Using derivatives of cysteine for preparing medicinal agent designated for treatment of pathology arising as result of formation of heterotrimeric protein g

FIELD: organic chemistry, amino acids, medicine, pharmacy.

SUBSTANCE: invention relates to using derivatives of cysteine for preparing a medicinal agent. The proposed agent is designated for treatment of diseases arising as a result of formation of heterotrimeric protein G, and to new derivatives of cysteine, and pharmaceutical composition based on thereof. Derivatives of cysteine, in particular, involve the following compounds: bis-1,1'-[7-(2-amino-1-oxo-3-thiopropyl)-8-(cyclohexylmethyl)-2-(2-methoxyphenyl)-5,6,7,8-tetrahydroimidazo-[2,2a]-pyrazine]-disulfide and bis-1,1'-[7-(2-amino-1-oxo-3-thiopropyl)-2-91-naphthyl)-8-(2-methylpropyl)-5,6,7,8-tetrahydroimidazo-[1,2a]-pyrazine-7-yl]-disulfide. Invention provides high effectiveness of treatment.

EFFECT: valuable medicinal properties of compounds.

6 cl, 7 dwg, 2 tbl, 7 ex

 

The present invention relates, in particular, to the use of derivatives of cysteine for obtaining a medicinal product intended for the treatment of pathologies caused by the formation of heterotrimeric protein G. These disorders are, in particular, diseases that are associated with the following biological functions or disorders: smell, taste, light perception, neurotransmission, neurodegeneration, the functioning of the endocrine and exocrine glands, autocrine and paracrine regulation of arterial blood pressure, embryogenesis, benign cell proliferation, oncogenesis, viral infection, immune function, diabetes, obesity, and benign and malignant proliferative diseases.

The G proteins are, in fact, the structural Association of three different subunits, called α, β and γbut function as dissociable particles formed, on the one hand, subunits α and, on the other hand, dimers β/γ.

The G proteins are involved in transmitting signals from the cell surface due to its interaction with receptors of the seven transmembrane domains to the internal parts through various effectors, including adenylate cyclase, phospholipase C, or ion channels. The enzyme adenyl cyclase generates cyclic monophosphate (camp) (see Gilman A.G., Biosci. Rep., 15, 65-97 (1995)). It is also known that activation of adenylate cyclase requires that the protein G was temporarily in heterotrimeric form, i.e. the form in which the monomer is formed by subunit α, associated with the dimer formed by the subunits β and γ. Only in this state, the signal from the cell surface may be activated subunit and protein G, which after dissociation can modulate adenylate cyclase and modulate products CATF.

It is also known that the dimers β/γ can directly activate effectors, leading to activation of kinases that are regulated by extracellular signals (ERKs), or MAR-kinases. It has been shown there is a direct connection between subunits β/γ and the kinases src, or src like (see Gut kind J.S., J.Biol. Chem., 273, 1839-1842 (1998)).

In addition, it is shown that bacterial toxins, such as Vibrio cholerae and Bortella pertussis, peptides, such as mastoparan and suramin, directly modulate the activity of protein G (see, Freissmuth M., Boehm, S., Beindl W., and others, Mol. Pharmacol., 49, 602-611 (1996); Boehm, S., Huck, S., Motejlek A., and others, Journal of Neurochemistry, 66, 1019-1026 (1966); Cachero T.G., R. Rigual, Rocher, A. and Gonzaiez S., Eur. J. Neurosci., 8, 2320-2327 (1996); M. Danilenko, Worland, P., Carlson C., Sausviile E.A. and Sharoni Y., Biochem. Biophys. Res. Commun., 196, 1296-1302 (1993); Beindl W., Mitterauer So, Hohenegger, M., A. P. Ijzerman, Nanoff C. and Freissmuth M., Mol. Pharmacol., 50, 415-423 (1996)).

Cholera toxin, for example, modifies subunit αsprotein DZA the joining of adenosinetriphosphatase (ADP-ribose), derived from nicotinamide-adenine-dinucleotide (ABOVE), to the specific arginine-acceptor site. It completely blocks the activity of guanozintrifosfata (GTPase)that provokes prolonged stimulation of its next-effector, adenylate cyclase and leads to overproduction camp.

The negative effect of increased amounts of camp are also known and, in particular, it is manifested at the level of the following biological functions and disorders: smell, taste, light perception, neurotransmission, neurodegeneration, the functioning of the endocrine and exocrine glands, autocrine and paracrine regulation of arterial blood pressure, embryogenesis, benign cell proliferation, oncogenesis, viral infection and immune function, diabetes, and obesity.

The applicant has now found that some derivatives of cysteine, namely compounds of General formula (A):

the corresponding partial formulas (A1) and (A2)

in which:

X means R12and Y means R8or X and Y are part of the six-membered cycle, and the set of X-Y represents a radical-CH(R8)-CH(R9)-;

R1means H, lower alkyl or lower allylthiourea;

R2and R3not avisio mean H or lower alkyl;

R4means N2or;

R5means N or one of the following radicals: lower alkyl, lower alkenyl, lower quinil, aryl, lower arylalkyl, heterocyclic or lower alkylchlorosilanes radical, these radicals can be substituted by radicals selected from the group consisting of lower alkyl, -O-R10, -S(O)mR10(where m means 0, 1 or 2), -N(R10)(R11), -N-C(O)-R10, -NH-(SO2)-R10, -CO2-R10C(O)-N(R10)(R11and -(SO2)-N(R10)(R11);

R6and R7independently mean H, the radical-C(O)-NH-CHR13-CO2R14or one of the following radicals: lower alkyl, aryl, lower arylalkyl, heterocyclic or lower alkylchlorosilanes radical, these radicals can be substituted by radicals selected from the group consisting of HE, lower alkyl or alkoxyl, N(R10)(R11), COOH, CON (R10)(R11and halogen;

or R6and R7together form an aryl or heterocyclic radical;

R8and R9independently denote H or one of the following radicals: lower alkyl, aryl, lower arylalkyl, heterocyclic or lower alkylchlorosilanes radical, these radicals can be substituted by the radicals, the choice is Emami from the group consisting of HE, lower alkyl or alkoxyl, N(R10) (R11), COOH, CON(R10)(R11and halogen;

or R8and R9together form an aryl or heterocyclic radical;

R10and R11independently mean H, aryl or heterocyclic radical, or a lower alkyl, lower arylalkyl or lower alkylchlorosilanes radical;

R12mean NR9, S, or O;

R13means lower alkyl, possibly substituted by a radical chosen from the following radicals: lower alkyl, -OR10, -S(O)mR10(where m means 0, 1 or 2) and-N(R10)(R11);

R14means H or lower alkyl;

or compounds of General formula (I):

in which:

W1mean residue derived from cysteine in restored or unrestored form;

Ar means a radical derived from aminobenzoic acid, the aromatic nucleus which may be substituted;

W2means the balance of amino acids, preferably aliphatic amino acids;

or compounds of the General formula (C):

in which:

Z1means lower alkyl;

Z2and Z3both mean N or Z2and Z3together form a chain of 2-4 units, chosen among the radicals-S(O)-, -CH2-, -CH(NH2)-and-S-, priruslovye, what two serial link cannot be-C(O)-;

provided that compounds of General formula (C) can also be in the form of dimers, when the radical Z2means a hydrogen atom that can be removed by oxidation;

or pharmaceutically acceptable salt of the compounds of General formula (A), (B) or (C);

can be used to produce medicines for the treatment of pathologies caused by the formation of heterotrimeric protein G.

Under lower alkyl understand linear or branched alkyl radical with 1-6 carbon atoms and in particular methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl and tert-butyl, pentyl, neopentyl, isopentyl, hexyl, isohexyl. Under the heterocyclic radical see radical formed by one or more cycles and containing at least one heteroatom. Under the lowest arylalkyl, alkylchlorosilanes radical, alkylthiol or alkoxyl understand radicals, the alkyl part of which has the above value.

The radical Ar, which is formula (C), preferably may be substituted by alkyl with 1-6 carbon atoms or aryl, and these alkyl and aryl radicals themselves can also be preferably substituted by alkoxyl with 1-4 carbon atoms, fluorine, chlorine, bromine. And the sludge preferably the phenyl itself may also be substituted by alkyl.

Compounds of General formula (I) preferably are those where Ar means a radical derived from aminobenzoic acid, the aromatic nucleus of which is substituted by phenyl, and W2means the residue of aliphatic amino acids.

To obtain drugs for the treatment of pathologies caused by the formation of heterotrimeric G protein, in particular, can be used the following compounds:

- 7-(2-amino-1-oxo-3-thiopropyl)-8-(cyclohexylmethyl)-2-(2-were)-5,6,7,8-tetrahydroimidazo[1,2A]pyrazin;

- 7-(2-amino-1-oxo-3-thiopropyl)-8-butyl-2-(2-methoxyphenyl)-5,6,7,8-tetrahydroimidazo[1,2A]pyrazin;

- 7-(2-amino-1-oxo-3-thiopropyl)-8-(1-methylpropyl)-2-(2-methoxyphenyl)-5,6,7,8-tetrahydroimidazo[1,2A]pyrazin;

- 1-[2(R)-amino-3-mercaptopropyl]-2(S)-n-butyl-4-(1-naphtol)piperazine;

bis-1,1'-[7-(2-amino-1-oxo-3-thiopropyl)-2-(methoxyphenyl)-8-(1-methylpropyl)-5,6,7,8-tetrahydroimidazo[1,2A]pyrazin]disulfide;

bis-1,1'-[7-(2-amino-1-oxo-3-thiopropyl)-8-(cyclohexylmethyl)-2-(2-methoxyphenyl)-5,6,7,8-tetrahydroimidazo[1,2A]pyrazin]disulfide;

bis-1,1'-[7-(2-amino-1-oxo-3-thiopropyl)-2-(1-naphthyl)-8-(2-methylpropyl)-5,6,7,8-tetrahydroimidazo[1,2A]pyrazin-7-yl]disulfide;

the compound of formula (VII):

connection f is rmula:

- 7-(2-amino-1-oxo-3-thiopropyl)-8-(cyclohexylmethyl)-2-phenyl-5,6,7,8-tetrahydroimidazo[1,2A]pyrazin;

- 7-(2-amino-1-oxo-3-thiopropyl)-2-(2-methoxyphenyl)-8-(phenylmethoxy)methyl-5,6,7,8-tetrahydroimidazo[1,2A]pyrazin;

- 7-(2-amino-1-oxo-3-thiopropyl)-2-(2-methoxyphenyl)-8-(1-phenylmethoxy)ethyl-5,6,7,8-tetrahydroimidazo[1,2A]-pyrazin;

- 7-(2-amino-1-oxo-3-thiopropyl)-2-(2-methoxyphenyl)-8-(phenoxyethyl)-5,6,7,8-tetrahydroimidazo[1,2A]pyrazin;

- 7-(2-amino-1-oxo-3-thiopropyl)-2-(2-methoxyphenyl)-8-(phenoxyethyl)-5,6,7,8-tetrahydroimidazo[1,2A]pyrazin in dimeric form;

- and 7-(2-amino-1-oxo-3-thiopropyl)-2-(2-methoxyphenyl)-8-(phenylsulfonyl)-5,6,7,8-tetrahydroimidazo[1,2A]-pyrazin;

or pharmaceutically acceptable salt of one of these compounds.

Preferably use one of the following compounds according to the invention:

bis-1,1'-[7-(2-amino-1-oxo-3-thiopropyl)-8-(cyclohexylmethyl)-2-(2-methoxyphenyl)-5,6,7,8-tetrahydroimidazo[1,2A]pyrazin]disulfide (I);

bis-1,1'-[7-(2-amino-1-oxo-3-thiopropyl)-2-(1-naphthyl)-8-(2-methylpropyl)-5,6,7,8-tetrahydroimidazo[1,2A]pyrazin-7-yl]disulphide (II);

- 7-(2-amino-1-oxo-3-thiopropyl)-8-(cyclohexylmethyl)-2-(2-were)-5,6,7,8-tetrahydroimidazo[1,2A]pyrazin (III);

the compound of formula (IV):

- 7-(2-amino-1-oxo-3-thiopropyl)-8-butyl-2-(2-methoxyphenyl)-5,6,7,8-tetrahydroimidazo,2A]pyrazin (V);

bis-1,1'-[7-(2-amino-1-oxo-3-thiopropyl)-2-(methoxyphenyl)-8-(1-methylpropyl)-5,6,7,8-tetrahydroimidazo-[1,2A]pyrazin]disulfide (VI);

the compound of formula (VII):

- 7-(2-amino-1-oxo-3-thiopropyl)-8-(cyclohexylmethyl)-2-phenyl-5,6,7,8-tetrahydroimidazo[1,2A]pyrazin;

- 7-(2-amino-1-oxo-3-thiopropyl)-8-(1-methylpropyl)-2-(2-methoxyphenyl)-5,6,7,8-tetrahydroimidazo[1,2A]pyrazin;

- 1-[2(R)-amino-3-mercaptopropyl]-2(S)-n-butyl-4-(1-naphtol)piperazine;

or pharmaceutically acceptable salt of one of these compounds.

More preferably, use one of the following compounds according to the invention:

bis-1,1'-[7-(2-amino-1-oxo-3-thiopropyl)-8-(cyclohexylmethyl)-2-(2-methoxyphenyl)-5,6,7,8-tetrahydroimidazo-zo[1,2A]pyrazin]disulfide (I);

bis-1,1'-[7-(2-amino-1-oxo-3-thiopropyl)-2-(1-naphthyl)-8-(2-methylpropyl)-5,6,7,8-tetrahydroimidazo[1,2A]pyrazin-7-yl]disulphide (II);

- 7-(2-amino-1-oxo-3-thiopropyl)-8-(cyclohexylmethyl)-2-(2-were)-5,6,7,8-tetrahydroimidazo[1,2A]pyrazin (III);

- 7-(2-amino-1-oxo-3-thiopropyl)-8-butyl-2-(2-methoxyphenyl)-5,6,7,8-tetrahydroimidazo[1,2A]pyrazin (V);

the compound of formula (VII):

or pharmaceutically acceptable salt of one of these compounds.

Finally, in highly preferred are the following compounds:

bis-1,1'-[7-(2-amino-1-oxo-3-thiopropyl)-8-(cyclo is exility)-2-(2-methoxyphenyl)-5,6,7,8-tetrahydroimidazo[1,2A]pyrazin]disulfide (I);

bis-1,1'-[7-(2-amino-1-oxo-3-thiopropyl)-2-(1-naphthyl)-8-(2-methylpropyl)-5,6,7,8-tetrahydroimidazo[1,2A]pyrazin-7-yl]disulphide (II);

or pharmaceutically acceptable salt of one of these compounds.

The invention therefore relates to the use of compounds of General formula (A), (B) or (C), such as those mentioned above, for obtaining a medicinal product intended for the treatment of pathologies caused by the formation of heterotrimeric protein G. In particular, the invention relates to the application of the aforementioned inhibitors to obtain drugs for treatment of diseases associated with the following biological functions or disorders: smell, taste, light perception, neurotransmission, neurodegeneration, the functioning of the endocrine and exocrine glands, autocrine and paracrine regulation of arterial blood pressure, embryogenesis, viral infection, immune function, diabetes, and obesity.

More preferably, the invention relates to the use of compounds of General formula (A), (B) or (C) for obtaining a medicinal product intended for the treatment of cholera, acquired immunodeficiency syndrome (AIDS), diarrhea traveler and hereditary male premature puberty.

The subject invention is also are the new products of General formula (A) under the numbers 1-7 and described below in the examples, namely:

- 7-(2-amino-1-oxo-3-thiopropyl)-8-(cyclohexylmethyl)-2-(2-were)-5,6,7,8-tetrahydroimidazo[1,2A]pyrazin;

- 7-(2-amino-1-oxo-3-thiopropyl)-8-(cyclohexylmethyl)-2-phenyl-5,6,7,8-tetrahydroimidazo[1,2A]pyrazin;

- 7-(2-amino-1-oxo-3-thiopropyl)-2-(2-methoxyphenyl)-8-(phenylmethoxy)methyl-5,6,7,8-tetrahydroimidazo[1,2A]pyrazin;

- 7-(2-amino-1-oxo-3-thiopropyl)-2-(2-methoxyphenyl)-8-(1-phenylmethoxy)ethyl-5,6,7,8-tetrahydroimidazo[1,2A]-pyrazin;

- 7-(2-amino-1-oxo-3-thiopropyl)-2-(2-methoxyphenyl)-8-(phenoxyethyl)-5,6,7,8-tetrahydroimidazo[1,2A]pyrazin;

- 7-(2-amino-1-oxo-3-thiopropyl)-2-(2-methoxyphenyl)-8-(phenoxyethyl)-5,6,7,8-tetrahydroimidazo[1,2A]pyrazin in dimeric form;

- and 7-(2-amino-1-oxo-3-thiopropyl)-2-(2-methoxyphenyl)-8-(phenylsulfonyl)-5,6,7,8-tetrahydroimidazo [1,2A]pyrazin.

The object of the present invention is also the above new products, or their pharmaceutically acceptable salts as drugs and their use for obtaining a medicinal product intended for the treatment of pathologies caused by the formation of heterotrimeric protein G. In particular, the invention relates to the use of the above products to obtain drugs for treatment of diseases associated with the following biological functions or disorders: the smell, the flavors of the e feeling light perception, neurotransmission, neurodegeneration, the functioning of the endocrine and exocrine glands, autocrine and paracrine regulation of arterial blood pressure, embryogenesis, benign cell proliferation, oncogenesis, viral infection, immune function, diabetes, obesity and benign and malignant proliferative diseases.

For use according to the invention particularly preferred products are thus the following:

bis-1,1'-[7-(2-amino-1-oxo-3-thiopropyl)-8-(cyclohexylmethyl)-2-(2-methoxyphenyl)-5,6,7,8-tetrahydroimidazo[1,2A]pyrazin]disulfide;

bis-1,1'-[7-(2-amino-1-oxo-3-thiopropyl)-2-(1-naphthyl)-8-(2-methylpropyl)-5,6,7,8-tetrahydroimidazo[1,2A]pyrazin-7-yl]disulfide;

the compound of the formula:

- 7-(2-amino-1-oxo-3-thiopropyl)-8-(cyclohexylmethyl)-2-(2-were)-5,6,1,8-tetrahydroimidazo[1,2A]pyrazin;

- 7-(2-amino-1-oxo-3-thiopropyl)-8-(cyclohexylmethyl)-2-phenyl-5,6,7,8-tetrahydroimidazo[1,2A]pyrazin;

- 7-(2-amino-1-oxo-3-thiopropyl)-2-(2-methoxyphenyl)-8-(phenylmethoxy)methyl-5,6,7,8-tetrahydroimidazo[1,2A]pyrazin;

- 7-(2-amino-1-oxo-3-thiopropyl)-2-(2-methoxyphenyl)-8-(1-phenylmethoxy)ethyl-5,6,7,8-tetrahydroimidazo[1,2A]-pyrazin;

- 7-(2-amino-1-oxo-3-thiopropyl)-2-(2-methoxyphenyl)-8-(phenoxyethyl)-5,6,7,8-tetrahydroimidazo[1,2A]pyrazin;

- -(2-amino-1-oxo-3-thiopropyl)-2-(2-methoxyphenyl)-8-(phenoxyethyl)-5,6,7,8-tetrahydroimidazo[1,2A]pyrazin in dimeric form;

- and 7-(2-amino-1-oxo-3-thiopropyl)-2-(2-methoxyphenyl) -8-(phenylsulfonyl)-5,6,7,8-tetrahydroimidazo[1,2A]pyrazin;

or pharmaceutically acceptable salt of one of these compounds.

Similarly, the invention relates in particular to the use of the above compounds for obtaining a medicinal product intended for the treatment of cholera, acquired immunodeficiency syndrome (AIDS), diarrhea traveler and hereditary male premature puberty.

Compounds of General formula (a) and obtaining them are described in International patent application 97/30053 or below in the examples. Compounds of General formula (I) and obtaining them are described in International patent application 96/21456. Finally, obtaining compounds of General formula (C) described in the International patent application PCT 95/00497 with the exception of the compounds of formula (VII), the synthesis of which is described in the experimental part of this application.

Pharmaceutical compositions comprising the compound according to the invention, can be in the form of solids, e.g. powders, granules, tablets, gelatin capsules, liposomes or suppositories. Appropriate solid supports can be, for example, calcium phosphate, magnesium stearate, talc, sugars, lactose, dextrin, starch, gelatin, cellulose, methyl cellulose, sodium alkarbilliards, polyvinylpyrrolidine and wax.

Pharmaceutical compositions comprising the compound according to the invention can also be in liquid form, e.g. as solutions, emulsions, suspensions or syrups. Appropriate liquid supports can be, for example, water, organic solvents such as glycerol or glycols, as well as their mixtures, in various proportions, in water.

The drug according to the invention can be entered by topical, oral, parenteral, by injection (intramuscular, subcutaneous, intravenous, etc.), etc. route of administration depends, of course, on the type of disease.

Provided for the medicinal product according to the invention injected dose of from 0.1 mg to 10 g, depending on the type of pathology.

If not stated otherwise, all technical and scientific terms used in this description have the meanings commonly understood by a specialist in the field that applies the present invention. Similarly, all publications, patent applications, all patents and all other the links included in the present description by reference.

EXAMPLES

Example 1

7-(2-Amino-1-oxo-3-thiopropyl)-8-(cyclohexylmethyl)-2-(2-were)-5,6,7,8-tetrahydroimidazo[1,2A]pyrazin: 1

Connection 1 receive according to the following scheme of synthesis:

Scheme 1

1.a) Carbobenzoxy-L-cyclohexylamin

10.0 g (to 60.6 mmol) of L-Phenylalanine mixed with 430 mg PtO2in 60 ml of acetic acid and the mixture hydronaut when the hydrogen pressure of 1.38-3.45 bar during the night. To the mixture of aqueous 5%HCl solution to obtain a clear solution and the hydrogenation continued until the cessation of the consumption of hydrogen. The catalyst is filtered off and the filtrate is concentrated under reduced pressure. The residue is treated with methanol and water and the pH value is set equal to 4.4 by addition of 10%NaOH solution. The resulting product is extracted by filtration and used without further purification.

to 60.6 mmol L-Cyclohexylamine suspended in 100 ml of water, add at 8.36 g (to 60.6 mmol) To a2CO3then a solution of 15.1 g (to 60.6 mmol) of N-(benzyloxycarbonyloxy)succinimide in 150 ml of CH3CN and the mixture was intensively stirred for 45 minutes. The mixture is concentrated to a final volume of about 100 ml and washed with 100 ml of diethyl ether, then acidified with concentrated hydrochloric acid and extracted with 2 times 50 ml of ethyl acetate. United an ethyl acetate phase is dried over sodium sulfate, filtered and concentrated, obtaining 17,27 g (93%) of a clear oil.

1H-NMR (DMSO-d6[hexacyanometallate]), δ (ppm): 7.5 to about 7.6 (d, 1H); 7,2-75 (m, 5H); a 5.0 to 5.1 (s, 2H); 3,9-4,1 (m, 1H); 0,7-1,8 (m, 13H).

1.b) 2-(1-(S)-((Phenylmethoxy)carbonyl)amino-2-(cyclohexyl)methyl)-4-(2-were)imidazol

4,58 g (15.0 mmol) of Cbz-(L)-Cyclohexylamine and 2,44 g (7,50 mmol) Cs2CO3bring in 75 ml of a mixture of dimethylformamide with water in the ratio 2:1. The resulting mixture is stirred until then, until it becomes homogeneous. The solvents are removed under reduced pressure, the residue is dissolved in 60 ml of dimethylformamide and add 3,20 g (15.0 mmol) of 2-bromo-2'-methylacetophenone in 30 ml of dimethylformamide. The mixture is stirred over night at room temperature, then filtered and concentrated under reduced pressure. The obtained complex ketoester dissolved in 100 ml of a mixture of xylene and added 19.5 g (0.25 mol) of ammonium acetate.

The mixture is refluxed for about three hours when you remove the excess ammonium acetate and water released when using traps Dean-stark. The reaction mixture was concentrated under reduced pressure, treated with ethyl acetate, the resulting solution is washed with 100 ml of a saturated solution of NaHCO3and 100 ml saturated NaCl solution. An ethyl acetate phase is dried over sodium sulfate, filtered and concentrated in vacuo. The resulting crude product was then purified by flash chromatography on silica gel using mixtures of chloroform with me what anulom in the ratio 98:2 as eluent. Containing pure product fractions are combined and concentrated, obtaining 2,52 g (40%) of product as a slightly brown foam, which is used without further purification in the next stage.

1.) 2-(1-(S)-((Phenylmethoxy)carbonyl)amino-2-(cyclohexyl)methyl)-1-((2-ethoxy-2-oxo)ethyl)-4-(2-were)imidazol

2,52 g (6.0 mmol) of the Intermediate 1.b) was dissolved in 20 ml of dimethylformamide, is treated with a rate of 1.67 g (12.1 mmol) To a2CO3and add to 1.34 ml (12.5 mmol) of ethylbromoacetate. The resulting mixture is heated at a temperature of 45°With over one and a half hours. The mixture is diluted with 50 ml diethyl ether and the solution is washed with 50 ml of a saturated solution of NaHCO3and then with 50 ml saturated NaCl solution. The ether layer is dried over sodium sulfate, filtered and concentrated, obtaining an oil which is used without further purification in the next stage.

Mass spectrometry: 504,3 MN+.

1.d) 8-(Cyclohexylmethyl)-6-oxo-2-(2-were)imidazo[1,2A]pyrazin

The crude intermediate compound obtained in stage 1., is dissolved in 50 ml of acetic acid containing 152 mg of 10%palladium-on-coal as a catalyst, then hydronaut when the hydrogen pressure of 3.45 bar for 18 hours at room temperature. The catalyst is filtered off and the filtrate is heated at a temperature of 70°With the two hours. The resulting mixture was concentrated under reduced pressure, dissolved in 100 ml of dichloromethane and washed with 100 ml of a saturated solution of NaHCO3. The dichloromethane layer is dried over sodium sulfate, filtered and concentrated, obtaining a viscous oil which is used without further purification in the next stage.

Mass spectrometry: 324,3 MN+.

1.E) 8-(Cyclohexylmethyl)-2-(2-were)-4,5,6,7-tetrahydroimidazo[1,2A]pyrazin

The crude intermediate compound obtained in stage 1.d, dissolved in 25 ml of tetrahydrofuran and, at room temperature, treated with 25 ml of 1M solution NR3in tetrahydrofuran within half an hour, then refluxed for 1 hour. The mixture is cooled in a bath with ice and the temperature 0°With added dropwise 40 ml of 4 N. hydrochloric acid. The mixture is brought to room temperature, then refluxed for 1 hour. After that the reaction mixture is cooled and concentrated under reduced pressure. The residue is treated with 50 ml of a saturated solution of NaHCO3and extracted three times with 50 ml dichloromethane. Dichloromethane phase is dried over sodium sulfate, filtered and concentrated, obtaining 1.63 g (the yield is 87% compared to stages 1.c, 1.d, and 1.e) oil lightly brown.

Mass spectrometry: 310,3 MN+.

1.f) 8-(Cyclohex ylmethyl)-7-[2-(((1,1-dimethylmethoxy)carbonyl)amino)-1-oxo-3-((triphenylmethyl)thio)propyl]-2-(2-were)-4,5,6,7-tetrahydroimidazo[1,2-a]-piperazine

908 ál (5,80 mmol) Diisopropylcarbodiimide and lower than the 5.37 g (11.6 mmol) BocCys(trt)-HE dissolved in 25 ml of dichloromethane, the mixture is stirred for 45 minutes. Then add 1.63 g (5,27 mmol) 8-(cyclohexylmethyl)-2-(2-were)-4,5,6,7-tetrahydroimidazo[1,2A]pyrazine. The reaction mixture was stirred over night at room temperature. The solvent is removed under reduced pressure and the obtained product was then purified by flash chromatography on silica gel using mixtures of dichloromethane with methanol in the ratio 98:2 as eluent. Pure fractions are concentrated, receiving a viscous oil which is used without further purification in the next stage.

Mass spectrometry: 755,6 MN+.

1.g) 7-(2-Amino-1-oxo-3-(mercaptopropyl))-8-(cyclohexylmethyl)-2-(2-were)-4,5,6,7-tetrahydroimidazo[1,2A]piperazine: 1:

of 3.54 g (4,69 mmol) Obtained in stage 1.f intermediate compounds are dissolved in 80 ml triperoxonane acid (TFUC)containing 1,92 ml (9,38 mmol) triisopropyl-silane, and the reaction mixture was stirred at room temperature under nitrogen atmosphere for one hour. The reaction mixture is filtered and the filtrate concentrated under reduced pressure. The residue is extracted by powdering with water of 0.1%solution triperoxonane acid (6 times in 65 ml) and filtered. The crude product is purified by drug who ate high-performance liquid chromatography (HPLC) on a column with C 18using a gradient of 0-20% CH3CN in 0.1%aqueous solution triperoxonane acid for 30 minutes. Pure fractions of product are combined and lyophilizers. The initial product lyophilizer twice from dilute HCl solution, getting the product in the form of its hydrochloride (740 mg; 32%).

Mass spectrometry: 413,2 MN+.

1H-NMR (DMSO-d6), δ (ppm): 8,5-9,0 (l ush., 3H); 7,8-8,0 (s, 1H); 7.5 to 7.7 (d, 1H); 7,2-7,5 (m, 3H); 5,8-6,1 (m, 1H); 4,65-4,8 (s, 1H); 4,5-4,7 (d, 1H); 4,1-4,4 (m, 2H); 3,8-4,0 (m, 1H); 3,2-3,7 (H2O); 2,8-3,1 (m, 2H); 2,35 of 2.5 (s, 3H); 2,0-2,2 (m, 1H); 1,8-2,05 (m, 2H); 1,25-1,4 (ush., 4H); from 1.3 to 0.9 (m, 6N).

Example 2

7-(2-Amino-1-oxo-3-thiopropyl)-8-(cyclohexylmethyl)-2-phenyl-5,6,7,8-tetrahydroimidazo[1,2A]pyrazin: 2:

Connection 2 receive according to scheme 1, stage b-g, according to the method similar to that of example 1, replacing in stage b) 2-bromo-2'-methylacetophenone 2-bromoacetophenone.

Mass spectrometry: 399,2 MN+.

1H-NMR (DMSO-d6), δ (ppm): 8,5-8,9 (l ush., 3H); 8,0-8,2 (s, 1H); 7,8-8,0 (d, 2H); 7,45-7,56 (t, 2H); of 7.35-7.5 (t, 1H); 5.9 to 6,05 (ush., 1H); 4,65-4,8 (s, 1H); 4.5 to the 4.65 (d, 1H); 4.1 and of 4.35 (m, 2H); 3,8-4,0 (m, 1H); 3,2-3,8 (H2O); of 3.25 to 3.4 (t, 1H); 2.8 to 3,05 (m, 2H); 2,05-2,2 (d, 1H); 1.85 to 2.05 is (t, 2H); 1,55-1,75 (ush., 4H); 1,15-1,3 (ush., 1H); 1,2-0,9 (m, 5H).

Example 3

7-(2-Amino-1-oxo-3-thiopropyl)-2-(2-methoxyphenyl)-8-(phenylmethoxy)methyl-5,6,7,8-tetrahydroimidazo[1,2A]pyrazin: 3:

Connection 3 receive according to scheme 1, stage b-g, according to the method similar to that of example 1, ENEA in stage b) Cbz-(L)-cyclohexylamin on Boc-(L)-Ser(Bzl)-HE and replacing stage d) the stage of removal of the protective groups when using triperoxonane acid and triisopropylsilane, according to the method similar to that stage 1.g. The product is obtained in the form of a pair of diastereoisomers in the ratio 2:3.

Mass spectrometry: 453,2 MN+.

The retention time for diastereoisomeric respectively to 6.58 and 7.07 minutes in the following HPLC system:

eluent:30-50% of CH3CN/0,1%TFUK
the duration of elution:24 minutes
detection:254 nm
column:a Vydac protein and peptide C18

Example 4

7-(2-Amino-1-oxo-3-thiopropyl)-2-(2-methoxyphenyl)-8-(1-phenylmethoxy)ethyl-5,6,7,8-tetrahydroimidazo[1,2A]-pyrazin: 4:

Connection 4 receive according to scheme 1, stage b-g, according to the method similar to that of example 3, replacing, in stage b) of Boc-(L)-Ser(Bzl)-OH for Boc-(L)-Thr(Bzl)-HE. Mass spectrometry: 467,3 MN+.

1H-NMR (DMSO-d6), δ (ppm): 8,5-8,9 (l ush., 3H); to 8.0 and 8.1 (s, 1H); 7.95 is to 8.1 (d, 2H); of 7.4-7.5 (t, 1H); 7,15 of 7.3 (d, 1H); 7,0-7,2 (m, 3H); 6,9-7,05 (m, 2H); 5,85-5,95 (d, 1H); 4.75 V-4,85 (ush., 1H); 4,65-4,8 (ush., 1H); 4,35 with 4.65 (m, 3H); 4.1 and 4,25 (K, 2N); 3,9-4,0 (s, 3H); 2,8-3,1 (m, 2H); 1,2-1,4 (d, 3H).

Example 5

7-(2-Amino-1-oxo-3-thiopropyl)-2-(2-methoxyphenyl)-8-(phenoxyethyl)-5,6,7,8-tetrahydroimidazo[1,2A]pyrazin: 5:

Connection 5 receive according to the following scheme of synthesis:

Scheme 2

5.h) 2-(1-(S)-((is, teletaxi)carbonyl)amino-2-(2-oxo-2-(phenylmethoxy)ethyl)-4-(2-methoxyphenyl)imidazol

to 5.00 g (14.0 mmol) of Cbz-(L)-Asp(Obzl)-HE and 2.28 g (7,00 mmol) Cs2CO3mixed with 75 ml of a mixture of dimethylformamide with water in the ratio 1:1. The resulting mixture is stirred until then, until it becomes homogeneous. The solvents are removed under reduced pressure, the residue is dissolved in 60 ml of dimethylformamide and add 3,21 g (14.0 mmol) of 2-bromo-2'-methoxyacetophenone in 30 ml of dimethylformamide. The resulting mixture is stirred for half an hour at room temperature, then filtered and concentrated under reduced pressure. The obtained complex ketoester proscout with a mixture of diethyl ether and hexane (2 x 40 ml), then suspended in 100 ml of a mixture of xylenes. Add to 17.5 g (0.23 mol) of ammonium acetate and the mixture is refluxed for about one and a half hours while removing excess ammonium acetate and water released using the trap Dean-stark. The reaction mixture is washed with 50 ml of a saturated solution of NaHCO3, dried over sodium sulfate, filtered and concentrated in vacuo, obtaining of 6.66 g (98%) of the target product.

Mass spectrometry: 486,3 MN+.

5.i) 2-(1-(S)-((Phenylmethoxy)carbonyl)amino-2-((2-phenylmethoxy-2-oxo)ethyl)-1-((2-ethoxy-2-oxo)ethyl)-4-(2-methoxyphenyl)imidazol

The intermediate connection 5.i get according to the method similar to that phase 1.c.

Mass-Spa is trometry: 572,3 MN+.

5.j) (6-Oxo-2-(2-methoxyphenyl)-5,6,7,8-tetrahydroimidazo[1,2A]pyrazin-8-yl)-acetic acid

The intermediate connection 5.j receive according to the method similar to that stage 1.d.

Mass spectrometry: 302,2 MN+.

1H-NMR (DMSO-d6), δ (ppm): 8,35 to 8.5 (l ush., 1H); to 8.0 and 8.1 (DD, 1H); 7,45-of 7.55 (s, 1H); 7,15-7,25 (m, 1H); to 7.0, and 7.1 (m, 1H); 6,9-7,0 (m, 1H); 4,85-5,0 (ush., 1H); 4,55-4,75 (K, 2N); 3,85-3,95 (C, ZN); 2,8-2,95 (d, 2H).

5.k) 8-Hydroxyethyl-2-(2-methoxyphenyl)-5,6,7,8-tetrahydroimidazo[1,2-a]pyrazin

The intermediate connection 5.k receive according to the method similar to that phase 1.E except that they use the molar ratio NR3with respect to the substrate, comprising 6:9.

Mass spectrometry: 274,3 MN+.

5.1) 7-((1,1-Dimethylmethoxy)carbonyl)-8-hydroxyethyl-2-(2-methoxyphenyl)-5,6,7,8-tetrahydroimidazo[1,2A]pyrazin

of 1.36 g (5.0 mmol) of Intermediate compounds 5.k suspended in 5 ml of water and add a mixture of 1.20 g (5.5 mmol) of di-tert-BUTYLCARBAMATE in 10 ml of p-dioxane. The reaction mixture is intensively stirred and support when the pH value is average of 8.0-8.4 and by adding dropwise a 2.5 n NaOH solution until the end of the reaction (reaction monitored by thin layer chromatography on silica gel, elwira a mixture of ethyl acetate with hexane in the ratio 3:2). The crude product is purified by flash chromatography on silica gel using CME and ethyl acetate with hexane in the ratio of 3:2 as eluent (Biotage system, pre-filled columns 4×15 cm). Containing the product fractions are combined and concentrated in vacuo, getting to 1.60 g (86%) of a white foam.

Mass spectrometry: 374,3 MN+.

1H-NMR (DMSO-d6), δ (ppm): 7.95 is-with 8.05 (DD, 1H); 7,45-of 7.55 (s, 1H); 7,10-7,25 (m, 1H); to 7.0, and 7.1 (m, 1H); 6,9-7,05 (m, 1H); of 5.05-of 5.15 (t, 1H); 4,25 is 4.35 (t, 1H); 4,05-4,2 (ush., 1H); 4,0-4,1 (m, 1H); 3,9-4,0 (s, 3H); 3,9-4,0 (m, 1H); of 3.25 to 3.35 (m, 2H); 1,9-2,1 (m, 2H); 1,15-1,25 (s, N).

5.m) 7-((1,1-Dimethylmethoxy)carbonyl)-8-phenoxyethyl-2-(2-methoxyphenyl)-5,6,7,8-tetrahydroimidazo[1,2A]pyrazin

746 mg (2.00 mmol) of Intermediate compounds 5.1 dissolved in 10 ml of tetrahydrofuran containing 550 mg (2.1 mmol) of triphenylphosphine and 198 mg (2.1 mmol) of phenol. The mixture is cooled to a temperature of 0°C in an atmosphere of nitrogen and added dropwise within 10 minutes of 330 μl (2.1 mmol) of diethylazodicarboxylate. The reaction mixture was then stirred for two hours at room temperature. After that, the reaction mixture is again cooled to a temperature of 0°and add 275 mg (1.05 mmol) of triphenylphosphine and 99 mg (1.05 mmol) of phenol. Then within 10 minutes is added dropwise 166 μl (1.05 mmol) of diethylazodicarboxylate, after which the mixture is stirred for 1 hour at room temperature. The solvents are removed under reduced pressure and the crude product purified by flash chromatography on silica gel using mixtures of ethyl acetate with GE is sanami in the ratio of 3:2 as eluent. Containing the product fractions are combined and concentrated in vacuo. After recrystallization from ethyl acetate and hexanol get 863 g (96%) of the desired product in the form of a solid white color.

Mass spectrometry: 450,4 MN+.

5.n) 7-(2-(((1,1-Dimethylmethoxy)carbonyl)amino)-1-oxo-3-((triphenylmethyl)thio)-propyl)-2-(2-methoxyphenyl)-8-(phenoxyethyl)-4,5,6,7-tetrahydroimidazo[1,2A]piperazine

850 mg (1,89 mmol) Intermediate compound t process using a mixture of 10 ml triperoxonane acid containing 387 μl (1,89 mmol) of triisopropylsilane at room temperature for 20 minutes. The solvents are removed under reduced pressure and the crude product partitioned between 15 ml of ethyl acetate and 15 ml of a saturated solution of NaHCO3. An ethyl acetate phase is dried over sodium sulfate, filtered and concentrated under reduced pressure. The product from which the protective group is removed, connect with Boc-(L)-Cys(Trt)-according to the method similar to that stage 1.f (1.26 g; 84%).

5) 7-(2-Amino-1-oxo-3-thiopropyl)-2-(2-methoxyphenyl)-8-(phenoxyethyl)-4,5,6,7-tetrahydroimidazo[1,2-a]-piperazine

The product 5 is obtained from the intermediate 5.n according to the method similar to that stage 1.g.

Mass spectrometry: 274/3 MN+.

1H-NMR (DMSO-d6at a temperature of 90°), δ (ppm): 8,5-9,2 (ush., 3H); 7.95 is to 8.1 (d, 1H); 7,85 to 8.0 (s, 1H); of 7.35-7.5 (m, 1H); 7,15-7,35 (m, 3H);7,0-to 7.15 (t, 1H); 6,85-7,0 (m, 3H); 5,9-6,1 (ush., 1H); 4,5-4,8 (m ush., 2H); 4,15 is 4.45 (m ush., 3H); 3,9-4,0 (s, 3H); 3.75 to 4.0 (with mush., 1H); 2.8 to 3,05 (m ush., 2H); 2,55-2,75 (mush., 2H).

Example 6

7-(2-Amino-1-oxo-3-thiopropyl)-2-(2-methoxyphenyl)-8-(phenoxyethyl)-5,6,7,8-tetrahydroimidazo[1,2A]pyrazin in the form of a dimer: 6:

467 mg (0,687 mmol) of the Compound 5.0 dissolved in 25 ml of water and the pH value of the solution set equal to 7.2 by adding a water-diluted solution of NH4OH. Add acetonitrile, obtaining a clear solution, and the mixture is stirred at room temperature overnight. The crude product is purified by preparative HPLC on a column of C18using a gradient of 15-40% acetonitrile in 0.1%triflorum-susei acid for 50 minutes. Pure fractions of product are combined and lyophilizers. The initial product lyophilizer twice from dilute HCl solution, receiving the product as hydrochloride (161 mg; 45%).

Mass spectrometry: 903,5 MN+.

1H-NMR (DMSO-d6at a temperature of 90°), δ (ppm): 8,7-9,3 (ush., 3H); 7.95 is to 8.1 (d, 1H); 7,85 to 8.0 (s, 1H); of 7.35-7.5 (t, 1H); 7,1-7,3 (m, 3H); 7,0-to 7.15 (t, 1H); 6,8-7,0 (m, 3H); 5,85-6,1 (ush., 1H); 4,7-4,9 (ush., 1H); 4,45-4,7 (m ush., 1H); 4.1 and 4.5 (m ush., 4H); of 3.85 to 4.0 (s, 4H); 3,3-3,5 (m ush., 2H); 2,5-2,8 (m ush., 2H).

Example 7

7-(2-Amino-1-oxo-3-thiopropyl)-2-(2-methoxyphenyl)-8-(phenylsulfonyl)-5,6,7,8-tetrahydroimidazo[1,2A]-pyrazin: 7:

Connection 7 receive according to the following scheme of synthesis:

figure 3

7.p) 7-((1,1-Dimethylmethoxy)carbonyl)-2-(2-methoxyphenyl)-8-(phenylthiomethyl)-5,6,7,8-tetrahydroimidazo[1,2A]-pyrazin

of 1.23 g (3,30 mmol) Intermediate compound 5.1, 1,64 ml (6,60 mmol) tri-n-butylphosphine and 1.44 g (6,60 mmol) phenoldisulfonic mixed in 10 ml of tetrahydro-furan. The mixture is stirred at room temperature in an argon atmosphere for 4 hours. The solvents are removed under reduced pressure and the crude product purified by flash chromatography on silica gel using mixtures of ethyl acetate with hexane in the ratio of 1:1 as the eluent Containing the product fractions are combined and concentrated in vacuo, getting 1,43 g (93%) of product as a white foam.

Mass spectrometry: 466/3 MN+.

7.q) 7-((1,1-Dimethylmethoxy)carbonyl)-2-(2-methoxyphenyl)-8-(phenylsulfonyl)-5,6,7,8-tetrahydroimidazo[1,2A]pyrazin

650 mg (1,40 mmol) of the Intermediate product R dissolved in 10 ml dichloromethane and in several portions over a period of 10 minutes add 483 mg (2,80 mmol) 3-chloroperoxybenzoic acid. The mixture contribute in a column with silica and elute with a mixture of hexanol with ethyl acetate in the ratio of 7:3, then a mixture of hexanol with ethyl acetate in the ratio of 1:1, getting 220 mg (32%) of pure product.

Mass spectrometry: 498,3 MN+.

1H-NMR (DMSO-d6at a temperature of 30°), δ (ppm): of 7.9 to 8.0 (m, 3H); 77-7,85 (m, 1H); of 7.6 to 7.75 (m, 2H); 7,45-of 7.55 (s, 1H); 7,15-7,25 (m, 1H); 6,9-7,1 (m, 2H); 5.1 to a 5.25 (t, 1H); 4,1-4,3 (l ush., 1H); 4,0-to 4.15 (m, 1H); 3,8-4,0 (m, 1H); 3,85-of 3.95 (s, 3H); 3,6-3,8 (m, 1H); 3,4-3,6 (m, 1H); 3,2-3,4 (H2O plus a fuzzy signal); of 1.9-2.3 (m, 2H); 1.3 to 1.5 (s, N).

Stage 7.n)

Stage 7.n carried out according to the method similar stage 5.n. The crude product is used without additional purification in the next stage.

Stage a)

Stage o carried out according to the method similar stage 5.o.

Mass spectrometry: 501,3 MN+.

Obtaining the compounds of formula (VII):

This compound is close to that described in International patent application PCT 97/30053 compounds can be obtained according to the following scheme of synthesis:

where

R'=H or CH3;

Cys=cysteine; Cys-al=cysteinyl;

Pen=penicillamine; Pen-al=penicillamine;

Boc=tert-butoxycarbonyl;

EDC=N-(3-dimethylaminopropyl)-N-ethylcarbodiimide;

HOBT=hydroxybenzotriazole;

Trt=triphenylmethyl.

PHARMACOLOGICAL

In order to clarify the usefulness of the invention to study the effect of processing lines of human MCF-7 cells using the following connections:

bis-1,1'-[7-(2-amino-1-oxo-3-thiopropyl)-8-(cyclohexylmethyl)-2-(2-methoxyphenyl)-5,6,7,8-tetrahydroimidazo[1,2-a]pyrazin]disulfide, referred to in this part as the compound (I);

bis-1,1'-[7-(2-amino-1-oxo-3-thiopropyl)-2-(3-naphthyl)-8-(2-methylprop who yl)-5,6,7,8-tetrahydroimidazo[1,2A]pyrazin-7-yl]disulfide, referred to in this part as the compound (II);

- 7-(2-amino-1-oxo-3-thiopropyl)-8-(cyclohexylmethyl)-2-(2-were)-5,6,7,8-tetrahydroimidazo[1,2A]pyrazin, referred to in this part as the compound (III);

the compound of the formula:

referred to in this part as the compound (IV);

- 7-(2-amino-1-oxo-3-thiopropyl)-8-butyl-2-(2-methoxyphenyl)-5,6,7,8-tetrahydro-imidazo[1,2A]pyrazin, referred to in this part as the compound (V);

bis-1,1'-[7-(2-amino-1-oxo-3-thiopropyl)-2-(methoxyphenyl)-8-(1-methylpropyl)-5,6,7,8-tetrahydroimidazo[1,2A]pyrazin]disulfide, referred to in this part as the compound (VI);

the compound of the formula:

referred to in this part as the compound (VII).

Methods

Cell line

Cell lines MCF-7 (human pleural cells; breast cancer) were purchased from the American collection of tissue culture (Rockville, Maryland, USA).

Determination of the intracellular amount of camp in the case of MCF-7 cell

Cells MCF-7 (2·104cells per well), the planting of which is performed in 24-hole plates, cultured for 5 days in a modified method of Dulbecco environment Needle (Gibco-Brl, Cergy-Pontoise, France)supplemented with 10% inactivated by heating fetal calf serum (Gibco-Brl, Cergy-Pontoise, France), 50000 ediniz penicillin and 50 mg/l streptomycin (Gibco-Brl, Cergy-Pontoise, France), and 2 mm glutamine (Gibco-Brl, Cergy-Pontoise, France). The culture medium was replaced after two washes with medium without serum, supplemented or not by certain agents at the time specified in the various figures. Then at a temperature of 37°With added agents that activate the production of camp. The reaction was stopped after 30 minutes by removing the medium and quickly add 100 μl of 0,1N. HCl solution. These extracts were frozen at a temperature of -80°to their use. The concentration cGMP was determined using the sales set to measure (number NEK033 NEN, Les Ulis, France), following the manufacturer's instructions. Radioactivity was determined using a gamma counter (Gamma Master-1277, LKB, Turku, Finland).

Determination of cell proliferation in vitro

Cells MCF-7 (3000 cells per well) were cultured in 96-well tablets in 80 μl of modified according to the method of Dulbecco eagle medium (Gibco-Brl, Cergy-Pontoise, France)supplemented with 10% inactivated by heating fetal calf serum (Gibco-Brl, Cergy-Pontoise, France), 50,000 units/l of penicillin and 50 mg/l streptomycin (Gibco-Brl, Cergy-Pontoise, France), and 2 mm glutamine (Gibco-Brl, Cergy-Pontoise, France), produced seeding in 96-well plate on day 0. Cells were treated on day 1 within 96 hours increasing concentrations up to 50 µm of each of the tested compounds is rd. At the end of this period the quantitative assessment of cell proliferation was performed by a colorimetric test based on the splitting of the salt of tetrazole WST1 by mitochondrial dehydrogenase in viable cells, leading to the formation of formazan (Boehringer Mannheim, Milan, France). These tests were carried out two times, with 8 definitions for the tested concentration. For each of the tested compounds were considered included in the linear part of the sigmoid values for analysis by linear regression and used to determine the inhibitory concentration (IC50).

The determination of the activity of MAR-kinase

Cells MCF-7 (5105 cells per well) were cultured in sectionone tablets in modified according to the method of Dulbecco environment Needle (Gibco-Brl, Cergy-Pontoise, France)supplemented with 10% inactivated by heating fetal calf serum (Gibco-Brl, Cergy-Pontoise, France), with a mixture of antibiotics from 50,000 units/l of penicillin and 50 mg/l streptomycin (Gibco-Brl, Cergy-Pontoise, France) and 2 mm glutamine (Gibco-Brl, Cergy-Pontoise, France). After culturing for 24 hours, the cells were incubated for 48 hours in medium containing no serum, to bring the cells in the quiescent state. Cells were then treated for 1 hour or compound (I)or PD98059 (Calbiochem, France Biochem, Meudon, France), specific ing is an inhibitor of activation of the MAR-kinase. Cells are then stimulated (or not) for 5 minutes using a 12.5 ng/ml epidermal growth factor (EGF). The reaction was stopped by two washes with phosphate buffered saline (Gibco-Brl, Cergy-Pontoise, France), containing neither calcium nor magnesium, at a temperature of 4°and by adding 150 ál of buffer for lysis at a temperature of 4°Since whose composition is the following: 10 mmol Tris, 150 mmol NaCl, 2 mmol of ethylenebis(oxyethylenenitrilo)tetraoxane acid, 2 mmol of dithiothreitol, 1 mmol phenyl-methylsulfonylamino, 2 mmol of orthovanadate, 10 μg/ml leupeptin and 10 µg/ml Aprotinin. The number of proteins present in the extracts was determined by the method of Bradford (Biorad reagents, Ivry-Sur-Seine, France). These extracts were frozen at a temperature of -80°to their use. Activity MAR-kinase was determined using the sales set to measure (number RPN 84, Amersham, Life Science, Les Ulis, France), following the manufacturer's instructions. Radioactivity was determined using a scintillation counter Packard company (Tricarb 5000 SA).

Material

Vasointestinal peptide (VIP) was purchased from company Bachem (Voisins le Bretonneux, France). Cholera toxin, Forskolin, isoproterenol, prostaglandin E2and PD 98059 were purchased from the firm Calbiochem (France Biochem, Meodon, France). Compounds of formula (I), (II), (III), (IV), (V), (VI) and (VII) were provided to encourage company is Biomeasure Inc. (Milford, MA, USA). All these compounds were used in accordance with the recommendations of their manufacturers.

Results

The figure 1 shows that the activation of adenylate cyclase by cholera toxin (200 ng/ml) or Forskolin (10 Microm) leads to a very significant increase in the number of camp. Pretreatment of cells for 30 minutes with 30 mmol of compound (I) does not change the production of camp induced a direct activator of adenylate cyclase, Forskolin. At the same time, the products cGMP stimulated direct activator subunit, by cholera toxin, is strongly inhibited by compound (I). This shows that the adenylate cyclase itself is not modified by the compound (I) and that it prevents the formation of heterotrimeric complex.

VIP presents as extracellular ligand receptor associated with protein G, which stimulates the synthesis of camp in human breast cancer cells. The figure 2 shows that treatment with VIP human breast cancer cells MCF-7 increases the intracellular amount of camp-dependent concentration. The concentration of VIP, component 10 nmol, which is almost optimal products cdmp use for the following tests. This concentration is consistent with already published data concerning the line of human cancer cells milk is cancer T47D.

The figure 3 shows that pre-treatment within 30 minutes of MCF-7 cells, derived from cultures in vitro, the compound of the formula (I) is sufficient for inhibition dependent on the concentration of the way of accumulation of camp stimulated with VIP. Almost complete inhibition was achieved with concentree 100 µmol the compounds of formula (I). These results show that treatment with compound (I) is sufficient to block signal transduction path, which are used heterotrimeric G proteins as mediators.

The figure 4 shows that treatment within one hour of the compound of the formula (I) is sufficient to modify the response to VIP. Treatment for longer periods of time (8 hours and 24 hours) continue to inhibit camp, but the main effect is achieved very quickly.

The compound (I) can also inhibit the formation of cyclic amp induced by other agents that stimulate the receptors of the seven transmembrane domains. In MCF-7 cells, for example, adenylate cyclase activity, greatly increase the prostaglandin E2that is inhibited by treatment for 30 minutes, compound (I). This suggests that the treatment of cells with compound (I) modifies the heterotrimeric form of protein G due to the dissociation of the a subunit dimer ² /γ.

Inhibition of stimulation with VIP was not reduced by the connection structure similar to that of the compounds of formula (I). As shown in table I, the compound (II), (III), (IV), (V), (VI) and (VII), tested using the same model, can also reduce induced with VIP number of camp.

Collectively, these results suggest that the tested compounds modulate the activity of adenylate cyclase through modification of heterotrimeric forms of protein G. However, it is known that the dimers β/γ can directly activate effectors, leading to activation of kinases that are regulated by extracellular signals (ERKs), or MAR-kinases.

The figure 6 shows that treatment of cells within 1 hour of compound (I) increases twice basal activity MAR-kinase. This suggests that by preventing the formation of the heterotrimeric complex of the compound (I) releases heterodimer, which itself remains bound to the membrane and activates the path of ras. However, figure 7 shows that after stimulation for 5 minutes MAR-kinase epithelial growth factor (EGF), enzyme activity increases by about 7 times. Pre-treatment for 1 hour cell or compound (I)or with PD98059, a specific inhibitor of the activation of map-kinase, reduces energy is 2 times the activity MAR-kinase. These results suggest that the compound (I) stimulates in the basal state-of-way ras and inhibits the same way, if it is stimulated, showing thus their antiproliferative action.

Indeed, table II shows that the compounds (I), (II), (III) and (IV) are able to inhibit proliferation in vitro human tumor cells MCF-7.

Table I

Effects of compounds (I), (II), (III) and (IV), inkubiruemykh for 30 minutes, then stimulated with VIP products cGMP in the MCF-7 cell
ConnectionInhibition at a concentration of 30 µmol
(I)86%
(II)71%
(III)59%
(IV)52%
(V)68%
(VI)52%
(VII)65%

Cells were incubated for 30 minutes in the presence or not of compounds (I), (II), (III) and (IV) at a concentration of 30 µmol, which are then stimulated with 10-8mol VIP. The amount of camp was determined by radioimmunoassay analysis. Data represent the average value of ± standard deviation (n=5 for control and n=1 for different connections).

Table II

Inhibition of growth in vitro MCF-7 cell compounds (I), (II), (III) and (IV)
Test connectionIR50(µmol)
the compound (I)9,4
the compound (II)15,0
the compound (III)16,1
the compound (IV)34,6

The results of IR50expressed in µmol and represent the average of two experiments.

FIGURES

Figure 1: Effect of compound (I) stimulated by cholera toxin or Forskolin production of camp in cells MCF-7.

Cells were incubated for 30 minutes in the presence or not of the compound (I) in a concentration of 30 µmol and then stimulated either by cholera toxin (200 ng/ml) for 90 minutes, or Forskolin (10-5mol) over 30 minutes. The amount of camp was determined by radioimmunoassay analysis. Data represent the average value of ± standard deviation (n=3).

Figure 2: Effect of increasing concentrations of VIP products cGMP in MCF-7 cells.

Cells were stimulated for 30 minutes with the VIP in these concentrations. The amount of camp was determined by radioimmunoassay analysis.

Figure 3: Effect of different concentrations of compound (I) on promoting the ing with VIP production of camp in cells MCF-7.

Cells were incubated for 24 hours in the presence of increasing concentrations of compound (I) and then stimulated for 30 minutes with 10-8mol VIP. The amount of camp was determined by radioimmunoassay analysis. Data represent the average value of ± standard deviation (n=3).

Figure 4: Effect of time of incubation of MCF-7 cells in the presence of compound (I) stimulated with VIP production of camp.

Cells were incubated for 1, 8 or 24 hours in the presence or not of the compound (I) in a concentration of 30 µmol and then stimulated with 10-8mol VIP. The amount of camp was determined by radioimmunoassay analysis. Data represent the average value of ± standard deviation (n=1).

Figure 5: Effect of compound (I) stimulated with VIP, isoproterenol or prostaglandin E2production of camp in cells MCF-7.

Cells were incubated for 30 minutes in the presence or not of the compound (I) in a concentration of 30 µmol and then stimulated for 30 min or with 10-8mol VIP, or with 10-6mol isoproterenol or with 10-6mol prostaglandin E2. The amount of camp was determined by radioimmunoassay analysis. Data represent the average value of ± standard deviation (n=3).

Figure 6:Effect of compound (I) on basal activity MAR-kinase in MCF-7 cells.

Cells deprived of serum for 48 hours, were treated for 1 hour compound (I) (10 and 50 Microm). Activity MAR-kinase activity was determined in cell lysates and expressed in pmol included Pi per minute and 4 µg of protein (n=3 for control samples and n=2 is treated with compound (I) cells).

Figure 7: Effect of compound (I), compared with PD98059, on the activity MAR-kinase in MCF-7 cells.

Cells deprived of serum for 48 hours, were treated for 1 hour or compound (I) (10 and 50 Microm)or with PD98059 (10 and 50 Microm). Cells are then stimulated by the addition of 12.5 ng/ml epidermal growth factor for 5 minutes. Activity MAR-kinase activity was determined in cell lysates and expressed in pmol included Pi per minute and 4 µg of protein (n=4).

1. The use of compounds of General formula

or

in which R1means cyclohexylmethyl, 2-methylpropyl, butyl, 1-methylpropyl;

R2represents 2-methoxyphenyl, 1-naphthyl, 2-were;

R3means a hydrogen atom or a group

or their pharmaceutically acceptable salts for obtaining a medicinal product intended for the treatment of Pato is Ogii, arising due to the formation of heterotrimeric protein selected among the pathologies associated with the following biological functions or disorders: smell, taste, light perception, neurotransmission, neurodegeneration, the functioning of the endocrine and exocrine glands, autocrine and paracrine regulation of arterial blood pressure, embryogenesis, viral infection, immune function, diabetes, and obesity.

2. The use according to claim 1, characterized in that the compound is chosen from one of the following connections:

7-(2-amino-1-oxo-3-thiopropyl)-8-butyl-2-(2-methoxyphenyl)-5,6,7,8-tetrahydroimidazo[1, 2A]pyrazin;

bis-1,1'-[7-(2-amino-1-oxo-3-thiopropyl)-2-(methoxyphenyl)-8-(1-methylpropyl)-5,6,7,8-tetrahydroimidazo[1, 2A]pyrazin]disulfide;

bis-1,1'-[7-(2-amino-1-oxo-3-thiopropyl)-8-(cyclohexylmethyl)-2-(2-methoxyphenyl)-5,6,7,8-tetrahydroimidazo[1, 2A]pyrazin]disulfide;

bis-1,1'-[7-(2-amino-1-oxo-3-thiopropyl)-2-(1-naphthyl)-8-(2-methylpropyl)-5,6,7,8-tetrahydroimidazo[1, 2A]pyrazin-7-yl]disulfide.

3. Product representing one of the following connections:

7-(2-amino-1-oxo-3-thiopropyl)-8-(cyclohexylmethyl)-2-(2-were)-5,6,7,8-tetrahydroimidazo[1, 2A]pyrazin;

7-(2-amino-1-oxo-3-thiopropyl)-2-(2-methoxyphenyl)-8-(phenylmethoxy)methyl-5,6,7,8-tetrahydroimidazo[1, 2A]pyrazin

7-(2-amino-1-oxo-3-thiopropyl)-2-(2-methoxyphenyl)-8-(1-phenylmethoxy)ethyl-5,6,7,8-tetrahydroimidazo[1, 2A]-pyrazin;

7-(2-amino-1-oxo-3-thiopropyl)-2-(2-methoxyphenyl)-8-(phenylsulfonyl)-5,6,7,8-tetrahydroimidazo[1, 2A]pyrazin,

or pharmaceutically acceptable salt of one of these compounds.

4. The product according to claim 3 or a pharmaceutically acceptable salt of the product as a medicinal product for the treatment of pathologies caused by the formation of heterotrimeric protein G.

5. The product according to claim 4, wherein the pathology is related to one of the following biological functions or disorders: smell, taste, light perception, neurotransmission, neurodegeneration, the functioning of the endocrine and exocrine glands, autocrine and paracrine regulation of arterial blood pressure, embryogenesis, benign cell proliferation, oncogenesis, viral infection, immune function, diabetes, obesity, and benign and malignant proliferative diseases.

6. Pharmaceutical composition for the treatment of pathologies caused by the formation of heterotrimeric G protein, including as applicable the beginning of the connection according to claims 1 to 4 or a pharmaceutically acceptable salt of the compound.



 

Same patents:

FIELD: organic chemistry, chemical technology, medicine, pharmacy.

SUBSTANCE: invention describes bicyclic N-acylated imidazo-3-amines or imidazo-5-amines salts of the general formula (I): wherein R1 means tert.-butyl, 1,1,3,3-tetramethylbutyl, (C4-C8)-cycloalkyl, phenyl disubstituted with (C1-C4)-alkyl, -CH2Ra wherein Ra means the group -CO(OR') wherein R' means (C1-C8)-alkyl; R2 means hydrogen atom, the group -CORb wherein Rb means (C1-C8)-alkyl or (C3-C8)-cycloalkyl; R3 means (C1-C8)-alkyl, (C3-C8)-cycloalkyl, phenyl, pyridyl, furfuryl or thiophenyl; A means tri-linked fragment of ring of the formula: wherein R6 and R7 mean hydrogen atom or tetra-linked fragment of ring of the following formulae: wherein R4' means hydrogen atom or benzyloxy-group; R5' means hydrogen atom; R6' means hydrogen atom, (C1-C8)-alkyl or nitro- (NO2)-group; R7' means hydrogen atom, (C1-C8)-alkyl, or R6' and R7' mean in common the following fragment of ring: -CRi=CRj-CH=CH- wherein Ri and Rj mean hydrogen atom; R5'' means hydrogen, chlorine atom or (C1-C8)-alkyl; R6'' means hydrogen atom; R7''n means hydrogen atom, amino- (NH2)-group or (C1-C8)-alkyl; R4''', R6''' and R7''' mean hydrogen atom; R8 means (C1-C8)-alkyl or (C3-C8)-cycloalkyl; X means anion of inorganic or organic acid, or their acid-additive compounds. Also, invention relates to a method for their preparing and a pharmaceutical composition based on thereof. These new compounds show affinity to opiate μ-receptor and can be used, in particular, as analgesic agents.

EFFECT: improved preparing method, valuable medicinal properties of compounds and pharmaceutical compositions.

12 cl, 2 dwg, 32 ex

FIELD: organic chemistry, medicine, hematology.

SUBSTANCE: invention elates to new compounds that inhibit activated blood coagulating factor X (Fxa factor) eliciting the strong anti-coagulating effect. Invention proposes compound of the formula (1): Q1-Q2-C(=C)-N-(R1)-Q3-N(R2)-T1-Q4(1) wherein R1, R2, Q1, Q2, Q4 and T1 have corresponding values, and Q2 represents the group of the formula: wherein R9, R10 and Q5 have corresponding values also, or its salt, solvate or N-oxide. Invention provides the development of a novel compound possessing strong Fxa-inhibiting effect and showing the rapid, significant and stable anti-thrombosis effectin oral administration.

EFFECT: valuable medicinal properties of compounds.

13 cl, 1 tbl, 195 ex

FIELD: organic chemistry, biochemistry, pharmacy.

SUBSTANCE: invention relates to new anellated carbamoyl azaheterocycles of the general formula (1)

or (2) possessing the inhibitory effect on protein kinase activity, a focused library comprising these compounds, and pharmaceutical composition based on thereof. In the general formula (1) or (2) R1 represents hydrogen atom or optionally substituted (C1-C6)-alkyl; R2 and R3 represent independently of one another hydrogen atom, inert substitute, optionally substituted (C1-C6)-alkyl, optionally substituted (C3-C8)-cycloalkyl, optionally substituted phenyl, optionally substituted aryl, optionally substituted heterocyclyl; R4 represents optionally substituted (C1-C6)-alkyl, optionally substituted (C3-C8)-cycloalkyl, optionally substituted phenyl, optionally substituted aryl, optionally substituted heterocyclyl; A and B in common with carbon and nitrogen atoms joined to the form an optionally substituted and optionally condensed azaheterocycle; D and F in common with carbon atoms joined form an optionally substituted and optionally condensed phenyl or aryl, optionally substituted and optionally condensed azaheterocycle. K and L in common with carbon and nitrogen atoms joined to them form an optionally substituted azaheterocycle. Also, invention related to methods for preparing compounds of the general formulae (1) or (2).

EFFECT: improved preparing methods.

10 cl, 2 sch, 25 tbl, 7 ex

FIELD: organic chemistry, biochemistry, pharmacy.

SUBSTANCE: invention relates to a heteroarylamino-substituted derivative of dihydropyrimido[4,5-d]pyrimidinone taken among of compounds order corresponding to the formula (I): wherein a subscript symbol n mans a whole number 1; R1 means (C1-C6)-alkyl (substituted with one or two substitutes taken among group involving hydroxy group, (C1-C6)-alkoxy group and others), piperidinyl-(C0-C4)-alkyl [wherein piperidinyl fragment is monosubstituted optionally with benzyl, carbamoyl, (C1-C4)-alkane sulfonyl, (C1-C6)-alkyl and so on], morpholinyl-(C0-C4)-alkyl, tetrahydropyranyl-(C0-C4)-alkyl, 2-oxoimidazolidinyl-(C0-C4)-alkyl, 2-oxopyrrolidinyl-(C0-C4)-alkyl or 1,1-dioxotetrahydrothienyl-(C0-C4)-alkyl, (C3-C6)-cycloalkyl (monosubstituted with monohydroxy group, (C1-C6)-alkoxy group and so on), 1,4-dioxaspiro[4,5]decane-8-yl, 2,4-dione-1,3-diazaspiro[4,5]decane-8-yl or (3-hydroxymethyl-3-methyl)-1,5-dioxaspiro[5,5]undecane-9-yl; R2 means (C1-C4)-alkyl, halogen atom; R3 means hydrogen atom, (C1-C6)-alkyl (optionally substituted with one or two substitutes taken among group involving (C1-C4)-alkoxy group, pyrrolidinyl, di-(C1-C4-alkyl)-amino-group and so on), phenyl, benzyl or piperidinyl (N-substituted optionally with (C1-C4)-alkyl); R4 means hydrogen atom, and also its individual isomers, racemic and nonracemic mixtures of isomers, prodrugs and its pharmaceutically acceptable salts. Also, invention proposes a pharmaceutical composition possessing inhibitory activity with respect to activity of p38 MAP kinase. The composition comprises a heteroalkylamino-derivative of dihydropyrimido[4,5-d]pyrimidinone of the formula (I), isomer, racemic or nonracemic mixture of isomers or its pharmaceutically acceptable salt in mixture with at least one pharmaceutically acceptable vehicle. Invention provides representing a heteroalkylamino-substituted derivative of dihydropyrimido[4,5-d]pyrimidinone possessing inhibitory activity with respect to activity of p38 MAP kinase.

EFFECT: valuable biochemical properties of compounds and composition.

14 cl, 4 tbl, 90 ex

FIELD: organic chemistry of heterocyclic compounds, biochemistry, pharmacy.

SUBSTANCE: invention describes alkylamino-substituted bicyclic nitrogen-containing heterocycles of the general formula (I):

wherein n = 1; R1 means (C1-C6)-alkyl; R2 means halogen atom; R3 means (C1-C6)-alkoxy-(C1-C6)-alkyl, (C1-C6)-alkylsulfonyl-(C1-C6)-alkyl, hydroxy-(C1-C6)-alkyl, dihydroxy-(C1-C6)-alkyl, N-heterocyclyl-(C1-C6)-alkyl or (C1-C6-alkylene)-C(O)R31 wherein R31 means hydroxy- or (C1-C6)-alkoxy-group, and its pharmaceutically acceptable salts. New compounds are inhibitors of protein kinase p38 and can be used in medicine.

EFFECT: valuable medicinal properties of compounds.

8 cl, 13 ex

FIELD: organic chemistry, medicine, pharmacy.

SUBSTANCE: invention relates to novel substituted 2-aryl-3-(heteroaryl)imidazo[1,2-a]-pyrimidines of the formula (I):

or to their pharmaceutically acceptable salts wherein: (a) R1 is taken among the group consisting of -NH2, C1-5-alkylamino-, di-C1-5-alkylamino-, phenylmethylamino-group; (b) Y is taken among the group consisting of hydrogen atom (H), halogen atom, piperidine, OR4, SR4, -SO2CH3, NHR4 and NR4R5 wherein R4 and R5 are taken independently among hydrogen atom (H), α-alkylphenyl-C1-5-alkyl, linear or branched alkyl substituted optionally with C3-5-carbocycle, phenyl or substituted phenyl wherein indicated phenyl can be substituted with one or some substituted taken among C1-5-alkoxy-group; (c) R2 represents from one to five members taken independently among the group including hydrogen atom (H), halogen atom, trifluoromethyl; (d) R3 represents hydrogen atom (H), or radicals R3 taken in common form aromatic ring; (e) X represents nitrogen atom (N) or -CH. Also, invention relates to methods for preparing indicated compounds and to a method for treatment based on these compounds. Invention provides preparing novel compounds that can be used in relief states by reducing the level of inflammatory cytokines, for example, the indicated state represents proliferative (rheumatic) arthritis.

EFFECT: valuable medicinal properties of compounds and compositions.

40 cl, 1 tbl, 4 ex

FIELD: organic chemistry, chemical technology, pharmacy.

SUBSTANCE: invention describes derivatives of imidazo-3-ylamine of the general formula (I):

wherein X and Y mean CH or nitrogen atom (N) under condition that X and Y don't mean nitrogen atom (N) simultaneously; R1 means tert.-butyl, (CH2)nCN wherein n means 4, 5 or 6, phenyl substituted optionally with (C1-C4)-alkyl, (C1-C4)-alkoxy-group, (C4-C8)-cycloalkyl, 1,1,3,3-tetramethylbutyl or CH2Ra wherein Ra represents hydrogen atom, branched or linear (C1-C8)-alkyl, phenyl substituted optionally with halogen atom, (C1-C4)-alkoxy-group, CO(OR') wherein R' means linear (C1-C4)-alkyl or branched (C3-C5)-alkyl, PO(OR')2 wherein R' means linear (C1-C4)-alkyl or branched (C3-C5)-alkyl; R2 means hydrogen atom, CORb wherein Rb represents branched or linear (C1-C4)-alkyl; R3 means methyl, ethyl, tert.-butyl, (C3-C8)-cycloalkyl, phenyl monosubstituted optionally at position 3, 5 or 6 or optionally multisubstituted at position 4 and additionally at position 2 and/or 3, and/or 5, and/or 6 with halogen atom, hydroxyl group (OH), (C1-C4)-alkyl or (C1-C4)-alkoxy-group, naphthyl, optionally substituted (C1-C4)-alkoxy-group, di-(C1-C4)-alkylamino-group, pyrrole substituted optionally with (C1-C4)-alkyl, benzylsulfonyl, COOCH3, pyridyl substituted optionally with (C1-C4)-alkyl, OH, hydroxy-(C1-C4)-alkyl, furan substituted optionally with (C1-C4)-alkyl, nitro-group (-NO2), halogen-substituted phenyl, CH2COOCH3, COOH, thiophene substituted optionally with halogen atom, (C1-C4)-alkyl, (C1-C4)alkylsulfanyl, -NO2, phenoxy-group, thiophene, alkynylphenyl, unsubstituted anthracene or quinoline substituted optionally with halogen atom under condition that R3 doesn't means cyclohexyl-unsubstituted phenyl or phenyl monosubstituted with carboxylic acid amide at position 3 if R1 means tert.-butyl, n-propyl, n-butyl, 1,1,3,3-tetramethylbutyl, cyclohexyl, monosubstituted phenyl, 2,6-dimethylphenyl or benzyl, and R2 means simultaneously hydrogen atom or -CO-(methyl) and under condition that R2 doesn't mean hydrogen atom if R1 means benzyl simultaneously and R3 means methyl or R1 means simultaneously CH2C(O)-tert.-butyl and R3 means unsubstituted phenyl, in forms of bases or pharmaceutically acceptable salts, and a method for their preparing and a medicinal agent based on thereof. Described compounds possess analgesic activity and can be used in medicine.

EFFECT: improved preparing method, valuable medicinal properties of compounds and agent.

7 cl, 2 tbl, 33 ex

FIELD: organic chemistry, chemical technology, medicine.

SUBSTANCE: invention relates to new derivatives of pyrrolopyrimidine of the formula (1) and their pharmaceutically acceptable salts possessing properties of selective inhibitor of specific cyclic guanosine 3',5'-monophosphate phosphodiesterase (specific cGMP PDE) (PDE V). In the formula (1) R1 represents hydrogen atom (H), (C1-C3)-alkyl substituted optionally with one or some fluorine atoms; R2 represents H, halogen atom, (C1-C6)-alkyl substituted optionally with hydroxyl group (-OH), (C1-C3)-alkoxy-group, (C3-C6)-cycloalkyl or one or some fluorine atoms, (C3-C6)-cycloalkyl; R3 represents (C1-C6)-alkyl substituted optionally with (C3-C6)-cycloalkyl or one or some fluorine atoms; R4 represents (C1-C6)-alkyl substituted optionally with one or some fluorine atoms; R5 represents -SO2NR6R, -NHSO2R8 or heterocyclyl such as tetrazolyl; each R6 and R7 represents independently H or (C1-C6)-alkyl substituted optionally with -CO2H or one or some fluorine atoms; or in common with nitrogen atom to which they are bound form monocylic ring, such as imidazole, pyrrolidine, piperidine, morpholine, piperazine and homopiperazine wherein indicated group is replaced optionally with R9 wherein R9 represents (C1-C6)-alkyl substituted optionally with one or some halogen atoms, hydroxyl group (OH), (C1-C3)-alkoxy-group that is replaced optionally with one or some fluorine atoms, -NR11R12, -C=NR13(NR14R15) or tetrazolyl group, 6-membered nitrogen-containing heteroaryl group; each R11 and R12 represents independently H or (C1-C4)-alkyl; R13represents H; each R14 and R15 represents independently H. Also, invention relates to intermediate compounds, methods for preparing compounds and pharmaceutical compositions. Proposed compounds can be used in treatment of impotency, sexual dysfunction in females, stable, nonstable and variant (Prinzmental) stenocardia and other diseases also.

EFFECT: improved preparing method, valuable medicinal properties of compounds.

15 cl, 1 tbl, 250 ex

FIELD: organic chemistry, medicine, hormones.

SUBSTANCE: invention describes imidazole derivatives of the formula (I) , racemic-diastereomeric mixtures and optical isomers, pharmaceutical salts wherein ---- represents an optional bond; R1 represents hydrogen atom (H), -(CH2)m-C(O)-(CH2)m-Z1, -(CH2)m-Z1; R2 represents hydrogen atom (H), or R1 and R2 are joined with nitrogen atoms to which they are bound forming compounds represented by formulae (Ia), (Ib) or (Ic) wherein R3 represents -(CH2)m-E-(CH2)m-Z2; R4 represents hydrogen atom (H) or -(CH2)m-A1; R5 represents (C1-C12)-alkyl, (C0-C6)-alkyl-C(O)-NH-(CH2)m-Z3 and optionally substituted phenyl; R6 represents hydrogen atom (H); R7 represents (C1-C12)-alkyl or -(CH2)m-Z4; m = 0 or a whole number from 1 to 6; n is a whole number from 1 to 5. Proposed compounds bind with subtypes of somatostatin receptors selectively.

EFFECT: valuable properties of compounds.

20 cl, 13776 ex

FIELD: organic chemistry, biochemistry, medicine, pharmacy.

SUBSTANCE: invention describes 2-phenyl-substituted imidazotriazinones of the general formula (I): wherein R1 and R2 mean independently linear (C1-C4)-alkyl; R3 and R4 are similar or distinct and represent hydrogen atom or linear or branched (C1-C4)-alkenyl or (C1-C4)-alkoxy-group, linear or branched (C1-C6)-alkyl chain that can be broken by oxygen atom, and/or it can comprise from to some similar or different the following substitutes: methoxy-, hydroxy-, carboxyl, linear or branched (C1-C4)-alkoxycarbonyl, and/or residues of formulae -SO3H, -(A)a-NR7R8, -O-CO-NR7'R8', and/or wherein A means a number 0 or 1; A means residue -CO or -SO2; R7 and R8 mean hydrogen atom (H), cyclopentyl, cyclohexyl, cycloheptyl, phenyl, piperidinyl or pyridyl that can be substituted with different substitutes, methoxy-, (C1-C6)-alkyl and others; R7' and R8' mean (C1-C6)-alkyl. Also, other values of radicals R3 and R4 are given, a method for their preparing and a pharmaceutical composition. Described compounds are inhibitors of phosphodiesterases and can be used in manufacturing agents showing an anti-thrombosis, anti-proliferative, anti-vasospastic and vasodilating effect.

EFFECT: improved preparing method, valuable biochemical and medicinal properties.

10 cl, 6 tbl, 337 ex

FIELD: medicine.

SUBSTANCE: method involves carrying out radiation treatment combined with textile material application on the tumor. Tumor-transformed vaginal uterus neck portion volume is determined with ultrasonic examination techniques. Koletex napkin impregnated with therapeutic cytostatic preparation dose is used as the textile material. The napkin pattern is produced on the basis of ultrasonic examination data. The napkin is quilted with ligature along the perimeter and fixed at the level of vagina fornix by drawing tightly in purse-string mode. Napkin is changed every 24 h within 10-20 days long treatment course.

EFFECT: improved life quality.

FIELD: medicine, biology.

SUBSTANCE: invention relates to application of tissue factor agonist, namely FVII or FVIIa to induce or increase cell journey, as well as to application tissue factor antagonist modified with FVII to reduce or avoid cell journey in treatment of pathological conditions associated with specific control of cell journey or chemotaxis.

EFFECT: method for treatment of improved effectiveness.

14 cl, 14 ex, 10 dwg

FIELD: medicine.

SUBSTANCE: disclosed are combine product and kits useful in treatment for solid tumor by application of ZD6126 compound having formula in combination with platinum antitumor drug (e.g. cysplatine) and/or taxane. Treatment methods may include ionizing radiation. In was discovered that certain doses of abovementioned combined substances according to method of present invention have synergic action on solid tumors (as well as on angiogenesis neoplasm).

EFFECT: new method for solid tumor treatment.

12 cl, 5 tbl, 9 dwg

FIELD: medicine, oncology.

SUBSTANCE: one should carry out chemoradiation therapy at applying a cytostatic preparation followed by distance and intracavitary irradiation. Depending upon development of tumor lesion during the first 3 or 6 d it is necessary to conduct monochemotherapy only due to introducing proxiphen together with dimethyl sulfoxide at weight ratio of 4.5-5.0 : 0.5-1.5, correspondingly by applications in "Coletex" napkins. Moreover, a napkin should be pre-impregnated in 20%-dimethylsulfoxide solution and fixed with a tough vaginal tamponade by changing napkins every 24 h. Then since the 4th d or the 7th d simultaneously with application it is necessary to carry out contact irradiation and distance impact onto minor pelvis every 4-6 h at single focal dosage (SFD) being 2 Gy at 10 seances 5 times/weekly with high-activity sources of SFD 2 Gy. The innovation provides tumor regress under conditions of no therapeutic complications, thus, improving patients' quality of life.

EFFECT: higher therapy.

3 ex

FIELD: medicine, in particular angiogenesis prophylaxis and treatment.

SUBSTANCE: invention relates to 2-cyclooxygenase inhibitors selected from group containing 4-[5-(4-chlorophenyl)-3-phenyl-1H-pyrazole-1-yl] benzenesulfonamide; 4-[5-(4-methylphenyl)-3-(trifluoromethyl)-1H-pyrazole-1-yl] benzenesulfonamide; 4-[5-methyl-3-phenyloxazole-4-yl] benzenesulfonamide or pharmaceutically acceptable salts thereof and pharmaceutical composition containing the same in therapeutically effective amount. Said composition are useful in treatment and/or prophylaxis of angiogenesis disorders such as metastasis, eye angiogenesis, diabetic retinopathy, etc. in subjects are needed in such treatment and/or prophylaxis.

EFFECT: new pharmaceuticals for angiogenesis treatment and/or prophylaxis.

5 cl

FIELD: medicine.

SUBSTANCE: method involves introducing antitumor chemo preparations with blood components. To do it, 300 ml of patient autoblood is subjected to centrifuging during 20 min at 2200 rpm. The produced 150-200 ml of autoplasma and 100 ml of packed red blood cells are placed into separate reservoirs. Cys-platinum as single dose of 100 mg is incubated with the autoplasma and cyclophosphane as single dose of 1 g is incubated with the packed red blood cells. Single doxorubicin dose of 30-50 mg is concurrently introduced with one of the preparations. When combined with cis-platinum, doxorubicin is incubated with the packed red blood cells. When combined with cyclophosphane, doxorubicin is incubated with the autoplasma. Reinfusion is carried out to bring total dose of the preparations to 150 mg of doxorubicin, 3-5 g of cyclophosphane and 200 mg of cis-platinum. Pause between the procedures is 3-4 days long.

EFFECT: avoided risk of adverse side effects; increased preparation activity; accelerated treatment course.

FIELD: medicine.

SUBSTANCE: method involves making experimental animal temperature increase by infecting it with spotted fever after having removed the main tumor. The temperature is to be increased to the level causing tumor cells destruction. The temperature is supported for 4-6 h and typhus treatment is started.

EFFECT: enhanced effectiveness of treatment; inhibited metastases development.

FIELD: organic chemistry, medicine, oncology, pharmacology.

SUBSTANCE: invention relates to a composition used in treatment of proliferative diseases and comprising the platinum coordinating complex with an anti-tumor agent and derivative of pentafluorobenzene sulfonamide. Also, invention relates to a method for treatment by using the indicated composition. The composition possesses the synergetic effect.

EFFECT: improved and valuable method of therapy.

13 cl, 5 dwg, 1 ex

FIELD: medicine, pharmacy.

SUBSTANCE: invention relates to lyophilized composition comprising epotilone in the effective amount and mannitol or cyclodextrin. The second variant of the lyophilized composition involves epotilone and hydroxypropyl-beta-cyclodextrin. The preferable content of epotilone in the lyophilized composition is from 0.1% to 1.5%, and cyclodextrin - from 90% to 99% as measured for the total mass of solid components. Epotilone-containing lyophilized compositions can be used fro preparing an anti-tumor medicinal agent useful for parenteral administration and the lyophilized composition can be reduced preferably before administration directly. Epotilone-containing lyophilized compositions show improved indices of epotilone solubility and can retain stability for 24-36 months at temperature from 2°C to 30°C being without change of the solubility index.

EFFECT: improved and valuable properties of composition.

10 cl, 4 tbl, 14 ex

Medicinal agent // 2268037

FIELD: medicine, chemical-pharmaceutical industry, pharmacy.

SUBSTANCE: invention relates to using 4-chloro-2-methylphenoxyacetic acid of the formula (I)

and its pharmacologically acceptable sodium, potassium and lithium salts (mixture of these salts, or mixture of salts and 4-chloro-2-methylphenoxyacetic acid) as a medicinal agent possessing immunomodulating, anti-inflammatory and antitumor properties, and antiviral activity also. 4-Chloro-2-methylphenoxyacetic acid and its mixtures with pharmacologically acceptable alkaline metal salts possess high effectiveness and enhanced bioavailability.

EFFECT: valuable medicinal properties of medicinal agent.

9 cl, 13 ex

FIELD: medicine, neurology, psychiatry.

SUBSTANCE: one should affect with amplipulsephoresis of "Berlition" preparation in rectified mode. One should apply types III and V of operations at modulation frequency being 130-150 Hz, modulation depth of 50-75%, impact duration for every type of operation lasts for 5-7 min. During the 1st d electrode-cathode should be located in inferior cervical - superior thoracic department of vertebral column, and electrode-anode - at anterior and posterior surfaces of forearms. During the 2nd d - in inferior thoracic - superior lumbar department of vertebral column and at anterior-lateral and posterior surfaces of shins, correspondingly. The present method improves clinico-electrophysiological and biochemical parameters.

EFFECT: higher efficiency of therapy.

5 ex, 8 tbl

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