RU2519218C2 - Selective anti-tuberculosis agents, representing 3-aminosubstituted 6-(3,5-dimethylpyrazol-1-yl)-1,2,4,5-tetrazines - Google Patents

Abstract

FIELD: chemistry.

SUBSTANCE: invention represents 3-aminosubstituted 6-(3,5-dimethylpyrazol-1-yl)-1,2,4,5-tetrazines, which applied as anti-tuberculosis medications make it possible to increase activity and specificity of antimicrobacterial action, extend its spectrum (impact on atypical strains of mycobacteria) as well as reduce toxicity in comparison with analogues.

EFFECT: toxicity of claimed compounds is 10-20 times lower than toxicity of anti-tuberculosis medications applied in medicine.

5 tbl, 16 ex

Description

The invention relates to asymmetric 3,6-disubstituted 1,2,4,5-tetrazines containing a 3,5-dimethylpyrazolyl group at position 6 of the tetrazine ring, and at position 3, an aromatic or aliphatic amine residue. These compounds can be used as anti-tuberculosis agents, primarily for the treatment of tuberculosis patients.

The structure and method for producing compounds claimed as selective anti-tuberculosis agents are described [ZhOKh, 1999, 35, p.1392; Abstracts of Second International Symposium of Molecular Design and Synthesis of Supramolecular Architectures, Kazan, 2002, p. 154], but their activity against tuberculosis mycobacteria is not known and is not described.

An analogue of the above substances in structure is methyl 2-N- [6- (3,5-dimethylpyrazol-1-yl) -1,2,4,5-tetrazin-3-yl] -aminopropionic acid of the formula (I), which exhibits a noticeable tuberculostatic activity against the mycobacterial strain H37Rv, however, other representatives of this series of compounds are characterized by the opposite effect - increased growth of colonies of mycobacteria in their presence, associated with the release of amino acids [Chemical Pharm. Journal, 39, No. 1, 2005, pp. 10-12].

The intended analogue, isoniazid of formula II, is a selective anti-tuberculosis drug, however, the disadvantages of this drug are its high toxicity and various side effects when used. Isoniazid causes allergies, negatively affects the function of the gastrointestinal tract, and has a side effect on the nervous system [M.D. Mashkovsky. Medicines, 15th ed., Revised., Rev. and add. -M .: RIA "New Wave": publisher Umerenkov, 2007, p.858 and Pharmacology and Toxicology, 1987, 50, (4), p.87]. A wide spread of isoniazid-resistant clinical strains of mycobacteria has been noted [Antibiotics and chemotherapy, 2002, 47, No. 6, p. 28-30].

Methods for preparing compounds A are described. Substituted 3- (NR ₁ -amino) -6- (3,5-dimethylpyrazol-1-yl) -1,2,4,5-tetrazines of the general formula A are prepared by reacting 3,6-bis (3,5- in acetonitrile) dimethylpyrazol-1-yl) -1,2,4,5-tetrazine with the corresponding substituted amine [ZhOKH, 2006, 42, No. 3, pp. 772-780].

The objective of the invention is the use of known substances as antimycobacterial agents with increased specificity while expanding the spectrum of antimycobacterial action.

The problem is solved in that asymmetric 3,6-disubstituted 1,2,4,5-tetrazines containing in the position 3 the residue of a substituted amine - 3-amino-substituted 6- (3,5-dimethylpyrazole-1, are used as anti-tuberculosis agents -yl) -1,2,4,5-tetrazines of the general formula A

where R ₁ = amino group, C ₁ -C ₁₂ -alkyl, 3-hydroxy-5-hydroxypentyl, propin-2-yl, ethylindolyl, adamantyl, aryl selected from optionally substituted phenyl, heteryl selected from indol-3-yl, quinolin-3-yl, pyridyl, wherein substituted phenyl has a substituent selected from the group consisting of methyl, bromo, hydroxy, and pyridyl has substituents selected from the group of chloro, methyl.

Unlike analogues in structure and purpose, these compounds are 3,6-disubstituted 1,2,4,5-tetrazines containing amino-derived fragments, as well as additional heteroaromatic residues, the presence of tuberculostatic activity of which was not obvious.

The structural features of the claimed compounds lead to an increase in the specificity of the antimycobacterial action and to an expansion of the spectrum of action — an increase in the activity of these compounds in relation to typical and atypical strains of mycobacteria (Tables 1, 3). Due to their specificity for mycobacteria, they do not inhibit the normal microflora of the body, they are practically non-toxic (Tables 2, 4, 5). The therapeutic effect of 3- (4-bromophenylamino) -6- (3,5-dimethylpyrazol-1-yl) -1,2,4,5-tetrazine (Example 9) was revealed in the treatment of experimental tuberculosis of white mice.

The study of the antimycobacterial activity of the claimed compounds in in vitro experiments is carried out bacteriologically, by the method of vertical diffusion on a solid nutrient medium of Livenstein - Jensen or "New". In a test tube with a seeded test microbe along the free edge, 0.3 ml of each dilution is instilled into the bottom. The tubes are placed in a thermostat in an upright position and incubated at 37 ° C, the results are taken into account for 10-12 days.

The antibacterial activity of the claimed compounds is studied by sowing drug solutions in wells. A test microbe is added to a Petri dish with simple nutrient agar. 0.02 ml of the drug in the required concentrations are added to the well located in the middle of the Petri dish. The results are taken into account after 24-hour incubation in a thermostat at 37 ° C, by determining the zone of growth inhibition of microorganisms at the edges of the well.

The study of the cytotoxicity of the claimed compounds is carried out on cell culture MT-4 (transplantable human cell line T lymphocytes). The toxicity of various concentrations of compounds is determined by the cell viability relative to the control using a spectrophotometer at a wavelength of 540 nm, dose-dependent curves are constructed from the obtained data and the concentrations are determined that reduce cell viability by 50% compared to the control (CD ₅₀ ).

The study of acute toxicity of the claimed compounds is carried out according to standard methods on white mice. Animals receive the drug per os in 10% starch solution. The study of the therapeutic effect of the compound (example 9) is carried out on white laboratory mice weighing 18-23 g, infected intravenously, in the tail vein at a dose of 0.5 mg of a two-week-old culture (strain H37R.V) in 0.5 ml of physiological saline. Treatment begins one week after infection and is carried out daily for 8 weeks. The studied drug, including the control isoniazid, is given to mice at a dose of 10 mg / kg of body weight.

The optimal ratio of tuberculostatic activity - toxicity has 3- (2- (1H-indol-3-yl) ethylamino) -6- (3,5-dimethylpyrazol-1-yl) -1,2,4,5-tetrazine. This compound has the lowest toxicity in the studied series. According to the data of intragastric administration, according to GOST 12.1.007-76 and the OECD classification project, it belongs to the IV class of low toxic and low hazard substances (DL ₅₀ > 5000.0 mg / kg). According to the data of subcutaneous administration, this drug is on the border between the III and IV hazard class (DL ₅₀ = 1949.8 (1183.09 - 3213.43) mg / kg (DL ₅₀ for compounds of hazard class IV with subcutaneous administration should exceed 2000.0 mg / kg).

The purity of the compounds obtained is controlled by TLC, R _f values are determined using Sorbfil plates (Silica gel STX-1A, UV) (Russia) or Silufol-UV (Czechoslovakia). The structure of the obtained compounds is confirmed by elemental analysis (C, H, N, O — Karlo Erba analyzer). IR spectra were recorded on a PerkinElmer Spectrum One Fourier transform IR spectrometer using a Diffuse Reflectance Sampling Accessory (DRA). ¹ H NMR spectra were recorded in DMSO-d ₆ on a Bruker-400 instrument with with a working frequency of 400 MHz and TMS as an internal standard. Melting points are determined on a Boetius heating table. For microwave irradiation of the studied reactions, a ProLabo microwave chemical reactor (frequency 2.45 Hz, radiation power 200 W) is used.

Examples of specific performance.

Example 1

3-Hydrazino-6- (3,5-dimethylpyrazol-1-yl) -1,2,4,5-tetrazine

(The compound of formula A, where R ₁ = amino group)

1.0 mmol of 3,6-bis- (3,5-dimethylpyrazol-1-yl) -1,2,4,5-tetrazine is suspended in 10 ml of acetonitrile and 1.1 mmol of hydrazine hydrate is added with stirring.

The reaction mixture was placed in a porcelain dish, the solvent was evaporated, the residue was recrystallized from aliphatic alcohol, and dried in air. Yield 91%, mp. 153-154 ° C (from CH ₃ CN). C ₇ H ₁₀ N ₈ . Calculated, (%): C, 40.77; H, 4.89; N, 54.34. Found, (%): C, 40.77; H, 4.98; N, 54.24. ¹ H NMR (CDCl ₃ ), δ): 2.37 (s, 3H, 3 ^Pz —CH ₃ ); 2.58 (s, 3H, 5 ^Pz -CH ₃ ); 6.13 (s, 1H, 4 ^Pz- CH); 4.18 (br s, 2H, NH ₂ ); 7.16 (br s, 1H, NH).

Example 2

3-Methylamino-6- (3,5-dimethylpyrazol-1-yl) -1,2,4,5-tetrazine

(The compound of formula A, where R ₁ = methyl)

Obtain analogously to example 1 of 1.0 mmol of 3,6-bis- (3,5-dimethylpyrazol-1-yl) -1,2,4,5-tetrazine and 1.1 mmol of methylamine.

Yield 90%. Mp 194-195 ° C. Calculated, (%): C, 46.83; H, 5.37; N, 47.80. C ₈ H ₁₁ N ₇ .

Found, (%): C, 47.16; H, 5.32; N, 48.13. ¹ H NMR Spectrum (CDCl ₃ , δ, ppm, J / Hz): 6.18 (s, 1H, 4Pz-CH); 8.71 (m, 1H, -NH-CH3), 2.23, 2.38 (both s, 6H, 2CH ₃ Pz); 3.01 (d, 3H, J = 4.9).

Example 3

3-Undecylamino-6- (3,5-dimethylpyrazol-1-yl) -1,2,4,5-tetrazine

(The compound of formula A, where R ₁ = undecyl)

Obtain analogously to example 1 of 1.0 mmol of 3,6-bis- (3,5-dimethylpyrazol-1-yl) -1,2,4,5-tetrazine and 1.1 mmol of undecylamine.

Yield (%): 91.4. Mp 57-58 ° C. Calculated, (%): C 62.57; H 9.05; N, 28.38. C ₁₈ H ₃₁ N ₇ . Found, (%): C 62.63; H 9.20; N, 28.59. NMR spectrum ^l H (CDCl ₃ , δ, ppm, J / Hz): 6.10 (s, 1H, 4 ^Pz -CH); 5.80 (t, 1H, -NH-CH ₂ ^- ), 2.56, 2.35 (both s, 6H, 2CH ₃ ^Pz ); 3.62 (q, 2H, -NH-CH ₂ ^- -), 1.76-1.69 (m, 2H, -NH- (CH ₂ ) ₉ CH ₂ CH ₃ ); 1.42-1.27 (m, 16H, -NH-CH ₂ - (CH ₂ ) ₈ -C ₂ H ₅ ); 0.88 (t, 3H, -NH- (CH ₂ ) ₁₀ CH ₃ J = 7.0 Hz).

Example 4

3-dodecyl-6- (3,5-dimethylpyrazol-1-yl) -1,2,4,5-tetrazine

(The compound of formula A, where R ₁ = dodecyl)

Obtain analogously to example 1 of 1.0 mmol of 3,6-bis- (3,5-dimethylpyrazol-1-yl) -1,2,4,5-tetrazine and 1.1 mmol of dodecylamine.

Yield 94%. Mp 71-72 ° C. Calculated, (%): C 63.47; H 9.25; N, 27.27. C ₁₉ H ₃₃ N ₇ . Found, (%): C 63.45; H 9.13; N, 27.32. ¹ H NMR Spectrum (CDCl ₃ , δ, ppm, J / Hz): 6.10 (s, 1H, 4 ^Pz —CH); 5.79 (t, 1H, -NH-CH ₂ ^- ), 2.56, 2.35 (both s, 6H, 2CH ₃ ^Pz ); 3.62 (q, 2H, -NH-CH ₂ ^- ), 1.76-1.69 (m, 2H, -NH- (CH ₂ ) ₁₀ CH ₂ CH ₃ ); 1.43-1.26 (m, 18H, -NH-CH ₂ - (CH ₂ ) ₉ -C ₂ H ₅ ); 0.88 (t, 3H, -NH- (CH ₂ ) ₁₁ CH _3. J = 7.0 Hz).

Example 5

2- {2- [6- (3,5-Dimethylpyrazol-1-yl) -1,2,4,5-tetrazin-3-ylamino] ethoxy} ethanol

(The compound of formula A, where R ₁ = 3-hydroxy-5-hydroxypentyl)

Obtain analogously to example 1 of 1.0 mmol of 3,6-bis- (3,5-dimethylpyrazol-1-yl) -1,2,4,5-tetrazine and 1.1 mmol of 3-hydroxy-5-hydroxy-heptylamine.

Yield 95%. Mp 88-89 ° C. Calculated, (%): C 47.30; H 6.14; N 35.11. C ₁₁ H ₁₇ N ₇ O ₂ . Found, (%): C 47.27; H 6.22; N 34.91. ¹ H NMR Spectrum (DMCO, δ, ppm, J / Hz): 8.83 (br s, 1H, -NH-); 6.19 (s, 1H, 4 ^Pz- CH); 4.62 (br s, 1H, -OH), 2.38, 2.22 (both s, 6H, 2CH ₃ ^Pz ); 3.67-3.65; 3.51-3.46 (both m, 8H, - (CH ₂ ) ₂ -O- (CH ₂ ) ₂ ).

Example 6

3-N (t-Butylamino) -6- (3,5-dimethylpyrazol-1-yl) -1,2,4,5-tetrazine

(The compound of formula A, where R ₁ = t-butyl)

Obtain analogously to example 1 of 1.0 mmol of 3,6-bis- (3,5-dimethylpyrazol-1-yl) -1,2,4,5-tetrazine and 1.1 mmol of t-butylamine.

Yield 93%. Mp 127 ° C. Calculated,%: C 53.42, H 6.93, N 39.65. C ₁₁ H ₁₇ N ₇ . Found, (%): C 53.18, H 6.50, N 39.83. ¹ H NMR spectrum (400 MHz, CDCl ₃ , δ, ppm, J / Hz): 6.09 (s, 1H at C (4) pyrazolyl), 5.85 (br s, 1H, NH), 2.57, 2.35 1.56 (all s, 15H, 5CH ₃ ).

Example 7

3- (Adamantan-1-ylamino) -6- (3,5-dimethylpyrazol-1-yl) -1,2,4,5-tetrazine

(The compound of formula A, where R ₁ = adamantyl)

Obtain analogously to example 1 of 1.0 mmol of 3,6-bis- (3,5-dimethylpyrazol-1-yl) -1,2,4,5-tetrazine and 1.1 mmol of adamantylamine.

Yield 92%. Mp 162-164 ° C. Calculated, (%): C 62.77; H 7.08; N 30.15. C ₁₇ H ₂₄ N ₈ . Found, (%): C 62.69; H 7.12; N 30.21. ¹ H NMR (CDCl ₃ ), δ): 8.48 (br s, 1H, NH); 6.17 (s, 1H, 4 ^Pz- CH); 2.05-2.08 (m, 9H, adamantyl), 2.21, 2.40 (both s, 6H, 2CH ₃ ).

Example 8

3-N- (2-Hydroxyphenylamino) -6- (3,5-dimethylpyrazol-1-yl) -1,2,4,5-tetrazine

(The compound of formula A, where R ₁ = 2-hydroxyphenyl)

Obtain analogously to example 1 of 1.0 mmol of 3,6-bis- (3,5-dimethylpyrazol-1-yl) -1,2,4,5-tetrazine and 1.1 mmol of 2-hydroxyaminobenzene.

Yield 94%. Mp 218-218.5 ° C. Calculated, (%): C 55.12, H 4.59, N 34.63. C ₁₃ H ₁₃ N ₇ O. Found, (%): C 55.23, H 4.50, N 34.43. ¹ H NMR spectrum (400 MHz, CDCl ₃ , δ, ppm, J / Hz): 8.16-8.12, 7.07-6.95 (both m, 5H, OH, Ar) 6.14 (s, 1H at C (4) pyrazolyl), 2.62, 2.38 (both s, 6H, 2CH ₃ at C (3) and C (5) pyrazolyl).

Example 9

3- (4-Bromophenylamino) -6- (3,5-dimethylpyrazol-1-yl) -1,2,4,5-tetrazine

(The compound of formula A, where R ₁ = 4-bromophenyl)

Obtain analogously to example 1 from 1.0 mmol of 3,6-bis- (3,5-dimethylpyrazol-1-yl) -1,2,4,5-tetrazine and 1.1 mmol of 4-bromo-3-aminobenzene.

Yield 95.3%. Mp 215-216 ° C. Calculated, (%): C 45.09; H 3.47; N, 28.56. d ₃ H ₁₂ BrN ₇ . Found, (%): C 45.21; H 3.56; N, 28.58. ¹ H NMR Spectrum (CDC13, δ, ppm, J / Hz): 6.23 (s, 1H, 4Pz-CH); 11.14 (s, 1H, -NH-Ar), 2.25, 2.47 (both s, 6H, 2CH ₃ Pz); 7.59, 7.74 (both d, 2H each, J = 6.8).

Example 10

3- (Indol-3-yl-ethylamino) -6- (3,5-dimethylpyrazol-1-yl) -1,2,4,5-tetrazine

(The compound of formula A, where R ₁ = 2- (indol-3-yl) ethyl)

Obtain analogously to example 1 of 1.0 mmol of 3,6-bis- (3,5-dimethylpyrazol-1-yl) -1,2,4,5-tetrazine and 1.1 mmol of indol-3-yl-ethylamine.

Yield 92%. Mp 197-198 ° C. Calculated, (%): C 61.09, H 5.67, N 33.64. C ₁₇ H ₁₇ N ₈ . Found, (%): C 61.08, H 5.39, N 33.53. ¹ H NMR Spectrum (400 MHz; CDCl ₃ ; δ, ppm): 8.93 (t, 1H, NH); 6.18 (s, 1H at C (4) pyrazolyl); 3.07 (t, 2H, CH ₂ ); 3.75 (q, 2H, CH ₂ ); 2.23, 2.38 (both s, 6H, 2CH ₃ ); 6.97-7.02, 7.05-7.09 (both m, 2H, 2CH); 7.25, 7.35, 7.59 (all d, 3H, 3 CH); 10.86 (s, 1H, NH).

Example 11

3- (6-Methylpyridin-2-yl-amino) -6- (3,5-dimethylpyrazol-1-yl) -1,2,4,5-tetrazine

(The compound of formula A, where R ₁ = 6-methylpyridin-2-yl)

Obtain analogously to example 1 of 1.0 mmol of 3,6-bis- (3,5-dimethylpyrazol-1-yl) -1,2,4,5-tetrazine and 1.1 mmol of 2-amino-6-methylpyridine.

Yield 93%. Mp 164-165 ° C. Calculated, (%): C 55.32, H 4.96, N 39.72. C ₁₃ H ₁₄ N ₈ . Found, (%): C 55.35, H 4.98, N 39.69. ¹ H NMR Spectrum (400 MHz; CDCl ₃ ; δ, ppm): 11.21 (br s, ¹ H, NH); 6.15 (s, 1H, 4 ^Pz- CH); 7.63, (q, 1H, pyridino); 7.96 (d, 1H, pyridino); 8.20 (t, 1H, pyridino), 2.27, 2.33, 2.52 (all s, 9H, 3CH ₃ ).

Example 12

3- (Pyridin-2-ylamino) -6- (3,5-dimethylpyrazol-1-yl) -1,2,4,5-tetrazine

(The compound of formula A, where R ₁ = pyridin-2-yl)

Obtain analogously to example 1 of 1.0 mmol of 3,6-bis- (3,5-dimethylpyrazol-1-yl) -1,2,4,5-tetrazine and 1.1 mmol of 2-aminopyridine.

Yield 90%. Mp 248 ° C. Calculated, (%): C 53.73, H 4.48, N 41.79. C ₁₂ H ₁₂ N ₈ . Found, (%): C 53.79, H 4.51, N 41.56. ¹ H NMR Spectrum (400 MHz; CDCl ₃ ; δ, ppm): 11.35 (br s, ¹ H, NH); 6.20 (s, 1H, 4 ^Pz- CH); 7.10-7.14, 7.83-7.86, 8.02-8.04, 8.37-8.39 (all m, 4H, pyridino) 2.27, 2.48 (both s, 6H, 2CH ₃ ).

Example 13

3- (4-methylpyridin-2-ylamino) -6- (3,5-dimethylpyrazol-1-yl) -1,2,4,5-tetrazine

(The compound of formula A, where R ₁ = 4-methylpyridin-2-yl)

Obtain analogously to example 1 of 1.0 mmol of 3,6-bis- (3,5-dimethylpyrazol-1-yl) -1,2,4,5-tetrazine and 1.1 mmol of 2-amino-4-methylpyridine.

Yield 92%. Mp 245 ° C. Calculated, (%): C 55.32, H 4.96, N 39.72. C ₁₃ H ₁₄ N ₈ . Found, (%): C 55.47, H 4.83, N 40.00. ¹ H NMR Spectrum (400 MHz; CDCl ₃ ; δ, ppm): 11.28 (br s, ¹ H, NH); 6.24 (s, 1H, 4 ^Pz -CH); 6.98-7.00, 7.83-7.85, 8.25-8.26 (all m, 3H, pyridino) 2.24, 2.36, 2.49 (all s, 9H, 3CH ₃ ).

Example 14

3- (5-chloropyridin-2-ylamino -) - 6- (3,5-dimethylpyrazol-1-yl) -1,2,4,5-tetrazine

(The compound of formula A, where R ₁ = 5-chloropyridin-2-yl)

Obtain analogously to example 1 of 1.0 mmol of 3,6-bis- (3,5-dimethylpyrazol-1-yl) -1,2,4,5-tetrazine and 1.1 mmol of 2-amino-5-chloropyridine.

Yield 90%. Mp 260 ° C. Calculated, (%): C 47.60, H 3.64, N 37.02. C ₁₂ H ₁₁ ClN ₈ . Found, (%): C 47.79, H 3.66, N 37.01. ¹ H NMR Spectrum (400 MHz; CDCl ₃ ; δ, ppm): 11.68 (br s, ¹ H, NH); 6.26 (s, 1H, 4 ^Pz- CH); 7.63, (q, 1H, pyridino); 7.96 (d, 1H, pyridino); 8.20 (t, 1H, pyridino), 2.26; 2.50 (both s, 6H, 2CH ₃ ); 8.00, 8.06, 8.45 (all k, 3H, pyridino).

Example 15

3- (Quinolin-3-ylamino) -3- (3,5-dimethylpyrazol-1-yl) -1,2,4,5-tetrazine

(The compound of formula A, where Ri = quinolin-3-yl)

Obtain analogously to example 1 of 1.0 mmol of 3,6-bis- (3,5-dimethylpyrazol-1-yl) -1,2,4,5-tetrazine and 1.1 mmol of 3-aminoquinoline.

Yield 92.5%. Mp 240-242 ° C. Calculated, (%): C 60.38, H 4.40, N 35.22. C ₁₄ H ₁₀ N ₈ . Found, (%): C 60.40, H 4.32, N 35.31. ¹ H NMR Spectrum (400 MHz; CDCl ₃ ; δ, ppm): 11.50 (br s, ¹ H, NH); 6.16 (s, 1H, 4 ^Pz- CH); 7.53-7.63 (m, 2H, quinolino); 7.90-7.91 (q, 2H, quinolino); 8.78; 9.12 (both d, 2H, quinolino), 2.29, 2.56 (both s, 6H, 2CH ₃ ).

Example 16

[6- (3,5-Dimethylpyrazol-1-yl) -1,2,4,5-tetrazin-3-yl] prop-2-ynylamine

(The compound of formula A, where R ₁ = prop-2-ynyl)

Obtain analogously to example 1 of 1.0 mmol of 3,6-bis- (3,5-dimethylpyrazol-1-yl) -1,2,4,5-tetrazine and 1.1 mmol of propargylamine.

Yield 94%. Mp 182 ° C. Calculated, (%): C 52.39; H 4.84; N, 42.77. C ₁₀ H ₁₁ N ₇ . Found, (%): C 52.55; H 4.90; N, 42.77. ¹ H NMR (DMSO-d ₆ ), δ: 2.23 s (3H, 3 ^Pz -CH ₃ ); 2.41 d (3H, 5 ^Pz -CH ₃ ) (J 0.7 Hz); 6.20 s (1H, 4 ^Pz -CH ₃ ); 3.22 t (1H, CH) (2.5); 4.28 dd (2H, CH ₂ ) (2.5); 9.19 t (1H, NH) (5.8).

Tuberculostatic activity (minimal inhibitory concentration) and toxicity of these compounds compared to the closest analogues are given in table 1.

Table 1 Tuberculostatic activity in vitro experiments (strain H37R _ν ) and toxicity of the claimed compounds (mouse) in comparison with analogues Compound Minimum Inhibitory Concentration (MIC), mcg / ml LD ₅₀ , mg / kg Isoniazid (II) - an analogue 0.03 200 2-N- [6- (3,5-Dimethylpyrazol-1-yl) -1,2,4,5-tetrazin-3-yl] -aminopropionic acid methyl ester (II) analogue 1.25 undefined 3- (Adamantan-1-ylamino) -6- (3,5-dimethylpyrazol-1-yl) -1,2,4,5-tetrazine (example 7) 0.37 > 2500 3- (Indol-3-yl-ethylamino) -6- (3,5-dimethylpyrazol-1-yl) -1,2,4,5-tetrazine (example 10) 0.37 > 5000 3- (4-Bromophenylamino) -6- (3,5-dimethylpyrazol-1-yl) -1,2,4,5-tetrazine (example 9) 0.15 2500

table 2 Antibacterial activity of the claimed compounds in experiments in vitro Compound MIC, μg / ml E.coli Klebsiella pneumoniae Streptococcus Group C S.aureus Pseudomon as aeruginosa 3- (6-Methylpyridin-2-ylamino) -6- (3,5-dimethylpyrazol-1-yl) -1,2,4,5-tetrazine (example 11) > 100 > 100 > 100 > 100 > 100 [6- (3,5-Dimethylpyrazol-1-yl) -1,2,4,5-tetrazin-3-yl] prop-2-ynylamine (example 16) > 100 > 100 > 100 > 100 > 100

Table 3 Tuberculostatic activity of the claimed compounds in relation to typical and atypical strains of mycobacteria, MIC, μg / ml Structural formula H ₃₇ Rν M. fortuitium Isoniazid (II) - an analogue 0.03 250 3-Hydrazino-6- (3,5-dimethylpyrazol-1-yl) -1,2,4,5-tetrazine (example 1) 1.25 1.25 3- (4-Bromophenylamino) -6- (3,5-dimethylpyrazol-1-yl) -1,2,4,5-tetrazine (example 9) 0.15 0.15 3- (Adamantan-1-ylamino) -6- (3,5-dimethylpyrazol-1-yl) -1,2,4,5-tetrazine (example 7) 0.37 undefined 3- (Indol-3-yl-ethylamino) -6- (3,5-dimethylpyrazol-1-yl) -1,2,4,5-tetrazine (example 10) 0.37 undefined 2- {2- [6- (3,5-Dimethylpyrazol-1-yl) -1,2,4,5-tetrazin-3-ylamino] ethoxy} ethanol (example 5) 6.25 undefined 3-Dodecyl-6- (3,5-dimethylpyrazol-1-yl) -1,2,4,5-tetrazine (example 4) 6.25 undefined 3- (Pyridin-2-ylamino) -6- (3,5-dimethylpyrazol-1-yl) -1,2,4,5-tetrazine (example 12) 12.5 undefined

Table 4 The study of acute toxicity of the claimed compounds in mice (per os) Structural formula Tuberculostatic activity, mcg / ml Acute toxicity, mg / kg Isoniazid (II) - an analogue 0.03 200 3- (Indol-3-yl-ethylamino) -6- (3,5-dimethylpyrazol-1-yl) -1,2,4,5-tetrazine (example 10) 0.37 > 5000 3- (Adamantan-1-ylamino) -6- (3,5-dimethylpyrazol-1-yl) -1,2,4,5-tetrazine (example 7) 0.37 > 2500 3- (4-Bromophenylamino) -6- (3,5-dimethylpyrazol-1-yl) -1,2,4,5-tetrazine (example 9) 0.15 1200

Table 5 Cytotoxicity of the claimed compounds Compound Cytotoxicity, CD ₅₀ , mcg / ml 3- (Adamantan-1-ylamino) -6- (3,5-dimethylpyrazol-1-yl) -1,2,4,5-tetrazine (example 7) thirty 3- (Indol-3-yl-ethylamino) -6- (3,5-dimethylpyrazol-1-yl) -1,2,4,5-tetrazine (example 10) 28.5 3- (Pyridin-2-ylamino) -6- (3,5-dimethylpyrazol-1-yl) -1,2,4,5-tetrazine (example 12) 17.5 3- (4-Methylpyridin-2-ylamino) -6- (3,5-dimethylpyrazol-1-yl) -1,2,4,5-tetrazine (example 13) 16 6- (5-Chloropyridin-2-ylamino) -6- (3,5-dimethylpyrazol-1-yl) -1,2,4,5-tetrazine (example 14) 18.5 [6- (3,5-Dimethylpyrazol-1-yl) -1,2,4,5-tetrazin-3-yl] prop-2-ynylamine (example 16) 3.1

Thus, the claimed compounds in their characteristics (high specificity for mycobacteria, high activity against typical and atypical strains of mycobacteria, low toxicity) surpass analogues in structure and action. These compounds can be used in practical medicine for the treatment of patients infected with both typical and atypical mycobacteria.

Claims (1)

Selective anti-tuberculosis agents representing 3-amino-substituted 6- (3,5-dimethylpyrazol-1-yl) -1,2,4,5-tetrazines of the general formula A

where R ₁ = amino group, C ₁ -C ₁₂ -alkyl, 3-hydroxy-5-hydroxypentyl, propin-2-yl, ethylindolyl, adamantyl, aryl selected from optionally substituted phenyl, heteryl selected from indol-3-yl, 3-quinolinyl, pyridyl, wherein the substituted phenyl has a substituent selected from the group consisting of methyl, bromo, hydroxy, and pyridyl has substituents selected from the group of chloro, methyl.