Pharmaceutical composition, containing n-acyl derivatives of amino acids, and their application as anti-allergic, anti-anaphylactic and anti-inflammatory medications

FIELD: medicine, pharmaceutics.

SUBSTANCE: claimed invention relates to application of N-acyl derivatives of amino acids of general formula

, where n equals 2 or 3 and their pharmaceutically acceptable salts as anti-allergic, anti-inflammatory and anti-anaphylactic medications.

EFFECT: obtaining pharmaceutical composition for treatment of allergic and inflammatory diseases, for instance such as bronchial asthma, allergic rhinitis, pollinosis, psoriasis.

11 cl, 12 tbl, 19 ex

 

The present invention relates to the field of Bioorganic chemistry and relates to pharmaceutical compositions containing N-acyl derivatives of amino acids and their pharmaceutically acceptable salts, and their use in medicine as anti-allergic, anti-inflammatory and antianaphylactic funds.

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 / M: New wave, 2005, s.285).

Antihistamines 1st generation penetrate the blood-brain barrier and can cause a blockade of H1receptor cells of the Central nervous system, which makes them undesirable sedative effect. To achieve the expressed antihistamine action is s 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, for example, bronchial asthma using 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 / M: Medicine, 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 by U. the Floor., M.: Mir, 1989, Vol.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 (kromolin-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, cough, choking (Mashkovsky PPM Medicines / M: New wave, 2005, s].

In connection with the above, it is important to search for new effective anti-allergic and antianaphylactic 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, for example, 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 Century is. and others, the Study antianaphylactic activity and mechanisms of action of γ-L-glutamylcysteine // Pathogenesis, 2003, Vol. 1, No. 2, S. 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 of γ-glutamylcysteine significantly reduced 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 comparedwith 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 antianaphylactic funds, the inventors have 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 physicochemical to scanty.

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 form USA2005079515. 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, describes how the acylation of amino acids in aqueous-organic, strongly 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 NaHCO 3to 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 pharmaceutical compositions containing the new effective N-acyl derivatives of amino acids having antiallergic and antianaphylactic effect in low doses and does not exhibit side effects.

Brief description of the invention

The present invention relates to the use of compounds of General formula I

where n is 2 or 3; and

R represents

and their pharmaceutically acceptable salts as anti-allergic, anti-inflammatory and antianaphylactic funds.

Further, the present invention relates to pharmaceutical compositions and means of having antiallergic, antianaphylactic and anti-inflammatory effect, containing an effective amount of the compounds of General formula I or its pharmaceutically acceptable salts, and also, 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, topices the th 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, comprising introducing an effective amount 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.

Table 1
ConnectionNo. of connectionsRn
Nα-succinyl-L-tryptophanII(Ind)2
Nα-glutaryl-L-tryptophanIIIInd3
Nα-glutaryl-L-histidineIV(Im)3
Nα-succinyl-L-histidineVIm2

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 its salts, where

R represents

glutaric or succinic anhydride in the form of a solid substance, followed by separation of the target 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 acid, taken from itke, 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 you to use excess Alliluyeva agent, to achieve a complete acylation of the 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.

The preferred solvents, and is used for crystallization of the desired product, are water-alcohol mixture, 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 antianaphylactic activity and can be used for treatment of allergic, anaphylactic, including inflamed diseases.

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.

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

For the treatment of allergic diseases, including the surrounding 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 lung diseases, namely chronic obstructive bronchitis, emphysema, obliterative bronchitis, cystic fibrosis, 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, oncrete used a combination of medicines, as well as the severity of the disease in 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, t is Alacant, methylcellulose, hypromellose, 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 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 a paraffin wax and a wax; absorptive ointment bases 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 relationalization 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 for inye the Nations, the ringer's solution, glucose solution, 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)

Individuality is received to connect the clusters was checked by TLC on plates “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”, 2×250 mm, 5 μm, elution with 0.1%TFA, the rate of elution of 0.25 ml/min (1); the rate of elution 1 ml/min, detection at 220 nm, column Luna 5 “Phenomenex, C18, 250×4.6 mm, elution with 25% acetonitrile in 0.05 M phosphate buffer (pH 3.0) (2).

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 PR is +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]D20+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, leave at 4°C for 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 time odergivaniya,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°C.

[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°C.

[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-Glutamylation (III)

To a suspension of 1.0 g (4.9 mmol) of tryptophan in 7 ml of water are added dropwise a solution of 1 N NaOH (4.9 mmol). To the resulting solution was added a solution of 0.56 g (4.9 mmol) glutaric anhydride in 3 ml of ethyl acetate. The reaction mixture is stirred for 3 hours at room temperature in argon atmosphere in the dark, leave in cashow at +4°. The solvent of the reaction mixture is removed under vacuum. The obtained oily residue is dissolved in 30 ml of water under stirring, cooled to 0°C, add a solution of 1 N HCl to pH 4. The product is extracted with ethyl acetate (3×25 ml). United an ethyl acetate extract is cooled to 0°, washed with water (4×25 ml) to pH 7, the solution of 5% HCl (5 ml), water (4×25 ml) to p H7, dried over anhydrous Na2SO4within 1 hour. Sediment Na2SO4filtered off, washed with ethyl acetate, the solvent is removed in vacuum. Get a grayish solid residue, which is dried in vacuum.

Yield 1.0 g (70%).

Rf0,54 (1).

TPL= 150-152°C.

[α]D20= +8,20° (C 0.5, methanol).

1H-NMR spectrum (CD3OD), δ, ppm: 1,75-of 1.84 (m, 2H, β-CH2-Glt), 2,15-of 2.30 (m, 4H, α,γ-CH2-Glt), 3,30 is 3.40 (m, 2H, β-CH2-Trp), 3,80-3,90 (m, 1H, α-CH-Trp), 6,97 (t, J=7 Hz, 1H, CH-6-Ind), 7,06 (t, J=7 Hz, 1H, CH-7-Ind)to 7.15 (d, J=7 Hz, 1H, CH-2-Ind), 7,33 (d, J=7 Hz, 1H, CH-5-Ind), 7,55 (d, J=7 Hz, 1H, CH-8-Ind).

HPLC under the conditions: (2) individual peak, retention time 6,77 minutes

Found, %: 60,07; H 5,65; N Is 8.75. C16H18N2O5Calculated, %: C 60,37; H 5,7; N 8,8.

Example 4

Nα-Succinyl-L-tryptophan (II)

The synthesis was carried out in accordance with the methodology described for compound III.

The output of 100.5 mg (67%).

Rf0,63 (1).

[α]D20= +21,05° (C 0,6, water).

1H-NMR spectrum (DMSO-d6, δ, ppm: 2,33-to 2.41 (m, 4H, α,β-CH2-Suc), 2,93-a 3.01 (m, 1H, β-CH2-Trp), 3,10 -, and 3.16 (m, 1H, β-CH2-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; H 5,65; N 9,35. C15H16N2O5Calculated, %: C 59,21; H 5,3; N Of 9.21.

Example 5

Monosodium salt 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°C. add 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 per cent).

TPL= 208-210°C.

[α]D20= +16,27° C 0,58, water).

Found, %: C 45,25; H 5,51; N 14,52. C11H15N3O5Na. Calculated, %: C 45,21; H 5,17; N 14,38.

Example 6

Monosodium salt 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%).

[α]D20= +40,21° (C 0,48, water).

Found, %: C 43,25; H 4,51; N 15,52. C10H13N3O5Na. Calculated, %: C 4,17; H 4,71; N 15,10.

Example 7

Monosodium salt 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°C.

[α]D20= +22,02° (C 0,39, water).

Found, %: C 55,25; H 4,51; N 8,32. C15H16N2O5Na. Calculated, %: C 55,05; H Is 4.93; N 8,56.

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

Example 8

Monosodium salt 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°C.

[α]D20= +representing 22.06° (C 0,34, methanol).

Found, %: C 56,15; H To 5.21; N By 8.22. C16H18N2O5Na. Calculated, %: C 56,30; H 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°C. add 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 vacuo the E. The solid residue is dried over granular alkali.

The yield of 1.15 g (99,0%).

[α]D20= +11,92° (C 0,57, water).

Found, %: C 41,25; H 4,51; N 13,52. C11H15N3O5Na2. Calculated, %: C 41,91; H 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).

The output of 1.16 g (99,0%).

TPL= 124 to 128°C.

[α]D20= +grade of 20.06° (C 0,67, water).

Found, %: C 39,55; H Or 4.31; N 13,52. C10H13N3O5Na2. Calculated, %: C 39,88; H 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 the disodium salt of Nα-glutaryl-L-histidine (IV) (example 9).

Yield 0.56 g (97.7 percent).

Found, %: C 51,35; H Or 4.31; N By 8.22. C15H16N2O5Na2. Calculated, %: C 51,43; H 4,60; N 8,0.

Example 12

The disodium salt of Nα-glutaryl-L-tryptophan (III)

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

Yield 0.56 g (98.5 per cent).

Found, %: C 52,55; H 4,71; N 7,52. C16H18N2O5Na2. Calculated, %: C 52,75; H To 4.98; N 7,69.

Tests for biological activity

p> 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 double deposition of 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 in leukocytes supernatant fraction was centrifuged for 7 min at 100 g. To the precipitate of leukocytes to allali of 0.85% NaCl solution and the cell concentration was brought to 30×10 3/µl.

Staging test degranulation of basophils in vitro(Handbook of clinical laboratory research methods/edited Eaast. M, Medicine, 1975, page 130).

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°C for 10 min. working concentration of leukocytes is with 104/µl. From each tube was sampled (100 μl) in a separate test tube for an assessment of 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 Fcε 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 COI is litovali 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),

the experts. - % 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. Staining was performed at room temperature of not less than 50 minutes counting the number of stained basophils was performed using a camera Fuchs-Rosenthal. Braking is full of degranulation (TPD) basophils was calculated by the formula:

TPD(%) = 1-[(M cf (K) - M cf (exp.)]/[M cf (K) - M cf (OA)] ×100, where

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

M cf (OA) - average (3 samples) the number of basophils in the test maximum antigen-induced degranulation;

M cf (exp) - average (3 samples) the number of basophils in the test degranulation after incubation with the test compounds is receiving.

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)
to 99.2±11,22,6±2,6*
4.Compound IV
(10-4M)
99,5±12,02,9±1,6*
5.Compound IV
(10-5M)
113,5±2,70
6.Connect the s IV
(10-6M)
90,0±1,83,9±0,6
7.Compound IV
(10-7M)
76,7±1,59,1±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±6.73 x
11.Control 21000
12.Ovalbumin 1%
(max degranulation)
038,9±8,43
13.Connection V
(10-3M)
10,6±7,635,3±7,29
14.Connection V
(10-4M)
18,9±11,831,2±6,8
15.Connection V
(10-5M)
44,0±11,2724,6±10,38
16.Hydrocortisone
(10-3M)
71,0±1,610,0±0,7
17.Hydrocortisone
(10-4M)
48,0±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 on the system infilex the Yu 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, M., 2000, S. 242-250).

Guinea pigs were senzibilizirani the ovalbumin method Andersson (Anderson P. Antigen-induced bronchial anaphulaxis in actively sensitized geinea-pigs//Allergy, 1980, Vol.35, p.63-71) and 1-2 months after sensitization induced bronchospasm aerosol introduction resolution doses of ovalbumin (3 mg/kg in 1 ml of 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 (in 1 ml saline) was administered inhalation (via nebulizer) within three days 1 time per day. The control group was administered saline. 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, sThe duration of the subacute phase, s
Control 1
(saline)
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, sThe total reaction time, sec
Control 2
(saline)
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 antigen-induced bronchoconstriction.

Thus, compound IV has a significant protective effect against systemic anaphylactic 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 a nebulizer equipment (Pari) a solution of ovalbumin in increasing concentrations, ranging from 0.1% to 1% of the 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 a nebulizer 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 μl) in natalem flush
GroupIntact controlModel + provocation (rhinitis)Compound IV
(0.1% of R-R)
N677
Lymphocytosis18,0±2,1067,2±6,47**43,7±6,65 ° *
* the difference from the intact control; ° - contrast model
(° - P< 0,05; * P < 0,01; ** - P< 0,01)

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 were collected 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 8
The effect of compound IV on lymphocytosis (absolute number of cells in 1 μl) in bronchoalveolar washings
GroupIntact controlModel + provocation (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

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 Sephadex-induced (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 (hydrofining the powder with a particle size of from 20 to 80 μm) in a dose of 5 mg/kg using the dosing device, which 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. Wistar rats after administration of Sephadex A-25 quickly came out of the anesthesia and outwardly any peculiarities in behavior, respiration were 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 histology the definition sections of the lungs of a thickness of 4-5 μm, which counted the number of neutrophils millionary partitions, and estimated volumetric density of alveolitis and emphysema using grid Avtandilov (Avtandil - Introduction to quantitative pathological morphology // M.: Medicine, 1980, S. 203). Also conducted a morphometric study of the lymphoid tissue of the lungs. To this end, micropreparative lungs were fixed by the method inenstock et al. (Bienenstock J., Johnson N., Perey D.Y.E. Bronchial lymphoid tissue 1. Morphologic characteristics. // 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 (×7) was performed 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 p. and 200 g for 10 minutes, and then stained by Romanovsky-Giemsa, and counted andpulmonary they (in percent) (Avtsin A.P., Lukomsky GI Romanova L.K. et al. Andpulmonary they // Sov. med., 1982, No. 7, p.8 14).

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

Table 10
Indicators 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 mm BALL
CriterionThe intact
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 status is VA 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 mm BALL
The subpopulationThe intact
control
Model
(Sephadex)
Compound IV
N456
Macrophagesof 124.8±16,35226,1*±30,83152,4±18,5
Lymphocytes15,5±4,2728,2±6,0122,0*±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 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

Studies anti-inflammatory activity of compounds of General formula I (1%gel) to model carragenine edema in vivo

Materials and methods

Tests carried out on non-linear white mice weighing 30 to 32, Using the model carrageenin-induced edema by 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 - gain and edema

On - volume feet after entered what I 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:

, (2)

where

(o) - treated animals,

(K) - untreated animals.

Table 12
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 (%)
Vin 0.104±0,009**60,043,8
V-1Na0,14±0,02**79,026,2
IV-1Na0,105±0,002**47,633,0
II-1 Na0,1±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 12, 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 anti-inflammatory activity of compounds of General formula I, as shown 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 provide the report is ivoa anti-inflammatory effect.

Examples of dosage forms

Example 19

A. Preformed shape

Tablet form is received, using the following ingredients:

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

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 pharmaceutically acceptable salt0.2 g
Lactoseto 1 g

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

That is, the Nasal Spra the

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. Pharmaceutical composition having anti-allergic, antianaphylactic and anti-inflammatory activity, including N-acyl derivatives of amino acids of General formula (I)

where n is equal is 2 or 3; and
R is
or
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, or pharmaceutically acceptable salt in an effective amount and a pharmaceutically acceptable additive.

2. Pharmaceutical composition for regulating the content of eosinophils and reduce the degranulation of basophils, including N-acyl derivatives of amino acids of General formula (I)

where n is 2 or 3; and
R is
or
or its pharmaceutically acceptable salt in an effective amount and a pharmaceutically acceptable additive.

3. The use of N-acyl derivatives of amino acids of General formula (I)

where n is 2 or 3; and
R is
or
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 or their pharmaceutically acceptable salts as a means possessing anti-allergic, antianaphylactic and anti-inflammatory activity.

4. The use according to claim 3 N-acyl derivatives of amino acids or their pharmaceutically acceptable salts as a means possessing the ability to reduce an antigen secretion of histamine and to regulate the content of neutrophils and lymphocytes.

5. The use according to claim 3 N-acyl derivatives of amino acids as tools that can eliminate the symptoms of bronchial asthma and psoriasis.

6. The use of N-acyl derivatives of amino acids of General formula (I)

where n is 2 or 3; and
R is
or
or their pharmaceutically acceptable salts as a means to regulate the content of eosinophils and reduce the degranulation of basophils.

7. The use of N-acyl derivatives of amino acids of General formula (I)

where n is 2 or 3; and
R is
or
or their pharmaceutically acceptable salts as a means to eliminate the signs of allergic rhinitis, allergic rhinitis, seasonal and year-round rhinitis, allergic inflammation of the lungs.

8. A means of having antiallergic, antianaphylactic and anti-inflammatory activity, which is an N-acyl derivatives of amino acids total HUF the uly (I)

where n is 2 or 3; and
R is
or
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, or their pharmaceutically acceptable salts.

9. A method of treating allergic, anaphylactic, including diseases involving inflammation, comprising the administration to a mammal an effective amount of N-acyl derivatives of amino acids of General formula (I)

where n is 2 or 3; and
R is
or
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, or their pharmaceutically acceptable salts.

10. The method according to claim 9 for the treatment of bronchial asthma and psoriasis.

11. The method of treatment of allergic rhinitis, allergic rhinitis, seasonal and year-round rhinitis, allergic inflammation of the lung, comprising the administration to a mammal an effective amount of N-acyl derivatives of amino acids of General formula (I)

where n is 2 or 3; and
R is
or
or their pharmaceutically acceptable salts.



 

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15 cl, 2 dwg, 6 ex

FIELD: medicine.

SUBSTANCE: invention refers to inhibitors of enzymes cleaving protein after proline, such as depeptidyl peptidase IV inhibitors, as well as to their pharmaceutical compositions, and methods of application of such inhibitors. Particularly, inhibitors under this invention are improved in comparison with those currently in use in the present art by selecting special classes of side chains in P1 and/or P2 positions of inhibitor which contains carboxylic acid grouping.

EFFECT: compounds of specified formulas I, II and III can have the improved therapeutic index, partially owing to reduced toxicity and improved specificity in relation to target protease.

15 cl, 2 dwg, 6 ex

FIELD: chemistry.

SUBSTANCE: present invention relates to novel methods (versions) of producing N-acyl derivatives of amino acids of general formula , where n equals 2 or 3; and R is or their salts, using anhydrides of glutaric or succinic acid.

EFFECT: advantages of the proposed methods lie in simplicity of their realisation, easy extraction of the end product and high output of the end products.

2 cl, 27 tbl, 22 ex

FIELD: pharmacology.

SUBSTANCE: there is produced synthetic derivative of phthalyl-glycyl-agginyl-piperidide peptide expressing anticoagulant activity estimated by inhibiting amidolytic thrombin activity in system in vitro and double prolongation of rats' blood plasma coagulation as compared with the control values in tests activated partial thromboplastin and thrombin time tests and having formula 1, including pharmaceutically acceptable salts.

EFFECT: new synthetic peptide derivative expressing anticoagulant activity.

1 dwg, 2 tbl, 4 ex

FIELD: medicine.

SUBSTANCE: application describes ACE inhibitor complexes of Perindopril Erbumine with cyclodextrins, polyvinylpyrrolidone or hydroxypropyl cellulose, methods for making thereof, pharmaceutical compositions.

EFFECT: application for treatment of cardiovascular diseases.

19 cl, 23 dwg, 13 ex

FIELD: chemistry; medicine.

SUBSTANCE: invention relates to derivatives of 2-hydroxytetrahydrofurane , of general formula (I) , which possess ability to inhibit calpaines and/or ability to catch active oxygen forms and can be used to obtain medication, intended for inhibiting calpaines and/or lipid peroxidation.

EFFECT: medications possess higher efficiency.

9 cl, 64 ex

FIELD: medicine.

SUBSTANCE: invention relates to novel compounds of formula or to its pharmaceutically acceptable salts, where n is 0 or 1; R1 represents H or F; R2 represents C1-4alkyl; R7 represents H or C1-4alkyl; and Z represents hydroxyl C1-6alkyl or C1-6alkoxycarbonyl, or 5- or 6-member heteroaromatic ring, which belongs to aromatic rings which have given number of atoms, of which at least one is N, O or S, the remaining being carbon atoms, and which also optionally has methyl substituting group. Invention also relates to pharmaceutical composition, to application of compounds, as well as to method of obtaining formula I compounds.

EFFECT: obtaining novel biologically active compounds, possessing activity of receptor 5-HT2A antagonists.

9 cl, 25 ex

FIELD: chemistry.

SUBSTANCE: present invention relates to novel methods (versions) of producing N-acyl derivatives of amino acids of general formula , where n equals 2 or 3; and R is or their salts, using anhydrides of glutaric or succinic acid.

EFFECT: advantages of the proposed methods lie in simplicity of their realisation, easy extraction of the end product and high output of the end products.

2 cl, 27 tbl, 22 ex

FIELD: chemistry.

SUBSTANCE: invention pertains to new compounds with general formula (I) , where: R and Ri stand for phenyl, which can be substituted with 1, 2, 3 or 4 substitutes, which can be identical or different and chosen from a group consisting of chlorine, iodine, bromine, fluorine, trifluoromethyl and cyano. R2 and R3 can be identical or different and represent C1-5 branched or straight alkyl group. The alkyl group can be substituted with 1-3 fluorine atoms, or R2 and R3 - together with a nitrogen atom, with which they are bonded to, form pyrrolidine or piperidine ring. R7 represents hydrogen, C3-8 cycloalkyl, pyrrolidinyl or piperidinyl. The invention also pertains to salts of these derivatives used in pharmacology, as well as to compounds with general formula (IV), their pharmaceutical compositions, and their use.

EFFECT: obtaining new biologically active compounds which can function as modulators of cannabinoid receptors.

9 cl, 3 ex, 2 tbl

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

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

Anxiolytics // 2398761

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to medicine, namely to new amino acid derivatives of 3,4-dimethoxyphenylethylamine 1-4 exhibiting anxiolytic activity and described by general formula (1-4). In the formula, R represents

NH represents (COOH)2 (1-4); Y = 1 (1, 2); Y = 3 (3); Y = 0 (4).

EFFECT: preparation of an anxiolytic compound.

2 cl, 6 tbl, 2 ex

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