Pathogenic bacteria-inhibiting biologically active substances and method of inhibiting pathogenic bacteria type iii secretion

FIELD: chemistry.

SUBSTANCE: invention relates to derivatives of 1,3,4-thiadiazolines (I), thiadiazinones (II) and thiadiazepines (III), obtained based on thiohydrazides of oxamic acids, which can be used to inhibit pathogenic bacteria, and can particularly affect type III secretion system in pathogens, having general formula:

, , ,

where R denotes H; R1 denotes H, pyridinyl; phenyl, substituted with alkyl C1-C5, Hal, CF3; a group , where X denotes S, substituted with alkyl C1-C5, COOR4; R2, R3 denotes alkyl C1-C5, pyridinyl, phenyl, substituted Hal, OH, OR4, a R4 denotes unsubstituted alkyl C1-C4.

EFFECT: obtaining compounds which can be used to inhibit pathogenic bacteria.

2 cl, 2 dwg, 6 tbl, 21 ex

 

The invention relates to derivatives of 1,3,4-thiadiazoline (I), thiadiazines (II) and thiadiazines (III), obtained on the basis of thiohydrazide examinavi acids, which can be used to suppress pathogenic bacteria, in particular to influence the secretion system 3 types of pathogens.

According to who, almost 60% of deaths related to chronic diseases. The General trend is the significant reduction of effectiveness used in the clinical practice of antibiotics for chronic infections. At the moment of effective antibiotics, have a pronounced therapeutic effect on chronic infection, on the market of medicinal products does not exist. This is due to the fact that during chronic infection is developed evolutionary adaptation of the pathogen, aimed at long-term survival in the host, including changes in metabolic activity. This reduces the sensitivity of microorganisms to antimicrobial agents effective in the acute stage of infection.

No less urgent is the problem of development of antibiotic resistance to the used antibiotics. The formation of microorganisms resistance and is tibeticum associated with a number of negative consequences for individual patients, and for society as a whole. When resistance to infectious disease to basic antibiotics dramatically increases the likelihood of failure of empirical treatment of the individual patient. At the same time, it is clear that each specific disease caused by resistant microorganism, is a manifestation of the process of formation and spread of resistance in microbial populations. The situation is compounded by the fact that large pharmaceutical companies have significantly reduced the development of antibiotics as growth of antibiotic resistance limits the ability of the drug and, consequently, reduces the profits from sales.

If you look at the economic side of the issue under consideration, the main motivating factor in the development of new antibacterial drugs is currently the multi-drug resistance. However, the need for drugs to treat chronic and recurrent infections covers a much larger contingent of patients, nedopoluchili need an effective treatment that provides a broad perspective in the field of development and introduction of new medicines.

In connection with the foregoing it is obvious the necessity of choice of other search strategies medicines, based on the detection of p is incipiente new targets in bacterial pathogens.

A detailed study and understanding of the mechanisms of interaction of pathogens with the host organism and identifying the most vulnerable links in the chain of dynamic events manifestations pathogenic potential may be the basis for effective selection of targets for the action of drugs of new generation aimed at the suppression of virulence, the so-called antiviolent medicines. These drugs should have a number of advantages compared with conventional antibiotics. First, the effect on the virulence properties of the pathogen, which are important in the interaction in the system host-pathogen without suppression of viability, provides a basis to predict the absence of selective pressure and the selection of resistant strains of bacteria. Second, drugs that act on specific target pathogenic bacteria will not be toxic to eukaryotic cells and for normal microflora of a man that is a major problem when using antibiotics. Indeed, microbial cells that form the microbiota of healthy humans outnumber human cells ten times, colonizer various organs and contributing to the realization of many vital processes, the role of the microbiota in which only begins to be recognized. Microbiota is a dynamic state is the and in violation of its population composition under the action of antibiotics leads to the loss of symbiotic benefits and the development of infections due to opportunistic pathogens.

The leading role in the implementation of the virulence properties of pathogenic bacteria belongs to the secretion system, through which the transport protein virulence factors in the cell of the microorganism. The process is carried out by secretion and export of effector molecules through the membrane into the extracellular space or directly into the cytoplasm of the host cell, which alters its normal physiological state and promotes adhesion, invasion and intracellular multiplication of the pathogen. In addition, through the secretion implemented a range of adaptive mechanisms aimed at long-term survival in the host organism that is associated with their ability to inactivate a protective reaction of the organism.

In the prior art known compounds influencing the virulence properties of pathogenic bacteria. These compounds inhibit the secretory function of some gram-negative bacteria, such as Yersinia pseudotuberculosis, Salmonella enterica, Pseudomonas aeruginosa, pathogenic strains of E. coli, Chlamydia spp and do not cause the development of resistance to drugs based on them. Such compounds include hydrazones derived from the Hydra is Sydow benzoic and pyridineboronic acids (FEBS Letters, 581, (2007) 587-595; Infection and Immunity, 2005, p.3104-3114, Vol.73, No.5; PNAS, 26, 2006, vol.103, No.39, 14566-14571).

However, the disadvantages of these compounds are significant toxicity and low solubility in organic solvents.

The objective of the invention is to provide compounds that are not toxic to normal microflora and host cells, in addition to which will not develop resistance, and also has good solubility in organic solvents.

Technical result achieved the claimed group of inventions is to obtain biologically active compounds in good yield with high efficacy against pathogenic bacteria, including in the treatment of chronic diseases caused by pathogenic bacteria, and is characterized by selectivity, i.e. the effects on pathogenic bacteria. Thus, the compounds are non-toxic to human and animal cells, they are well soluble in organic solvents, and are characterized by a specific activity under conditions of biological systems effects on pathogenic bacteria and does not cause the development of resistance of pathogenic bacteria.

The technical result is ensured by the fact that biologically active compounds are substituted derivatives of 1,3,4-thiadiazoles is in (I), thiadiazines (II) and thiadiazines (III), obtained on the basis of thiohydrazide examinavi acids of the General formula:

where R represents H; R1represent H, pyridinyl;

phenyl substituted by alkyl of C1-C5, Hal, CF3;where X represents S, substituted with alkyl of C1-C5, COOR4; R2, R3represent alkyl C1-C5, pyridinyl, phenyl, substituted by Hal, HE, OR4and R4represents an unsubstituted alkyl C1-C4.

The method of inhibition of type III secretion in pathogenic bacteria is exposed to bacteria an effective amount of a compound according to claim 1.

According to the claimed invention substituted thiadiazoline, thiadiazine and thiadiazine calculated on the basis of thiohydrazide examinavi acids of General formula (I) and include the cooperation of relevant chloracetamide with a pre-prepared solution of elemental sulfur with morpholine, the transmittance of the obtained solution monetization through the layer of sorbent, followed by the interaction of monetization with hydrazine hydrate, the reaction of the compounds with aldehydes or ketones, the recovery of the sodium borohydride in methanol or tetrahydrofuran and subsequent interaction with various what legitime or their derivatives in ethanol or isopropanol under conditions of acid catalysis (thiadiazoline), halogenases acid (thiadiazine), β-kalogeropoulou acid (thiadiazine) under alkaline catalysis. Substituted thiadiazoline, thiadiazine and thiadiazine based derivative thiohydrazide examinavi acids can be synthesized according to the following scheme:

The method of inhibition of type III secretion in pathogenic bacteria is exposed to bacteria an effective amount of a compound according to claim 1.

This system type III secretion detected in taxonomically distant organisms (pathogenic bacteria) - causative agents of especially dangerous infections, such as Yersinia, Brucella, and pathogens socially significant infections such as Chlamydia, Salmonella, Shigella, Helicobacter, etc. This system is more conservative and much less prone to mutation as one of the factors in the development of antibiotic resistance. Therefore, inhibitors of type III secretion in pathogens will have a directional effect on the mechanisms responsible for the process of chronicity of infection. In addition, the system III secretion is present only in pathogenic bacteria, hence its inhibitors are not toxic to normal human microflora and the host cells, i.e. are characterized by selectivity.

Specific examples of the group of inventions.

Obtaining initial N'-benzyl(who iridis) substituted thiohydrazide examinavi acids.

2-[2-(3-ethoxy-2-hydroxybenzyl)hydrazino]-N-(4-forfinal)-2-dioxazine (1)

N-chloroacetyl-4-ftoranila (2).

A General method of obtaining chloracetamide.

To a solution of (0.1 mol) of the corresponding aniline in 100 ml of DMF is added at the cooling rate of 8.75 ml (0.11 mol) of chloroacetanilide, ensuring that the temperature did not exceed 20°C. After completion of addition, stirring at room temperature the solution for another 2 hours. Then pour the reaction mixture into 600 ml of cold water and filter out sediment. Wash it on the filter with water and dry in air.

Output (2) 80% TPL 110-112°C. NMR1N DMSOd6(δ, ppm, J, Hz): 4,20 (s, 2H, ArCH2Cl); 7,21 (m, 2H, Ar); to 7.61 (m, 2H, Ar); of 10.21 (s, 1H, Ar-NH-CO). Found (%): 51.17, H 3.73, N, 7.44. Calculated (%): 51.22, H 3.76, N, 7.47. Mass spectrum, m/z: 187

2-[hydrazino(thioxo)-acetylamino]-4-ftoranila (3).

A General method of obtaining thiohydrazide.

Prepare a solution of 0.15 mol of elemental sulfur in 50 ml of DMF and added to 0.06 mol of the research. The resulting mixture was stirred for 20-30 minutes and add to it a solution of 0.05 mol of the corresponding chloracetamide in minimum amount of DMF, cooled, ensuring that the temperature did not exceed 15°C. the resulting mixture was stirred 3-6 hours (control by TLC). After the reaction reactionuses pour in the water, extragere substance with ethyl acetate, skip the organic layer through a column of silica gel, upriver on a rotary evaporator, and the resulting residue dissolved in a small amount (5-6 ml) and DMF and added 6 ml (0.12 mol) of hydrazine hydrate and stirred at room temperature for 2 hours. Then pour in the water and podkisst diluted hydrochloric acid to pH 7. Filter out the precipitate and dry it in the air.

Output: (3) 6.4 g, 60%. TPL 154-155°C. NMR1N DMSOd6(δ, ppm, J, Hz): 7,22 (m, 2H, Ar); 7,63 (m, 2H, Ar); 10,17 (s, 1H, Ar-NH-CO). Found (%): 45.01, H 3.85, N 19,80. Calculated (%): 45.06, H 3.78, N 19,71. Mass spectrum, m/z: 213.

2-[2-(3-ethoxy-2-hydroxybenzyl)hydrazino]-N-(4-forfinal)-2-dioxazine (1)

A General method of obtaining N-benzylamine of thiohydrazide.

To 10 mmol of the corresponding thiohydrazide examinados acid in 5 ml of methanol added 11 mmol of the corresponding aldehyde and boil while stirring for 5 minutes. The cooled reaction mixture. Under ice cooling (0-5°C) and stirring was added 55 mg (1.5 mmol) sodium borohydride in small portions. The reaction mixture is left for 2 hours under cooling and stirring, then added 25 ml of water and neutralized with diluted hydrochloric acid to neutral environment. The precipitation was filtered and, if necessary, recrystallized the C isopropanol.

Output: (1) 272 mg, 75%. TPL 232-233°C. NMR1N CDCl3(δ, ppm, J, Hz): 1.35 (t, 3H, -O-CH2CH3, J=7,21); of 3.96 (q, 2H, -O-CH2CH3, J=7,00 (13,94)); 4,32 (s, 1H, -NH-CH2-Ar); 6,74 to 7.62 (m, 7H, Ar); 9,12 (s, 1H, ArOH); 10,02 (s, 1H, Ar-NH-CO). Found (%): 56.43, H 4.62, N, 11.78. Calculated (%): 56.19, H 4.99, N 11.56. Mass spectrum, m/z: 363.

2-[2-(2-hydroxy-5-terbisil)hydrazino]-N-(4-(trifluoromethyl)phenyl)-2-dioxazine (4).

N-chloroacetyl-4-(trifluoromethyl)-aniline (5).

The output of chloracetamide (5) 19 g of 80% TPL 115-116°C. NMR1N DMSOd6(δ, ppm, J, Hz): 4,20 (s, 2H, Ar-CH2-Cl); 7,07 (d, 2H, arene); 7,21 (d, 2H, arene);. Found (%): 45.38, H 2.81, N, 5.96. Calculated (%): 45.49, H 2.97, N, 5.89. Mass spectrum, m/z: 237.

2 hydrazino-2-thioxo-N-[4-(trifluoromethyl)phenyl]ndimethylacetamide (6).

Output thiohydrazide (6) 6.4 g, 60%. TPL 154-155°C. NMR1N DMSOd6(δ, ppm, J, Hz): was 7.08 (d, 2H, arene); from 7.24 (d, 2H, arene); 10,59 (s, 1H, NH). Found (%): 41.16, H 3.13, N 15.83. Calculated (%): 41.06, N 3.06, N 15.96. Mass spectrum, m/z: 263.

2-[2-(2-hydroxy-5-terbisil)hydrazino]-N-(4-(trifluoromethyl)phenyl)-2-dioxazine (4).

The output of the hydrazide (4) 290 mg, 75%. TPL 212-213°C. NMR1N CDCl3(δ, ppm, J, Hz): 4,32 (s, 1H, -NH-CH2-Ar); 6,70 to 7.62 (m, 7H, Ar); of 9.30 (s, 1H, ArOH); 10,12 (s, 1H, Ar-NH-CO). Found (%): 49.50, H 3.43, N 10.74. Calculated (%): 49.61, H 3.38, N 10.85. Mass spectrum, m/z: 387.

N-pyridin-2-yl-2-[2-(pyrid is n-4-ylmethyl)hydrazino]-2-dioxazine (7)

N-chloroacetyl-2-aminopyridine (8).

Output: N-chloroacetyl-2-aminopyridine (8) 11.9 g, (70%). TPL 140-141°C. NMR1H CDCl3(δ, ppm, J, Hz): 4,20 (s, 2H, CH2Cl); 7,41 was 7.45 (m, 2H, Ar); 8,07-8,11 (m, 2H, arene); 10,31 (s, 1H, NH). Found (%): 49.20, H 4.03, N 16.21. Calculated (%): 49.28, H 4.14, N 16.42. Mass spectrum, m/z: 170.

2 Hydrazino-N-pyridin-2-yl-2-thioxo-ndimethylacetamide (9).

Output: 2-hydrazino-N-pyridin-2-yl-2-thioxo-ndimethylacetamide (9) of 1.2 g (60%). TPL 164-165°C. NMR1N DMSOd6(δ, ppm, J, Hz): 7,40-the 7.43 (m, 2H, Ar); 8,07-8,10 (m, 2H, arene); 10,59 (s, 1H, NH). Found (%): 42.76, H 4.01, N 28.44. Calculated (%): at 42.85, H 4.11, N 28.55. Mass spectrum, m/z: 196.

N-pyridin-2-yl-2-[2-(pyridine-4-ylmethyl)hydrazino]-2-dioxazine (7).

The output of the hydrazide (7) 200 mg (70%), TPL 231-232°C. NMR1N DMSOd6(δ, ppm, J, Hz): NMR1N DMSOd6(δ, ppm, J, Hz): 4,05 (s, 1H, NH-CH2-Ar); 7,40-to 7.50 (m, 4H, Ar); 8.07-a 8,23 (m, 4H, arene); 10,33 (s, 1H, Ar-NH-CO). Found (%): 54.18, H 4.68, N at 24.23. Calculated (%): 54.34, H 4.56, N 24.37. Mass spectrum, m/z: 287.

5-ethyl-2-{[1-(2-hydroxy-benzyl)-hydrazino]-2-thioxo-N-[amino}-thiophene-3-ethylcarboxylate]ndimethylacetamide (10)

N-chloroacetyl-3-carbethoxy-5-ethyl-thiophene (11).

The output of chloracetamide (11) and 23.4 g, 85%. TPL 123-125°C. NMR1N CDCl3(δ, ppm, J, Hz): of 1.27 (t, 3H, CH2CH3 , J=7,07); of 1.30 (t, 3H, CH2CH3, J=7,21); of 2.45 (d, 2H, CH2CH3, J=7,11); or 4.31 (s, 2H, CH2Cl); 4,43 (d, 2H, CH2CH3, J=7,17); 7,53 (s, 1H, Thiofen); 10,65 (s, 1H, NH). Found (%): 47.85, H 5.21, N 5.01. Calculated (%): 47.91, H 5.12, N, 5.08. Mass spectrum, m/z: 275.

5-ethyl-2-[hydrazino]-2-thioxo-N-[amino-thiophene-3-ethylcarboxylate]ndimethylacetamide (12).

Output thiohydrazide (12) 8.6 g, 57%. TPL 152-154°C. NMR1N CDCl3(δ, ppm, J, Hz): of 1.27 (t, 3H, CH2CH3, J=7,07); of 1.30 (t, 3H, CH2CH3, J=7,21); of 2.45 (d, 2H, CH2CH3, J=7,11); or 4.31 (s, 2H, CH2Cl); 4,43 (d, 2H, CH2CH3, J=7,17); 7,53 (s, 1H, Thiofen); 10.40 (s, 1H, NH). Found (%): 43.75, H 5.14, N 13.90. Calculated (%): 43.84, H 5.02, N 13.94. Mass spectrum, m/z: 301.

5-ethyl-2-{[1-(2-hydroxy-benzyl)-hydrazino]-2-thioxo-N-[amino}-thiophene-3-ethylcarboxylate]ndimethylacetamide (10)

The output of the hydrazide (10) 364 mg, 90%. T square 159-160°C. NMR1N CDCl3(δ, ppm, J, Hz): a 1.25 (t, 3H, CH2CH3, J=7,07); to 1.31 (t, 3H, CH2CH3, J=7,21); 2,44 (d, 2H, CH2CH3, J=7,11); is 4.21 (s, 1H, -NH-CH2-Ar); 4,43 (d, 2H, CH2CH3, J=7,17); 6,97-7,11 (m, 4H, Ar); 7,52 (s, 1H, Thiofen); 9,10 (s, 1H, ArOH); 10,22 (s, 1H, Ar-NH-CO). Found (%): 53.23, H 4.83, N, 10.24. Calculated (%): 53.32, H 4.72, N 10.36.

2-[2-(2-hydroxybenzyl)hydrazino]-N-(2-were)-2-dioxazine (13)

N-chloroacetyl-2-methylaniline (14).

The output of chloracetamide 14) 15.5 g, 85%. TPL 124-126°C. NMR1N CDCl3(δ, ppm, J, Hz): 1,74 (s, 3H, ArCH3); 4,20 (s, 2H, CH2Cl); 7,07 (m, 1H, Ar); 7,21 (m, 1H, Ar); 7,41 (m, 1H, Ar); 7,63 (m, 1H, Ar); 10,57 (s, 1H, Ar-NH-CH2Cl). Found (%): 58.86, H 5.47, N, 7.64. Calculated (%): 58.87, H 5.49, N, 7.63. Mass spectrum, m/z: 183.

2-[hydrazino(thioxo)-acetylamino]-2-methylaniline (15).

Output thiohydrazide (15) 10.4 g, 60%. TPL 154-157°C. NMR1N CDCl3(δ, ppm, J, Hz): 1,74 (s, 3H, ArCH3); 7,07-to 7.61 (m, 4H, Ar); 10,32 (s, 1H, Ar-NH-CO). Found: 51.41, H 5.53, N 20,40. Calculated: 51.66, H 5.30, N 20.08. Mass spectrum, m/z: 209.

2-[2-(2-hydroxybenzyl)hydrazino]-N-(2-were)-2-dioxazine (13)

Output thiohydrazide (13) 236 mg, 60%. TPL 204-206°C. NMR1N CDCl3(δ, ppm, J, Hz): 1,74 (s, 3H, ArCH3); 4,25 (s, 1H, -NH-CH2-Ar); 7,89-7,58 (m, 8H, Ar); 9,18 (s, 1H, ArOH); 10,32 (s, 1H, Ar-NH-CO). Found: C 60.87, H 5.51, N 13.23. Calculated: 60.93, H 5.43, N 13.32. Mass spectrum, m/z: 315.

2-[2-(3-ethoxy-4-hydroxybenzyl)hydrazino]-N-(2,4-differenl)-2-dioxazine (16)

N-chloroacetyl-2,4-diptiranjan (17).

The output of chloracetamide (17) 87% TPL 101-102°C. NMR1N CDCl3(δ, ppm, J, Hz): 4,20 (s, 2H, ArCH2Cl); 7,10-7,25 (m, 2H, Ar); 7,53 (m, 1H, Ar); of 10.25 (s, 1H, Ar-NH-CO). Found (%): at 46.62, H 2.99, N, 6.84. Calculated (%): 46.74, H 2.94, N, 6.81. Mass spectrum, m/z: 205.

2-[hydrazino(thioxo)-acetylamino]-2,4-diptiranjan (18).

Output: (18) of 8.1 g, 70%. TPL 154-156°C. NMR1N CDCl3(δ, ppm, J, Hz): 7,10-7,26 (m, 2H, Ar); to 7.50 (m, 1H, Ar); 10,30 (s, 1H, Ar-NH-CO). Found (%): 41.40, H 3.18, N 18.25. Calculated (%): at 41.56, H 3.05, N 18.17. Mass spectrum, m/z: 231.

2-[2-(3-ethoxy-4-hydroxybenzyl)hydrazino]-N-(2,4-differenl)-2-dioxazine (16)

The output of the hydrazide (1) 272 mg, 75%. TPL 238-239°C. NMR1N CDCl3(δ, ppm, J, Hz): 1.35 (t, 3H, -O-CH2CH3, J=7,22); 3,96 (KD, 2H, -O-CH2CH3, J=7,00 (13,90)); 4,32 (s, 1H, -NH-CH2-Ar); 6,98 to 7.62 (m, 6N, Ar); of 9.02 (s, 1H, ArOH); 10,12 (s, 1H, Ar-NH-CO). Found (%): at 53.45, H 4.58, N, 10.94. Calculated (%): 53.54, H 4.49, N 11.02. Mass spectrum, m/z: 381.

Getting thiadiazoline (I).

A General method of obtaining thiadiazoline (I).

1 mmol of the corresponding N'-benzyl(pyridyl) substituted thiohydrazide examinados acid is added to a solution of 1.05 mmol of the corresponding aldehyde in 5 ml of ethanol. Added to the reaction mixture a catalytic amount of concentrated hydrochloric acid and leave for 2 days at room temperature. Loose sediment filter out and washed with cold methanol.

Example 1, group I, (CL022).

4-(3-ethoxy-2-hydroxybenzyl)-5-(3-ethoxy-2-hydroxyphenyl)-N-(4-forfinal)-4,5-dihydro-1,3,4-thiadiazole-2-carboxamide

As a source of N'-benzyl thiohydrazide was used for compound (1)as the aldehyde used is also commercially available 3-toxicology aldehyde.

Output: CL022 332 mg, 65%. TPL >300°C decomp. NMR1N CDCl3(δ, ppm, J, Hz): 1,30-1,40 (m, 6N, -O-CH2CH3); 3,90-3,98 (m, 4H, -O-CH2CH3); to 4.41 (d, 1H, CH2, J=15,3); of 4.67 (d, 1H, CH2, J=15,2); 6,62 (1H, S-CH-N-); 6,72-7,20 (m, 8H, Ar); 7,71 for 7.78 (m, 2H, Ar); charged 8.52 (s, 1H, ArOH); of 8.95 (s, 1H, ArOH); 10.24 (s, 1H, CO-NH). Found (%): 60.98, H 5.18, N 8.11. Calculated (%): 61.04, H 5.12, N 8.21. Mass spectrum, m/z: 511.

Example 2, group I, (CL024).

4-(2-hydroxy-5-forfinal)-5-(2-hydroxy-5-terbisil)-N-(4-(trifluoromethyl)phenyl)-4,5-dihydro-1,3,4-thiadiazole-2-carboxamide.

As a source of N'-benzyl thiohydrazide used the compound (4)as the aldehyde used commercially available 5-fluoro-salicylic aldehyde.

Output (CL024) 321 mg, 70%. So pl. >300°C decomp. NMR1N CDCl3(δ, ppm, J, Hz); of 4.45 (d, 1H, N-CH2-Ar, J=15,33); 4,71 (d, 1H, N-CH2-Ar, J=15,22); to 6.43 (1H, S-CH-N-); 6,70 to 7.62 (m, 10H, Ar); 9,12 (s, 1H, AGON); of 9.30 (s, 1H, AGON); 10,10 (s, 1H, Ar-NH-CO). Found (%): 57.41, H 3.55, N, 9.10. Calculated (%): at 57.51, H 3.51, N, 9.15. Mass spectrum, m/z: 459.

Example 3, group I, (CL025).

N-pyridin-2-yl-5-pyridin-4-yl-4-(pyridine-4-ylmethyl)-4,5-dihydro-1,3,4-thiadiazole-2-carboxamide.

As a source of N'-benzyl thiohydrazide used the compound (7)as the aldehyde used commercially available 4-pyridylcarbonyl.

Output (CL025) 225 mg (60%), So pl. >300°C decomp. NMR1H DMSOd6 (δ, ppm, J, Hz); NMR1H DMSOd6(δ, ppm, J, Hz): 4,62 1 (d, 1H, CH2, J=15,12); 4.72 in (d, 1H, CH2, J=15,32); 6,32 (1H, S-CH-N-); 7,38-of 7.55 (m, 6N, AG); 8.07-a of 8.33 (m, 6N, arene); accounted for 10.39 (s, 1H, Ar-NH-CO). Found (%): at 60.51, H 4.38, N 22.23. Calculated (%): 60.62, H 4.28, N 22.32. Mass spectrum, m/z: 376.

Example 4, group I, (CL021).

ethyl 2-({[4-(2-hydroxyphenyl)-5-(2-hydroxybenzyl)-4,5-dihydro-1,3,4-thiadiazole-2-yl]carbonyl}amino)-5-ethylthiophen-3-carboxylate

As a source of N'-benzyl thiohydrazide used the compound (10)as the aldehyde used commercially available salicylic aldehyde.

Output (CL021) 306 mg, 60%. So pl. >300°C decomp. NMR1N CDCl3(δ, ppm, J, Hz): a 1.25 (t, 3H, CH2CH3, J=7,07); to 1.31 (t, 3H, CH2CH3, J=7,21); 2,44 (d, 2H, CH2CH3, J=7,11); to 4.38 (d, 1H, CH2, J=15,3); 4,51 (d, 1H, CH2, J=15,2); 6,02 (C. 1H-S-CH-N-); 6,95-to 7.15 (m, 8H, Ar); 7,52 (s, 1H, Thiofen); of 8.95 (s, 1H, AGON); 9,10 (s, 1H, AGON); of 10.25 (s, 1H, Ar-NH-CO). Found (%): 58.58, H 5.03, N 8.11. Calculated (%): 58.69, H 4.93, N 8.21.

Example 5, group I, (CL020).

4-(2-hydroxyphenyl)-5-methyl-N-(2-were)-4,5-dihydro-1,3,4-thiadiazole-2-carboxamide

As a source of N'-benzyl thiohydrazide used the compound (13)as the aldehyde used commercially available paraldehyde is recommended.

Output (CL020) 130 mg, 40%. So pl. >250°C. with decomp. NMR1N Dl3(δ, ppm, J, Hz): 1,53 (d, 3H, CH the H 3J-14,23); 1,74 (s, 3H, Agsn3); to 4.41 (d, 1H, CH2, J=15,3); of 4.67 (d, 1H, CH2, J=15,2); 5,11 (KD, 1H, -S-CH-N-, J=7,12); 7,89-7,58 (m, 8H, Ar); 9,18 (s, 1H, ArOH); 10,32 (s, 1H, Ar-NH-CO). Found: 62.30, H 5.34, N 12.73. Calculated: 62.37, H 5.23, N, 12.83. Mass spectrum, m/z: 327.

Example 6, group I, (CL023).

4-(3-ethoxy-4-hydroxyphenyl)-5-(3-ethoxy-4-hydroxybenzyl)-N-(2,4-differenl)-4,5-dihydro-1,3,4-thiadiazole-2-carboxamide

As a source of N'-benzyl thiohydrazide used the compound (16)as the aldehyde used commercially available 4-hydroxy-3-ethoxybenzaldehyde.

Output (CL023) 272 mg, 75%. TPL >300°C decomp. NMR1N CDCl3(δ, ppm, J, Hz): 1.35 (t, 3H, -O-CH2CH3, J=7,22); 3,96 (KD, 2H, -O-CH2CH3, J=7,00 (13,90)); of 4.44 (d, 1H, CH2, J=15,3); 4,63 (d, 1H, CH2, J=15,2); 6,53 (s, 1H, -S-CH-N-); 6,98 to 7.62 (m, N, Ar); of 8.92 (s, 1H, ArOH); of 9.02 (s, 1H, ArOH); 10,19 (s, 1H, Ar-NH-CO). Found (%): 61.26, H 4.98, N, 5.21. Calculated (%): 61.35, H 4.96, N, 5.30. Mass spectrum, m/z: 528.

Getting thiadiazines (II).

A General method of obtaining thiadiazines (II).

1 mmol of the corresponding N'-benzyl(pyridyl) substituted thiohydrazide examinados acid is added to a solution of 1.2 mmol Chloroacetic (or bromoxynil) acid in 5 ml of isopropanol. Added to the reaction mixture a catalytic amount of ammonium acetate and boil for 2-5 hours (monitoring by thin-layer the chromatogram). After the reaction is elevem the reaction mixture 50 ml of water and extragroup substance with ethyl acetate, wash the organic layer with water 2*50 ml Then separated the organic layer, dry over sodium sulfate and remove the solvent on a rotary evaporator. The remainder of precrystallization of the minimum amount of ethanol.

Example 7, group II, (CL019)

4-(3-ethoxy-2-hydroxybenzyl)-N-(4-forfinal)-5-oxo-5,6-dihydro-4H-1,3,4-thiadiazin-2-carboxamide

As a source of N'-benzyl thiohydrazide was used for compound (1).

Output: (CL019) 302 mg, 75%. TPL 274-275°C. NMR1N CDCl3(δ, ppm, J, Hz): 1.35 (t, 3H, -O-CH2CH3, J=7,21); 3,92 (KD, 2H, -O-CH2CH3, J=7,00 (13,94)); 3,98 (s, 2H, S-CH2-CO) 5,12 (s, 1H, -NH-CH2-Ar); 6,74-of 7.60 (m, 7H, Ar); 9,10 (s, 1H, ArOH); 10,02 (s, 1H, Ar-NH-CO). Found (%): 56.50, H 4.62, N 10.31. Calculated (%): 56.57, H 4.50, N, 10.42. Mass spectrum, m/z: 403.

Example 8, group II, (CL029)

4-(5-fluoro-2-hydroxybenzyl)-N-(4-(trifluoromethyl)phenyl)-5-oxo-5,6-dihydro-4H-1,3,4-thiadiazin-2-carboxamide

As a source of N'-benzyl thiohydrazide used connection (4)

Output (CL029) 320 mg, 75%. TPL 243-244°C. NMR1N CDCl3(δ, ppm, J, Hz): of 3.94 (s, 2H, S-CH2-CO); 5,22 (s, 1H, -NH-CH2-Ar); 6,70 to 7.62 (m, 7H, Ar); of 9.30 (s, 1H, ArOH); 10,29 (s, 1H, Ar-NH-CO). Found (%): at 50.51, H 3.17, N, 9.74. Calculated (%): 50.59, H 3.07, N, 9.83. Mass spectrum, m/z: 427.

Example 9, group II, (CL030)

5-oxo-N-pyridin-2-yl-4-(pyridine-4-ylmethyl)-5,6-dihydro-4H-1,3,4-thiadiazin-2-ka is boxlid

As a source of N'-benzyl thiohydrazide used connection (7)

Output (CL030) 228 mg (70%), TPL 273-274°C. NMR1N DMSOd6(δ, ppm, J, Hz): NMR1N DMSOd6(δ, ppm, J, Hz): to 3.92 (s, 2H, S-CH2-CO)to 5.17 (s, 1H, -NH-CH2-Ar); 7,42-to 7.61 (m, 4H, Ar); 8.07-a 8,23 (m, 4H, arene); 10,33 (s, 1H, Ar-NH-CO). Found (%): 54.93, H 4.10, N 21.31. Calculated (%): 55.04, H 4.00, N 21.39. Mass spectrum, m/z: 327.

Example 10, group II, (CL027)

Ethyl-5-ethyl-2-({[4-(2-hydroxybenzyl)-5-oxo-5,6-dihydro-4H-1,3,4-thiadiazin-2-yl]carbonyl}amino)thiophene-3-carboxylate

As a source of N'-benzyl thiohydrazide used connection (10)

Output (CL027) 380 mg, 85%. T square >200°C. with decomp. NMR1N CDCl3(δ, ppm, J, Hz): a 1.25 (t, 3H, CH2CH3, J=7,07); to 1.31 (t, 3H, CH2CH3, J=7,21); 2,44 (d, 2H, CH2CH3, J=7,11); 3,99 (s, 2H, S-CH2-CO); 4,43 (d, 2H, CH2CH3, J=7,17); 5,14 (s, 1H, -NH-CH2-Ar); 6,97-7,11 (m, 4H, Ar); 7,52 (s, 1H, Thiofen); 9,10 (s, 1H, ArOH); 10,22 (s, 1H, Ar-NH-CO). Found (%): 53.54, H 4.82, N, 9.32. Calculated (%): 53.68, H 4.73, N 9.39.

Example 11, group II, (CL026)

4-methyl-N-(2-were)-5-oxo-5,6-dihydro-4H-1,3,4-thiadiazin-2-carboxamide

As a source of N'-benzyl thiohydrazide used connection (13)

Output (CL026) 213 mg, 60%. TPL 232-233°C. NMR1N CDCl3(δ, ppm, J, Hz): 1,74 (s, 3H, ArCH3); 3,90 (s, 2H, S-CH2-WITH); to 5.08 (s, 1H, -N-CH 2-Ar); 7,89-7,58 (m, 8H, Ar); 9,18 (s, 1H, ArOH); 10,32 (s, 1H, Ar-NH-CO). Found: 60.74, H 4.94, N 11.73. Calculated: 60.83, H 4.82, N, 11.82. Mass spectrum, m/z: 355.

Example 12, group II, (CL028)

4-(3-ethoxy-4-hydroxybenzyl)-N-(2,4-differenl)-5-oxo-5,6-dihydro-4H-1,3,4-thiadiazin-2-carboxamide

As a source of N'-benzyl thiohydrazide used connection (16)

Output (CL028) 316 mg, 75%. TPL >250°C. with decomp. NMR1N CDCl3(δ, ppm, J, Hz): 1.36 (t, 3H, -O-CH2CH3, J=7,22); 3,90 (KD, 2H, -O-CH2CH3, J=7.03 is (13,90)); 3,90 (s, 2H, S-CH2-WITH); to 5.21 (s, 1H, -NH-CH2-Ar); 6,98 to 7.62 (m, 6N, Ar); 9,04 (s, 1H, ArOH); 10,10 (s, 1H, Ar-NH-CO). Found (%): 54.05, H 4.14, N, 9.83. Calculated (%): 54.15, H 4.07, N 9.97. Mass spectrum, m/z: 421.

Getting thiadiazines (III).

A General method of obtaining thiadiazines (III).

1 mmol of the corresponding N'-benzyl(pyridyl) substituted thiohydrazide examinados acid is added to a solution of 1.2 mmol of β-bromopropionic acid in 5 ml of isopropanol. Added to the reaction mixture a catalytic amount of ammonium acetate and boil 4-7 hours (monitoring by thin-layer the chromatogram). After the reaction, pour the reaction mixture into 50 ml of water and extragroup substance with ethyl acetate, wash the organic layer with water 2*50 ml Then separated the organic layer, dry over sodium sulfate and remove the solvent on a rotary evaporator. The remainder of the cross is stylizowane of the minimum amount of ethanol.

Example 13, group III, (CL023)

4-(3-ethoxy-2-hydroxybenzyl)-N-(4-forfinal)-5-oxo-4,5,6,7-tetrahydro-1,3,4-thiadiazin-2-carboxamide

As a source of N'-benzyl thiohydrazide was used for compound (1).

Output: (CL023) 290 mg, 70%. TPL >240°C. with decomp. NMR1N CDCl3(δ, ppm, J, Hz): 1.36 (t, 3H, -O-CH2CH3, J=7,21); was 1.58 (m, 2H, -S-CH2-CH2-CO)to 2.18 (m, 2H, -S-CH2-CH2-CO); 3,90 (KD, 2H, -O-CH2CH3, J=7,00 (13,94)); 5,12 (s, 1H, -NH-CH2-Ar); 6.73 x-7,60 (m, 7H, Ar); 9,10 (s, 1H, ArOH); 10,00 (s, 1H, Ar-NH-CO). Found (%): 57.46, H 4.92, N, 10.01. Calculated (%): 57.54, H 4.83, N, 10.07. Mass spectrum, m/z: 417.

Example 14, group III, (CL035)

4-(5-fluoro-2-hydroxybenzyl)-N-(4-(trifluoromethyl)phenyl)-5-oxo-4,5,6,7-tetrahydro-1,3,4-thiadiazin-2-carboxamide

As a source of N'-benzyl thiohydrazide used connection (4)

Output (CL035) 352 mg, 80%. TPL >200°C. with decomp. NMR1N CDCl3(δ, ppm, J, Hz): 1.55V (m, 2H, -S-CH2-CH2-CO)of 2.16 (m, 2H, -S-CH2-CH2-CO); 5,28 (s, 1H, -NH-CH2-Ar); 6,70 to 7.62 (m, 7H, Ar); to 9.32 (s, 1H, ArOH); 10,08 (s, 1H, Ar-NH-CO). Found (%): 51.59, H 3.54, N, 9.41. Calculated (%): 51.70, H 3.43, N 9.52. Mass spectrum, m/z: 441.

Example 15, group III, (CL036)

5-oxo-N-pyridin-2-yl-4-(pyridine-4-ylmethyl)-4,5,6,7-tetrahydro-1,3,4-thiadiazin-2-carboxamide

As a source of N'-benzyl thiohydrazide used the ü connection (7)

Output (CL036) 204 mg (60%), TPL 293-294°C. NMR1N DMSOd6(δ, ppm, J, Hz): NMR1N DMSOd6(δ, ppm, J, Hz): 1,53 (m, 2H, -S-CH2-CH2-CO)to 2.18 (m, 2H, -S-CH2-CH2-CO); 5,14 (s, 1H, -NH-CH2-Ar); 7,42-rate of 7.54 (m, 4H, Ar); 8.07-a 8,23 (m, 4H, arene); 10,42 (s, 1H, Ar-NH-CO). Found (%): 56.26, H 4.52, N 20.43. Calculated (%): 56.29, H 4.43, N 20.51. Mass spectrum, m/z: 341.

Example 16, group III, (CL032)

ethyl 5-ethyl-2-({[4-(2-hydroxybenzyl)-5-oxo-4,5,6,7-tetrahydro-1,3,4-thiadiazin-2-yl]carbonyl}amino)thiophene-3-carboxylate

As a source of N'-benzyl thiohydrazide used connection (10)

Output (CL032) 370 mg, 80%. T square >200°C. with decomp. NMR1N CDCl3(δ, ppm, J, Hz): a 1.25 (t, 3H, CH2CH3, J=7,07); to 1.31 (t, 3H, CH2CH3, J=7,21); of 1.55 (m, 2H, -S-CH2-CH2-CO); of 2.16 (m, 2H, -S-CH2-CH2-CO)2,44 (d, 2H, CH2CH3, J=7,11); 4,43 (d, 2H, CH2CH3, J=7,17); 5,19 (s, 1H, -NH-CH2-Ar); 6,97-7,11 (m, 4H, Ar); 7,49 (s, 1H, Thiofen); to 9.15 (s, 1H, ArOH); 10,28 (s, 1H, Ar-NH-CO). Found (%): 54.52, H 5.12, N, 9.01. Calculated (%): 54.65, H 5.02, N 9.10.

Example 17, group III, (CL031)

4-methyl-N-(2-were)-5-oxo-4,5,6,7-tetrahydro-1,3,4-thiadiazin-2-carboxamide

As a source of N'-benzyl thiohydrazide used connection (13)

Output (CL031) 221 mg, 60%. TPL >200°C. with decomp. NMR1N CDCl3(δ, ppm, J, Hz): 1.55V (m, 2H, -S-CH2-CH2-CO)of 1.74 (s, 3H, ArCH3); of 2.16 (m, 2H, -S-CH2-CH2-CO)5,08 (s, 1H, -NH-CH2-Ar); 7,89-7,58 (m, 8H, Ar); 9,18 (s, 1H, ArOH); 10,32 (s, 1H, Ar-NH-CO). Found: 61.70, H 5.24, N 11.29. Calculated: 61.77, H 5.18, N 11.37. Mass spectrum, m/z: 369.

Example 18, group III, (CL034)

4-(3-ethoxy-4-hydroxybenzyl)-N-(2,4-differenl)-5-oxo-4,5,6,7-tetrahydro-1,3,4-thiadiazin-2-carboxamide

As a source of N'-benzyl thiohydrazide used connection (16)

Output (CL034) 348 mg, 80%. TPL >250°C. with decomp. NMR1N CDCl3(δ, ppm, J, Hz): 1.34 (t, 3H, -O-CH2CH3, J=7,22); was 1.58 (m, 2H, -S-CH2-CH2-CO)to 2.18 (m, 2H, -S-CH2-CH2-CO); 3,96 (KD, 2H, -O-CH2CH3, J=7,06 (13,90)); 5,20 (s, 1H, -NH-CH2-Ar); 6,98 to 7.62 (m, 6N, Ar); 9,00 (s, 1H, ArOH); 10,27 (s, 1H, Ar-NH-CO). Found (%): 55.04, H 4.52, N, 9.52. Calculated (%): 55.17, H 4.40, N, 9.65. Mass spectrum, m/z: 435.

Biological examples.

To determine the toxicity and biological activity of substituted derivatives of thiadiazoles, thiadiazines and thiadiazines obtained on the basis of thiohydrazide examinavi acids, were used the following methods:

1. Methods for the determination of cytotoxic effect

A) To determine the toxicity against eukaryotic cells used method of staining cells with methylene blue (standard method), with subsequent spectrometric based results. The work was carried out in the format of 96-well plates.

In su is full monolayer of cells MSO (hybrid line synovial cells of human and murine fibroblasts) and HL (epithelial lung cells) was replaced by the cultural medium with a fresh medium with cycloheximide (1 μg/ml) and made of different doses of the investigated chemical compounds. Cells were incubated for 24 and 48 hours in CO2incubator at 37°C. After 24 and 48 hours from the wells were collected culture medium and washed cells 0.1 mm solution of phosphate-saline buffer (FSB). The cells were fixed with chilled methanol (20 ml) for 15 min at 4°C. fixed cells were added 40 μl of 0.5% methylene blue, and incubated 20 min at room temperature. After incubation methylene blue were taken from the wells and washed cells FSB 4 times. To the wells were added with 100 μl of 5% sodium dodecyl sulfate (SDS) in the FSB, and incubated for 1 hour at room temperature until complete lysis of the cells. The number of living cells was determined spectrometrically at a wavelength of 540 nm at fluorimetry MultiscanEX.

B) the Method is intended for determining the metabolic activity of cells the MTT-test (Niks, M., Otto, M. Towards an optimized MTT assay. // J Immunol. - 1900. - V.130, no. 1. - p.149-151)based on the reduction of a colorless salt of tetrazole (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide, MTT) mitochondrial and cytoplasmic dehydrogenase living metabolically active cells with the formation of blue crystals formazan, the amount of which is measured spectrometrically.

The method is conducted in the format of a 96-well culture plate. In the daily monolayer cells Msso and HL C is changed environment culturing on fresh medium without cycloheximide and made of different doses of the investigated chemical compounds. Cells were incubated for 48 hours in CO2incubator at 37°C. For 4 hours before the end of the experiment was made 1:10 volume of culture medium 10x solution of MTT (5 mg/ml). Incubated 4 hours at 37°C, 5% CO2. Selected culture medium, washed once FSB. Was added to each well 100 ál of isopropanol (propanol-2). Incubated at room temperature for 30 minutes. Was assessed by optical density at a wavelength of 540 nm at fluorimetry Multiscan EX. Substrate uptake was assessed at 405 nm.

C) To evaluate the toxic effect of these compounds in vitro was used calcinosis test, which is based on a dual color live, metabolically active cells, and dead cells with damaged cytoplasmic membrane. As the first dye is used effluorescence acetoxystyrene ether calcein, which under the action of intracellular esterases in living cells becomes fluorescent anions calcein, what causes green glow of living cells with fluorescence microscopy. The second dye is of ethidium glycosilated, which penetrates into the cell only in terms of the integrity of its membrane and binding to nucleic acids, stains the cell nucleus in orange. When viewing drugs using fluorescent m is croscope differentiate and determine the number of live and dead cells in the tested conditions.

The method is conducted in the format of a 24-hole culture plate with the glass. The analysis was performed according to the Protocol attached to the commercial kit LIVE/DEAD Viability/Cytotoxicity Kit for mammalian cells (Invitrogen, USA). To do this, in the daily monolayer cells HL replaced the cultural medium with fresh medium without cycloheximide and made of different doses of the investigated chemical compounds. Cells were incubated for 48 hours in CO2incubator at 37°C. Then selected the medium, washed once FSB and without drying glasses, made a mixture of reagents: 20 ál of 2 mm ethidium of glycosilated and 5 μl of 2 mm Calcein AM, dissolved in 10 ml of sterile FSB, in a volume of 150 μl per well. Incubated for 20 min at 37°C. Then the slides were mounted on a glass slide when applied to him several 15-20 µl of the same solution. Analysis (determining the ratio of live and dead cells) was carried out by fluorescence microscopy.

2. The method of suspension of infection of eukaryotic cells.

The method of suspension of infection of eukaryotic cells by chlamydia allows you to standardize the conduct biological experiments in a screening format 96-hole dies. To obtain a cell suspension used daily monolayer cells Msso and HL who were treated with trypsin solution and versene (ratio 1:3 respectively) claudepelanne cells from the surface of the bottle. The vial was placed in a thermostat for 5 min Then took the trypsin solution and versene and cells suspended in the cultural medium (RPMI 1640 with 10% fetal serum).

To obtain a monolayer of the prepared cell suspension was made a count of the cells in the cell Goryaeva at the rate of 1.5×105cells/ml of Infected cell strain Bu-434 Chlamydia trachomatis serovar L2 was made in the ratio of bacteria:cell 1:1 in the required volume of transport medium (RPMI medium with 5% fetal serum, 25 mm glucose solution, 5 μg/ml amphotericin b and 4 μg/ml gentamicin), which provides 80-90% of infected cells. The prepared suspension was introduced into the wells of 96 - or 24-well plates in a volume of 100 μl or 1000 μl, respectively. For the deposition of cells and stimulation interact with chlamydia tablets centrifuged at 3000 rpm for 1 hour at 25°C. After that, the tablet was placed in CO2incubator for 48 h at 37°C.

3. Determination of the influence of substituted derivatives of thiadiazoles, thiadiazines and thiadiazines obtained on the basis of thiohydrazide examinavi acid on the viability of chlamydia.

The analyzed chemical compounds in varying concentrations were made in cell culture, simultaneously with the pathogen to assess their impact on the intracellular multiplication of chlamydia. The effect was evaluated by direct immunofluorescence and p is the seeding material, obtained from a lysate of cells infected in the presence of an inhibitor of the cell.

The method of direct immunofluorescence.

Methods immunofluorescence aimed at identifying objects containing a certain antigen, and based on the treatment of the corresponding antibody labeled with a fluorochrome, followed by microscopy in the ultraviolet range.

In this paper, we used the method of direct immunofluorescence (standard method)that allows for a semi-quantitative account of the development of chlamydial infection using monoclonal antibodies to species-specific protein antigen C.trachomatis.

The work was carried out on cell lines of Msso and HL infected by the method of suspension of infection .trachomatis serovar L2 in the format of 96-well plates or 24-hole with glass. The analyzed chemical compounds were added at various doses directly in infected cells. After 48 h from the wells were collected adosados and fixed cells. When you work with 96-well plates fixation was carried out by ice 72° ethanol with subsequent location of the tablet for 30-40 min at -20°C. When working with 24-hole tablets, the slides were washed in a 0.1 mm solution of the FSB and dried. After that, the cells were fixed with acetone for 15 min at room temperature. Fixed cells were inflicted 30-50 ál of the monoclone is selected, labeled FITZ antibodies to protein antigen C.trachomatis (Humanskin-2, LLC "NEARMEDIC PLUS"), and incubated for 30 min in a humid chamber at 37°C. After incubation, the cells were washed 2 times with a solution of the FSB. The product is fully dried. In the format of 24-well plates thus prepared glass was mounted on a glass slide using a mounted liquid (glycerin in the FSB). Preparations were examined in a fluorescent microscope.

4. Determination of the influence of substituted derivatives of thiadiazoles, thiadiazines and thiadiazines obtained on the basis of thiohydrazide examinavi acids, III transport system of chlamydia.

Assessing the impact of these compounds on III transport system of chlamydia was carried out on the basis of detection of translocation of the effector protein IncA C.trachomatis on the membrane of intracellular chlamydial inclusions. Protein IncA were detected using the method of indirect immunofluorescence (standard method) using a commercial antibody to the effector protein TTC IncA C.trachomatis (polyclonal rabbit serum specific for IncA, "Innovagen, Sweden) followed by detection of anti-rabbit antibodies labeled FITZ.

For this daily monolayer was infected to obtain 80-90% of infected cells, was centrifuged at 3000 rpm for 30 min at 25°C. After 8 h after infection (in EMA the beginning of the translocation of the effector protein into the membrane of the inclusion) was added to different doses of the investigated chemical compounds. After 24 h from the wells were collected culture medium, the slides were washed in a 0.1 mm solution of the FSB and dried. After that, the cells were fixed with acetone for 15 min at room temperature. Fixed cells were inflicted 50 ál of primary anti-IncA antibodies, and incubated for 30 min at 37°C. Then the slides were washed in this solution FSB and inflicted antibodies labeled FITZ. Incubated for 30 min in a humid chamber at 37°C. After incubation, the glass cells were washed 2 times with a solution of the FSB. The product is fully dried. The thus prepared glass was mounted on a glass slide with the help of Assembly fluids. Preparations were examined in a fluorescent microscope. There was also the color of monoclonal antibodies to protein MOMR outer membrane of the cell wall, which allowed to assess the development of chlamydial infection.

Example 19. The influence of compounds of class thiadiazoline (I) on the viability of eukaryotic cells and the development of chlamydial infection

The compounds are characterized by a high degree of solubility in DMF, do not form a precipitate during storage and when converting to the transport medium.

Analysis of the toxicity of 6 of the studied compounds of this group in vitro when added to the environment of the cultivation of eukaryotic cells at doses of 12.5, 25 and 50 μm and sleduushem incubation for 24 and 48 hours at 37°C, were performed using 1. As In, revealed differences in the effect of the compounds on the viability of eukaryotic cells. The results of the evaluation of the toxicity of thiadiazoline for eukaryotic cells presented on digramma, table 1.

The lowest toxic effect was shown for compounds CL020, CL022, CL023, CL024 and CL025. Connection CL021 was characterized by severe toxicity, which limits its further tested as an inhibitor of chlamydial infection.

The ability of compounds to inhibit the intracellular life cycle of chlamydia in vitro was assessed using methods 2-4. On a chart, table 2, presents the results of the characteristics of thiadiazoline on the development of chlamydial infection.

The best effect is exerted connection CL023, suppressing the viability of chlamydia by 80% at a concentration of 12.5 μm and completely at a concentration of 25 μm when added to the transport medium simultaneously with infectious material (strain .trachomatis L2) and subsequent cultivation for 48 hours. It is shown that under these conditions, infected cells in the analyzed monolayer missing. Figure 1 shows the dose-dependent suppression of infections caused .trachomatis, under the action of selected chemical compounds. Figure 1 shows the effect of compounds CL023 on nutriclean the e development of chlamydial infection in vitro after 48 hours. On a chart, table 2, presents the results of the characteristics of thiadiazoline on the development of chlamydial infection.

Connection CL022, CL024 and CL025 effectively inhibited the development of chlamydia dose of 50 μm. Analysis of the activity of these compounds at a dose of 25 μm under the same conditions as the experiment has revealed a pronounced decrease in the size of the inclusions and the reduction of their number up to 40, 30 and 20%.

Connection CL020 and CL021 showed the lowest inhibitory effect even at a concentration of 50 μm, suppressing the development of infection is only 90 and 50%.

Example 20. The influence of compounds of class thiadiazines (II) on the viability of eukaryotic cells and the development of chlamydial infection

The compounds are characterized by a high degree of solubility in DMF, do not form a precipitate during storage and when converting to the transport medium.

The toxicity of compounds to eukaryotic cells was estimated by 1. And In adding compounds to the environment of the cultivation of cells at doses of 12.5, 25 and 50 μm and subsequent incubation for 48 hours at 37°C. toxicity Analyses revealed differences in the effect of the compounds on the viability of eukaryotic cells. In the diagram, table 3, presents the results of the evaluation of the toxicity of thiadiazines for eukaryotic cells

The lowest toxic effect was pok is Zan for connections CL026 and CL030 at concentrations of 25 and 50 ám.

Connection CL027, CL019, CL028 and CL029 showed more pronounced toxicity at concentrations of 25 and 50 ám.

The ability of compounds to inhibit the intracellular life cycle of chlamydia in vitro was assessed using methods 2-4. In the diagram, table 4, presents the results of the characteristics thiadiazine on the development of chlamydial infection.

The best effect is exerted connection CL027, suppressing the viability of chlamydia by 80% at a concentration of 25 μm and 90% at a concentration of 50 μm when added to the transport medium simultaneously with infectious material (strain C.trachomatis L2) and subsequent cultivation for 48 hours. It is shown that under these conditions in the analyzed monolayer was present significantly fewer infected cells, and inclusions were smaller (see Figure 1).

Connection CL028, CL029 and CL030 effectively inhibited the development of chlamydia dose of 50 μm. Analysis of the activity of the compounds CL029 and CL030 at a dose of 25 μm under the same conditions as the experiment has revealed a pronounced decrease in the size of the inclusions and the reduction of their number to 50 and 30%.

Connection CL019 and CL026 showed the lowest inhibitory effect even at a concentration of 50 μm, suppressing the development of infection is only 50 and 80%.

Example 21. The effect of compounds of the class of tial is azepino (III) on the viability of eukaryotic cells and the development of chlamydial infection.

The compounds are characterized by a high degree of solubility in DMF, do not form a precipitate during storage and when converting to the transport medium.

Analysis of the toxicity of 6 of the studied compounds of this group in vitro when added to the environment of the cultivation of eukaryotic cells at doses of 12.5, 25 and 50 μm and subsequent incubation for 24 and 48 hours at 37°C, were performed using 1. As In, revealed differences in the effect of the compounds on the viability of eukaryotic cells. In the diagram, table 5, presents the results of the evaluation of the toxicity of thiadiazines for the eukaryotic cell.

The lowest toxic effect was shown for compounds CL031, CL033 and CL036. Connection CL032, CL034 and CL035 was associated with greater toxicity, however, even at a concentration of 50 μm toxicity did not exceed 40%.

The ability of compounds to inhibit the intracellular life cycle of chlamydia in vitro was assessed using methods 2-4. On a chart, table 6 presents the results of the characteristics of thiadiazines on the development of chlamydial infection.

The best effect is exerted connection CL032, suppressing the viability of chlamydia by 80% at a concentration of 25 μm and 94% at a concentration of 50 μm when added to the transport medium simultaneously with infectious material (strain C.trachomatis L2) and sleduushem cultivation for 48 hours. It is shown that under these conditions, infected cells in the analyzed monolayer virtually absent (see figure 1).

Connection CL034, CL035 and CL036 effectively inhibited the development of chlamydia dose of 50 μm. Analysis of the activity of these compounds at a dose of 25 μm under the same conditions as the experiment has revealed a pronounced decrease in the size of the inclusions and the reduction of their number to 60, 40 and 30%.

Connection CL031 and CL033 showed the lowest inhibitory effect even at a concentration of 50 μm, suppressing the development of infection is only 10%and 55%.

The characteristic action of substituted derivatives of thiadiazoles, thiadiazines and thiadiazines obtained on the basis of thiohydrazide examinavi acids, III transport system of chlamydia.

Research on the specific activity of the substituted derivatives of thiadiazoles, thiadiazines and thiadiazines obtained on the basis of thiohydrazide examinavi acids in relation to inhibition of the function of the third transport system (TTS) was carried out according to method 4.

When immunochemical detection of effector protein IncA on the surface of chlamydial inclusions in the control preparations revealed inclusions, corresponding in size to the time of infection. In the case of drug action on TTC specific staining should not be detected.

The nature of the stick of action of the compounds CL023, CL027 and CL032 on TTC showed that making these connections after 8 hours after infection inhibited the translocation of the effector protein IncA.

Another test for the specificity of the compounds in respect of TTC is that translatively using TTC chlamydial protein IncA involved in the process of merging the individual, developing inside the cell inclusions. At colouring of infected cells by antibodies to the protein of MOMS in the format of the experiment: infected - 8 hours making connections - incubation up to 24 hours from the moment of infection in the control cells was observed coarse inclusions, one in each cell, and in the case of the connections on the TTC - some small, not-fused inclusions in the cytoplasm of the cell.

Specific inhibitory TTC effect was shown for compounds CL023, CL027 and CL032. Figure 2. presents an analysis of the specific activity of selected compounds against III transport system of chlamydia. Figure 2 shows the inhibition of translocation of the effector protein IncA C.trachomatis under the action of these compounds.

Tests indicate that a number of biologically active compounds, which are compounds of General formula I, II, III, may show specific activity against TTC C.trachomatis, inhibiting the process of translocation of the effector protein IncA and preventing process homotypically primary inclusions i.e. to inhibit the system type III secretion in pathogenic bacteria, without causing the development of resistance to drugs in the selection of resistant mutants, as well as providing directional effect on the mechanisms responsible for the process of chronicity of infection.

The above leads to the conclusion that the technical problem solved.

1. Biologically active substances that inhibit pathogenic bacteria, represents a substituted derivatives of 1,3,4-thiadiazoline (I), thiadiazines (II) and thiadiazines (III), obtained on the basis of thiohydrazide examinavi acids of General formula

where R represents H; R1represents H, pyridinyl; phenyl substituted by alkyl of C1-C5, Hal, CF3;where X represents S, substituted with alkyl of C1-C5, COOR4; R2, R3represent alkyl C1-C5, pyridinyl, phenyl, substituted by Hal, HE, OR4, a R4represents an unsubstituted alkyl C1-C4.

2. The method of inhibition of type III secretion in pathogenic bacteria, is the impact on bacteria an effective amount of a compound according to claim 1.



 

Same patents:

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to a compound of formula I wherein the substitutes A, B, B', Q and R1-R5 in formula I are specified as follows: A and B' are one of the following groups: (i) (R6)N(CH2)n, wherein n is 0 or 1; (ii) (CH2)n, wherein n is 0, 1 or 2; (iii) C(O)(CH2)n, wherein n is 0 or 1; or provided each of A and B' represents nitrogen, together they can form a bivalent radical of formula: -(CH2)s-X1-(CH2)t- (a), wherein each s and t is independently 1 or 2, and X1 represents (CH2)n, wherein n is 0 or 1; B is one of the following groups: (i) (R6)N; (ii) oxygen; (iii) C=δ, wherein δ represents oxygen or sulphur; (iv) C(R6)=C(R7); each R6 and R7 independently represent hydrogen, C1-4-alkyl; R1 is specified in the following groups: (i) phenyl group substituted by one or more substitute such as: - halogen specified in F, CI, Br or I, or alkyl1 group; aryl1 or heteroaryl group1; cyano, NH-alkyl1, N(alkyl1)(alkyl1) and amino; - NHCO-R or NHCOO-R, or COO-R, or CONH-R, wherein R represents hydrogen or alkyl group, or (ii) pyridinyl group which can be substituted by one substitute, such as halogen specified in I, F, Cl or Br; alkyl1 group; aryl1 group; cyano, NH-alkyl1, N(alkyl1)(alkyl1), and amino; -NHCO-R or NHCOO-R, or COO-R, or CONH-R, wherein R represents hydrogen or alkyl1 group; each R2, R3, R4 and R5 are independently specified in hydrogen or linear or branched alkyl group containing 1 to 10 carbon atoms; Q is specified in the following groups: (i) alkyl1; (ii) aryl1; (iii) heteroaryl1. The compounds of formula (I) are used for preparing a drug showing the c-kit inhibitor properties and aiming at treating a disease specified in neoplastic, allergic, inflammatory and autoimmune diseases.

EFFECT: use of oxazole derivatives as tyrosine kinase inhibitors.

13 cl, 1 tbl, 31 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to compounds of formula

and ,

where the ring X represents benzole or pyridine; R1 represents substituted alkyl; R2 represents optionally substituted aryl or optionally substituted 4-7-member monocyclic heterocyclic group or optionally substituted condensed group of heterocyclic group with the benzole ring where the substitutes of optionally substituted aryl, optionally substituted 4-7-member monocyclic heterocyclic group and optionally substituted condensed group of heterocyclic group with the benzole ring are selected from a group consisting of; (1) alkyl optionally substituted by a group selected from halogen and alkoxycarbonyl, (2) alkoxy optionally substituted by halogen, (3) halogen, (4) 4-7-member monocyclic heterocyclic group or (5) amino, optionally mono- or disubstituted alkyl, and (6) hydroxyl, R3 represents hydrogen or alkyl: R4 represents hydrogen, halogen or alkyl; R5 represents hydrogen or alkyl; R6 and R7 are identical or different, and each represents hydrogen or halogen; or pharmaceutically acceptable salt. Also, the invention refers to a IKur blocker containing the compounds described above as an active ingredient, and also to a preventive and therapeutic agent for cardiac arrhythmia and atrial fibrillation.

EFFECT: there are produced and described new compounds applicable as a IKur blocker effective for preventing or treating cardiac arrhythmia, such as atrial fibrillation.

12 cl, 13 ex

FIELD: chemistry.

SUBSTANCE: invention relates to a novel compound of general formula I

,

and a pharmaceutically acceptable salt thereof, where X denotes CH2, CHF or S, Y denotes CN, R1, R2, R3 and R4 denotes hydrogen, n equals 1, m equals 0 or 1, R denotes R11, R12 or R13, where R11 includes at least one group selected from the following b) or c), where optionally substituted heterocyclic and heteroaryl groups are bonded with a noradamantyl part either directly or through a methylene adjacent group or a C-C bond or C-N bond; b) the substituted 5-member heteroaryl group, in which the heteroaryl ring is a monocyclic aromatic ring system, includes two or more heteroatoms selected from nitrogen and oxygen; c) the heterocyclic group is optionally substituted with a C1-C3 alkyl or oxo group, where the heterocyclic ring system is a 5-9-member mono- or bicyclic ring system with one or more heteroatoms selected from a group consisting of nitrogen and sulphur, where heteroatoms can also be present as functional groups, where the heterocyclic ring system can contain one or two double bonds, and where the monocyclic heterocyclic ring can be condensed with a phenyl ring, R12 is selected from hydrogen, halogen, hydroxy, amino and C1-C4 alkoxy; R13 is a substituted phenyl, where the substitutes, which can be identical or different, include at least one group selected from a) hydrogen; b) nitro, amino; c) the saturated or unsaturated monocyclic heterocyclic ring system is optionally substituted with one or more groups selected from C1-C3 alkyl and oxo, where the heterocyclic ring system is a 5-member ring with one or more heteroatoms selected from a group consisting of nitrogen and sulphur, where the heteroatoms can also be present as functional groups. The present invention also relates to a pharmaceutical composition having dipeptidyl peptidase IV inhibiting activity, methods of obtaining the novel compound of formula I and use in treating type II diabetes and diabetic complications as well as for treating dyslipidaemia, hypercholesteremia, obesity and hyperglycaemia.

EFFECT: novel dipeptidyl peptidase IV inhibitors.

10 cl, 1 tbl, 43 ex

FIELD: chemistry.

SUBSTANCE: invention refers to new indazole derivants with the formula (1.0) or to their pharmaceutically acceptable salts and isomerides that act as inactivators in relation to ERK2. In formula (1.0): meanings of the chemical groups Q, R1, R2 are given in the invention formula. The invention also refers to the pharmaceutical composition containing the mentioned compounds and to application of the compounds with the formula (1.0) for production of crude drugs used in malignant growth treatment.

EFFECT: application of the compounds for production of crude drugs used in malignant growth treatment.

65 cl, 611 ex, 27 tbl

FIELD: chemistry.

SUBSTANCE: compounds can be used to treat such diseases as hypertension, congestive heart failure, cardiac hypertrophy and others. In formula I R1 denotes a) cyclohexyl or trifluoromethyl; or b) phenyl, 2-thienyl, 3-thienyl, 2-pyridyl, 2-imidazolyl, 2-thiazolyl, 2-benzothienyl, 4-benzofuryl, 4-benzothienyl, 7-benzofuryl, 2,3-dihydro-7-benzofuryl, 7-benzothienyl, 1,3-benzodioxol-4-yl, 7-indazolyl, or 8-quinolinyl, optionally substituted with 1-3 substitutes, and X and Y each denotes a single bond; R2 denotes methyl, ethyl, propyl, butyl, pentyl, hexyl, 5-pentenyloxy, 3,33-trifluoropropyl, 4,4-difluoropentyl, 3-(cyclopropyl)propyl, 4-(cyclopropyl)butyl, 3-hydroxypropyl, 4-hydroxybutyl, 4-hydroxypentyl, 4-hydroxyhexyl, 5-hydroxyhexyl, 2-hydroxyethoxy etc, given in the claim; R3 denotes H, F, OH, methoxy, ethoxy, 3-hydroxypropoxy, acetylamino, propionylamino, (2-methylpropionyl)amino, or butanoylamino; A denotes 2,4-disubstituted morpholine with R1XCR2R3Y, bonded on the second position and Q bonded on the fourth position, 1,3-disubstituted piperidine with R1XCR2R3Y bonded on the third position and Q bonded on the first position, 1,3-dibustituted-3-methylpiperidine with R1XCR2R3Y bonded on the third position and Q bonded on the first position, 1,3-disubstituted benzene or 1,3-disubstituted cyclohexane; Q denotes Q1, Q2, Q4, Q5, Q9, or Q10 given in the claim, to which A and N are bonded on cut-off bonds, R4 denotes H or methyl.

EFFECT: obtaining novel compounds having aspartic protease inhibitor properties, particularly renin inhibitor.

10 cl, 1 tbl, 166 ex

FIELD: chemistry.

SUBSTANCE: invention relates to compounds of formula or pharmaceutically acceptable salt thereof, synthesis methods thereof, pharmaceutical compositions containing said compounds, and use thereof to prepare a medicinal agent having mTOR kinase and/or PI3K kinase inhibiting action.

EFFECT: improved properties of the derivatives.

15 cl, 72 ex

FIELD: chemistry.

SUBSTANCE: invention relates to a compound of general formula where: R1 denotes COORa1, CONRa2Ra2', CONRa4ORa4', where: each of Ra1 and Ra4 denotes a hydrogen atom; each of Ra2 and Ra2' denotes a hydrogen atom; Ra4' denotes a lower alkyl; or R1 denotes a heterocyclic group selected from the following groups, where Y2 denotes a hydrogen atom or a lower alkyl: R2 denotes O, S, SO, SO2; R3 denotes a phenyl which is substituted with 2 substitutes selected from halogen, CF3; X2 denotes CH or N; W denotes the following residue: where: W1 denotes CH or S; W2 denotes CH; W3 denotes C or N; and at least one of W1, W2 and W3 denotes a carbon atom; or pharmaceutically acceptable salt or ester thereof. The invention also relates to a pharmaceutical composition having Avrora A selective inhibitory action, which, along with a pharmaceutically acceptable carrier or diluent, contains at least one compound of formula I a an active ingredient.

EFFECT: aminopyridine or aminopyrazine derivatives which inhibit growth of tumour cells based on Avrora A kinase selective inhibitory action.

11 cl, 3 tbl, 24 ex

FIELD: chemistry.

SUBSTANCE: invention relates to novel organic compounds of formula where R1 denotes H; halogen; -C0-C7-alkyl-O-R3; -NR4R5; R2 denotes phenyl, substituted with one or two substitutes selected from a group consisting of C1-7alkyl, halogen-C1-7alkyl, C1-7alkoxy, halogen-C1-7alkoxy, phenoxy, halogen, C1-7alkylpiperazinyl-C1-7alkyl, C3-C8-cyclalkyl, C1-7alkylpiperidinyl-C1-7alkyl and C1-7alkylimidazolyl; R3 denotes H or phenyl-lower alkyl; R4 and R5 are independently selected from a group consisting of H; lower alkyl; lower alkoxy-carbonyl and amino; A, B and X are independently selected from C(R7) or N, provided that not more than one or A, B and X denotes N; R7 denotes H; R8 denotes hydrogen; n equals 0; Y denotes O; Z denotes C; W is absent; K denotes N or C, and either a) if K denotes C, the bond shown by a wavy line () is a double bond, Q is selected from O-N, S-N, O-CH and S-CH, where in each case, the left-hand O or S atom is bonded through a bond shown in formula I to K, the right-hand N or carbon (CH) atom is bonded to C through a bond shown by a dotted line () in formula I, provided that said bond, which is shown by the dotted line, is a double bond with C; and the bond shown by a thick line () is a single bond; or b) if K denotes N, the bond shown by a wavy line () is a single bond; Q denotes N=CH, where the left-hand N atom is bonded through a bond shown in formula I to K, the right-hand carbon (CH) atom is bonded to C through a bond shown by a dotted line () in formula I, provided that said bond, which is shown by a dotted line, is a single bond with C; and the bond shown by thick line () is a double bond; or salt thereof (preferably pharmaceutically acceptable salt). The invention also relates to a pharmaceutical composition, having inhibiting action on protein kinase, containing a compound of formula I or salt thereof in an effective amount and at least one pharmaceutically acceptable carrier material.

EFFECT: heterocyclic carboxamides as kinase inhibitors.

12 cl, 25 ex

FIELD: chemistry.

SUBSTANCE: present invention relates to a compound of formula I or a pharmaceutically acceptable salt thereof, where R1 denotes C1-C8-alkylaminocarbonyl, which is optionally substituted with a 5- or 6-member heterocyclic ring containing 3-4 ring heteroatoms selected from a group consisting of oxygen, nitrogen and sulphur, where the ring can be optionally substituted with C1-C8-alkyl or C1-C8-alkoxy group ; R2 denotes C1-C3-alkyl or a halogen; one of R3 and R4 denotes R6, and the other denotes R7; R5 denotes hydrogen or halogen; R6 denotes hydrogen, hydroxy group amino group, -SO2R8, -SO2NH2, -SO2NR9R10, -COR8, -CONHR8, -NHSO2R8, nitrile, carboxy, -OR8 or C1-C8-halogenalkyl; R7 denotes hydrogen, OR11, halogen, carboxy, -SO2R8, cyanogroup or C1-C8-halogenalkyl, or when R4 denotes R7, then R7 can also denote -NR12 R13 ; R8 R11 independently denote C1-C8-alkyl or C3-C8-cycloalkyl, which can be optionally substituted with hydroxy group, C1-C8-alkoxy group, nitrile, amino group, C1-C8-alkylamino group or di-C1-C8-alkyl)amino group; any R9 denotes C1-C8-alkyl or C3-C8-cycloalkyl, which can optionally be substituted with hydroxy group, C1-C8-alkoxy group, nitrile, amino group, C1-C8-alkylamino group, di(C1-C8-alkyl)amino group or a 5- or 6-member heterocyclic ring containing one or two ring heteroatoms selected from a group consisting of oxygen and nitrogen, where the ring can optionally be substituted with C1-C8-alkyl, and R10 denotes hydrogen or C1-C8-alkyl; or R9 and R10 together with a nitrogen atom with which they are bonded form a 5- or 6-member heterocyclic ring which can contain one or two additional nitrogen heteroatoms, where the ring can be optionally substituted with C1-C8-alkyl; any R12 denotes C1-C8-alkyl or C3-C8-cycloalkyl which can be optionally substituted with di(C1-C8-alkyl)aminogroup, and R13 denotes hydrogen or C1-C8-alkyl; or R12 and R13 together with a nitrogen atom with which they are bonded form a 5- or 6-member heterocyclic ring which contains one or two additional nitrogen heteroatoms, where the ring can optionally be substituted with C1-C8-alkyl.

EFFECT: possibility of using the compounds to produce a pharmaceutical agent for treating diseases mediated by phosphatidylinositol-3 kinase.

6 cl, 3 tbl, 181 ex

FIELD: chemistry.

SUBSTANCE: invention relates to hydroximoyl-tetrazole derivatives of formula (I), , where T is a tetrazole substitute, A is a phenyl or heterocycle, L1 and L2 are different linker groups, and Q is a carbocycle, use thereof as fungicide active agents, particularly in form of fungicide compositions, and methods of controlling phytopathogenic fungi, especially plants, using said compounds or compositions.

EFFECT: more effective use of the compounds.

13 cl, 2 tbl, 2 ex

FIELD: chemistry.

SUBSTANCE: invention relates to a compound of formula

,

where R denotes a substituted or unsubstituted thiazolyl group of formula or ; R4 and R5, each independently, are selected from i) hydrogen; ii) a substituted or unsubstituted C1-C6 linear, C3-C6 branched or C3-C6 cyclic alkyl; iii) a substituted or unsubstituted phenyl; iv) a substituted or unsubstituted heteroaryl containing 5 or 6 ring atoms and 1 or 2 heteratoms, where the heteroatoms are selected from nitrogen, oxygen, sulphur and combination thereof; or R4 and R5 can be taken together to form a saturated or unsaturated ring, having 5-7 atoms; said substitutes are independently selected from one or more groups, selected from C1-C6 linear, C3-C6 branched or C3-C6 cyclic alkyl, halogen, hydroxyl or cyano; R6 denotes a group selected from i) hydrogen; ii) a substituted or unsubstituted C1-C6 linear, C3-C6 branched or C3-C6 cyclic alkyl; iii) a substituted or unsubstituted phenyl or iv) a substituted or unsubstituted heteroaryl containing 5 or 6 ring atoms and 1 or 2 heteroatoms, where the heteroatoms are selected from nitrogen, oxygen, sulphur and combination thereof; where said substitutes are independently selected from one or more groups selected from C1-C6 linear, C3-C6 branched or C3-C6 cyclic alkyl, halogen, hydroxyl or cyano; R1 is selected from i) hydrogen; ii) C1-C6 linear or C3-C6 branched alkyl; iii) a substituted or unsubstituted phenyl or iv) a substituted or unsubstituted benzyl; where said substitutes are independently selected from one or more groups selected from C1-C6 linear, C3-C6 branched or C3-C6 cyclic alkyl, halogen, hydroxyl or cyano; R2 is selected from i) C1-C6 linear or C3-C6 branched alkyl or ii) C1-C6 linear or C3-C6 branched alkoxy; R3 denotes hydrogen or C1-C4 linear or C3-C6 branched alkyl.

EFFECT: compounds of formula (I) are effective as human protein tyrosine phosphatase beta (HPTP-β) inhibitors.

20 cl, 10 tbl, 8 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: present inventions refers to a new crystalline form of tetomilast hydrate of the X-ray powder diffraction spectrum having characteristic peaks at the angle 20=10.6°, 12.9°, 21.1°, 22.3° and 25.0°, to a new crystalline form of anhydrous tetomilast type C of the X-ray powder diffraction spectrum having characteristic peaks at the angle 2θ=4.2°, 8.2°, 12.0°, 16.4°, 24.7° and 25.9°, to a new crystalline form of acetonitrile tetomilast solvate of the X-ray powder diffraction spectrum having characteristic peaks at the angle 2θ=3.6°, 7.1°, 10.6°, 14.2° and 24.8°, to based pharmaceutical compositions and to methods for preparing.

EFFECT: new crystalline forms shows useful processing characteristics with relation to preparing pharmaceutical drugs of them.

13 cl, 14 dwg, 8 ex

FIELD: chemistry.

SUBSTANCE: invention relates to indole and indazole compounds of formula in which n equals a whole number from 1 to 3, m equals 0 or 1, A denotes phenyl, X denotes C or N, R1 denotes hydrogen, alkyl, -(CH2)rNR7R8, where r equals a whole number from 1 to 5, and R7 and R8 independently denote hydrogen, alkyl or alkylcarbonyl, or can together form an optionally alkyl-substituted alkylene chain, where optionally one methylene is substituted with a N atom, R2 denotes hydrogen, halogen, cyano, nitro, hydroxy, alkyl, alkoxy or trialkylsilyl, denotes -(CH2)pCO2R7, -(CH2)pOR7, -(CH2)pNR7R8, -NHR10, -N(H)S(O)2R7, -NHC(O)R10, -(CH2)pS(O)2R7 or (CH2)p-heterocycle-R10, where p equals a whole number from 0 to 3, R7 and R8 are as defined above, R10 denotes hydrogen, oxo, alkylsulphonyl, alkylcarbonyl, alkyloxycarbonyl, alkoxy, alkyl or heterocycle, R3 denotes hydrogen, cyano, halogen, alkyl or phenyl, or denoes -(CH2)n-heterocycle or -(CH2)n-aryl, where n equals a whole number from 0 to 3, provided that R3 denotes phenyl when X denotes C and m=0, R4 denotes -YR11, where Y denotes a direct bond or -(CR7R8)pY′-, where p equals a whole number from 0 to 3, R7 and R8 are as defined above, Y′ is selected from a group consisting of -O-, -S-, -NR12-, -NR12C(O)-, -C(O)-, -C(O)O-, -C(O)NR12-, -S(O)q- and -S(O)qNR12-, where R12 denotes hydrogen, alkyl, aryl or heteroaryl, q equals a whole number from 0 to 2, R11 is selected from a group consisting of hydrogen, cyano, halogen, hydroxy, thiol, carboxy, alkyl and -(CH2)tB-R13, where t equals a whole number from 0 to 3, B denotes heterocycle, heteroaryl or aryl, R13 denotes hydrogen, cyano, halogen, hydroxy, oxo, thiol, carboxy, carboxyalkyl, alkylcarbonyloxy, alkyl, alkoxy, alkylthio, alkylcarbonyl or alkylsulphonyl, R5 denotes hydrogen, alkyl, cycloalkyl, heterocycle or heterocyclylalkyl, R6 denotes (CR7R8)p-Z-D-W-R14, where Z denotes a direct bond, or is selected from a group consisting of -C(O)-, -C(O)O, -C(O)NR12- and -S(O)y-, y equals a whole number from 1 or 2, D denotes a direct bond, or denotes cycloalkyl, heteroaryl or heterocycle, W denotes a direct bond, or denotes -NR -, -C(O)-, -C(O)O-, -C(O)NR12-, -S(O)y-, -S(O)yNR12- or -NR12S(O)y, wherein R14 denotes hydrogen, hydroxy, alkyl, alkoxy, heterocycle, heteroaryl, aryl or aralkyl, R5 and R6 together denote an alkylene chain, provided that R6 denotes cycloalkyl or heterocyclyl when X denotes N, where the heteroaryl is a 5-6-member aromatic ring containing 1-2 heteroatoms selected from N, O and S, the heterocycle is a 3-8-member ring containing 1-3 heteroatoms selected from N, O and S, where the alkyl, alkoxy, aryl, cycloalky, heterocycle and heteroaryl can be optionally substituted, and the substitutes, one or more, are selected from a group consisting of hydroxy, halogen, nitrile, amino, alkylamino, dialkylamino, carboxy, alkyl, alkoxy, carboxyalkyl, alkylcarbonyloxy, alkylthio, alkyloxycarbonyl, alkylaminocarbonyl, arylalkoxy and oxo, and pharmaceutically acceptable salts or stereoisomers thereof. The invention also relates to a composition, as well as a method of preparing said composition.

EFFECT: obtaining novel biologically active compounds for preventing or treating necrosis and necrosis-associated diseases.

40 cl, 162 ex, 2 tbl

FIELD: chemistry.

SUBSTANCE: invention relates to compounds of general formula where R denotes a thiazolyl group of formula R2 and R3 are selected from: hydrogen, C1-C3linear alkyl; R4 is selected from: C1-C3linear or C3cyclic alkyl, phenyl and thiophenyl; Z denotes a group of formula: -(L)n-R1; R1 is selected from: i) C1-C3linear or branched alkyl, optionally substituted with C1-C4alkoxycarbonyl, halogen; ii) substituted phenyl or substituted with one or two substitutes selected from halogen, methoxy- or hydroxy group, C1-C4alkoxycarbonyl; iii) dioxopiperazinyl and 2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl, substituted with C1-C3alkyl; or iv) heteroaryl rings containing 5-10 atoms selected from thiazole, triazole, 1H-imidazole, thiadiazole, oxazole, isoxazole, oxadiazole, benzodioxole, benzo(1,4)dioxepanyl, pyridine, pyrimidine, 1H-indole, 2,3-dihydrobenzo[b][1,4]dioxynil, which can be substituted with oine or two substitutes selected from: a) hydroxy; b) C1-C3alkyl (which can be substituted with one more two substitutes selected from: ) phenyl; ii) C1-C4alkoxycarbonyl; iii) naphthalenyl; iv) 2-methylthiazolyl) ; c) NHC(O)C1-C3alkyl; d) C1-C4alkoxycarbonyl; e) 1 -(tert-butoxycarbonyl)-2-phenylethyl; f) methoxybenzyl; g) phenyl which can be substuted with C1-C4alkoxy, halogen, methoxycarbonyl or >NHC(O)CH3; h) (methoxy-2-oxoethyl)carbamoyl; L denotes a group selected from: i) C(O)NH[C(R5aR5b)]w-; ii) -C(O)[C(R6aR6b)]x-; iii) -C(O)[C(R7aR7b)]yC(O)-; iv) -SO2[C(R8aR8b)]z-; R5a, R5b, R6a, R6b, R7a, R7b, R8a and R8b, each independently denotes: i) hydrogen; ii) C1-C3 linear alkyl which can be substituted with 1 or 2 halogen atoms; iii) phenyl which can be substituted with 1-2 substitutes selected from halogen and lower alkoxy; iv) heteroaryl rings selected from imidazolyl, imidazolyl substituted with methyl, benzo(1,4)oxazinyl, oxadiazolyl substituted with methyl; index n equals 0 or 1; indices w, x, y and z are each independently equal to a number from 1 to 3. The invention also relates to pharmaceutically acceptable salts of compounds of formula (I) and use of compounds of formula (I) to prepare a medicinal agent for treating protein tyrosine phosphatase beta-mediated conditions.

EFFECT: obtaining compounds of formula (I) as human protein tyrosine phosphatase beta (HPTP-β) inhibitors.

15 cl, 17 dwg, 13 tbl, 16 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention relates to derivatives of 5-amino-3-(2-nitroxipropyl)-1,2,4-thiadiazoles of general formula , where R1, R2 can be similar or different and independently represent hydrogen, substituted or non-substituted aryl or heteroaryl or aralkyl, alkyl, cycloalkyl, and R1 + R2 can represent heteroaryl (optionally substituted piperasin and piperidin).

EFFECT: obtained are novel compounds, which can be applied in medicine for treatment of neurodegenerative diseases.

1 cl, 3 ex

FIELD: chemistry.

SUBSTANCE: invention relates to novel carbostyril compounds of general formula (1) or salts thereof with common pharmaceutically acceptable acids or pharmaceutically acceptable basic compounds, having activity on promotion of TFF2 production, a pharmaceutical composition based on said compounds, an agent based on disclosed compounds used in case of a disorder where up-regulation of TFF has a prophylactic and/or therapeutic effect, use of disclosed compounds to prepare said agent and a method of producing disclosed compounds. The invention also relates to novel specific carbostyril compounds or salts thereof with common pharmaceutically acceptable acids or pharmaceutically acceptable basic compounds. In structural formula (1), A is a direct bond, a lower alkylene group or lower alkylidene group, X is an oxygen or sulphur atom, the bond between positions 3 and 4 of the carbostyril backbone is a single bond or a double bond, R4 and R5 each denotes a hydrogen atom provided that, when the bond between positions 3 and 4 of the carbostyril backbone is a double bond, R4 and R5 can instead be bonded to each other in form of a -CH=CH-CH=CH- group, and R1, R2 and R3 assume values given in the claims.

EFFECT: high efficiency of compositions based on said compounds.

32 cl, 23 dwg, 184 tbl, 1535 ex

Iap inhibitors // 2425838

FIELD: chemistry.

SUBSTANCE: invention relates to novel compounds of formula

, which can inhibit binding of protein Smac with apoptosis protein inhibitor (IAP).

EFFECT: improved properties of the inhibitor.

4 cl, 198 ex

Heterocompound // 2425832

FIELD: chemistry.

SUBSTANCE: invention relates to a compound of formula

or pharmaceutically acceptable salt thereof, where symbols assume the following values; ring denotes

or , X denotes a single bond, -CH2-, -NR3-, -O-, -S-, R1 denotes a halogen; phenyl; pyridyl; (C3-C8)cycloalkyl; or (C1-C6) alkyl or (C2-C6) alkenyl, each of which can contain a halogen, -CONH2, phenyl or (C3-C8)cycloalkyl as a substitute, R2 denotes CN, -O-(C1-C6)alkyl, -C(=O)H, halogen; or (C1-C6)alkyl, which can be substituted with a halogen or -OH, R3 can form morpholino or 1-pyrrolidinyl together with R1 and nitrogen, and when X denotes a single bond, R1 and R2 can jointly form a 5-member ring and additionally contain -(C1-C6)alkyl as a substitute, R4 denotes the following ring: , , , , , , , , , , or , where any one of the bonds in the ring is linked to an oxazole ring, R5 denotes -H, (C1-C6)alkyl, which can be substituted by not less than one group selected from: -C(=O)NRXRY, -NHRX and -ORX- (C2-C6)alkenyl-; -C(=O)H; -C(=O)NRXRY, RX and RY can be identical or different and denote -H; or (C1-C6)alkyl. The invention also relates to a pharmaceutical composition based on said compounds, having SlP1 agonist activity.

EFFECT: compounds and compositions can be used in medicine for preventing and treating rejection during organ transplant, bone marrow or tissue transplant and autoimmune diseases.

16 cl, 84 tbl, 198 ex

FIELD: medicine.

SUBSTANCE: compounds can be used for treating neurological conditions, more specifically for treating neurodegenerative conditions, such as Alzheimer's disease. In a compound of formula I R2 represents H or CH2NR1R4 where R1 and R4 are independently selected from H, unsubstituted C1-6alkyl, substituted or unsubstituted C3-6 cycloalkyl, R3 represents H; substituted or unsubstituted C1-4alkyl; substituted or unsubstituted C2-4alkenyl; substituted or unsubstituted 6-members aryl condensed or uncondensed with substituted or unsubstituted 6-members aryl or 5-6-members heteroaryl, containing 1-2 nitrogen atoms in a cycle; substituted or unsubstituted saturated or unsaturated 5 or 6-members N-containing heterocycle which can additionally contain nitrogen, oxygen or the sulphur atom condensed or ucondensed with substituted or unsubstituted 6-members aryl or 5-6-members heteroaryl containing nitrogen in a cycle; (CH2)nR6 where n is an integer from 1 to 6, and the values of R6 and the values of other radicals are specified in the patent claim.

EFFECT: increased antiamyloidogenic action.

20 cl, 20 tbl, 6 dwg, 7 ex

FIELD: chemistry.

SUBSTANCE: invention relates to compounds of formula , where R1 is a 3-7-member carbocyclic ring and n is a number ranging from 1 to 8, and the rest of the radicals are described in the claim.

EFFECT: possibility of using such compounds and compositions in therapy as metabotropic glutamate receptor modulators.

33 cl, 367 ex

FIELD: chemistry.

SUBSTANCE: N,S-containing heterocycle obtained using the disclosed method can be used as a selective sorbent and extraction agent for precious metals, and as a special reagent for inhibiting bacterial action in different media. The method involves reaction of hydrogen sulphide-saturated aqueous formaldehyde solution with o-aminothiophenol in the presence of EtOH at 20-80°C and stirring for 3 hours. Output of the desired product is equal to 79% at 20°C and 83% at 80°C.

EFFECT: high output.

1 cl, 1 ex

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