The present invention relates to the new antituberculosis agents based on the nitro group- substituted phenyltetrazole, which are active against drug-susceptible and multidrug-resistant strains of mycobacteria.

BACKGROUND ART

Increasing occurrence of bacterial resistance to antibiotic therapy is one of the main reasons for the development of new antimicrobial active substances. Tuberculosis (TB) is considered as a major global health problem, especially due to the increasing emergence of its multidrug resistant (MDR-TB) and extensively drug resistant (XDR-TB) forms. TB is highly infectious disease caused by Mycobacterium tuberculosis {M.tb.), which is spread by air from the patients with pulmonary form of TB.

Tuberculosis (TB) caused by Mycobacterium tuberculosis complex (MTB), belongs for many years to the most widespread infectious diseases in the world. Nowadays, one third of the world's population is infected by M.tb. In 2013, approximately 9 million people fell ill with TB and 1.5 million people died due to TB (WHO - Global Tuberculosis Report 2014), ranking TB as the second greatest killer worldwide due to a single infectious agent.

Combinations of drugs with antimycobacterial effect are used for the treatment and they are administered usually for 6 - 9 months. Long period of the treatment leads to the adverse effects, non-compliance of patients and can be unavailable due to its price. Treatment regimen of drug susceptible TB consist of the administration of isoniazid, rifampicin, pyrazinamide a ethambutol for 2 month followed by the administration of isoniazid and rifampicin for 4-6 months. Resistant forms of TB must be treated by second and third line antiTB drugs, such as fluoroquinolones, amikacin, kanamycin, streptomycin, cycloserine, ethionamide, p-aminosalicylic acid, for the period of 18 - 24 months. Another serious complication is a combination of TB with HIV/ AIDS, which is usually lethal.

Therefore the development of the compounds that will be active against MDR-TB and latent forms of TB is essential. New antiTB drugs should possess new mechanism of action, which would overcome the possibility of cross resistances with common antiTB drugs. Some of new active molecules, which are in preclinical and clinical development, contain nitro group. This moiety is essential for their antimycobacterial activity, but several mechanism of action is connected with it. Nitroimidazole-oxazine PA-824 (Stover, C.K.; Warrener, P.; VanDevanter, D. R; Sherman, D.R.; Arain, T. .; Langhorne, M.H.; Anderson, S.W.; Towell, J.A.; Yuan, Y.; McMurray, D.N.; Kreiswirth, B.N.; Barry, C.E; Baker W.R. A small-molecule nitrimidazopyran drug candidate for the treatment of tuberculosis. Nature 2000, 405, 962-

nitro-dihydro-imidazooxazole OPC-67683 (Matsumoto, M.; Hashizume, H.; Tomishige, T.; Kawasaki, M.; Tsubouchi, H.; Sasaki, H.; Shimokawa, Y.; Komatsu, M. OPC-67683, a nitro- dihydro-imidazooxazole derivative with promising action against tuberculosis in vitro and in mice. PLOS Medicine 2006 3, 2131-2143),

PBT2169 dinitrobenzamides

and benzothiazinone PBTZ169 (Makarov, V.; Manina, G.; Mikusova, K.; Mollmann, U.; Ryabova, O.; Saint- Joanis, B.; Dhar, N.; Pasca, M.R.; Buroni, S.; Lucarelli, A.P.; Milano, A.; De Rossi, E.; Belanova, M.; Bobovska, A.; Dianiskova, P.; Kordulakova, J.; Sala, C; Fullrrm, E.; Schneder, P.; McKinney, J.D.; Brodin, P.; Christophe, T.; Waddell, S.; Butcher, P.; Albrethesen, J.; Rosenkrands, I.; Brosch, R.; Nandi, V.; Bharath, S.; Gaonkar, S.; Shandil, R.K.; Balasubramanian, V.; Balganesh, T.; Tyagi, S.; Grosset, J.; Riccardi, G.; Cole, S.T. Benzothiazinones kill Mycobacterium tuberculosis by blocking arabinan synthesis. Science 2009, 324, 801-804) can be mentioned as the expamples.

Another antiTB active nitro group-bearing compounds are dinitrobenzamides (Christophe, T.; Jackson, M.; Jeon, H.K.; Fenistein, D.; Contreras-Dominguez, M.; Kim, J.; Genovesio, A.; Carralot, J.P.; Ewarm, F.; Kim, E.H.; Lee, S.Y.; Kang, S.; Seo, M.S.; Park, E.J.; Skovierova, H.; Pham, H.; Riccardi, G.; Nam, J.Y.; Marsollier, L.; Kempf, M.; Joly-Guillou, MX.; Oh, T.; Shin, W.K.; No, Z.; Nehrbass, U.; Brosch, R.; Cole, S.T.; Brodin, P. High content screening identifies decaprenyl-phosphoribose 2'epimerase as a target for intracellular antimycobacterial inhibitors. PLOS Pathog 2009, 5, 1-10) and nrtroaromates derived from benzothiazinones (Tiwari, R.;M6llmann, U.; Sanghyun, Ch.; Franzblau, S.G.; Miller, P.A.; Miller MJ. Design and syntheses of anti-tuberculosis agents inspired by BTZ043 using a

nitroaromates R = CF ₃ , N0 ₂

SUMMARY OF THE INVENTION

New substituted phenyltetrazoles of general formula (I) show significant activity against Mycobacterium tuberculosis, against non-tuberculous mycobacteria and also against clinically isolated multidrug resistant strains of M. tuberculosis.

Compounds of general formula (I)

I

wherein

R is selected from the group consisting of: H, Q-Cn alkyl, phenyl-, or phenyl- substituted in positions 2, 3, 4 or 5, with one or more electron-acceptor groups comprising -N0 ₂ ,

-N ⁺ (Ci-C ₄ alkyl) ₃ , -CF ₃ , CC1 _3? -CN, -COOH, -COO(d-C ₄ alkyl), -COOAryl, -CHO,

-CO(C ₁ -C ₄ alkyl), -COAryl, -F, -CI, -Br, -I, and/or by one or more electron-donor groups comprising -N¾, -NH(d-C ₄ alkyl), -N(C C ₄ alkyl) ₂ , -OH, -0(d-C ₄ alkyl), -OAryl, -NHCOCHs, -NHCO(d-C ₄ alkyl), -NHCOaryl, -(C _r C ₄ alkyl), -phenyl or -naphtyl.

The term„electron-donor groups" as used herein, refers to substituents that increase electron density on the phenyl substituent R. Examples of electron- donor groups include especially -NH _2> -NHAlk, -NAlk ₂ , -OH, -OAlk, -OAr, -NHCOCH ₃ , -NHCOAlk, -NHCOAr, -Alk, -Ar, wherein Alk = alkyl, Ar = aryl, wherein aryl = phenyl or phenyl substituted in position 2, 3, 4, and 5 by one or more electron-acceptor groups and/or by one or more electron-donor groups, naphtyl or pyridyl.

The term„electron-acceptor groups" as used herein, refers to substituents that decrease electron density on the phenyl substituent R. Examples of electron-donor groups include - N0 ₂ , -NAlk ₃ , -CF ₃ , CC1 ₃ , -CN, -COOH, -COOAlk, -COOAr, -CHO, -COAlk, -COAr, -F _s -CI, -Br, -I, wherein Alk = alkyl, Ar = aryl, wherein aryl = phenyl or phenyl substituted in position 2, 3, 4, and 5 by one or more electron-acceptor groups and/or by one or more electron-donor groups, naphtyl or pyridyl. (Source: (a) John McMurry: Organic Chemistry, Sixth edition, 2004, Brooks/Cole, a Thomson Learning Company; b) L. G. Wade, Jr.: Organic Chemistry, Sixth edition, 2006, Pearson Prentice Hall Inc.; c) J. Clayden, N. Greeves, S. Warren, P. Wothers: Organic Chemistry, 2001, Oxford University Press).

Another aspect of the invention is the use of the above mentioned nitro group-substituted phenyltetrazole of general formula (I) according to the current mvention as antituberculosis agent.

Further aspect of the invention also relates to a pharmaceutical preparation containing the nitro group-substituted phenyltetrazole of general formula (I) as the active ingredient.

Compounds of general formula (I) can be obtained by routine methods of organic synthesis. The starting 5-(3,5-dinitrophenyl)-lH-tetrazole for synthesis of the compounds of general formula (I) were prepared by synthetic procedures as described in„Roh, J.; Artamonova, T. V.; Vavrova, K.; Koldobskii, G. I.; Hrabalek, A.-.Synthesis 2009,(13), 2175-2178." (Scheme

1)

Scheme 1. The final products of general formula (I) were prepared by Williamson synthesis from the 5- (3,5-dinitrophenyl)-lH-tetrazole with the appropriate alkylating agent (Scheme 2). The synthetic route is very simple and raw materials for them are easily accessible and also cheap.

Scheme 2.

The prepared compounds corresponding to general formula (I) were evaluated in Regional Institute of Public Health, Ostrava (Department for Diagnostic of Mycobacteria, Partyzanske namesti 7, 702 00 Ostrava) in in vitro conditions in Sula's semisynthetic liquid medium (SEVAC, Prague) and the minimum inhibitory concentrations (MIC) were determined. The antimycobacterial activity was tested against Czech National Collection strains Mycobacterium tuberculosis CNCTC My 331/88, M. avium CNCTC My 330/88, and M kansasii CNCTC My 235/80, and a clinical isolate M. kansasii 6509/96. Isoniazid (ΓΝΗ) was used as a standard in each assay. The results are shown in Table 2.

The most active compounds of general formula (I) were also evaluated for their activity against multidrug resistant strains of Mycobacterium tuberculosis (MDR strains). The strains are labelled as M. tuberculosis 234/2005, M. tuberculosis 9449/2007, M. tuberculosis 8666/2010, M. tuberculosis Praha 1, M. tuberculosis Praha 4 and M. tuberculosis Praha 131 M. tuberculosis 7357/1998. These strains were clinically isolated from patients and are deposited in Regional Institute of Public Health, Ostrava (Department for Diagnostic of Mycobacteria, Partyzanske namesti 7, 702 00 Ostrava). The sensitivity/resistance of the mentioned clinically isolated strains to common antituberculotics and antibiotics used are summarized in Table 3. The activity of compounds of general formula (I) was expressed as minimum inhibitory concentration (MIC). The results were obtained under the same conditions as described above. Isoniazid (INH) was used as a standard in each assay. The results are shown in Table 4. Substituent -CH -R on the heterocycle plays a crucial role in the antimycobacterial activity. When the substituent -CH ₂ -R is replace by hydrogen atom - i.e. in the case of 5-(3,5- dinitrophenyl)-lH-tetrazole - the antimycobacterial activity disappear.

Accordingly, the subject matter of the present invention comprises a highly antimycobacterially active low molecular weight tetrazole based compounds with a combination of a substituent -C¾-R in the position 2 and 3,5-dinitrophenyl moiety bound in the position 5 of tetrazole.

Examples:

The following examples describe how to prepare nitro group-substituted phenyltetrazoles of general formula (I)

(I) wherein R has the above mentioned meaning.

Example 1 : 5 -(3 ,5-dinitrophenyl)-2-methyl-2H-tetrazole (1)

Compound 1 was prepared according to Scheme 2. The starting 5-(3 ₅ 5-dinitrophenyl)-lH- tetrazole (0.2 g, 0.85 mmol ) was stirred with dimethyl sulfate (0.083 mL, 0.76 mmol) in the system CH2CI2 (10 mL) and water (10 mL) in the presence of tetrabutylammonium bromide (0.01 g; 0.032 mmol) and NaOH (0.038 g, 0.85 mmol) at room temperature for 50 hours. Upon completion, the reaction was diluted with 30 mL of CH ₂ C1 ₂ . The organic phase was washed with 5% Na ₂ C0 ₃ (3 x 20 mL) and 20% NaCl (1 x 20 mL). The organic phase was dried over Na ₂ S0 ₄ and evaporated. Compound 1 was isolated and purified by column chromatography (mobile phase: hexane/ethyl acetate 2.5:1).

The starting dimethyl sulfate is a commercially available compound. The starting 5-(3,5- dinitrophenyl)-lH-tetrazole was prepared according to published procedure shown in Scheme 1 (Roh, J.; Artamonova, T. V.; Vavrova, K.; Koldobskii, G. I.; Hrabalek, A.Synthesis 2009, (13), 2175-2178).

Example 2: 2-benzyl- 5 -(3 , 5 -dinitrophenyl)-2H-tetrazole (2)

2

Compound 2 was prepared according to Scheme 2. The starting 5-(3,5-dinitrophenyl)-lH- tetrazole (0.2 g ₅ 0.85 mmol ) was stirred with benzyl bromide (0.091 mL, 0.76 mmol) and triethylamine (0.12 mL, 0.85 mmol) in acetonitrile (10 mL) at 90 °C for 4 hours. Upon completion, the reaction mixture was evaporated and diluted with 30 mL of ethyl acetate. The organic phase was washed with 10% Na ₂ C0 ₃ (2 x 20 mL) and 20% NaCl (1 x 20 mL). The organic phase was dried over Na ₂ S0 ₄ and evaporated. Compound 2 was isolated and purified by column chromatography (mobile phase: hexane/ethyl acetate 6:1).

The starting benzyl bromide is a commercially available compound. The starting 5 -(3, 5- dinitrophenyl)-lH-tetrazole was prepared according to published procedure shown in Scheme 1 (Roh, J.; Artamonova, T. V.; Vavrova, K.; Koldobskii, G. I.; Hrabalek, K.:Synthesis 2009, (13), 2175-2178).

Example 3: 5 -(3 ,5 -dinitrophenyl)-2-(4-methylbenzyl) -2H-tetrazole (3)

Compound 3 was prepared according to Scheme 2. The starting 5-(3,5-dinitrophenyl)-lH- tetrazole (0.1 g, 0.42 mmol ) was stirred with 4-methylbenzyl bromide (0.050 mL, 0.38 mmol) and triethylamine (0.058 mL, 0.42 mmol) in acetonitrile (10 mL) at 90 °C for 5 hours. Upon completion, the reaction mixture was evaporated and diluted with 30 mL of ethyl acetate. The organic phase was washed with 10% Na ₂ C0 ₃ (2 x 20 mL) and 20% NaCl (1 x 20 mL). The organic phase was dried over Na ₂ S0 ₄ and evaporated. Compound 3 was isolated and purified by column chromatography (mobile phase: hexane/ethyl acetate 15:1).

The starting 4-methylbenzyl bromide is a commercially available compound. The starting 5- (3,5-dinitrophenyl)-lH-tetrazole was prepared according to published procedure shown in Scheme 1 (Roh, J.; Artamonova, T. V.; Vavrova, K.; Koldobskii, G. L; Hrabalek, A.:Synthesis 2009, (13), 2175-2178).

Numerous other compounds of general formula (I) (compounds 4 - 19) can be prepared using the above mentioned synthetic procedures.

Compound 4 was prepared using the above mentioned synthetic procedures from 5-(3,5- dinitrophenyl)-lH-tetrazole and 4-bromobenzyl chloride. The starting 5-(3,5-dinitrophenyl)- lH-tetrazole was prepared according to published procedure shown in Scheme 1. 4- Bromobenzyl chloride is a commercially available compound.

Compound 5 was prepared using the above mentioned synthetic procedures from 5-(3,5- dinitrophenyl)-lH-tetrazole and 3,5-dinitrobenzyl chloride. The starting 5-(3,5-dinitrophenyl)- lH-tetrazole was prepared according to published procedure shown in Scheme 1. 3,5- Dinitrobenzyl chloride is a commercially available compound.

Compound 6 was prepared using the above mentioned synthetic procedures from 5-(3,5- dinitrophenyl)-lH-tetrazole and 4-chlorobenzyl chloride. The starting 5-(3,5-dinitrophenyi)- lH-tetrazole was prepared according to published procedure shown in Scheme 1. 4- Chlorobenzyl chloride is a commercially available compound.

Compound 7 was prepared using the above mentioned synthetic procedures from 5-(3,5- dinitrophenyl)-lH-tetrazole and 4-nitrobenzyl chloride. The starting 5-(3,5-dinitrophenyl)- lH-tetrazole was prepared according to published procedure shown in Scheme 1. 4- Nitrobenzyl chloride is a commercially available compound.

Compound 8 was prepared using the above mentioned synthetic procedures from 5 -(3, 5- dinitrophenyl)-lH-tetrazole and 3,4-dichlorobenzyl chloride. The starting 5 -(3, 5- dinitrophenyl)-lH-tetrazole was prepared according to published procedure shown in Scheme 1. 3,4-Dichlorobenzyl chloride is a commercially available compound.

Compound 9 was prepared using the above mentioned synthetic procedures from 5-(3 _; 5- dinitrophenyl)-lH-tetrazole and 4-methoxybenzyl chloride. The starting 5-(3,5- dinitrophenyl)-lH-tetrazole was prepared according to published procedure shown in Scheme 1. 4-Methoxybenzyl chloride is a commercially available compound.

Compound 10 was prepared using the above mentioned synthetic procedures from 5-(3,5- dinitrophenyl)-lH-tetrazole and propyl bromide. The starting 5-(3,5-dinitrophenyl)-lH- tetrazole was prepared according to published procedure shown in Scheme 1. Propyl bromide is a commercially available compound.

Compound 11 was prepared using the above mentioned synthetic procedures from 5-(3,5- dinitrophenyl)-lH-tetrazole and dodecyl bromide. The starting 5-(3,5-dinitrophenyl)-lH- tetrazole was prepared according to published procedure shown in Scheme 1. Dodecyl bromide is a commercially available compound.

Compound 12 was prepared using the above mentioned synthetic procedures from 5-(3,5- dinitrophenyl)-lH-tetrazole and 4-fluorobenzyl chloride. The starting 5-(3,5-dinitrophenyl)- lH-tetrazole was prepared according to published procedure shown in Scheme 1. 4- Fluorobenzyl chloride is a commercially available compound.

Compound 13 was prepared using the above mentioned synthetic procedures from 5 -(3 ,5- dinitrophenyl)-lH-tetrazole and 3,4-difluorobenzyl chloride. The starting 5-(3,5- dinitrophenyl)-lH-tetrazole was prepared according to published procedure shown in Scheme 1 , 3,4-Difluorobenzyl chloride is a commercially available compound.

Compound 14 was prepared using the above mentioned synthetic procedures from 5-(3,5- dinitrophenyl)-lH-tetrazole and 3,5-difluorobenzyl chloride. The starting 5-(3,5- dinitrophenyl)-lH-tetrazole was prepared according to published procedure shown in Scheme 1. 3,5-Difluorobenzyl chloride is a commercially available compound.

Compound 15 was prepared using the above mentioned synthetic procedures from 5-(3,5- dinitrophenyl)-lH-tetrazole and 2-chloro-6-fluorobenzyl chloride. The starting 5-(3,5- dinitrophenyl)-lH-tetrazole was prepared according to published procedure shown in Scheme 1. 2-Chloro-6-fluorobenzyl chloride is a commercially available compound. Compound 16 was prepared using the above mentioned synthetic procedures from 5-(3,5- dinitrophenyl)-lH-tetrazole and 3-fluorobenzyl chloride. The starting 5-(3,5-dinitrophenyl)- lH-tetrazole was prepared according to published procedure shown in Scheme 1.

3-Fluorobenzyl chloride is a commercially available compound.

Compound 17 was prepared using the above mentioned synthetic procedures from 5-(3,5- dinitrophenyl)-lH-tetrazole and 3-bromobenzyl chloride. The starting 5-(3 _? 5-dinitrophenyl)- lH-tetrazole was prepared according to published procedure shown in Scheme 1.

3-Bromobenzyl chloride is a commercially available compound.

Compound 18 was prepared using the above mentioned synthetic procedures from 5-(3,5- dinitrophenyl)-lH-tetrazole and 3-methoxybenzyl chloride. The starting 5-(3,5- dinitrophenyl)-lH-tetrazole was prepared according to published procedure shown in Scheme 1. 3-Methoxybenzyl chloride is a commercially available compound.

Compound 19 was prepared using the above mentioned synthetic procedures from 5-(3,5- dinitrophenyl)-lH-tetrazole and 3-chlorobenzyl chloride. The starting 5-(3,5-dinitrophenyl)- lH-tetrazole was prepared according to published procedure shown in Scheme 1.

3-Chlorobenzyl chloride is a commercially available compound.

Table 1. Examples of compounds of general formula (I) (compounds 4 - 19)

2-(3-chlorobenzyl)-5-(3,5-dinitrophenyl)- 2H-tetrazole

Table 2. The minimum inhibitory concentration in vitro (expressed in μπιοΙ.Γ ¹ ) of compounds of general formula (I) (micromethod for determination of minimum inhibitory concentration in Sula's semisynthetic medium on plastic P-microplates; MICs determined after incubation at 37 °C for 14 and 21 days for M. tuberculosis and M. avium, for 7, 14 and 21 days for M. kansasii).

M. tuberculosis M. avium M. kansasii M. kansasii My 331/88 My 330/88 My 235/80 6509/96

1 32/32 62,5 / 125 62,5 / 125 / 125 62,5 / 62,5 / 62,5

2 0.5/1 8/8 1/2/2 1/2/2

3 0.125/0.125 8/16 0.5/1/1 0.25/0.5/0.5

4 0.25 / 0.25 4/8 0.25 / 1 / 1 0.25 / 1 / 1

5 1/1 4/4 1/2/2 2/4/4

6 0.25 / 0.25 8/8 0.25/1/2 0.5/1/2

7 1/1 4/4 0.5/1/2 2/4/4

8 0.125/0.125 4/8 0.25 / 0.5 / 1 0.25/0.5/0.5

9 0.03 / 0.03 4/8 0.125/0.125/0.25 0.06/0.06/0.125

10 8/16 32/62.5 16/32/32 16/32/32 11 1/2 8/16 4/8/8 2/4/4

12 0.25/0.5 8/8 1/2/2 1/2/4

13 0.25 / 0.5 4/8 1/2/4 1/2/2

14 0.25 / 0.5 8/16 1/2/4 1/2/4

15 0.5/0.5 4/8 0.5/1/1 0.5/1/1

16 1/1 8/16 1 /2/2 0.5/1/2

17 0.25 / 0.25 4/8 1/2/2 0.5/1/2

18 1/1 4/8 1/2/4 1/2/2

19 0.5 / 0.5 4/8 1/2/2 0.5/1/2

INH 0.5/1 >250 >250 4/4/4

Table 3. The minimum inhibitory concentration in vitro (expressed in μηιοΙ.Γ ) of selected antibiotics and antituberculotics (micromethod for determination of minimum inhibitory concentration in Sula's semisynthetic medium on plastic P-microplates after incubation for 14 and 21 days) for multidrug resistant strains of M. tuberculosis

R - strain resistant to mentioned antituberculosis drug

C - strain sensitive to mentioned antituberculosis drug

Table 4. The minimum inhibitory concentration in vitro (expressed in μηιοΙ.Γ ¹ ) of compounds by general formula (I) (micromethod for determination of minimum inhibitory concentration in Sula's semisynthetic medium on plastic P-microplates) after incubation at 37 °C for 14 and 21 days for multidrug resistant strains of tuberculosis

Tabulka 5. Melting points and NMR spectra of compounds of general formula (I).

Melting 1HNMR ^1J C NMR point [°C]

1 144-145 1H NMR (300 MHz, CDC1 ₃ ) δ ^li CNMR(75MHz,CDCI ₃ )6

9.30 (d,J= 2.1 Hz, 2H), 9.12 (t, 161.76, 149.07, 130.99, 126.55, J =2.1 Hz, 1H), 4.50 (s, 3H). 119.76,39.99.

2 113-114 1H NMR (300 MHz, CDC1 ₃ ) 6 ^li C NMR (75 MHz, CDC1 ₃ ) 6

9.29 (d,J=2.1 Hz,2H),9.10(t, 161.94, 148.99, 132.47, 131.01, J=2.1 Hz, 1H) _; 7.51- 7.38 (m, 129.38, 129.20, 128.61, 126.62, 5H), 5.87 (s, 2H). 119.74, 57.47.

3 109-110 1H NMR (500 MHz, Acetone) δ C NMR (126 MHz, Acetone) δ

9.18 (d, J = 2.1 Hz, 2H), 9.04 (t, 162.74, 150.09, 139.64, 131.62, J=2.1 Hz, 1H), 7.41 (d,J=8.1 131.54, 130.38, 129.49, 126.96, Hz,2H), 7.23 (d,J=8.1Hz, 120.55, 57.67,21.09.

2H), 6.00 (s,2H), 2.31 (s, 3H) 144-145 1H NMR (500 MHz, Acetone) δ ¹ C NMR (126 MiferAcetone 6- 9.18 (d,J= 2.1 Hz, 2H), 9.05 (t, 162.90, 150.13, 133.96, 132.91, J=2.1 Hz, 1H), 7.62 (d,J= 8.4 131.65, 131.45, 127.02, 123.46, Hz,2H), 7.51 (d,J=8.4Hz, 120.65, 57.09

2H), 6.08 (s, 2H).

136-138 Ή NMR (300 MHz, Acetone) δ C NMR (75 MHz, Acetone) δ

9.19 (d, J =2.1 Hz,2H), 9.06 (t, 163.14, 150.16, 149.75, 130.35 (2C), J=2.1 Hz, 1H), 8.98 (t,J=2.1 127.07 (2C), 120.83, 119.95, 56.06 Ηζ,ΙΗ), 8.89 (d,J=2.1Hz,

2H), 6.51 (s, 2H).

136-137 Ή NMR (500 MHz, Acetone) δ ^1J C NMR (126 MHz, Acetone) δ

9.19 (d, J =2.1 Hz, 2H), 9.05 (t, 162.90, 150.14, 135.30, 133.51, J=2.1Hz, 1H), 7.58 (d,J= 8.5 131.46, 131.39, 129.91, 127.02, Hz,2H), 7.46 (d,J=8.5Hz, 120.66, 57.04

2H), 6.09 (s, 2H)

168-169 Ή NMR (500 MHz, Acetone) δ ¹³ C NMR (126 MHz, Acetone) δ

9.20 (d,J=2.1 Hz,2H), 9.06 (t, 163.08, 150.16, 149.19, 141.57, J=2.1Hz, 1H) _S 8.30 (d,J= 8.7 131.35, 130.70, 127.06, 124.84, Hz, 2H), 7.82 (d,J= 8.7 Hz, 120.76, 56.82

2H), 6.30 (s, 2H)

162-163 1H NMR (500 MHz, DMSO) δ ^lj C NMR (126 MHz, DMSO) δ

9.03(d,J=2.1Hz, 2H), 8.94 (t, 161.72, 148.93, 134.60, 131.83, J= 2.1 Hz, 1H), 7.80 (d, J= 2.1 131.59, 131.29, 130.95, 129.55, Hz, 1H), 7.69 (d,J=8.3Hz, 129.22, 126.34, 120.15,55.27 1H),7.46 (dd,J=8.3,2.1 Hz,

1H), 6.13 (s, 2H)

104-105 1H NMR (500 MHz, Acetone) δ ¹³ C NMR (126 MHz, Acetone) 6

9.18 (d, J = 2.1 Hz, 2H), 9.04 (t, 162.74, 161.19, 150.14, 131.60, J=2.1 Hz, 1H), 7.49 (d,J= 8.7 131.16, 126.97, 126.48, 120.56, Hz,2H), 6.97(d,J=8.7Hz, 115.14, 57.50,55.62

2H), 5.98 (s, 2H), 3.79 (s, 3H) 103-104 Ή NMR (500 MHz, Acetone) δ ^Vi C NMR (126 MHz, Acetone) δ 9.21 (d,J=2.1 Hz, 2H), 9.06 (t, 162.47, 150.16, 131.74, 126.94, J=2.1 Hz, 1H), 4.81 (t,J=7.0 120.52,55.92, 23.40, 11.08 Hz, 2H), 2.19-2.07 (m,2H),

1.00(t,J=7.4Hz,3H)

68-69 'HNMR (500 MHz, Acetone) δ ^lj C NMR (126 MHz, Acetone) δ

9.21 (d, J= 2.1 Hz, 2H), 9.06 (t, 162.44, 150.17, 131.74, 126.92, J=2.1 Hz, 1H), 4.85 (t,J=7.0 120.52, 54.40, 32.59, 30.29, 30.18, Hz, 2H), 2.15- 2.07 (m, 2H), 30.05, 30.02, 29.95, 29.88, 29.54, 1.45 - 1.34 (m, 4H), 1.34 - 1.21 26.93, 23.28, 14.30.

(m, 14H), 0.88- 0.84 (m, 3H)

151-152 ¾ NMR (500 MHz, Acetone) S ⁱ³ C NMR (126 MHz, Acetone) δ

9.18(d,J=2.2Hz,2H), 9.05 (t, 163.84 (d,J= 245.9 Hz), 162.86, J - 2.2 Hz, 1H), 7.65 - 7.60 (m, 150.14, 131.94 (d,J= 8.6 Hz), 2H), 7.23- 7.16 (m, 2H), 6.08 131.49, 130.80 (d,J=3.2 Hz), (s,2H). 127.01, 120.64, 116.62 (d,J=22.0

Hz), 57.07.

213-214 1H MR (500 MHz, Acetone) δ ¹³ C NMR (126 MHz, Acetone) δ

9.19 (d,J = 2.1 Hz, 2H), 9.05 (t, 162.95, 151.29 (dd,J = 247.8, 12.2 J=2.1 Hz, 1H), 7.61- 7.54 (m, Hz), 150.94 (dd,J= 247.0, 12.5 Hz), 1H), 7.48- 7.35 (m, 2H), 6.11 150.14, 132.01 (dd, J = 6.1, 3.9 Hz), (s, 2H). 131.44, 127.05, 126.71 (dd,J=6.9,

3.7 Hz), 120.68, 119.07- 118.66 (m, 2C), 56.62 (d,J= 1.6 Hz).

122-123 1H NMR (500 MHz, Acetone) δ ⁱ³ C NMR (126 MHz, Acetone) δ

9.69 - 9.62 (m, 2H), 9.53 - 9.48 164.01 (dd,J= 248.2, 13.0 Hz),

163.05,150.15,138.65 (t,J=9.8 (m, 1H), 7.68-7.64 (m, 2H),

Hz), 131.43, 127.11, 120.71, 112.65 7.57 - 7.49 (m, 1H), 6.61 (s, 2H) (dd, J= 20.1, 6.4 Hz), 105.02 (t, J=

25.8 Hz), 56.60 (t,J= 2.1 Hz).

137-138 1H NMR (500 MHz, Acetone) δ ^U C NMR (126 MHz, acetone) δ

9.16 (d, J =2.1 Hz, 2H), 9.05 (t, 162.06 (d,J= 251.6 Hz), 161.81, J= 2.1 Hz, 1H), 7.62 - 7.53 (m, 149.28, 135.87 (d,J=4.5Hz), 1H), 7.46 - 7.42 (m, 1H), 7.36 - 132.38 (d,J= 10.0 Hz), 130.51, 7.28 (m, 1H), 6.23 (d,J=1.5 126.16, 125.95 (d,J=3.4 Hz), Hz, 2H). 119.81, 119.32 (d,J= 17.3 Hz),

114.82 (d, J= 22.2 Hz), 48.34 (d, J = 4.4 Hz).

145-148 1H NMR (500 MHz, Acetone) δ ^U C NMR (126 MHz, acetone) δ

9.20 (d,J=2.1 Hz,2H), 9.05 (t, 163.66 (d, J= 245.3 Hz), 162.93, J- 2.1 Hz, 1H), 7.53 - 7.45 (m, 150.14, 137.14 (d,J= 7.8 Hz), 1H), 7.38 - 7.35 (m, 1H), 7.34 - 131.85 (d,J= 8.3 Hz), 131.47, 7.30 (m, 1H), 7.20-7.15 (m, 127.06, 125.45 (d,J=3.1 Hz), 1H), 6.12 (s, 2H) 120.66,116.58 (d, J= 21.1 Hz),

116.32 (d,J= 22.7 Hz), 57.10

128-129 1H NMR (500 MHz, Acetone) δ ^U C NMR (126 MHz, Acetone) δ

9.19(d,J=2.1Hz,2H), 9.06 (t, 162.94, 150.13, 137.05, 132.82, J= 2.1 Hz, 1H), 7.77 - 7.73 (m, 132.44, 131.83, 131.44, 128.53, 1H), 7.61- 7.57 (m, 1H), 7.57- 127.06, 123.13, 120.66, 56.98. 7.52 (m, 1H), 7.42-7.38 (m,

1H), 6.11 (s, 2H).

92-94 1H NMR (500 MHz, acetone) 6 ^1J C NMR (126 MHz, acetone) δ

9.19 (d, J = 2.1 Hz, 2H), 9.05 (t, 162.81, 161.01, 150.12, 135.98, J = 2.1 Hz, 1H),7.33 (t, J = 7.9 131.52, 130.95, 127.01, 121.41, Ηζ,ΙΗ), 7.10-7.05 (m,2H), 120.59, 115.15, 115.01,57.74, 6.97-6.93 (m, 1H), 6.03 (s, 55.60.

2H),3.79 (s, 3H).

140-143 1H NMR (500 MHz, acetone) δ C NMR (126 MHz, acetone) δ

9.19 (d, J- 2.1 Hz, 2H), 9.05 (t, 162.94, 150.13, 136.82, 135.06, J =2.1 Hz, 1H), 7.60- 7.59 (m, 131.58, 131.44, 129.84, 129.49, 1H), 7.51- 7.42 (m, 3H), 6.11 128.09, 127.06, 120.66, 57.03 (s,2H) Tabulka 6. Elemental analysis of compounds of general formula (I). calculated found

1 C, 38.41; H, 2.42; N, 33.59; C, 38.15; H, 2.33; N, 33.78;

2 C, 51.54; H, 3.09; N, 25.76; C, 51.81; H, 3.26; N, 25.48;

3 C, 52.94; H ₅ 3.55; N, 24.70; C, 53.17; H, 3.19; N, 24.50;

4 C, 41.50; H, 2.24; N, 20.74; C, 41.39; H, 2.07; N, 21.03;

5 C, 40.40; H, 1.94; N, 26.92; C, 40.08; H, 2.14; N, 27.06;

6 C, 46.62; H, 2.52; N, 23.30; C, 46.34; H, 2.41; N, 23.07;

7 C, 45.29; H, 2.44; N, 26.41; C, 44.98; H, 2.27; N, 26.17;

8 C, 42.55; H, 2.04; N, 21.27; C, 42.19; H, 1.87; N, 20.92;

9 C _f 50.57; H, 3.39; N, 23.59; C, 50.35; H, 3.57; N, 23.48;

10 C, 43.17; H, 3.62; N, 30.21; C, 42.95; H, 3.32; N, 30.01;

11 C, 56.42; H, 6.98; N, 20.78; C, 56.80; H, 7.21; N, 21.59;

12 C, 48.84; H, 2.64; N, 24.41; C. 48.75; H ₅ 2.70; N ₅ 24.40;

13 C, 46.42; H, 2.23; N ₅ 23.20; C, 46.30; H, 1.92; N, 23.10;

14 C, 46.42; H, 2.23; N, 23.20; C, 46.78; H, 2.32; N, 23.07;

15 C, 44.40; H, 2.13; N, 22.19; C, 44.52; H, 2.19; N, 22.15;

16 C, 48.84; H ₅ 2.64; N, 24.41; C, 48.97; H, 2.80; N, 24.49;

17 C, 41.50; H, 2.24; N, 20.74; C, 41.29; H, 2.47; N, 21.02;

18 C, 50.57; H, 3.39; N, 23.59; C, 50.51; H, 3.52; N, 23.93;

19 C, 46.62; H, 2.52; N, 23.30; C, 46.92; H, 2.29; N ₅ 23.15;

Examples of the pharmaceutical compositions - tablets

In manufacture of solid dosage forms, the technology common in the given art is used, i.e., dry or wet granulation, which is well-known to a person skilled in the art. There are used routine and well proven excipients and suitable additives that give to the dosage form the desired physical characteristics.

Examples for dry granulation:

Example 1 ( ^" content of active substance 100 rug)

Active ingredient of general formula (I) 1 100.0 mg

Cellulose, microcrystaline 75.0 mg

Sodium carboxymethylstarch 3.5 mg

Magnesium Stearate 0.5 mg

Silicon Dioxide, Colloidal 0.5 mg

Example 2 (content of active substance 200 mg ^" )

Active ingredient of general formula (I) 13 200.0 mg

Cellulo se, microcrystaline 95.0 mg

Sodium carboxymethylstarch 7.0 mg

Magnesium Stearate 1.0 mg

Silicon Dioxide, Colloidal 1.0 mg

Example 3 (content of active substance 300 mg)

Active ingredient of general formula (I) 9 300.0 mg

Cellulose, microcrystaline 115.0 mg

Sodium carboxymethylstarch 10.5 mg

Magnesium Stearate 1.5 mg

Silicon Dioxide, Colloidal 1.5 mg

Example 4 (content of active substance 400 mg )

Active ingredient of general formula (I) 5 400.0 mg

Cellulose, microcrystaline 130.0 mg

Sodium carboxymethylstarch 14.5 mg Magnesium Stearate 2.0 mg Silicon Dioxide, Colloidal 2.0 mg

Example 5 (content of active substance 500 mg

Active ingredient of general formula (I) 2 500.0 mg

Cellulose, microcrystaline 140.0 mg

S odium carboxymethy lstarch 17.5 mg

Magnesium Stearate 2.5 mg

Silicon Dioxide, Colloidal 2.5 mg

The active ingredient is mixed together with other individual excipients and the obtained mixture is compressed by regular manner using a conventional tablet machine.

Examples for wet granulation

Example 6 (content of active substance 100 mg

Active ingredient of general formula (I) 19 100.0 mg

Potato starch 48.0 mg

Lactose 27.0 mg

Povidone 3.0 mg

Sodium carboxymethylstarch 4.0 mg

Magnesium Stearate 0.2 mg

Talc 1.8 mg

Example 7 (content of active substance 200 mg

Active ingredient of general formula (I) 6 200.0 mg

Potato starch 60.8 mg

Lactose 34.2 mg

Povidone 6.0 mg

Sodium carboxymethylstarch 8.0 mg

Magnesium Stearate 0.4 mg

Talc 3.6 mg

Example 8 (content of active substance 300 mg)

Active ingredient of general formula (I) 7 300.0 mg 73.6 mg

41.4 mg

9.0 mg

12.0 mg

0.6 mg

5.4 mg

Example 9 (content of active substance 400 mg)

Active ingredient of general formula (I) 3 400.0 mg

Potato starch 82.3 mg

Lactose 46.8 mg

Povidone 12.0 mg

S odium carboxymethylstarch 16.0 mg

Magnesium Stearate 0.8 mg

Talc 7.2 mg

Example 10 (content of active substance 500 mg)

Active ingredient of general formula (I) 1 500.0 mg

Potato starch 96.0 mg

Lactose 54.0 mg

Povidone 15.0 mg

Sodium carboxymethylstarch 20.0 mg

Magnesium Stearate 1.0 mg

Talc 9.0 mg

The therapeutically effective compound is mixed with lactose, potato starch and this mixture is granulated with povidone. The dried granulate is mixed then with sodium carboxymethylstarch, magnesium stearate, and talc. The obtained mixture is compressed by regular manner using a conventional tablet machine.