N-acyl derivatives of aminoacids, their pharmaceutically acceptable salts, pharmaceutical composition and application as hypolipidemic preparations

FIELD: chemistry.

SUBSTANCE: invention relates to bioorganic chemistry and concerns N-acyl derivatives of aminoacids of the general formula I , where n is 2 or 3, and R is , as well as their pharmaceutically acceptable salts and their application as hypolipidemic preparations. Another subject of invention is pharmaceutical composition containing claimed compounds in pharmaceutically effective quantity and method of treatment of lipidosis, atherosclerosis, cardiac and cerebral ischemia, cardiac infarction, stroke.

EFFECT: obtaining aminoacid derivatives with hypolipidemic effect.

8 cl, 28 tbl, 1 dwg, 22 ex

 

The present invention relates to the field of Bioorganic chemistry and relates to N-acyl derivatives of amino acids, their pharmaceutically acceptable salts, of new methods of their synthesis and pharmaceutical compositions on their basis and application in medicine as anti-allergic, anti-inflammatory and lipid-lowering means.

Prior art

As you know currently allergic diseases and disorders of lipid metabolism are very common due to poor environmental conditions, changing eating patterns and lifestyle of the population. Therefore, the problem of drugs to combat these diseases, as well as with inflammatory processes, generally accompanying allergies, continues to be relevant.

The most widespread group of anti-allergic drugs are antagonists of H1-histamine receptors. Currently, there are 2 generation antihistamines [Mashkovsky PPM Medicines. / Moscow: the New wave. 2005. s.285].

Antihistamines 1st generation penetrate the blood-brain barrier and can cause a blockade of N1receptor cells of the Central nervous system, which makes them undesirable sedative effect. To achieve verazanno the antihistamine action required high concentrations of these drugs in the blood, that requires the use of them in large doses. A negative feature of these drugs is frequent development of tahiphylacsii, the effect on the Central nervous system, manifested by loss of coordination, dizziness, feeling of lethargy, decreased ability to concentrate. Notwithstanding the above, antihistamines are the first generation still apply, especially in those situations when you need a very fast effect from treatment, such as anaphylaxis. To antihistamine preparations of the first generation include diphenhydramine, pipolphen, peritol, suprastin, clemastin, tavegil, fenkarol.

Antihistamines 2nd generation has received in recent years a wide application in Allergy practice, because it does not have side effects inherent in the drugs of the 1st generation. In particular, the preparations of the 2nd generation does not penetrate the blood-brain barrier, does not have a sedative and hypnotic effects. They are characterized by fast and long-lasting antihistamine effect. To antihistamines means of the second generation are claritin (loratadine), zyrtec (cetirizine), Kesten (Avastin). However, clinical trials have revealed side effects and these drugs, due to their interaction with other drugs or a violation of their metabolic the ISM by the cytochrome P 450. Thus, the identified potentially sedative (cetirizine, loratadine) and potentially cardiotoksicnae (terfenadine, astemizole, Avastin) effects of antihistamines 2nd generation.

In some cases, such as in bronchial asthma, use corticosteroids, providing a powerful anti-allergic effects. However, their use is accompanied by systemic manifestations in the form of Itsenko-Cushing syndrome, hypertension, hyperglycemia, osteoporosis and other [Mashkovsky PPM Medicines. / Medicine. Moscow. 1993. vol. 1, s].

Of particular importance in the development of allergic diseases has pathogenicity stage of allergic reactions, which is largely determined by the degree of activation of the target cells allergies 1-th order (basophils and mast cells). Their important feature is the ability to accumulate and release under the action of stimulus (allergen) biologically active compounds, primarily histamine. When IgE and/or IgG-mediated response to the antigen, these cells determine the severity of the clinical picture of immediate Allergy [Parker CW / Mediators: the release and function. // In the book: Immunology. Edited Upolu. - M - World. - 1989. - v.3. - s-247; Chakravarty N.K. // In: The mast cell: Its role in health and disease, ed. J Pepys. - 1979. - p.38-46].

There is a group of drugs (cromoly is-sodium, ketotifen, oxatomide), used in bronchial asthma and bronchospastic conditions, the basis of which lies the ability to inhibit the degranulation of mast cells and to delay the release of these substances mediator contributing to the development of bronchospasm, Allergy and inflammation (bradykinin, histamine). As side effects may cause irritation of mucous membranes, headache, and edema of the larynx, coughing, choking [Mashkovsky PPM Medicines./Moscow: the New wave. 2005. s].

It is known that the most common manifestation of atherosclerosis is coronary heart disease, standing on the first place among the causes of mortality of the adult population of the planet. One of the leading disorders in this disease is recognized as lipid metabolism, expressed in increased levels in plasma cholesterol, including the composition of lipoproteins, low (LDL) and very low density (VLDL), called "atherogenic", with a simultaneous decrease in the number of "antiatherogenic" high density lipoprotein (HDL).

It is shown that the changes in the content and ratio of lipids in plasma reflects the change in membrane structures parenchymatous organs. The composition of cell membranes, for example, microsomal, directly depends on the composition of the diet of experimental animals [ade A., Harred W. // Feder. Proct.- 1976.- vol. 55.- pp. 2475-2479]. Introduction animal cholesterol causes its accumulation in cell membranes, reducing its fluidity, which in turn leads to changes in the functional state of enzymes [J.T.M. Buters, Zysset T., Reichen J. // Biochem. Pharmacol.- 1993.- vol. 46.- Iss 6.- pp. 983-991.

Lipid-lowering means lowers cholesterol and triglycerides in the blood, can be used for the treatment and prevention of diseases associated with disorders of lipid metabolism. The latter are characterized by increased levels of triglycerides, total cholesterol, cholesterol in the lipoproteins of low and very low-density (LDL and VLDL) and decreased content of cholesterol in the high density lipoprotein in such diseases as atherosclerosis, obesity, coronary heart disease and cerebral, myocardial infarction, stroke, and which serve as a risk factor for the manifestation of diabetes and blood clots.

Known clinical application of the so-called statins, inhibitors of cholesterol biosynthesis. This group of drugs in doses of 80 mg/day is quite effective mainly against lowering total cholesterol in the blood, while the inaccessible roads and are alien to the body chemical compounds. In addition, their use may be accompanied by the full-time effects: changes of the liver with increased levels of transaminases in the blood, dyspepsia [Mashkovsky PPM Medicines./Medicine. Moscow. 1993. vol. 1, s].

In connection with the above it is important to search for new effective anti-allergic and lipid funds with alternative mechanisms of action that can be active at low concentrations and devoid of side effects. In this regard, of particular interest are compounds comprising residues of substances of natural origin, as they can be predicted lower toxicity and frequency of side effects.

In the published international application WO 99/01103 described antiallergic and hypolipidemic action of N-acyl derivatives of biogenic amines, such as γ-glutamylcysteine and its nearest analogue of glutamylcysteine that are most similar in structure and action of the claimed compounds. In article Krzeczkowska CENTURIES, Zheltukhina GA, Nebolsin V.E. and other Study antianaphylactic activity and mechanisms of action γ-L-glutamylcysteine. // Pathogenesis. 2003. Vol. 1. No. 2. C. 60-64 shown that γ-glutamylcysteine has expressed antianaphylactic activity using different animal species and routes of administration. The results obtained indicate that in the fat cells of animals under the action γ-glutamylcysteine significantly reduced the tsya the content of histamine and antigen-stimulated secretion. In the test on investigating the influence of glutamylcysteine on the severity of antigen-induced bronchoconstriction has been shown to reduce the magnitude of bronchoconstriction by more than 50% compared with control. This effect was manifested as oral and intratracheal method of its introduction in a low dose of 50 mcg/kg. Glutamylcysteine has the ability to reduce the symptoms of passive cutaneous anaphylaxis from 34 to 42% and thus exceeds the efficiency of suprastin, but inferior to claritin. In WO 99/01103 shown that the introduction of animals glutamylcysteine at doses of 50 and 500 mg/kg demonstrated a significant decrease in the intensity of delayed-type hypersensitivity. In addition, glutamylcysteine at doses of 50 and 500 mg/kg also had some anticholesterolemic activity, reducing the total cholesterol compared to animals with atherogenic load of 5-7%.

Lack of glutamylcysteine is its relatively high cost and low availability of raw materials to receipt of histamine. In addition, the substance is not effective in these tests.

With the aim of expanding Arsenal of technical tools and create a more effective and affordable anti-allergic, anti-inflammatory and lipid-lowering means, the authors izopet the deposits have been identified some specific N-acyl derivatives of amino acids of General formula (I), disclosed in published international application WO 99/01103, but specifically it is not described, is not received and is not described.

So, the General formula (I) above international application fall compounds of the present invention Nα-succinyl-L-tryptophan (II), Nα-glutaryl-L-tryptophan (III), Nα-glutaryl-L-histidine (IV) and Nα-succinyl-L-histidine (V). However, in this publication is not given any specific structural formulas of these compounds, nor any physico-chemical characteristics, and also does not describe the methods for their preparation. In the published international application WO 03/072124 formulae Nα-glutaryl-L-tryptophan (II), Nα-glutaryl-L-histidine (IV) and Nα-succinyl-L-histidine (V), but not described method of their synthesis and not given their physico-chemical constants.

One of the connections glutamylcysteine is mentioned only in the form of a methyl ester at the C-end of His [Glt-His(OMe)] in U.S. patent 3963691, as intermediate compounds in the synthesis of peptide Glt-His-Trp-Ser-Tyr-Gly-Leu-Arg-Pro-Gly-poly-Lys.

Connection succinylated mentioned in the published international application WO 93/04690. In this publication indicated that adding free imidazole or succinyldicholine accelerates the interaction of carnosine with dihydroxyacetone. Methods of synthesis of succinyldicholine and its physico-chemical constants are not shown.

The structural formula of succinyldicholine mentioned in the application U.S. 2005079515. In this publication, neither the methods of synthesis of succinyldicholine nor its physico-chemical constants are not shown.

Obtaining Nα-glutaryl-L-histamine described in published international application WO 99/01103 and represents the N-acylation of biogenic amine glutaric anhydride in the medium of anhydrous N,N-dimethylformamide. In addition, the publication herskowitz A.A., Kibirev VK // Chemical synthesis of peptides / Kiev, Naukova Dumka, 1992, 360 described method of acylation of amino acids in aqueous organic alkaline environment.

In F. Sorm, Pravda Z. Proteins and amino acids. X. Synthesis of two peptide analogs. // Chemicke Listy pro Vedu a Prumysl. 1951. V.45. P.423-425 described method of synthesis of succinylcholine ethyl ester in a mixture of water and ethyl acetate at a ratio of (1:1) on the basis of the hydrochloride of the ethyl ester of tyrosine and glutaric anhydride in the presence of NaHCO3to maintain a slightly alkaline pH.

However, the acylation with anhydrides of dicarboxylic acids free of histidine are not described in literature.

The aim of the present invention are effective new N-acyl derivatives of amino acids having antiallergic and hypolipidemic effect in low doses and does not exhibit side effects, the pharmaceutical compositions based on them, their use as a more e is effective antiallergic, anti-inflammatory and lipid, as well as new methods of synthesis of N-acyl derivatives of amino acids.

The inventors have developed a simple preparative method of synthesis of compounds of General formula (I), namely, that the glutaric anhydride or succinic acid in the form of a solid substance is added to aqueous solution of amino acids in the absence of inorganic and organic bases to obtain the target product with a sufficiently high yield of 55-60%.

The inventors have also developed another method for the synthesis of compounds of General formula (I), including N-acyl derivative of histidine, comprising conducting the reaction in a biphasic system consisting of an aqueous solution of histidine and solution of the anhydride in a suitable organic solvent using an excess Alliluyeva agent.

Brief description of the invention

The present invention relates to new N-acyl derivatives of amino acids of General formula I:

where n is 2 or 3; and

R represents

and their pharmaceutically acceptable salts having anti-allergic, anti-inflammatory and hypolipidemic effect.

The present invention also relates to a method for producing N-acyl derivatives of amino acids on the formula I and their salts, including the addition of glutaric anhydride or succinic acid in the form of solids to aqueous solution of amino acids of General formula:

or its salts, where

R represents

and, optionally, the transformation of the target product in its salt.

The present invention further relates to a method for producing N-acyl derivatives of amino acids of General formula I and their salts, comprising conducting the reaction in a biphasic system with glutaric anhydride or succinic acid in a water-immiscible organic solvent with an aqueous solution of the amino acid of General formula:

or a salt thereof, where R represents

and, optionally, the transformation of the target product in its salt.

The present invention relates also to the use of compounds of General formula I and their pharmaceutically acceptable salts as anti-allergic, anti-inflammatory and lipid-lowering means.

Further, the present invention relates to pharmaceutical compositions and means of having antiallergic, antianaphylactic and anti-inflammatory and hypolipidemic effect, containing an effective amount of the compounds of General formula I or its pharmaceutically p is yimlamai salt, and if you want pharmaceutically acceptable carrier.

Another object of the invention is a method of treatment of allergic diseases, including bronchial asthma, allergic rhinitis, allergic rhinitis, seasonal rhinitis, perennial rhinitis, atopic dermatitis, psoriasis, urticaria, allergic (including anaphylactic) reactions to insect stings and medications, cold Allergy, allergic conjunctivitis, chronic obstructive pulmonary diseases, namely chronic obstructive bronchitis, emphysema, obliterative bronchitis, cystic fibrosis and diseases associated with lipid metabolism disorders: atherosclerosis, obesity, coronary heart disease and cerebral, myocardial infarction, stroke, involving the introduction of effective the number of compounds of General formula I or its pharmaceutically acceptable salt.

A detailed description of the invention

Preferred compounds of General formula I are presented in table 1.

The synthesis of compounds of General formula I can be carried out in two ways. The first way is to gradually added to the aqueous solution of amino acids of General formula

or a salt thereof, where R represents

glucurono is about or succinic anhydride in the form of solids, with subsequent isolation of the desired product, ion exchange chromatography, preferably by passing the reaction mixture through a column of cation exchange resin and subsequent crystallization from aqueous solution. The obtained crystals of the target product is washed with a suitable solvent, preferably methanol. The main advantage of the proposed method consists in the absence of alkali in the aqueous solution of amino acids, which prevents inactivation of the anhydride of dicarboxylic acid by hydrolysis. In addition, the balance of imidazole in the composition of amino acid molecules may carry out acid-base autocatalyst reaction of acylation of the amino group of amino acids. Relatively high yields (55-60%) when using the proposed method is achieved, in particular, due to gradual addition of an anhydride of dicarboxylic acids taken in excess, and intensive mixing of the reaction mass.

Compounds of General formula I can also be obtained in an alternative way in the two-phase system, including the addition of glutaric anhydride or succinic acid in a water-immiscible organic solvent to aqueous solution of amino acids of General formula:

or its salts, where

R represents

This method allows the use of the SQL excess Alliluyeva agent, to achieve complete acylation α-amino group of the amino acid and the yield of the target product is about 70%. To maintain the required pH instead of inorganic alkalis are used, an organic base is pyridine, which is not hydrolyzes anhydride and, in addition, as is known, is a catalyst for acylation. The use of pyridine avoids contamination of the final product inorganic salts, which, together with the reaction product remains in the aqueous layer. Used approaches can simplify the separation of the target product from the unreacted anhydride and the appropriate amino acids and select the target product by simple crystallization.

Preferred water-immiscible organic solvents are butanol, ethyl acetate, chloroform.

Preferred solvents used for crystallization of the desired product are water-alcohol mixtures, in particular water-ethanol.

Compounds of General formula I can also be obtained in the form of pharmaceutically acceptable salts with sodium hydroxide, potassium hydroxide, magnesium carbonate, lithium hydroxide, calcium carbonate routine methods widely described in the literature.

Compounds of General formula I possess anti-allergic, anti-inflammatory and hypolipidemic activity and can be used too for the treatment of allergic, anaphylactic, including inflamed diseases and disorders of lipid metabolism.

In particular, the compounds of the present invention can be used to treat the following allergic diseases: bronchial asthma, allergic rhinitis, allergic rhinitis, seasonal rhinitis, perennial rhinitis, atopic dermatitis, psoriasis, urticaria, allergic (including anaphylactic) reactions to insect bites and medications, cold Allergy, allergic conjunctivitis, chronic obstructive pulmonary diseases, namely chronic obstructive bronchitis, emphysema, obliterative bronchitis, cystic fibrosis and diseases associated with lipid metabolism disorders, such as atherosclerosis, obesity, coronary heart disease and cerebral, myocardial infarction the stroke.

Compounds of the present invention are introduced in an effective amount that provides the desired therapeutic result.

For the treatment of allergic diseases, including bronchial asthma, allergic rhinitis, allergic rhinitis, seasonal rhinitis, perennial rhinitis, atopic dermatitis, psoriasis, urticaria, allergic (including anaphylactic) reactions to insect stings and medications, cold Allergy, allergic shall conjuctiva, chronic obstructive lung diseases, namely chronic obstructive bronchitis, emphysema, obliterative bronchitis, cystic fibrosis and diseases associated with lipid metabolism disorders, such as atherosclerosis, obesity, coronary heart disease and cerebral, myocardial infarction, stroke, compounds of General formula I can be administered orally, topically, parenterally, intranasally, inhalation and rectal in the form of standard dosage forms containing non-toxic pharmaceutically acceptable carriers. Used in the present description, the term "parenteral" refers to subcutaneous, intravenous, intramuscular or intrathoracic injection or infusion.

Compounds of the present invention can be administered to the patient in doses, comprising from 0.01 to 10 mg/kg of body weight per day, preferably at doses from 0.05 to 5 mg/kg once or more than once a day.

It should be noted that the specific dose for each particular patient will depend on many factors, including the activity of this used compound, the age, body weight, sex, General health status, and diet of the patient, time and route of administration of drugs, the rate of its excretion from the body, specifically used a combination of drugs and the severity of the disease in Yes the tion of the individual, being treated.

The pharmaceutical compositions of the present invention contain a compound of General formula (I) in an amount effective to achieve the desired result, and can be entered as standard medicinal forms (for example, in solid, semisolid, or liquid form), containing compounds of the present invention as an active ingredient in a mixture with a carrier or excipient suitable for intramuscular, intravenous, oral, sublingual, inhalation, intranasal, and intrarectal administration. The active ingredient can be included in a composition together with commonly used non-toxic pharmaceutically acceptable carriers suitable for the manufacture of solutions, tablets, pills, capsules, pills, suppositories, emulsions, suspensions, ointments, gels or any other medicines.

As fillers can be used in a variety of substances such as sugars, for example glucose, lactose or sucrose, lures or sorbitol, cellulose derivatives and/or calcium phosphates, for example tricalcium phosphate or acid phosphate of calcium, as a binder component can be used such as starch paste, for example corn, wheat, rice, potato starch, gelatin, tragakant, methylcellulose, hydroxypropylmethyl llulose, sodium carboxymethylcellulose and/or polyvinylpyrrolidone. If necessary, can be used loosening agents, such as the abovementioned starches and carboximetilkrahmal, transversely crosslinked polyvinylpyrrolidone, agar or alginic acid or its salt, such as sodium alginate.

Can be used optional additives, such as agents that regulate the fluidity and lubricating agents, such as silica, talc, stearic acid and its salts, such as magnesium stearate or calcium stearate and/or propylene glycol.

The core tablets generally covered with a layer, which is resistant to gastric juice. For this purpose, can be used in concentrated solutions of sugars, which may not necessarily contain the Arabian gum, talc, polyvinylpyrrolidone, polyethylene glycol and/or titanium dioxide, and suitable organic solvents or mixtures thereof.

The additives can also be used stabilizers, thickeners, dyes and fragrances.

As ointment bases may be used hydrocarbon ointment bases such as petrolatum, white and yellow (Vaselinum album, Vaselinum flavum), vaseline oil (Oleum Vaselini), white ointment and liquid (Unguentum album, Unguentum flavum), and as additives to give a more solid consistency, such as paraffin wax and wax; absorptive motivaional, such as hydrophilic petrolatum (Vaselinum hydrophylicum), lanolin (Lanolinum), cold cream (Unguentum leniens); ointment bases, water washable, such as hydrophilic ointment (Unguentum hydrophylum); water-soluble ointment bases, such as polietilenglikolja ointment (Unguentum Glycolis Polyaethyleni), bentonite foundations and others.

As the basis for gels can be used methyl cellulose, sodium carboxymethyl cellulose, oxypropylation, polyethylene glycol or polyethylene oxide, carbopol.

As the basis for the suppository can be used bases, insoluble in water, such as cocoa butter, principles, soluble in water or miscible with water, such as gelatin-glycerin or polietilenoksidnoy; combined basics - soap-glycerin.

In the preparation of standard dosage forms, the amount of active ingredient used in combination with a carrier, may vary depending on the recipient treated, the specific method of administration of the drug.

For example, when using the compounds of the present invention in the form of solutions for injection, the concentration of active agent is from 0.01 to 5%. As diluents can be used with 0.9% sodium chloride solution, distilled water, a solution of novocaine injection, ringer's solution, the solution is lukose, specific additives to dissolve. When introduced into the body of the compounds of the present invention in the form of tablets and suppositories their number is 5.0-500 mg standard dosage form.

Dosage forms of the present invention receive by standard techniques, such as, for example, the processes of mixing, granulating, the formation of drops, dissolution and lyophilization.

It should be noted that the compounds of the present invention show a biological activity in doses of two to three orders of magnitude lower in comparison with the known compounds used for comparison, with almost the same efficiency, and there are no identified negative side effects and no contraindications. In this case study, the toxicity of the compounds of the present invention at a dose of 3000 mg/kg, oral, has not registered the death of experimental animals.

Detailed description of the compounds of the present invention, their preparation and study of pharmacological activity are presented in the following examples are intended to illustrate the preferred variants of the invention and are not limiting its scope.

Examples of the synthesis of N-acyl derivatives of amino acids of General formula (I)

The identity of the compounds was checked by TLC on what Laschenko "Kieselgel 60 F 254" Merck (Germany) systems: methanol (1), chloroform-methanol-ammonia(4:3:1) (2).

The chromatogram showed chlorthalidone reagent, ninhydrin, iodine to glow in UV light.

The angles of the optical rotation was measured on a polarimeter "Perkin Elmer 341 (Sweden).

1H-NMR were recorded on a device "AMX-400 Bruker (Germany).

The melting point was determined on the device "Boetius (Germany).

Analytical HPLC sang and danced on the device "System Gold" ("Beckman, USA), the rate of elution of 0.25 ml/min, detection at 214 nm in the conditions: column Ultrasphere ODS "Beckman", C mm, 5 μm, elution with 0.1%TFA, the rate of elution of 0.25 ml/min (1).

Example 1

Nα-Glutaryl-L-histidine (IV)

Method And

To a solution of 103,4 g (0.67 mol) of histidine in 400 ml of water is added to 83.7 g (0.73 mol) of glutaric anhydride. The suspension is stirred for 1 hour, the resulting solution is evaporated to a volume of 150 ml, stored in the refrigerator for 16 hours. The precipitation is filtered off, washed with 150 ml of methanol and dried. Cleaning is performed by ion-exchange chromatography on resin Purolite N+form elwira water. The fractions containing the desired product are pooled, evaporated prior to the deposition of sediment and left for 16 hours at +4°C. the precipitation is filtered off, washed with 200 ml of methanol and dried to constant weight. Output 98,8 g (55%). Rf0,55 (1), 0,37 (2). TPL= 222-224°C. [α]DȊ 20+15, 95° (C 0,53, water). [M+H]+270,1.1H-NMR spectrum (D2O), δ, ppm: 1,60-1,80 (m, 2H, β-CH2-Glt), 2,10-of 2.25 (m, 4H, α,γ-CH2-Glt), 2,90-of 3.25 (m, 2H, β-CH2-His), 4,40-4,50 (m, 1H, α-CH-His), to 7.15 (s, 1H, CH-4-Im), and 8.50 (s, 1H, CH-2-Im).

Found, %: C 49,18; H 5,91; N 15,42.

C11H15N3O5.

Calculated, %: C 49,07; H 5,62; N 15,61.

Method B

To a suspension of 0.3 g (of 1.93 mmol) of histidine in 5 ml of water with vigorous stirring of 0.44 g (3,86 mmol) of glutaric anhydride, dissolved in 2.5 ml of ethyl acetate. Stirred for 2 hours, the pyridine was adjusted pH to 7 and stirred for further 1 hour. An ethyl acetate and the aqueous layer was separated. The aqueous layer was washed twice with ether, the ether layer discarded. Water is removed in vacuum, the residue is dissolved in a minimum amount of water and add ethanol until the beginning of precipitation of a white precipitate, leaving at +4°C 20 hours the Precipitate was separated by filtration, dried in vacuum. Yield 0.36 g (70%). Rf0,56 (1), 0,35 (2). TPL= 219-221°C. [α]D20= +15,71° (C 0,56, water). [M+H]+270,1.1H-NMR spectrum (D2O), δ, ppm: 1,40-of 1.55 (m, 2H, β-CH2-Glt), 1,90-2,0 (m, 4H, α, γ-CH2-Glt), of 2.7-3.0 (m, 2H, β-CH2-His), 4,20-4,30 (m, 1H, α-CH-His), 6,95 (c, 1H, 4-CH-Im), 8,30 (c, 1H, 2-CH-Im). HPLC under the conditions: (1) individual peak, retention time 14,55 minutes

Found, %: C 49,07; H 5,65; N 15,65.

C11H15N3O5.

Calculated, %: C 49,07; H 5,62; N 15,61.

Example 2

Nα-Succinyl-L-histidine (V)

The synthesis was carried out in accordance with methods As described for compound IV.

Yield 0.08 g (57%).

Rf0,44 (1), 0,25 (2).

TPL= 179-181°

[α]D20= +30,71° (C 0,56, water)

[M]+255,2

1H-NMR spectrum (D2O), δ, ppm: 2,15-of 2.30 (m, 4H, (CH2)2-Suc), of 2,75 2,95 (m, 2H, β-CH2-His), 4,25 (USS, 1H, α-CH-His), 6,95 (c, 1H, 4-CH-Jm), 8,25 (c, 1H, 2-CH-Jm).

Found, %: C 47,09; H 5,04; N 16,40.

C10H13N3O5.

Calculated, %: C 47,06; H 5,13; N 16,46.

The synthesis was carried out according to method B described for compound IV.

Output 0,101 g (67%).

Rf0,45 (1), 0,27 (2).

TPL= 178-180°

[α]D20= +30,8° (C 0,57, water)

HPLC under the conditions (1) - individual peak, retention time 7,54 minutes

Found, %: C 47,15; H 5,2; N 16,50.

C10H13N3O5.

Calculated, %: C 47,06; H 5,13; N 16,46.

Example 3

Nα-Succinyl-L-tryptophan (II)

The synthesis was carried out according to method B described for compound IV.

The output of 100.5 mg (67%).

Rf0,39 (1).

1H-NMR spectrum (CD3OD), δ, ppm: 1,87-of 1.93 (m, 2H, β-CH2-Glt), 2,25-is 2.37 (m, 4H, α,γ-CH2-Glt), 3,37-3,47 (m, 2H, β-CH2-Trp), 3,81-of 3.94 (m, 1H, α-CH-Trp), of 6.99 (t, J 7 Hz, 1H, CH-6-Ind), 7,07 (t, J 7 Hz, 1H, CH-7-Ind), from 7.24 (d, J 7 Hz, 1H, CH2-Ind), to 7.35 (d, J 7 Hz, 1H, CH-5-Ind), 7,55 (d, J 7 Hz, 1H, CH-8-Ind).

[M]+304,3

Found, %: C 59,07; H 5,65; N 9,35.

C15H16N2O5.

Calculated, %: C 59,21; H 5,3; N Of 9.21.

Example 4

Glutamylation (III)

The synthesis was carried out according to method B described for compound IV.

The output of 100.5 mg (67%).

Rf0.63 (1).

=+21,05° (0,6, water)

1H-NMR spectrum (DMSO-d6), δ, ppm: 2,33-to 2.41 (m, 4H, α,βCH2-Suc), 2,93-a 3.01 (m, 1H, βCH2-Trp), 3.10-3.16 (m, 1H, βCH2-Trp), 4,39-4,47 (m, 1H, α-CH-Trp), 6,93-7,06 (m, 2H, CH-6,7-Ind), 7,11 (d, J 2.2 Hz, 1H, CH-2-Ind), 7,30-7,32 (m, 1H, CH-5-Ind), 7,44-7,47 (m, 1H, CH-8-Ind). [M]+304,3.

HPLC under the conditions: (2) individual peak, retention time 6,35 minutes

Found, %: C 59,07; N The 5.65; N 9,35.

C15H16N2O5.

Calculated, %: C 59,21; N 5,3; N Of 9.21.

Example 5

Monosodium salt of Nα-glutaryl-L-histidine (IV)

To a solution of 1.0 g (3.7 mmol) of Nα-glutaryl-L-histidine in 15 ml of water under stirring and cooling to +5°added a solution of 0.15 g (3.7 mmol) of NaOH in 20 ml of water. The solution is stirred for 30 min, the solvent is removed in vacuum. To the oily residue are added in several portions of benzene, the solvent is removed in vacuum. The solid residue is dried over granular alkali.

Yield 1.07 g (99.7%).

TPL=208-210°

=+16.27° (0.58, water)

Found, %: C 45,25; N 5,51; N 14,52.

C11H15N3O5Na.

Calculated, %: C 45,21; N. Of 5.17; N 14,38.

Example 6

Monosodium salt of Nα-succinyl-L-histidine (V)

The synthesis was carried out in accordance with the methodology described for the monosodium salt of Nα-glutaryl-L-histidine (IV) (example 5).

Yield 1.06 g (97.0%).

=+40.21° (C 0.48, water)

Found, %: C 43,25; N 4,51; N 15,52.

C10H13N3O5Na.

Calculated, %: C 43,17; N 4,71; N 15,10.

Example 7

Monosodium salt of Nα-succinyl-L-tryptophan (II)

The synthesis was carried out in accordance with the methodology described for the monosodium salt of Nα-glutaryl-L-histidine (IV) (example 5).

Yield 0.21 g (98.0%).

TPL=147-150°

=+22.02° (C 0.39, water)

Found, %: C 55,25; N 4,51; N 8,32.

C15H16N2O5Na.

Calculated, %: 55,05; N Is 4.93; N 8,56.

HPLC under the conditions: (2) individual peak, retention time 6,56 minutes

Example 8

Monosodium salt of Nα-glutaryl-L-tryptophan (III)

The synthesis was carried out in accordance with the methodology described for the monosodium salt of Nα-glutaryl-L-histidine (IV) (example 5).

Yield 0.11 g (99.0%).

TPL=128-130°

=+22.06° (C 0.34, methanol)

Found, %: C 56,15; N To 5.21; N by 8.22

C16H18N2O5Na.

Calculated, %: C 56,30; N 5,32; N 8,21.

HPLC under the conditions: (2) individual peak, retention time of 6.96 minutes

Example 9

The disodium salt of Nα-glutaryl-L-histidine (IV)

To a solution of 1.0 g (3.7 mmol) of Nα-glutaryl-L-histidine in 15 ml of water under stirring and cooling to +5°added a solution of 0.3 g (7,44 mmol) of NaOH in 15 ml of water. The solution is stirred for 30 min, the solvent is removed in vacuum. To the oily residue are added in several portions of benzene, the solvent is removed in vacuum. The solid residue is dried over granular alkali.

Yield 1.15 g (99.0%).

=+11.92° (C 0.57, water)

Found, %: C 41,25; N 4,51; N 13,52.

C141H15N3O5Na2.

Calculated, %: C 41,91; N 4,80; N 13,3.

Example 10

The disodium salt of Nα-succinyl-L-histidine (V)

The synthesis was carried out in accordance with the methodology described for the disodium salt of Nα-glutaryl-L-histidine (IV) (example 9).

Yield 1.16 g (99.0%).

TPL=124 to 128°

=+20.06° (C 0.67, water)

Found, %: C 39,55; N Or 4.31; N 13,52.

C10H13N3O5Na2.

Calculated, %: C 39,88; N. Of 4.35; N 13,95.

Example 11

The disodium salt of Nα-succinyl-L-tryptophan (II)

The synthesis was carried out in accordance with the methodology described for dinat what Ieva salt N α-glutaryl-L-histidine (IV) (example 9).

Yield 0.56 g (97.7%).

Found, %: C 51,35; N Or 4.31; N By 8.22.

C15H16N2O5Na2.

Calculated, %: 51,43; N 4,60; N 8,0.

Example 12

Di-sodium salt of Nα-glutaryl-L-tryptophan (III)

The synthesis was carried out in accordance with the methodology described for the di-sodium salt of Nα-glutaryl-L-histidine (IV) (example 9).

Yield 0.56 g (98.5%).

Found, %: C 52,55; N 4,71; N 7,52.

C16H18N2O5Na2.

Calculated, %: C 52,75; N To 4.98; N 7,69.

Tests for biological activity

Example 13

The effect of compounds of General formula I at allergic reactions of immediate type (test induced by ovalbumin (OA) degranulation of basophils in the blood of immunized Guinea pigs in vitro.

The selection of leukocytes from the blood of Guinea pigs was carried out according to the method of Primes [Immunological methods / edited Grimes /, M.: Medicine, 1987, str in our modification].

For the production test was used Guinea pigs of both sexes weighing 600-800, Animals were immunized once with a mixture of 10 μg of ovalbumin and 100 mg of aluminium hydroxide on animal Andersson [Anderson P. Antigen-induced bronchial anaphulaxis in actively sensitized geinea-pigs. // Allergy. - 1980. - Vol.35. - P.63-71].

Under ether anesthesia from the heart of the Guinea pig were collected 15 ml of blood. For selection of basophils in the composition of the leukocyte suspension used in isovale dual deposition cells by EDTA and using nitratsoderzhaschie precipitating liquid.

The blood was mixed with 5% EDTA·Na22H2O (Sigma) in a ratio of 9:1 and after 30 minutes, gently centrifuged (12 min at 80 g). The supernatant was collected and centrifuged 15 min at 500 g.

The remaining blood cells were added nitratsoderzhaschie precipitating liquid (3) in a ratio of 3:10 (thermostatically at 37°C for 30 min). Enriched leukocytes supernatant fraction was centrifuged for 7 min at 100 g. To the precipitate of cells was added to a 0.85% NaCl solution and the cell concentration was brought to 30×103/µl.

Staging test degranulation of basophils in vitro [Handbook of clinical laboratory research methods / edited Eaast /, M.: Medicine, 1975, str].

For the production test in the centrifuge tube (used 3 tubes per sample) were placed 300 µl of cell suspension was then added 300 μl of salt solution of tested compound (or saline in the control of spontaneous and maximal degranulation) and preincubated at 37°C for 15 min, then was added 300 μl of 1% saline OA in each tube (in the control of spontaneous degranulation was added to the saline solution in the same amount) and again was preincubated at 37°within 10 minutes working concentration of leukocytes is when 104/µl. From each tube was sampled (100 μl) in the Department of the major vessels to assess the full degranulation of basophils, and the remaining cells were added to the cooled saline solution (5 ml in each tube) to stop reaction degranulation, and then were centrifuged for 7 min at 100 g, and the precipitate was preparing preparations for microscopic examination. Fixation and coloration of the drugs was performed by the method Seder et al. [R.A. Seder et al. Mouse splenic and bone marrow cell populations that express high - affinity Fes receptors and produce interleukin-4 are highly enriched in basophils. // Proc. Natl. Acad. USA - 1991 - V.88 - P.2835-2839].

To identify specific grain basophils used dye 0,5% Allenby blue (pH 1.0)and kernel they finished painting areas safranin (0.1% solution in 1% acetic acid). Drugs used to estimate total inhibition of degranulation.

Inhibition of total degranulation (TG) (%) was calculated by the formula

where the Mach - % degranulated basophils at maximum degranulation (OA);

spent. - % degranulated basophils in spontaneous degranulation (control);

exp. - % degranulated basophils after exposure to the compounds.

A full assessment of degranulation of basophils

Selected after setting the test degranulation of basophils samples (100 μl) were placed in tubes with dye (0.5% of Allenby blue, pH 1.0) 1:1 ratio. The painting was made at a room temperature not less than 50 minutes counting the number of stained basophils held the camera Fuchs-Rosenthal. Braking is full of degranulation (TPD) basophils was calculated by the formula

TPD(%)=1-[(MCP-MCP(exp.)]/[MCP-MCP(OA)]×100,

where MCP(K) - average (3 samples) the number of basophils in the test spontaneous degranulation;

MCP(OA) - average (3 samples) the number of basophils in the test maximum antigonadotropin degranulation;

MCP(exp) - average (3 samples) the number of basophils in the test degranulation after incubation with the test compound.

Table 2
Inhibition of OA-induced degranulation of basophils in the blood of immunized Guinea pigs in vitro under the influence of the compounds of General formula I
no experience n/pGroupBraking is full of degranulation (TPD), (%)The number of completely degranulated basophils, (%)
1.Control 1 (spontaneous degranulation)1000
2.Ovalbumin 1% (max degranulation)035,2±0,8
3.Compound IV (10-3M)99,2±11,22,6±2,6*
4.Compound IV (10-4M)99,5±to 12.02,9±1,6*
5.Compound IV (10-5M)113,5±2,70
6.Compound IV (10-6M)90,0±1,83,9±0,6
7.Compound IV (10-7M)76,7±1,59,11±0,8
8.Glutamylcysteine (10-3M)9,3±5,531,6±6,33
9.Glutamylcysteine (10-4M)24,1±1,125,3±3,38
10.Glutamylcysteine (10-5M)029,8±of 6.73
11.Control 21000
12.Ovalbumin 1% (max degranulation)038,91±8,43
13.Compound V (10-3M)10,6±7,635,31±7,29
14.Compound V (10-4M)18,9±11,831,2±6,8
15.Compound V (10-5M)44,01±11,2724,61±10,38
16.Hydrocortisone (10-3M)71,0±1,610,0±0,7
17.Hydrocortisone (10-4M)480± 0,817,3±0,9
18.Hydrocortisone (10-5M)40,0±0,620,0±1,3
(* - P<0,001)

The data in table 2 show that, compared with glutamylcysteine compound (IV) has expressed antianaphylactic effect, manifested almost 100% inhibition of the degranulation test full OA-induced degranulation of basophils blood actively immunized Guinea pigs (reaction anaphylaxis in vitro in ascallaway environment). Significant antianaphylactic the effect of compound IV is manifested in the decrease in the number degranulating cells, especially pronounced at a concentration of 10-5M (no degranulating cells).

Example 14

To study the effect of compounds of General formula I in systemic anaphylaxis in vivo

Used the model of bronchospasm in generalizirovanny actively sensitized Guinea pigs with aerosol exposure of ovalbumin as an antigen [V.L. Kovalev "guidelines for the study of bronchodilators, mucolytic and anti-inflammatory drugs. // Guidance on experimental (preclinical) study of new pharmacological substances. - Moscow. - 2000. - S-250].

Guinea pigs were senzibilizirani the ovalbumin according to the method of Andersson [ndersson P. Antigen-induced bronchial anaphulaxis in actively sensitized geinea-pigs. // Allergy. - 1980. - Vol.35. - P.63-71] and in 1-2 months after sensitization induced bronchospasm aerosol introduction resolution doses of ovalbumin (3 mg/kg in 1 ml of physiologic saline).

In the experimental groups of Guinea pigs within three days intragastric tube has introduced the compounds in doses of 10 ág/kg 50 ág/kg 150 ág/kg In another series of experiments the compounds at a dose of 50 mcg/kg (1 ml physiologic saline) was administered inhalation (via nebulizer) within three days 1 time per day. The control group was injected physiologic solution. Within 1 hour after the last injection of substances was ingalirovti using the nebulizer ovalbumin and estimated duration (in seconds) and the intensity of bronchospastic reactions of animals.

Table 3
The braking system anaphylactic reaction Guinea pigs inhalation introducing the compound IV at a dose of 50 mg/kg
GroupThe duration of the acute phase, withThe duration of the subacute phase, with
Control 1

(physiologic solution)
180±6650±34
Compound IV

50 ág/kg
0400±25

Table 4
The braking system anaphylactic reactions of Guinea pigs after intragastric administration of compound IV in doses of 10 and 150 µg/kg (M±m)
GroupThe duration of the acute phase, withThe total reaction time,
Control 2 (physiologic solution)296,7±104,6628,3±80,6
Compound IV

10 ug/kg
68,0±54,7*428,0±75,0
Compound IV

150 ág/kg
72,0±42,1*337,0±78,5*
* - P<0,001

The results of the experiments are presented in tables 3 and 4 show that compound IV after intragastric administration at doses of 10 and 150 mcg/kg and inhalation dose of 50 mg/kg showed antianaphylactic activity. Inhalation introduction of a substance at a dose of 50 mg/kg blocked the development of acute phase bronchoconstricting reaction, which is causing asphyxia, is a cause of death. After intragastric administration of compound IV in doses of 10 mg/kg and 150 mg/kg was found a significant protective effect against antigonadotropin bronchospasm.

Thus, compound IV has a significant protective effect against systemelectrical reactions in vivo.

Example 15

Anti-allergic activity of the compounds of General formula I on the model of allergic rhinitis in Guinea pigs

Used model of allergic rhinitis in Guinea pigs.

Guinea pigs were immunized according to a certain scheme for 1.5-2 months (Hutson P.A., Church, M.K. et al. 1988): initially, animals were immunized by intraperitoneal injection of ovalbumin in the dose of 10 mg/kg at 7-day intervals (twice), and then the pigs were ingalirovti using nebulizers technology (Pari) a solution of ovalbumin in increasing concentrations, ranging from 0.1% to 1%, with an interval of 4 days between inhalations. The last dose of ovalbumin was administered in the nasal passages using a micropipette. 24 hours after the last injection of ovalbumin produced the fence nasal rinse (through the system of special tubes) and changes in the nasal mucosa was assessed by using a range of methods: histological and cytological. The compounds (0.1% solution) was administered daily by inhalation using nebulizers equipment for 6 days, on the 6th day of injection caused the provocation antigen (OA). Nasal wash was received the next day after the provocation.

Table 5
The effect of compound IV on lymphocytosis (absolute number of cells in 1 ml) in nasal is flush
GroupIntact.

control
Model + provokes.

(rhinitis)
Compound IV

(0.1% of R-R)
N677
Lymphocytosis18,012,1067,2±6,47**43,7±6,65°*
* the difference from the intact control; ° - contrast model

(° - P<0,05; * P<0,01; ** - P<0,001)

Table 6
The effect of compound IV on indicators cytogram (%) nasal rinse Guinea pigs
Group

The subpopulation
The intact

control

n=6
Model

(rhinitis)

n=7
Compound IV

(0.1%of R-R) n=7
Macrophages21,8±1,6412,7*±11,5714,6*±1,27
Lymphocytes4,7±0,886,1±1,425,7±1,54
Neutrophils0,3±0,213,3*±10,807,8**°±1,35
Eosinophils7,0±2,4547,9***±shed 15.376,9°°°±1,32
The epithelial cells66,2±3,09 27,1***±15,0163,6°°°±3,02
* the difference from the intact control; ° - contrast model (rhinitis)

(° - P<0,05; ** - P<0,01; ***, °°° - P<0,001)

From the data of tables 5-7, it follows that in terms of modeling allergic rhinitis compound IV significantly suppresses eosinophilic inflammation. This is confirmed by the reduction to normal absolute and relative number of eosinophils, as well as reduction of lymphocytosis in natalem flush.

Example 16

Anti-allergic activity of the compounds of General formula I on the model of allergic lung inflammation in Guinea pigs

Used model of allergic lung inflammation in Guinea pigs.

Immunization of animals similar to that described in example 14.

24 hours after the last injection of ovalbumin produced the fence bronchoalveolar flushing (via a cannula inserted into the trachea), and changes in the bronchial mucosa was assessed by using a range of methods: histological and cytological.

The compounds (0.1% of R-R) was administered daily by inhalation using a nebulizer equipment for 6 days, on the 6th day of injection caused the provocation antigen (OA).

Table 7
The effect of compound IV on the absolute number of cell subpopulations nasal rinse Guinea pigs (1 ál)
Group

The subpopulation
Intact.

control

n=6
Model (rhinitis)

n=7
Compound IV

(0.1% of R-R)

n=7
Macrophages6,2±1,297,9±1,886,1±0,75
Lymphocytes0,9±0,244,1*±1,163,2*±52
Neutrophils0,03±0,0212,1±0,673,1±1,02
Eosinophils1,2±0,4626,5***±4,473,2°°±0,97
The epithelial cells11,8±1,411,3±2,1026,3**°°±or 4.31
Note: * - the difference from the intact control; ° - contrast model

(* - P<0,05; **, °° - P<0,01; ***, °°° - P<0,001)
Table 8
The effect of compound IV on lymphocytosis (absolute number of cells in 1 μl) in bronchoalveolar washings
GroupIntact. controlModel + provokes.
(rhinitis)
Compound IV

(0.1% of R-R)
N657
Lymphocytosis726,8±82,41849,4±287,3*1331,3±277,4
* P<0,01 - contrast to the intact control

Table 9
The effect of compound IV on the absolute number of cell subpopulations bronchoalveolar flushing Guinea pigs (1 ál)
Group

The subpopulation
Intact.

counter.
ModelCompound IV

(0.1% of R-R)
Macrophages502,6±60,31

(n=6)
680,3±105,0

(n=5)
640,1±57,98

(n=6)
Lymphocytes101,4±24,3

(n=6)
328,3**±49,18

(n=5)
233,0*±45,77

(n=7)
Neutrophils0

(n=6)
56,8±17,08

(n=5)
16,9°±4,22

(n=7)
Eosinophils37,0±9,04

(n=5)
773,0**±171,7

(n=5)
487,4**±98,70

(n=6)
The epithelial cells16,9±6,09

(n=6)
0

(n=5)
2,48*±1,84

(n7)
* the difference from the intact control; °- contrast model

(*, ° - P<0,05; ** - P<0,01)

From the data of tables 8 and 9 it follows that the compound IV in the model of allergic inflammation of the lungs significantly inhibits the inflammatory process, which is a reduction of lymphocytosis, a decrease in the content key cells of allergic inflammation, eosinophils, a sharp decline in neutrophils and a decrease in the number of lymphocytes.

Example 17

The study of anti-inflammatory action of compounds of General formula I in models of inflammation of the lungs in rats induced by Sephadex

Model cephalexindiovan (6-day) pneumonia in rats

In experiments were used rats male Wistar breed weighing 270-300 g

Inflammation in the lungs induced single inhalation introduction of Sephadex A-25 (hydrophilic powder with a particle size of from 20 to 80 μm) in a dose of 5 mg/kg with a measuring device, which is the laboratory analogue inhaler "Ziklohaler" (research Institute of pulmonology of the RF).

The technique of inhalation of Sephadex and pharmacological substances

Rats with the original dosing device for inhalation dry powders under ether anesthesia was introduced Sephadex A-25 in the dose of 5 mg per 1 kg of body weight. Rats vist the R after the introduction of Sephadex A-25 quickly came out of the anesthesia and outwardly any peculiarities in the behavior, the nature of the breath they were not observed. Substances in the form of dry powder was introduced inhalation at a dose of 500 mcg/kg over 1 hour after administration of Sephadex, then for 5 consecutive days daily 1 time per day in the same morning. The control is represented by 2 groups: the group of intact animals, a group of rats that inhaled once introduced Sephadex.

The results of therapeutic effects of pharmacological substances on the development of the inflammatory process in the lungs was assessed 6 days after aerosol exposure Sephadex using morphological and morphometric parameters (bulk density emphysema and alveolitis).

The methods used in the work

Histologic

Conducted histological examination of the lungs stained with hematoxylin and eosin.

Morphometric

Prepared histological sections of the lungs of a thickness of 4-5 microns, which counted the number of neutrophils in millionary partitions, and estimated volumetric density of alveolitis and emphysema using grid Avtandilov [Avtandil GG, Introduction to quantitative pathological morphology. // M: Medicine. - 1980. - s]. Also conducted a morphometric study of the lymphoid tissue of the lungs. To this end, micropreparative lungs were fixed by the method Beinenstock et al [Bienenstock J., Johnson N., Perey D.Y.E. Bronchial lymphoid tissue 1. Morphologic chaacteristics // Lab. Invest. - 1973. - v.28 - p.693-698]. Lungs with trachea was removed from the thoracic cavity, and microreport were placed in a 2% aqueous solution of acetic acid. After 18-24 hours, the trachea, the main and lobar bronchi were cut off; the method of point account under a magnifying glass (UV) conducted a morphometric assessment of bulk density of lymphoid tissue associated with the bronchi. Method point account determined bulk density of alveolitis and emphysema.

Cytological

Bronchoalveolar wash in rats and Guinea pigs received under geksenalovy anesthesia by twice washing the lungs through the trachea with 10 ml of saline. Cell viability was determined in the test with Trifanova blue. In bronchoalveolar fluid flush (BASS) with the camera Goryaeva determined the absolute number of cells in 1 ml (lymphocytosis). In smears of the sediment liquid BASS, obtained by centrifugation at 200 g for 10 minutes and then stained by Romanovsky-Giemsa, and counted andpulmonary they (in percent) [Avtsin A.P., Lukomsky GI, Romanov, L.K. et al. Andpulmonary they. // Sov. the honey. - 1982. No. 7. - p.8-14].

The research results are processed by methods of variation statistics using t-student test.

Table 10
parameters of bronchoalveolar lymphocytosis flushing of Wistar rats after aerosol exposure Sephadex A-25 and treatment with pharmacological agents (M± m)
Lymphocytosis
The absolute number of cells in 1 μl of BAL
CriterionIntact.

n=5
The Sephadex

(model)

n=6
Compound IV

n=6
Lymphocytosisof 160.4±20,65259,2*±32,42178,6*±20,4
P0,050,05
Note: * - the difference from the intact control

Table 11
Indicators of cellular composition of bronchoalveolar flushing of Wistar rats after aerosol exposure Sephadex A-25 and the treatment of the compounds (M±m)
Lymphocytosis
The absolute number of cells of different subpopulations in 1 μl of BAL
The subpopulationIntact. counter.Model (Sephadex)Compound IV
N456
Macrophagesof 124.8±16,35226,1*±30,83152,4±18,5
Lymphocytes15,5±4,2728,2±6,0122,0*�B1; 3,5
Neutrophils020,4*±6,385,1*±0,6
Note: * - the difference from the intact control (P<0,05)

Histological examination of lungs

Compound IV caused a distinct anti-inflammatory effect: the incidence of alveolitis was significantly lower compared with the model group of animals; emphysema is almost not detected; not marked infiltration of neutrophils millionary partitions. The level of lymphocytosis and the number of neutrophils in BAL inflammatory process is also much less pronounced than in animals treated with Sephadex within 5 days.

Thus, the whole complex used experimental models shows a significant antiallergic, antianaphylactic and anti-inflammatory activity of compounds of General formula I, as shown in in vitro tests, and modeling, allergic and inflammatory pathology in vivo.

Example 18

The study of the hypolipidemic action of compounds of General formula (I) models of hypercholesterolemia in rats

The study was performed on rats male Wistar mass 200±20, Hyperlipidemia caused by oral administration of cholesterol load - oil suspension cholesterol:

olive oil (Acorsa, And the company) - 5 ml/kg of animal weight;

cholesterol (Sigma, USA), 1 g/kg;

Holt sodium (Sigma, USA), 100 mg/kg of body weight.

As the comparison drug used drug from the group of statins "mevacor" (lovastatin) Merck Sharp & Dohn dose 4 0 mg/kg Cholesterol suspension was injected in the morning daily for 10 days. The compounds (at a dose of 500 mcg/kg) and the reference product (at a dose of 40 mg/kg) was administered to animals with cholesterol suspension within 10 days. All animals received standard briquetted feed.

Animals were divided into following groups:

"Control" - intact animals (n=6)

"Cholesterol" - rats treated per os cholesterol loading (n=10)

"Lovastatin" - rats treated per os cholesterol loading and lovastatin (n=10)

"compound IV" - rats treated per os cholesterol loading and investigational compound IV (n=10).

Blood samples were taken at 5, 8, 10 day experiment.

Statistical processing of the hypolipidemic action of the compounds under investigation conducted in relation to the group of "cholesterol" (table 13).

Table 12
Cholesterol and triglycerides in the serum and liver of rats treated for 10 days per os olive oil, cholesterol and compound IV at the same time holesterinovmi load
Control

(n=6)
Cholesterol

(n=10)
Lovastatin

(n=10)
compound IV

(n=9)
Total cholesterol
Serum (mg/100 ml)67,2±6,1120,0±8,4100,5±6,7*96,6±5,7**
HDL (mg/100 ml)41,6±1,354,3±1,750,6±1,350,5±1,3*
LDL + VLDL (mg/100 ml)25,6±0,865,7±1,2to 49.9±0,9***46,1±0,9***
Liver (mg/g tissue)2,29±0,133,6±0,42,41±0,17***2,78±0,28***
Triglycerides
Serum (mg/100 ml)85,7±9,299,2±8,793,6±7,595,1±7,9
Liver (mg/g tissue)3,89±0,146,83±0,396,25±0,13of 5.53±0,19***
* - p<0,1
** p<0,05
*** - p<0,01

The rst is the group of compounds IV in a dose of 500 mg/kg resulted in a significant decrease in total serum cholesterol by 19.5%, liver 22.7%, LDL cholesterol by 29.8%, triglycerides liver by 19%. Connection glutamylcysteine, reducing total cholesterol by only 9%, influenced only on LDL and VLDL (lipoproteins of low and very low density), as shown in a publication of the international application WO 99/01103, and obviously less effective than compound IV, in similar biological experiment.

Below are the results of other studies of the claimed compounds.

The experimental group consisted of the following connections:

1) "Cholesterol" - treated for 10 days per os oil suspension cholesterol;

2) "compound IV" - rats treated per os oil suspension of cholesterol and investigational compound IV;

3) "compound IV-INa" - rats treated per os oil suspension of cholesterol and analyzed monosodium salt of compound IV;

4) "compound IV-2Na" - rats treated per os oil suspension of cholesterol and studied the disodium salt of compound IV;

5) connection V-INa" - rats treated per os oil suspension of cholesterol and analyzed monosodium salt of compound V;

6) the compound III-INa" - rats treated per os oil suspension of cholesterol and studied mono-sodium salt of compound III;

7) "compound II-INa" - rats treated per os oil suspension of cholesterol and studied monolatry the first salt of compound II;

8) "Sim" - rats treated per os oil suspension of cholesterol and simvastatin;

9) control - intact rats before the start of the experiment.

On the 10th day experience blood samples were taken after decapitate animals. Before decapitation, the animals were starved for 12 hours.

In serum measured total cholesterol, triglyceride, cholesterol, high density lipoprotein (HDL). Cholesterol of lipoproteins of low and very low density was calculated by the difference between total cholesterol and HDL cholesterol.

The content of total cholesterol and triglycerides in serum were determined by enzymatic method.

Determination of cholesterol in high density lipoprotein (α-PL) was performed by precipitation of LDL and lonp phosphomevalonate acid and magnesium ions.

Statistical processing.

Data in tables are presented as the mean ± standard error of the mean. The significance of differences between groups "cholesterol" and "the drug..." was assessed by two-sample t-test t-test. The probability of error (p) are indicated in the columns of the table.

Data on the effect of the studied compounds on cholesterol and triglycerides in serum of rats fed cholesterol load, are presented in tables 13-20.

Table 13
Experimental hypercholesterolemia
The lipid metabolism of rats (mg/100 ml)
before the start of the experiment

(n=50)
10 days of cholesterol load

(n=20)
Total cholesterol
Serum59,9±1,4135,7±10,9
HDL38,2±1,232,5±1,1
LDL + lamp20,3±1,3103,2±11,5
Triglycerides
Serum83,6±4,6of 156.6±11,4
HDL19,0±1,424,3±1,8
LDL + VLDL58,7±6,2to 132.6±12,8

Ten rats to an oil suspension of cholesterol resulted in significant increase of total cholesterol in serum by 2.3 times, triglycerides 1.9 times. During the development of induced hyperlipidemia cholesterol HDL was reduced by 15%. Cholesterol LDL + VLDL increased in 5 times. Triglycerides LDL + lonp increased 2.3 times.

Ten rats to an oil suspension of cholesterol resulted in significant increase of total cholesterol in serum 1.7 times, triglycerides 1.6 times (table 17). The HDL-cholesterol was decreased by 28% from the original 66,7 to 48 mg/100 ml during the development of induced hyperlipidemia. Cholesterol LDL + VLDL increased 4.4 times from 22.5 to 99 mg/100 ml

Table 18
Cholesterol and triglyceride levels (mg/100 ml) in the serum of rats on day 5 of the experiment
Indicatorsgroup
"Cholesterol" (n=12)"compound IV-1Na (n=12)"connection V-1Na (n=12)the compound III-1Na (n=12)"compound II-1Na (n=12)
Total serum cholesterol112,9±9,294,4±3,4

p=0,079
96,9±4,589,0±5,0

p=0,035
109,7±9,4
HDL47,5±3,760,7±3,1

p=0,012
55,1±2,8

p=0,11
49,0±2,149,5±3,8
LDL + VLDL65,4±10,633,7±4,5

p=0.015/td>
41,7±6,5

p=0,073
40,0±5,4

p=0,048
60,2±10,6
Triglycerides
Serum94,5±9,283,2±6,363,3±5,4

p=0.009
87,4±9,480,9±5,5
HDL35,0±3,139,4±3,530,9±1,830,0±2,232,6±2,5
LDL + VLDL59,5±7,643,7±4,6

p=0,09
32,4±4,4

p=0.007
57,4±9,148,2± and 4.9

Table 19
Cholesterol and triglyceride levels (mg/100 ml) in the serum of rats on day 8 of the experiment
Indicatorsgroup
"Cholesterol"

(n=12)
"compound IV-1Na"

(n=12)
"connection V-1Na"

(n=12)
the compound III-1Na"

(n=12)
"compound II-1Na"

(n=12)
Total cholesterol
Serum153,0±14,7120,8±4,3

p=0.07
of 124.8±5,8

P=0,11
108,8±8,2

p=0.03
140,8±13,6
HDL50,3±5,154,9±2,454,2±3,448,4±2,752,9±4,1
LDL + VLDL100,9±17,365,0±6,0

p=0,09
70,6±6,8

p=0,14
60,4±8,3

p=0.06
87,9±14,5
Triglycerides
Serum129,7±17,9116,3±10,595,4±5,7

p=0.009
to 124.4±8,9111,6±17,2
HDL

LDL + VLDL
76,0±14,770,5±9,053,2±4,679,2±and 7.773,9±15,4

Table 20
Cholesterol and triglyceride levels (mg/100 ml) in the serum of rats on the 10th the Yan experiment
Indicatorsgroup
"Cholesterol"

(n=12)
"compound IV-1Na (n=12)"connection V-1Na"

(n=12)
the compound III-1Na (n=12)"compound II-1Na (n=11)
Total cholesterol
serum144,6±12,8122,1±9,8

p=0,18
123,3±7,3

p=0,18
to 102.3±6,9

p=0.014
RUR 134.4±12,5
HDL46,7±2,953,4±2,2

p=0,09
52,9±3,848,8±2,454,7±2,2

p=0,046
LDL + VLDL97,9±14,068,7±10,1

p=0.11
70,5±8,9

p=0,12
53,5±7,3

p=0.02
79,7±12,6
Serum triglycerides115, 8mm±18,9109,3±5,181,9±4,4

p=0.11
109,3±10,5108,5± and 11.3
HDL35,5±3,136,2±2,631,5±1,834,3±2,3 38,9±2,4
LDL + VLDL80,3±16,273,1±5,50,4±4,8

p=0,1
75,0±8,569,6±11,1

Introduction animals monosodium salt of compound III was significantly reduced total cholesterol (LDL) serum 2 9% and cholesterol fractions VLDL + LDL 4 0%, but did not change the level of atherogenic LDL, HDL and total serum triglycerides.

The results obtained indicate that the mono - and disodium salt was superior to compound IV by the dynamics of steps on total serum cholesterol and LDL + VLDL and other lipid metabolism. While the 10-th day experience reliable and comparable decrease in these indicators occurred under the action of the aforementioned compounds and drug comparisons "Zocor (simvastatin), salts of compound IV was launched earlier experiment (5 and day 8). Both sodium salt of compound IV was increased HDL already to 5 days of the experiment, while compound IV was increased this figure is only 8 days. Comparison drug simvastatin had no effect on HDL-cholesterol. In addition, the disodium salt of compound IV reduced the levels of total serum triglycerides at 5 and 10 days of the experiment.

A distinctive property of the monosodium salt of compound V was the way the spine to reduce the levels of total serum triglycerides, while a reduction in total cholesterol, VLDL cholesterol and increased levels of HDL was lower than the monosodium salt of compound III and mono-sodium salt of compound IV.

Thus, salts of compounds II, III, IV and V have a high lipid-lowering activity compared with the activity of the compounds described in published international application WO 99/01103 and connections proposed in the present invention, including the ability to reduce the level of triglycerides, serum cholesterol, including LDL, and increase HDL cholesterol.

Example 19

The study of the hypolipidemic action of compounds of General formula (I) on the model of "endogenous" hypercholesterolemia Guinea pigs

The study was performed on Guinea pigs males (breed Agouti) initial mass 304±25 year duration of the experiment 31 days. The control group, 6 pigs - intact animals. The studied compounds were introduced from the first day of the experiment (from the first day of the introduction of the fat load).

Animals of the experimental groups every day for 31 days received per os investigated the connection and the fat load. Analyzed the connection in the following doses were administered in aqueous solution (0.5 ml per animal); fat load is a mixture of pork fat/pre-warmed corn oil, 4:1 by volume, at a rate of 5 ml/kg of body weight is via 0.5 hours after injection of the analyte.

Experimental group:

1) "control" - intact animals;

2) "fat" - animals that received only fat load;

3) "compound IV" - the animals treated with the fat load + compound IV at a dose of 500 mcg/kg;

4) "compound V" - animals that received fat load + connection V at a dose of 500 mcg/kg

Data on the content of cholesterol and triglycerides in the serum of Guinea pigs treated with the fat load and the compounds presented in tables 22-25.

Table 21
The content of total cholesterol in the serum of Guinea pigs treated with the fat load and test connection
Total serum cholesterol
28 D.31 D.
Control37,4±3,3
Fat71,7±14,5

n=9
74,4±11,4
Compound IV55,5±6,3

n=10
49,2±3,9

p=0,064
Connection V52,9±6,5

n=9
46,3±4,1

p=0,043

Statistical data processing was performed using one-way ANOVA.

Table 22
The content of total triglycerides in the serum of Guinea pigs treated with the fat load and test connection
Total serum triglycerides
28 D.31 D.
Control60,1±2,4
Fat89,7±14,0123,1±35,6
Compound IV66,8±6,769,0±13,4
Connection V62,3±5,456,0±6,1

Table 23
The total cholesterol content on the 31st day in the lipoprotein fractions of serum of Guinea pigs treated with the fat load and test connection
Cholesterol, mmol/100 ml
TotalVLDLLDL
Control38,4±3,31,6±0,0732,2±2,8
Fat74,4±11,44,1±1,267,6±10,2

p=0.009
Compound IV49,2±3,93,0±0,7142,1±3,5

p=0,040
Connection V46,3±4,12,1±0,441,9±2,7

p=0.038

Table 24
The content of total triglycerides on the 31st day in the lipoprotein fractions of serum of Guinea pigs treated with the fat load and test connection
Triglycerides, mg/100 ml
TotalVLDLLDL
Control60,1±2,3638,16±2,8616,13±0,97
Fatof 123.2±35,659,5±19,742,7±8,62

p=0,015
Compound IV69,0±13,433,2±12,329,6±2,4
Connection V56,0±6,1

P=0.1
22,5±4,4

P=0.1
23,1±1,9

p=0,053

The compounds IV and V were significantly reduced total cholesterol only 31 days of the experimental the NTA 33.9 and 37.8%, respectively. However, they significantly reduced the LDL cholesterol 37.7%and 38%.

An advantage of the claimed compounds, in particular compounds IV is a wide range of effective doses that provides the breadth of its therapeutic action. For example, compound IV is almost equally effectively reduced the total cholesterol within 20 days in doses ranging from 50 to 1500 μg/kg, characterized in 30 times.

Thus, the claimed compounds corresponding to General formula (I)have significant hypolipidemic activity, significantly improving the lipid metabolism in serum and in the liver.

Example 20

A comparative study of the hypolipidemic action of compounds IV and glutamylcysteine (YY, compound III WO 99/01103) in experimental models of hypercholesterolemia rats Wistar.

Materials and methods research.

Presents data obtained after combining the results of 3 experiments.

The study was performed on 118 rats male Wistar weighing 180-230 g Hyperlipidemia caused by oral administration (via stomach tube) cholesterol load - oil suspension cholesterol:

olive oil (refined, pre-heated 1 h 80° (C) 5 ml/kg;

cholesterol (Sigma, USA), 1 g/kg;

Holt sodium (Sigma, USA) 120 mg/kg of body weight.

Cholesterol is a new suspension was injected in the morning daily for 14 days. Animals received the compounds (at a dose of 500 mcg/kg of body weight) from day experience in the morning. All drugs were administered to the animals 30 minutes before cholesterol load.

Experimental group:

"Cholesterol" - rats treated for 14 days per os oil suspension cholesterol (n=37)

1) "IV rats treated per os oil suspension of cholesterol and Na salt of the compounds IV (n=37);

2) "YY" - rats treated per os oil suspension of cholesterol and investigated the connection YY/(n=22)

Blood samples (from the tail) were taken for analysis at 5, 8 and 10 days of the experiment. On day 14 blood samples were obtained after decapitate. Before blood sampling, the animals were fasted for 10 hours.

In serum measured total cholesterol, triglycerides, total cholesterol, high density lipoprotein (HDL). Cholesterol of lipoproteins of low and very low density was calculated by the difference between total serum cholesterol and HDL cholesterol.

The content of total cholesterol and triglycerides in serum were determined by enzymatic methods using test sets "CHOL L 250 S" (No. 10003268) and TG L 250 S" (No. 10003267) production company "Lachema (Czech Republic). The density measurement was performed on a spectrophotometer Shimadzu" at 500 nm, the calibration had to be attached to the test sets of standard samples.

Identify the bookmark cholesterol in high density lipoprotein (α -PL) was performed using test kits "HDL CHOL E" (10003202) and "CHOL L 250 S" (No. 10003267) firm "Lachema (Czech Republic) by precipitation of LDL and lonp phosphomevalonate acid and magnesium ions.

The data in tables 25-27 presented as the average value of ± standard error of the mean. The significance of differences between groups "cholesterol" and "the drug..." was assessed by two-sample t-test t-test. The probability of error (p) are listed in columns of tables.

The results of the study.

Experimental hypercholesterolemia.

The initial level of total cholesterol and triglycerides in the blood plasma are presented in table 25. Dynamics of changes of lipid metabolism in table 26.

Table 25
The lipid metabolism of rats (n=24) before the start of the experiment.
Initial biochemical parameters, mg/100 ml (n=30)
Total cholesterol
Serum72,3±1,8
LHP36,1±1,7
LDL + lamp36,2±2,2
Cholesterol atherogenic index1,07±0,05
Triglycerides
Serum69,0±5,7
LHP15,4±1,3
LDL + lamp53,6± and 4.9
Triglyceride indexto 3.58±0,22

The influence of the studied compounds on the development of hypercholesterolemia.

Data on the effect of the studied compounds on cholesterol and triglycerides in serum of rats fed cholesterol load, are presented in tables 26 and 27.

Cholesterol atherogenic index (total hol.)
Table 27
Significant (p≤0,05) differences changes in parameters of lipid metabolism between groups IV and GG on a two-sample t-test t-test.
Differences of changes of parameters of lipid metabolism in groups
daysGG (n=22)IV
Cholesterol HDL
+22%
+24%
+26%
+15,7%
- 15%
- 11%

Designation: - 10% - significantly below 10%. For 100% passed level in the group "Cholesterol".

Presented in tables 25-27 data clearly indicate that compound IV increases levels of cholesterol atherogenic HDL 10-14% compared with control, while under the influence of glutamylcysteine was significant lowering of HDL cholesterol.

As a result, the reduction of cholesterol atherogenic index by glutamylcysteine owing to lower cholesterol all fractions of plasma and HDL, and LDL + lamp.

Compound IV differs from glutamylcysteine significant significant decrease in cholesterol atherogenic index by 20-40% during the experiment due to the increase of the HDL fraction.

Comparison of lipid-lowering actions of glutamylcysteine with the appropriate activity IV showed that compound IV according to the invention is significantly superior to glutamylcysteine in raising HDL cholesterol and reducing the atherogenic index (table 27).

Example 21

Research protivovospalitelnoe activity of the compounds of General formula I (1% gel) to model carrage the new edema in vivo

Materials and methods

Tests carried out on non-linear white mice weighing 30 to 32, Using the model carrageenaninduced edema according to the method described in Winter et al. In: De Rosa M., Giroud J.P., D.A. Willoughby Studies of the mediators of the acute inflammatory response induced in rats in different sites by carrageenan and turpentine. // J. Phamacol. 1971. V.104. P.15-29. In the right paw of the mouse subplantar impose a 1% solution carragenin (SERVA) in a volume of 0.05 ml of a Substance in a 1% gel applied on the paw 2 times: the first time immediately after the introduction carragenine; the second time is 1.5 hours. The measurement of the volume of the right and left (intact) paws carried out with the help of Vernier caliper after 3 h after injection carragenin. Inflammatory response and therapeutic effect of the impact estimate by the formula.

where P is the increase in swelling,

On - volume feet after the introduction of logogen,

And - the value of the volume of the paws before the introduction of logogen.

The effect of therapeutic effects estimated by the degree of inhibition of the inflammatory response compared to the control and calculated by the formula:

where (o) - treated animals

(K) - untreated animals

Table 28
The effect of compounds of General formula I (1% gel) on the development of carragenine swelling of the paws of mice
AnalyteThe difference in the volume of pawIncrease (%) Inhibition of the inflammatory response (%)
V0.104 g±0,009**60.043.8
V - 1Na0,14±0,02**79.026.2
IV - 1Na0,105±0,002**47,633.0
II - 1Na0,1±0,009**32,351,6
III - 1Na0,15±0,01*50,025,0
Voltaren0,08±0,006**25,861,3
* - statistically significant relative to the model * P<0,01 ** P<0,01

The results of the experiments are presented in table 28 show a pronounced anti-inflammatory activity of the monosodium salt of compound (II), comparable with the activity of the comparison drug - voltaren.

Connection V and its monosodium salt show moderate activity in this model carragenine swelling of the paws of mice, inhibition of edema is 43.8% and 26.2%, respectively.

Thus, the whole complex used experimental models shows a significant antiallergic, antianaphylactic and protivovospalitel is through the activity of the compounds of General formula I, manifested in in vitro tests, and modeling, allergic and inflammatory pathology in vivo. It is shown that the described compounds corresponding to General formula I show significant protective effect against systemic anaphylactic reactions in vivo, under conditions modeling of allergic rhinitis, in addition, these compounds suppress eosinophilic inflammation and have a distinct anti-inflammatory effect.

Examples of dosage forms

Example 22

A. Preformed shape

Tablet form is received, using the following ingredients:

The compound corresponding to General formula

1-100 mg (I),

or its pharmaceutically acceptable salt

Potato starch20-50 mg
Magnesium stearate3 mg
Aerosil1 mg
Lactoseup to 300 mg

The components are mixed and pressed to form tablets weighing 300 mg each.

B. Suppositories

An example of the composition of the suppository:

The compound corresponding to General formula
(I)or its pharmaceutically acceptable salt1-100 mg
Cocoa butterthe amount needed to obtain suppository.

If necessary, it is possible to manufacture rectal, vaginal and urethral suppositories with appropriate fillers.

Century Ointment

An example of the composition of the ointment:

The compound corresponding to General formula
(I)or its pharmaceutically acceptable salt0,01-0,1 g
Vaseline10 g

Ointments are made by well-known technology.

, Gels

An example of the structure of the gel:

The compound corresponding to General formula
(I)or its pharmaceutically acceptable salt1-100 mg

The carbopol200 mg
Benzyl alcohol : 20 mg
Ethanol300 mg
Waterto 10 g

D. Dry powder for inhalation

An example of the composition of the powder:

The compound corresponding to General formula
(I)or its pharmaceutical is acceptable salt 0.2 g
Lactoseto 1 g

The powder is placed in a special device (container) or in a gelatin capsule.

E. Nasal spray

An example of the composition of the spray:

The compound corresponding to General formula
(I)or its pharmaceutically acceptable salt1.5 to 150 mg
Purified waterto 15 ml

E. Eye drops

An example of the composition drops:

The compound corresponding to General formula
(I)or its pharmaceutically acceptable salt0.5 to 50 mg
Preservative10 mg
Purified waterto 5 ml

E. injection

An example of the composition of the solution for injection:

The compound corresponding to General formula
(I)or its pharmaceutically acceptable salt0.2-20 mg
water for injection2 ml

1. 1. N-acyl derivatives of amino acids of General formula (I)

where n p is VNO 2 or 3 and

R represents

or

and their pharmaceutically acceptable salts, provided that the compound of General formula (I) is not succinyl-L-histidine, succinyl-L-tryptophan, succinyl-D-tryptophan, succinyl-D,L-tryptophan and Pikalevo salt.

2. The compound according to claim 1 where the pharmaceutically acceptable salt is a mono - or disodium salt.

3. Pharmaceutical composition having hypolipidemic activity, including N-acyl derivatives of amino acids according to claim 1 or its pharmaceutically acceptable salt in an effective amount and a pharmaceutically acceptable additive.

4. The use of N-acyl derivatives of amino acids according to claim 1,

or their pharmaceutically acceptable salts as a means of having hypolipidemic activity.

5. The use of N-acyl derivatives of amino acids according to claim 1 or their pharmaceutically acceptable salts as a means to eliminate the signs of atherosclerosis, obesity, coronary heart disease and cerebral, myocardial infarction, stroke.

6. A means of having hypolipidemic activity, which is an N-acyl derivatives of amino acids according to claim 1 or their pharmaceutically acceptable salts.

7. A method of treating diseases associated with violations is receiving lipid metabolism, includes introduction to the mammal an effective amount of N-acyl derivatives of amino acids according to claim 1 or its pharmaceutically acceptable salts.

8. A method of treating atherosclerosis, obesity, coronary heart disease and cerebral, myocardial infarction, stroke, comprising the administration to a mammal an effective amount of N-acyl derivatives of amino acids according to claim 1 or its pharmaceutically acceptable salts.



 

Same patents:

FIELD: organic chemistry, medicine, pharmacy.

SUBSTANCE: invention relates to imidazoline-2-yl-substituted derivatives of arylalkyl sulfonamide possessing agonistic activity with respect to α1A/L adrenoceptors, and to pharmaceutical compositions comprising these derivatives. Invention relates to compound of the formula: or its pharmaceutically acceptable salts wherein R1 represents (C1-C12)-alkyl; R2 represents hydrogen atom; each radical among R3, R4, R5 and R6 represents independently hydrogen atom, halogen atom, (C1-C12)-alkyl, -OR9 wherein R9 represents hydrogen atom, (C1-C12)-alkyl under condition that R3, R4, R5 and R6 don't represent (C1-C12)-alkyl simultaneously; or R3 and R4 in common with atoms to which they are bound form aromatic 5-membered ring comprising heteroatom, such as oxygen atom, and R14 represents hydrogen atom.

EFFECT: valuable medicinal properties of compounds and pharmaceutical compositions.

10 cl, 1 tbl, 8 ex

FIELD: bioorganic chemistry, biochemistry, pharmacy.

SUBSTANCE: invention relates to novel aspartyl derivatives of histamine of the general formula (I): , wherein R means hydrogen atom (H), or , or that are able to modulate activity of enzymes of antioxidant protection - superoxide dismutase (SOD) and catalase. Also, invention relates to using the known compounds of the general formula (I) for the same designation wherein at the same values of X the value R represents acetyl group, and to their pharmaceutically acceptable salts. Also, invention relates to a pharmaceutical composition possessing capacity to modulate activity of SOD and catalase and comprising the effective amount of compound of the general formula (I), and to a method for synthesis of compounds of the general formula (I). Method involves interaction of pentafluorophenyl ester Nα-Z-, β- or α-benzyl ester of aspartic acid with histamine followed by hydrogenolysis without isolation of intermediated protected derivatives of aspartyl histamine.

EFFECT: improved method of synthesis, valuable biochemical properties of derivatives.

12 cl, 3 tbl, 2 sch, 2 dwg, 8 ex

FIELD: organic chemistry, medicine, pharmacy.

SUBSTANCE: invention relates to derivatives of adamantine of the formula (I): wherein D represents -CH2 or -CH2-CH2; E represents -C(O)NH or -NHC(O); each R1 and R2 represents independently hydrogen atom or halogen atom but R1 and R2 can't mean hydrogen atom simultaneously; R3 represents group of the formula: -R4-X-R5 (II) wherein R4 represents (C1-C6)-alkyl group; X represents oxygen or sulfur atom or the group NR13; R5 represents (C1-C6)-alkyl or (C2-C6)-alkenyl and each of them can be optionally substituted with at least one substitute taken among halogen atom, hydroxyl, di-(C1-C6)-alkylamino-group, -Y-R6, , and 5- or 6-membered heteroaromatic ring comprising 1-4 heteroatoms taken independently among nitrogen atom wherein heteroaromatic ring can be optionally substituted with at least one (C1-C6)-alkyl; Y represents oxygen or sulfur atom or the group -NH; R6 represents the group -R7Z wherein R7 represents (C2-C6)-alkyl group and Z represents -OH; when Y represents oxygen or sulfur atom or the group -NH then R6 represents additionally hydrogen atom, (C1-C6)-alkyl, (C1-C6)-alkylcarbonyl; R13 represents hydrogen atom, (C3-C8)-cycloalkyl, (C3-C8)-cycloalkylmethyl; or R13 represents (C1-C6)-alkyl group optionally substituted with at least one hydroxyl, or to its pharmaceutically acceptable salts or solvates. These compounds are effective antagonists of P2X7 receptors and can be used in treatment of rheumatic arthritis or lung chronic obstructive disease. Also, invention describes methods for preparing these compounds, pharmaceutical composition comprising indicated compounds, method for preparing pharmaceutical composition and their using in therapy.

EFFECT: improved preparing and treatment methods, valuable medicinal properties of compounds and composition.

19 cl, 1 tbl, 77 ex

FIELD: organic chemistry, medicine, cardiology, biochemistry.

SUBSTANCE: invention relates to benzoyl guanidines of the formula (I): wherein R1 means -CF3; R2 means -Y-para-(C6H4)-R11, -Y-meta-(C6H4)-R11 or -Y-ortho-(C6H4)-R11 wherein R11 means (C1-C9)-heteroaryl comprising two or more nitrogen atoms adjoining across nitrogen (N) atom; Y means oxygen atom; R3 means hydrogen atom; R4 means (C1-C4)-alkyl, and to their pharmaceutically acceptable salts. Indicated compounds elicit very high activity with respect to inhibition of Na+/H+ exchange and improved water solubility and therefore they can be used as anti-arrhythmic medicinal agents with cardioprotective component for prophylaxis of infarction and treatment of infarction and for treatment of stenocardia. Also, proposed compounds inhibit pathophysiological processes in arising disorders induced by ischemia, in particular, in treatment of cardiac arrhythmia induced by ischemia.

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

17 cl, 2 ex

FIELD: organic chemistry, medicine, pharmacy.

SUBSTANCE: invention relates to derivatives of imidazole of the formula (I):

or its pharmaceutically acceptable salts wherein X represents -CH2-(CH2)p-, -O-; R1 represents phenyl, naphthyl, 1,2,3,4-tetrahydronaphthyl, (C3-C7)-cycloalkyl wherein indicated phenyl, naphthyl, 1,2,3,4-tetrahydronaphthyl, (C3-C7)-cycloalkyl are substituted optionally with 1-3 substitutes taken independently among halogen atom, -OH, halogen-(C1-C6)-alkyl, (C1-C6)-alkyl, (C1-C6)-alkoxy group and OH-(C1-C6)-alkyl; R2 represents hydrogen atom (H) or (C1-C6)-alkyl; R3 represents H or (C1-C6)-alkyl; R4 represents H or (C1-C6)-alkyl; R5 represents H, or R5 and R7 form in common a bond; each R6 represents independently halogen atom, -OH, halogen-(C1-C6)-alkyl, (C1-C6)-alkyl, (C1-C6)-alkoxy group or OH-(C1-C6)-alkyl; R7 represents H, or R7 and R5 form in common a bond; each R8 represents independently -OH, (C1-C6)-alkyl, halogen-(C1-C6)-alkyl or (C1-C6)-alkoxy group; m = 0, 1, 2 or 3; n = 0 or 1; p = 0 or 1; r = 0 or 1; t = 0. Also, invention relates to a method for preparing compounds of the formula (I) and to a pharmaceutical composition showing affinity to alpha-2-adrenoceptors based on these compounds. Invention provides preparing new compounds and pharmaceutical composition based on thereof used in aims for treatment of neurological disturbances, psychiatric disorders or disturbances in cognitive ability, diabetes mellitus, lipolytic diseases, orthostatic hypotension or sexual dysfunction.

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

25 cl, 1 tbl, 14 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

The invention relates to new imidazole derivative of the formula (I):where R1represents phenyl or pyridinyl, substituted by substituents selected from the group comprising (1) phenyl, (2) furyl, thienyl, (3) halogen, (4) halogen(lower)alkyl, (5) lower alkylthio, (6) nitro, (7) lower alkenyl, optionally substituted phenyl, (8) lower quinil, optionally substituted phenyl, (9) lower alkoxy, optionally substituted cyclo(lower)alkyl or phenyl, (10) lower alkyl, optionally substituted, phenyloxy or (11) amino, optionally substituted protected carboxyla; R2represents lower alkyl; R3represents halogen or lower alkyl; R4represents (1) lower alkenyl, optionally substituted phenyl, (2) phenyl, optionally substituted lower alkyl or lower alkenyl, (3) lower alkyl or (4) thienyl, optionally substituted with halogen; a represents a lower alkylene and L represents a simple bond, a lower albaniles or lower alkylene, optionally substituted phenyl or pyridinyl, or-X-CH2- where X represents O or NR5where R5represents hydrogen or n is

New drug substances // 2237657
The invention relates to organic chemistry and can find application in medicine

New drugs // 2237057
The invention relates to organic chemistry and can find application in medicine

The invention relates to new and nitrate salts of heterocyclic compounds of formulas (a) and (b), where R is hydrogen, alkoxyl, R1- alkyl, alkoxyl, R2is hydrogen, alkyl, R3- alkyl, alkoxyl, X denotes N-R11or oxygen, R11means the free valence, Y represents N-R16, sulfur or alkyl, R16means hydrogen; other values radicals presented in the description of the invention

FIELD: organic chemistry, food industry.

SUBSTANCE: invention relates to crystals of ammonium salt or alkaline metal salt of non-natural stereoisomers of monatine - (2R,4R)-stereoisomer, (2S,4R)-stereoisomer, (2R,4S)-stereoisomer or mixture of non-natural (2S,4R)- and (2R,4S)-stereoisomers of monatine. These substances possess the high sweet taste degree and can be used as sweetening agents. Except for, invention relates to a sweetening agent based on abovementioned crystals of non-natural stereoisomers of monatine and to a product, such as beverage or foodstuff prepared by using abovementioned crystals of non-natural stereoisomers of monatine.

EFFECT: valuable properties of crystals.

16 cl, 4 tbl, 52 dwg, 43 ex

FIELD: organic chemistry, medicine, oncology.

SUBSTANCE: invention relates to organic amine salts, amino acid salts and combrestatin A-4 phosphate amino acid ester salts. Invention describes compound of the general formula (I):

wherein one of substitute -OR1 or -OR represents -O-QH+ and another one represents hydroxyl or -O-QH+; Q represents (A) optionally substituted aliphatic organic amine comprising at least one nitrogen atom that in common with proton forms quaternary ammonium cation QH+; (B) amino acid comprising at least two nitrogen atoms wherein one of nitrogen atoms in common with proton forms quaternary ammonium cation QH+; (C) amino acid comprising one or some nitrogen atoms wherein one of nitrogen atoms in common with proton forms quaternary ammonium cation QH+ and wherein, except for, all carboxyl groups of amino acids are in ester form. Also, invention describes pharmaceutical compositions used in modulation of tumor or metastasis proliferation and growth of benign vascular proliferative disorders, using compound of the formula (I) and a method for synthesis of compound of the formula (I). Invention provides preparing new combrestatin A-4 salts showing useful physicochemical properties that enhance solubility of combrestatin A-4.

EFFECT: valuable medicinal properties of compounds and pharmaceutical compositions.

27 cl, 13 dwg, 5 ex

The invention relates to the field of medicine and relates to new N-pinakamaraming tryptophanase of dipeptides of the formula

C6H5-(CH2)n-CO-NH-(CH2)m-CO-X-Trp-R,

where n=1-5;

m=1-3;

X=L or D-configuration;

R=OH, OCH3OC2H5, NH2, NHCH3,

as well as pharmaceutical compositions containing them

The invention relates to medicine and relates to means for inhibiting endothelialised enzymes, which is a product of the formula (I), method of its production and pharmaceutical compositions containing the product of formula (I) as active principle

The invention relates to new derivatives thioamides formula (I), where X denotes the group-CO2H or -- CONHOH, Y and Z denotes sulfur or oxygen, R1denotes hydrogen, hydroxy, alkyl, alkenyl, R2denotes alkyl, phenylalkyl, phenylalkyl-alkyl, R3means defining a side chain of natural-amino acid in which any functional group may be protected, alkyl, cycloalkyl, R4denotes alkyl, phenylalkyl, optionally substituted phenyl, or a group of the formula -(Q-O - Q, where Q denotes alkyl

The invention relates to acylaminocinnamic derivative of the formula (I), where R denotes phenyl which is not substituted or may be substituted with halogen, alkyl, trifluoromethyl, hydroxy and alkoxygroup, R1is hydrogen, alkyl, R2is hydrogen, alkyl or phenyl which is not substituted or may be substituted with halogen, alkyl, trifluoromethyl, hydroxy and alkoxygroup, R3is phenyl which is not substituted or may be substituted with halogen, alkyl, trifluoromethyl, hydroxy and alkoxygroup, or represents naphthyl, lH-indol-3-yl or 1-alcheringa-3-yl, R4' and R4"is hydrogen, alkyl, and one of the radicals R4' and R4"is hydrogen, and R5- cycloalkyl, D-azacycloheptan-2-he-3-yl or L-azacycloheptan-2-he-3-yl, or its salt

The invention relates to derivatives of heterocyclic compounds, as well as agricultural and horticultural fungicides containing these compounds as active ingredients

FIELD: chemistry.

SUBSTANCE: invention refers to of serotonin receptor 5-NT6 antagonists, simultaneously regulating calcium ions homeostasis in cells, representing substituted 2,3,4,5-tetrahydro-1H-pyrido[4,3-b]indole of general formula I, its pharmaceutically acceptable salt and/or hydrate. In general formula I where: R1 is amides substitute selected from optionally substituted C1-C5 alkyl; R2i represents one or number of identical or various substitutes selected from hydrogen, halogen, C1-C3 alkyl, CF3, OCF3; Ar represents halogen unsubstituted or substituted C1-C6 alkyl, C1-C6 alkoxy, phenyl substituted with amides or trifluoromethyl or optionally substituted aromatic hexamerous heterocycle containing 1-2 nitrogen atoms per one cycle, W represents ethyl group-CH2CH2 - vinyl group or ethynyl group. Invention also concerns new compounds selected from group of compounds of formula 1, methods of production thereof, pharmaceutical compositions and methods of their use.

EFFECT: production of composition that simultaneously regulates calcium ions homeostasis in cells.

34 cl, 7 dwg, 4 tbl, 9 ex

FIELD: medicine.

SUBSTANCE: device includes core containing at least one pharmaceutically active agent. Core is made from elastomeric composition selected from the group consisting of poly(dimethylsiloxane), siloxane-based elastomer including 3,3,3-trifluoropropyl groups attached to Si atom of siloxane units, siloxane-based elastomer, which includes poly(alkyleneoxide) groups, and their mixtures. The said core is encased by membrane. Membrane is made from the same elastomeric composition as core. Pharmaceutically active agent is selected from the group, which includes anti-allergic agents, anti-infection agents, anti-asthmatic agents, anti-inflammatory agents, anti-virus agents, anti-bacterial agents, anti-histamine means, anti-fungal agents, anesthetics, opioids, vasorelaxants, muscarinic agents, sympathomimetics, corticosteroids and their mixtures. Delivering device, according to invention, for nasal or otological application ensures preliminary set, constant rates of release of one or more pharmaceutically active agents.

EFFECT: device is safe and does not cause discomfort to patients.

8 cl, 6 dwg, 1 tbl, 5 ex

Up!