta-phenyl-3-quinolineethanol, or an N-oxide thereof. Use according to claim 21, wherein the compound is (aS, pK) -6-bromo-alpha- [2 (dimethylamino) ethyl] -2-methoxy-alpha-1-naphthalenyl-beta-phenyl-3-quinolinethanol, or a a pharmaceutically acceptable acid addition salt. Use according to claim 25, wherein the compound is (aS, pK) -6-bromo-alpha- [2 (dimethylamino) ethyl] -2-methoxy-alpha-1-naphthalenyl-beta-phenyl-3-quinolinetanol. Application: 3 figures Literature 1. Corbett, E. L. et al. The growing burden of tuberculosis: global trends and interactions with the HIV epidemic. Atch. Intern. Med. 163, 10091021 (2003). 2. Dye, C., Scheele, S., Dolin, P., Pathania, V. & Raviglione, M. C. Consensus Statement. Global burden of tuberculosis: estimated incidence, prevalence, and mortality by country. WHO Global Surveillance and Monitoring Project. JAMA 282, 677-686 (1999). 3. Targeted tuberculin testing and treatment of latent tuberculosis infection. This official statement of the American Thoracic Socoety was adopted by the ATS Board of Directors, July 1999. This is a Joint Statement of the American Thoracic Society (ATS) and the Centers for Disease Control and Prevention (CDC). This statement was endorsed by the Council of the Infectous Disease 45 Society of America (IDSA), September 1999, and the sections of this statement. Am. J. Respir. Crit. Care Med. 161, S221-S247 (2000). 4. Halsey, N. A. et al. Randomised trial of * ^^^ * «». Ша «» ЬД, »дишШИ || 1 ^ УЙИТИ1ЙТ1 | Й1ППмТ liHiHWIirWT — rfrtli ·· теД .'- Д- |., 4Д« | 1 | амдмми,> ·! .. ^ - ίί τι. ». ,, i 1.,. 14 66437 Bl 17. Lalande, V. et al. Powerful bacterial activity of sparfloxacin (AT-4140) against Mycobacterium tuberculosis in mice. Antimicrobial. Agents Chemother. 37, 407-413 (1993). 18. Zhang, Y. Persistent and dormant tubercle 5 bacilli and latent tuberculosis. Front Biosci. 9,11361156 (2004). 19. Boon, C. & Dick, T. Mycobacterium bovis BCG response regulator essential for hypoxic dormancy. J. Bacteriol. 184, 6760-6767 (2002). Yu 20. Peh, H. L., Toh, A., Murugasu-Oei, B. & Dick, T. In vitro activities of mitomycin C against growing and hypoxic dormant tubercle bacilli. Antimicrobial. Agents Chemother. 45, 2403-2404 (2001). 15 21. Hutter, B & Dick, T. Increased alanine dehydrogenase activity during dormancy in Mycobacterium smegmatis. FEMS Microbiol. Lett. 167, 7-11 (1998). 22. Wayne, L. G. & Hayes, L. G. An in vitro model for a sequential study of the shiftdown of Mycobacterium tuberculosis through two stages of nonreplicant persistence. Infect. Immun. 64, 2062-2069 (1996). 23. Antia, R., Koella, J. C. & Perrot, V. Models of the intra-host dynamics of persistent mycobacterial infections. Proc. Biol. Sci. 263,257263 (1996).