Amino derivatives, method for the inhibition of the synthesis of nitric oxide, a method of selective inhibition of the synthesis of nitric oxide produced by inducible no-synthase, the method of reducing levels of nitric oxide, a pharmaceutical composition
(57) Abstract:The invention relates to new derivatives of formula (I), where R1- R4- hydrogen atoms; X - alkylene with 1 to 6 carbon atoms; Y is lower alkyl; B is - NR5R11where R5is a hydrogen atom, R11selected from 5 - to 6-membered heterocyclic radical, in which one ring member is a carbon and 1 to 4 members of the heteroatoms nitrogen, or sulfur, or their pharmaceutically acceptable salts, are useful as inhibitors of the synthesis of nitric oxide. Also proposed a method of inhibiting the synthesis of nitric oxide, a method of selective inhibition of the synthesis of nitric oxide produced by inducible NO-synthase, the method of reducing nitrogen oxide and pharmaceutical composition. 5 C.p. f-crystals, 1 PL.The invention relates to the derivatives of aminotetrazole and their use in therapy, in particular to their use as inhibitors synthase nitric oxide.Since the 1980-ies it is known that the vascular relaxation induced by acetylcholine, depends on the presence of endothelium, and this activity is attributed to the labile humoral factor, named secreted by the endothelium relaxing factor (EDRF). The activity of nitric oxide (NO) as vazo the other nitrovasodilators. The recent identification of EDRF as NO coincided with the opening of the biochemical pathway by which NO is synthesized from the amino acid L-arginine by the enzyme NO synthase.NO is an endogenous stimulator of soluble guanylate cyclase and in addition to endothelium-dependent relaxation is involved in many biological activities, including cytotoxicity phagocytic cells and the communication cell - to-cell in the Central nervous system (see Moncada et al. Biochemical Pharmacology, 38, 1709-1715 (1989) and Moncada et al. Pharmacological Reviews, 43, 109-142 (1991). Currently, it is believed that excessive production of NO may affect many States, especially States, which include systemic hypotension, such as toxic shock syndrome and the treatment of certain cytokines.The synthesis of NO from L-arginine may be Engibarov analogue of L - arginine, L-N-monomethyl-arginine (L-NMMA), and it was suggested that therapeutic use of L-NMMA for the treatment of toxic shock and other types of systemic hypotension (WO 91/04024 and GB-A-2240041). Therapeutic use some other inhibitors of NO synthase, excluding L-NMMA, for the same purpose has also been proposed in WO 91/04024 and EP-A-0446699.Recently, it has become apparent that there are at m is calitanii in the endothelium and produce NO in response to receptor or physical stimulation.(ii) a constitutive, Ca++/calmodulin - dependent enzyme that is localized in the brain and produce NO in response to receptor or physical stimulation.(iii) Ca++- independent enzyme, which is induced after activation of vascular smooth muscle, macrophages, endothelial cells and other cells by endotoxin and cytokines. Once originated, this inducible NO synthase synthesizes NO in for extended periods.NO, the selected constitutive enzymes, acts as a transduction mechanism, emphasizing different physiological reactions. NO produced induced enzyme, is a cytotoxic molecule for tumor cells and invading microorganisms. It also appears that the harmful effects of excessive NO production, in particular pathological vasodilatation and tissue damage, may be largely a result of the effects of NO synthesized induced by NO synthase.There is increasing evidence that NO may be involved in the degeneration of cartilage, which takes place under certain conditions, such as arthritis, and it is well known that the synthesis of NO is enhanced when revmote is of NO from L-arginine, include autoimmune and/or inflammatory condition affecting the joints, such as arthritis, inflammatory bowel disease, cardiovascular ischemia, diabetes, hyperalgesia (allodynia), cerebral ischemia (local ischemia, thrombotic lesion and extensive ischemia, secondary stopping of the heart), and other disorders of the Central nervous system, mediated by NO, and other disorders mediated by NO.In addition, the conditions under which it is advisable to inhibit the production of NO from L - arginine include systemic hypotension associated with septic and/or toxic shock caused by a wide variety of agents; therapy with the use of cytokines, such as TNF, IL-1 and IL-2, and used as adjuvant to short term immunosuppression in transplantation.Some of the inhibitors of NO synthase proposed for therapeutic use long enough, and in particular L-NMMA, are indiscriminate in that they inhibit both constitutive and inducible NO synthase. The use of such a non-selective inhibitor of NO synthase requires great care when taking in order to avoid potentially seriously the thrombosis and tissue damage. In particular, in the case of therapeutic use of L-NMMA for the treatment of toxic shock it is recommended that continuous monitoring of blood pressure of the patient during treatment. Thus, although non-selective inhibitors of NO synthase have therapeutic utility, provided appropriate precautions, inhibitors of NO synthase, which are selective in the sense that they inhibit inducible NO synthase to a much greater extent than constitutive isoforms of NO synthase, would be even more therapeutically useful and more convenient to use.W094/12165, W094/14780, W093/13055, EP0446699A1 and U.S. patent N 5132453 disclose compounds that inhibit the synthesis of nitric oxide and preferably inhibit the induced synthase isoforms of nitric oxide. Messages, therefore, are by reference in its entirety, as described here.In a broad aspect, the present invention is directed to the inhibition or modulation of the synthesis of nitric oxide in the body of a subject in need of such inhibition or modulation, by introducing a compound that mainly inhibits or modulates the induced synthase isoforms of nitric oxide compared to constata the subject, in need of such reduction.Compounds of the present invention represented by the following chemical formula
< / BR>and their pharmaceutically acceptable salts;
where R1, R2independently selected from the group consisting of hydrogen, lower alkyl, lower alkenyl and lower quinil;
R3, R4independently selected from the group consisting of hydrogen, lower alkyl, lower alkenyl, lower quinil, aryl, COR7or SO2R8where R7and R8independently selected from the group consisting of lower alkyl, lower alkenyl, lower quinil and aryl;
X is independently selected from the group consisting of lower alkyl, lower alkenyl and lower quinil, and all of them can be substituted by lower alkyl, lower alkoxygroup, hydroxy, halogen, trifluoromethyl, nitro, cyano, amino;
or X is selected from the group of the formula -(CH2)pQ(CH2)r- where p = 1-3, r=1-3, and Q is oxygen, C= 0, S(0)twhere t = 0-2, or NR12where R12is hydrogen or lower alkyl which may be substituted by lower alkyl, lower alkoxygroup, hydroxy, halogen, trifluoromethyl, nitro, cyano, amino; or
X is selected from the group of the formula -(CH2)
B-NR5R11where R5selected from the group consisting of hydrogen, lower alkyl, lower alkenyl, lower quinil and aryl and R11selected from a 3-8 - membered heterocyclic radical, in which at least one member of the ring is hydrogen and in which from 1 to about 4 members are heteroatoms independently selected from oxygen, nitrogen and sulfur, and the heterocyclic radical may be substituted by hydroxyl, lower alkoxygroup, lower alkyl, halogen, nitro, carboxyla, S02R13where R13selected from lower alkyl, lower alkoxygroup, NR1R2, amino, acyloxy, trifloromethyl, phenyl and naphthyl which may be substituted with halogen, nitro, lower alkoxygroup and lower alkyl.
< / BR>2S-amino-6-[(1-iminoethyl)amino]-N-(1H-tetrazol-5-yl) hexanamide, hydrate, N,N - diisopropylethylamine (DIPEA) (5,1 g, 6.9 ml, 39,54 mmol) in 20 ml of dimethylformamide (DMF) at ambient temperature add hexaphosphate benzotriazol-1-yl-oxy-Tris(dimethylamino)phosphonium (THIEF) (6.4 g, 14,49 mmol).After stirring for 1 h the reaction mixture was concentrated under vacuum. The residue is partitioned between 60 ml of ethyl acetate (EtOAc) and 50 ml of water. The layers are separated. The organic layer is washed with 50 ml of 1 M solution of KHSO4and 2 times 50 ml of water. The product begins to precipitate, and the suspension was concentrated in vacuo, receiving 9 g of crude compound. After drying the product was then purified by boiling in methylene chloride, followed by filtration, gaining 3.7 g 1A (62,7%). Connection characterize1H YARM.1B 1A (2 g, 4.5 mmol) restore under conditions of catalytic hydrogenation using Pd mobiles when 0,352 kg/cm250% solution of EtOH/AcOH within 12 hours, getting 1,55 r (100%) 1B. Connection characterize1H YARM.1C To a stirred solution of 1B (1.55 g, 4,15 mmol) and hydrochloride of methylacetamide (0,91 g, 8,31 mmol) in 25 ml DMF added triethylamine (tea) (1.26 g, of 1.74 ml, 12,45 mmol). After stirring for 16 h at ambient temperature the reaction mixture is filtered off from triethylamine hydrochloride and the filtrate kontsentriruemoy by phase C-18 column, obtaining 0.9 g (52,3%) 1C. The product is characterized1H YARM.1 1C (0.9 g, 2,17 mmol) dissolved in 30 ml of acetic acid and add 3 ml of 4 N HCL/dioxane. The reaction mixture was stirred for 20 min at ambient temperature, then add 150 ml simple ethyl ester. After 2 h the precipitate is filtered off, washed simple with ethyl ether and dried, obtaining 0,78 g 1 (96%). Anal. Rasch. for C9H18N8O, 2HCl, 1,25 H2O: C, 30,91; H, 6,48; N, 32,04; Cl, 20,27. Found: C, 31,64; H, To 6.43; N, 32,19; Cl, 20,19. DSC so pl. 144,9oC.The compound of example 1 is also more selective than NIL. The compound of example 1 is finely crystalline product as well as all intermediate compounds. On the contrary, NIL is glass, which makes handling them.Example 2
< / BR>2S-amino-5-[[amino(nitramino)methyl]amino]-N-(1H-tetrazol-5 - yl)pentanone, hydrochloride
< / BR>2A Sample t-Boc of nitroarginine (5.0 g, 15.6 mmol) and N - methylmorpholine (1.6 g, 15.6 mmol) dissolved in a mixture of methylene chloride (CH2Cl2, 25 ml) and DMF (25 ml) cooled to -78oC. To this stirred reaction mixture in the atmosphere of nitrogen (N2) add isobutylparaben (Aldrich, 2.2 g, of 16.6 mmol). Then allow the reaction cm is t to -78oC. Sample monohydrate 5-aminotetrazole (Aldrich, of 1.62 g, 15.8 mmol) is added to the reaction mixture. The reaction mixture was allowed to warm to room temperature and stirred for 48 hours All the solvent is removed under reduced pressure and the residue distributed between ethyl acetate (EtOAc) and water. The aqueous layer was free from all water and specified in the header of the material isolated from the crude residual product (9.3 g) by chromatography.2 Specified in the title material was obtained from 2A in the manner described in Example 1.Example 3
< / BR>2S-amino-6-[(1-iminoethyl)amino] -N-(1H-imidazol-2 - yl)-hexanamide, dihydrochloride
3 Specified in the title material was obtained in the same way as 1, based on 2-aminoimidazole.Example 4
< / BR>2S-amino-6-[(1-iminoethyl)amino] -N-(1H-1,2,4-triazole-3 - yl)-hexanamide, dihydrochloride
4 is Listed in the title material was obtained in the same way as 1, based on 3-aminotriazole.Example 5
< / BR>2S-amino-6-[(1-iminoethyl)amino] -N-(5 - pyrimidinyl)hexanamide, hydrate, dihydrochloride
5 is Listed in the title material was obtained in the same way as 1, starting from 5-aminopyrimidine.Example 6
< / BR>2S-amino-6-[(1-iminoethyl)amino the way that and 1, based on 3-aminopyrazole.Example 7
< / BR>2S-amino-6-[(1-iminoethyl)amino] -N-(thiazol-2 - yl)hexanamide, dihydrochloride
7 is Listed in the title material was obtained in the same way as 1, based on 2-aminothiazole.Biological data
The activity of the above compounds as inhibitors of NO synthase define the following tests:
Citrullinemia analysis synthase nitric oxide
Synthase activity of nitric oxide measured by monitoring the conversion of L-[2,3-3H] -arginine to L-[2,3-3H]-citrulline (1,2). Inducible NOS man (hiNOS), endothelial constitutive NOS man (hecNOS) and neuronal constitutive NOS man (hncNOS), each clone from RNA extracted from human tissues. Recombinant enzymes Express in insect cells using a baculovirus vector. The active enzyme is extracted from cell extracts and partially purified by DEAE chromatography-Sepharose (2). The enzyme and inhibitors added to a volume of 50 μl in 50 mm Tris (pH 7.6) and initiate the reaction by adding 50 µl of a solution containing 50 mm Tris (pH 7,6), 2.0 mg/ml bovine serum albumin, 2.0 mm DTT, 4.0 mm CaCl2, 20 μm FAD, 100 μm tetrahydrobiopterin, 0.4 to 2.0 mm NADPH and 60 μm L-arginine, soderzhashchaya at 37oC for 15 min the reaction is stopped by adding 300 ál of cold buffer containing 10 mm EGTA, 100 mm HEPES (pH 5.5) and 1.0 mm L-citrulline. [3H]-Citrulline was separated by chromatography on a cation-exchange resin Dowex 50W X-8 and quantify the radioactivity in a liquid scintillation counter.1. Bredt, D. S. and Snyder, S. H. (1990) Proc. Natl. Acad. Sci. U. S. A. 87, 682-685.2. Misko, T. P., Moore, W. M., Kasten, T. P., Nickols, G. A., Corbett, J. A. , Tilton, R. G., McDaniel, M. L, Williamson, J. R. and Currie, M. G. (1993) Eur. J. Pharm. 233, 119-125.Compounds according to the invention are non-toxic, as evidenced by tests, for example, the compounds of example I in vivo for 0.1, 0.3, 1.0, 3.0, 10.0, and 600.0 mpk, showed the following results:
Analysis: MTOL po for example 1.5/5 0.1 mpk 5/5 0.3 mpk 5/5 1.0 mpk
Portable = 0.1 mpk Portable = 0.3 mpk Portable = 1.0 mpk
5/5 3.0 mpk 5/5 10.0 mpk 5/5 600 mpk
Portable = 3.0 mpk-Tolerance = 10.0 mpk Portable = 600 mpk
From the preceding description specialist in this field can easily set the essential characteristics of this invention and, without departing from the scope of its content and scope, can make various changes and modifications of the invention to adapt it to different needs and conditions. 1. Aminophosphonate;
Y is lower alkyl;
B - NR5R11where R5is a hydrogen atom, R11selected from 5 - to 6-membered heterocyclic radical, in which at least one ring member is a carbon and 1 to 4 member heteroatoms independently selected from nitrogen and sulfur,
or their pharmaceutically acceptable salts.2. Amino derivatives of formula (I) under item 1, where the specified connection selected from the group of 2S-amino-6-[(1-iminoethyl)amino]-N-(1H-tetrazol-5-yl)hexanamide, hydrate, dihydrochloride; 2S-amino-6-[(1-iminoethyl)amino]-N-(1H-imidazol-2-yl)hexanamide, dihydrochloride; 2S-amino-6-[(1-iminoethyl)amino]-N-(thiazol-2-yl)hexanamide, dihydrochloride.3. The method of inhibition of the synthesis of nitric oxide from the subject in need of such inhibition, by introducing the inhibitor, characterized in that as an inhibitor use a connection on p. 1 or 2 in an effective amount.4. The method of selective inhibition of the synthesis of nitric oxide produced by inducible NO-synthase, relative to the nitric oxide produced by the constitutive forms of NO synthase, the subject in need of such selective inhibition by injection of the inhibitor, characterized in that as an inhibitor opolska, in need thereof, by introducing a therapeutic ingredient, characterized in that the quality of therapeutic ingredient is used as a compound under item 1 or 2 in an effective amount.6. Pharmaceutical composition having inhibitory effect on the synthase nitric oxide in the subject in need of such inhibition, containing therapeutic ingredient together with one or more pharmaceutically acceptable carriers, characterized in that as the quality of therapeutic ingredient it contains a connection on p. 1 or 2 in an effective amount.
R-NH--Nwhere R is phenyl, unsubstituted or mono - or multiply substituted by halogen atom, lower alkyl, lower alkoxygroup, arroceros, trifluoromethyl, and unsubstituted or once substituted lower alkoxygroup benzothiazolyl, or benzoxazolyl, R1and R2means a hydrogen atom or lower alkyl, and R1and R2cannot simultaneously denote a hydrogen atom
R(I) where R1and R2the same or different and represent aliphatic group1-C3, optionally substituted by a methoxy group, cyclohexyl, or R1-R2together with the nitrogen atom to which they are attached, form a heterocyclic ring of formula II
-(II) where R8hydrogen, Allenova group, optionally interrupted by oxygen or sulfur, which may be substituted by lower alkyl or by Deputy, at two adjacent carbon atoms alkalinous group form a benzene ring or In-Allenova group with three carbon atoms,
R3straight or branched alkyl radical containing 1-5 carbon atoms or cyclohexyl, or a group of the formula
-N(III) where R4and R4' are the same or different and represent a hydrogen atom or an alkyl group containing 1-4 carbon atoms, R5a hydrogen atom or a straight or branched aliphatic Radiatsionnaya aliphatic group with 1-5 carbon atoms, optionally substituted by actigraphy, acetoxypropionyl, alkoxygroup containing 1-3 carbon atoms, alkylthiol containing 1-3 carbon atoms, optional alkilirovanny amino group, phenyl group or cycloalkenyl ring containing 6 carbon atoms or the radicals R3and R5together with the carbon atoms and the nitrogen to which they are attached, form a heterocyclic ring of formula IV
(IV) in which R6has the specified values, R9and R10the same or different, are hydrogen atom or alkyl group with 1-4 carbon atoms, D Allenova group with 2-5 carbon atoms or the radicals R3and R5together with the carbon atoms and the nitrogen to which they are attached, form a heterocyclic ring of the formula V
(V) where R6has the specified values, R11a hydrogen atom, an alkyl group with 1-2 carbon atoms,Allenova group with 2 carbon atoms, or R5and R6together with the nitrogen atom to which they are attached, form a heterocyclic ring of the formula VI
-Nthen, at least one of R4, R4', R5or R6different from hydrogen
FIELD: medicine, oncology.
SUBSTANCE: the present innovation deals with treating patients with uterine cervix cancer with relapses in parametral fiber and in case of no possibility for radical operative interference and effect of previous radiation therapy. During the 1st d of therapy one should intravenously inject 30 mg platidiam incubated for 1 h at 37 C with 150 ml autoblood, during the next 3 d comes external irradiation per 2.6 G-r. During the 5th d of therapy one should introduce the following composition into presacral space: 60 ml 0.5%-novocaine solution, 1 ml hydrocortisone suspension, 2 ml 50%-analgin solution, 1 ml 0.01%-vitamin B12 solution, 1.6 g gentamycine, 800 mg cyclophosphan, 10 mg metothrexate. These curative impacts should be repeated at mentioned sequence four times. The method enables to decrease radiation loading and toxic manifestations of anti-tumor therapy at achieving increased percent of tumor regression.
EFFECT: higher efficiency of therapy.
FIELD: medicine, cardiology.
SUBSTANCE: the suggested method should be performed at the background of medicinal therapy with preparations out of statins group, tevetene, polyoxidonium and conducting seances of plasmapheresis by removing 800 ml plasma twice weekly with N 5 due to additional intramuscular injection of immunophan 0.005%-1.0 with N 10 and fluimucyl 300 mg intravenously daily with N 5-10, total course of therapy lasts for 2 mo. The method provides modulation of leukocytic functional activity, moreover, due to altered cytokine profile and, thus, through disintegration of protein-lipid complexes participating in the development of atherosclerotic platelets.
EFFECT: higher efficiency of therapy.