Dipeptidyl peptidase iv inhibiting agent and pharmaceutical composition thereof

FIELD: medicine, pharmaceutics.

SUBSTANCE: there are presented: agent applicable for treating or preventing diseases involving the enzyme dipeptidyl peptidase-IV (DPP-IV) representing Limiglidol (the same 9-diethylaminoethyl-2,3-dihydroimidazo[1,2-a]benzimidazole dihydrochloride, the same diabenol), a pharmaceutical composition thereof for the same application, using Limiglidol for preparing the pharmaceutical composition for the same application and using Limiglidol for normalising blood incretins (e.g., GLP-1, GIP decomposed under action of DPP-IV). What is shown is a high selectivity of Limiglidol in relation of DPP-IV.

EFFECT: invention enables using Limiglidol for preventing, delaying progression and/or treating conditions the pathogenesis of which involves the enzyme DPP-IV, including sterility, polycystic ovary syndrome, growth disorder, asthenia, arthritis, allograft rejection, autoimmune diseases (sclerodermia and multiple sclerosis), a variety of immunomodulatory diseases (lupus erythematosus and psoriasis), AIDS, intestinal diseases (necrotic enteritis, microvilli involving diseases or abdominal diseases), intestinal inflammatory syndrome, atrophy, etc.

10 cl, 1 dwg, 2 tbl, 1 ex

 

The invention relates to medicine, specifically to the means used to prevent, slow the progression and/or treatment of conditions mediated by the enzyme dipeptidyl peptidase IV, i.e. diseases, disorders or conditions that can be treated or prevented by inhibition of this enzyme.

The dipeptidyl peptidase IV (according to the nomenclature of enzymes IUBMB EC, hereinafter referred to DPP-4) is a membrane-bound serine peptidase, which is localized in certain tissues (intestine, liver, lungs, kidneys), as well as T-lymphocytes, where the enzyme known as CD-26. In the literature this enzyme has a wide variety of names, including DPP4, DP4, DAP-IV, FAPS, protein 2, forms a complex with adenoidectomies, protein linking adenoidectomies (ADAbp), dipeptidylpeptidase IV; Xaa-Pro-dipeptidylpeptidase; Gly-Pro-naphthylamide; postpoliomyelitis IV; lymphocytic antigen CD26; glycoprotein GP110; glycyrrhizinate; glycyrrhizinate; X-polydimethylaminoethyl; pep X; leukocyte antigen CD26; glycolpolyvinylpyrrolidone; dipeptidylpeptidase; picimplementation; dipeptidylpeptidase IV; aminoethylethanolamine; initiating T-cell molecule Tp103; X-PDAP.

DPP-4 is a non-classical is eminovoy aminopeptidases, which it dipeptides XAA-Pro with aminocore (N-Terminus) of polypeptides and proteins. It is also known that dependent on DPP-4 slow release of dipeptides type X-Gly or X-Ser happens in the case of some naturally occurring peptides.

DPP-4 is constitutively expressed on epithelial and endothelial cells of many different tissues (intestine, liver, lung, kidney and placenta), and also detected in the body fluids. DPP-4 is also expressed on circulating T-lymphocytes, and it has been shown that DPP-4 is identical to a cell surface antigen CD-26.

It is known that DPP-4 in vivo easy inactivates glucagon-like peptide-1 (GLP-1) and polypeptide ugnetajushe gastric secretion (GIP) and shows proteolytic activity against a number of immunoregulatory, endocrine and neurological peptides (GHRH, NPY, GLP-2, VIP).

GLP-1 and GIP are incriminati and produced after consumption of food. 29-amino acid peptide GLP-1 (7-36) is obtained by post-translational processing of proglucagon in the small intestine. GLP-1 (7-36) has many effects in vivo, including the inhibition of secretion of glucagon stimulation of insulin secretion, increased satiety and slow gastric emptying. However, GLP-1 (7-36) is very rapidly decomposed in vivo and, as shown, has a short half-life in vivo (t1/2) - 15 minutes therefore, in clinical practice it cannot be used as a medicine.

The study of genetically bred mice knocked out for DPP-4 and in vivo/in vitro tests with the use of selective inhibitors of DPP-4 have shown that DPP-4 is the primary enzyme GLP-1 (7-36) in vivo. In this regard, it was proposed that inhibition of the enzyme DPP-4, breaking incretin in the body. The main function of the enzyme is rapid inactivation of oligopeptides in different tissues and organs by removal of two amino acids from the end portion. GLP-1 is cleaved using DPP-4 to GLP-1 (9-36), which behaves as a physiological antagonist of GLP-1 (7-36). Studied that the enzyme DPP-4 rapidly inactivates GLP-1 and GIP immediately after their secretion in the body. When the inhibition of DPP-4 level and activity of incretins increased and extended. Therefore, inhibition of DPP-4 in vivo increases the levels of endogenous GLP-1 (7-36) and reduces the formation of his antagonist GLP-1 (9-36). As a result of this normalized ratio imbalance glucagon/insulin. As a result, the DPP-4 inhibitors can be used as medicines for preventing, slowing the progression and/or treatment of conditions mediated by DPP-4. The studies identified a number of compounds inhibiting DPP-4, which were applied IU izinski practice. However, there is still a high need for new DPP-4 inhibitors.

The purpose of this connection is to develop tools with which inhibit DPP-4 activity and the creation of a pharmaceutical composition for preventing, slowing the progression and/or treatment of conditions, the pathogenesis of which involved the enzyme DPP-4.

To solve this problem is proposed a means of having inhibiting DPP-4 activity, which is Limpida.

In another aspect of the invention features a pharmaceutical composition having inhibiting DPP-4 activity, which contains Limpida in a therapeutically effective amount and a pharmaceutically acceptable excipient. Preferably the pharmaceutical composition is intended for normalization of the level of incretins in the blood and/or normalize the function of endothelial cells.

In another embodiment, the invention features the use of Somigliana for the treatment and/or prophylaxis of diseases which can be treated or prevented by inhibiting the enzyme dipeptidyl peptidase IV, it is preferable to normalize the level of incretins in the blood and/or normalize the function of endothelial cells.

Also according to the invention features the use of Somigliana for the manufacture of a pharmaceutical is tion of the composition, possessing inhibiting DPP-4 activity. In a preferred variant the use of Somigliana for the manufacture of pharmaceutical compositions for the normalization of the level of incretins in the blood and/or normalize the function of endothelial cells, more preferably for the manufacture of a composition in the form of solid dosage forms.

The basis of the invention lies unexpectedly discovered by the authors of a new property of a known connection Somigliana regarding the key enzyme in the metabolism of DPP-4. As a result of the research Limpida was identified as a means for inhibiting the enzyme DPP-4.

Limpida (or dihydrochloride 9-diethylaminoethyl-2,3-dihydroimidazo[1,2-a] benzimidazole) was known as the connection with antiaggregatory, reducing blood viscosity and reduces insulin-resistance effect (RF patents 2061481, 2386634). Information about the availability of other types of activity, in particular the inhibition of dipeptidylpeptidase in the literature are missing.

Activity Somigliana according to the present invention as an inhibitor of DPP-4 can be demonstrated using methods known in the art. The inhibition constants determined in the following way. Apply a continuous fluorescence assay with the substrate Gly-Pro-AMC, which is cleaved p and using DPP-4 for release of fluorescent AMC leaving group. Kinetic parameters that describe this reaction are as follows: Km=50 µm; k cat=75 s-1; kcat / Km=1.5 x 106 M-1s-1. The reaction usually contains approximately 50 PM enzyme, 50 μm Gly-Pro-AMC, and buffer (100 mm HEPES, pH 7.5, 0.1 mg/ml BSA, 0.1 mm DTT and 1 mm EDTA) in a total reaction volume of 100 μl. The fluorescence of AMC continuously recorded using a microplate fluorometer in 96-well-plate at the wavelength of excitation 380 nm and emission wavelength of 460 nm. Under these conditions, about 0.8 μm AMC is formed for 30 minutes at 37°C. the Enzyme used in these studies was purified to a homogeneous state from human blood plasma methods precipitation of the sulfate pmoney, hydrophobic chromatography on phenyl-sepharose, gel filtration on Sephadex G-100 and affinity chromatography on concanavalin sepharose. Pelicanry drug DPP-4 was homogeneous, was a glycosilated by weight of the monomer 110 KD.

To measure the dissociation constants of the compounds solutions of inhibitor in DMSO was added to the reaction mixture containing the enzyme and the substrate (final concentration of DMSO was 1%). All experiments were performed at 37°C, using standard reaction conditions. To determine the IC50 was determined reaction rate using nonlinear regression according to the equation of Cheng-Prussia for competitive inhibition. Error when you play the AI dissociation constant is generally no more than two.

Limpida shows high selectivity for inhibiting DPP-4, which is confirmed by the experimental data presented below. The dipeptidyl peptidase-IV (all work on the production of enzyme preparations was carried out at 4°C).

The blood plasma was diluted 5 times with 0.02 M Tris-HCl buffer with pH 7.5, containing 0.1 mm dithiothreitol and 1.0 mm EDTA. To the diluted plasma with stirring was added a crystal (NH4)2SO4up to 70% of saturation (560 g/l). The precipitate was collected by centrifugation, the precipitate was dissolved in 0.05 M Na-phosphate buffer with pH 7.0, containing 1.5 M (NH4)2SO4, and was applied to a column (2.6×15 cm) phenyl-sepharose (Amersham-Pharmacia). Proteins were suirable declining gradient (NH4)2SO4in phosphate buffer. The fractions containing the activity of DPP-4, were collected, concentrated by the addition of (NH4)2SO4and the resulting residue was dissolved in 0.02 M Tris-HCl buffer pH 7.5, containing 0.1 mm dithiothreitol and 1.0 mm EDTA and applied to a column (2.6×90 cm) of Sephadex G-100 (Amersham-Pharmacia). Proteins were suirable with the same buffer at 30 ml/hour. Active fractions were collected, concentrated by ultrafiltration on a membrane UM-30 (Amicon) and chromatographically on column concanavalin-a-sepharose (Amersham-Pharmacia). Final purification was performed by ion-exchange chromatography on a MonoQ column (Amersham-Pharmacia). Received ven the rat DPP-4 was a dimer with a molecular weight in gel chromatography on ultragel ACA-34 220 Kd. The enzyme was homogeneous during electrophoresis in polyacrylamide gel in the presence of Na dodecyl sulfate and had a value of molecular weight of a monomer of approximately 110 Kd.

Purified DPP-4 was concentrated by ultrafiltration and stored at

-80°C.

The dipeptidyl peptidase-II.

The enzyme was purified from placenta of cattle. The division of activities DPP-2 and DPP-4 in the allocation process is followed by hydrolysis of the substrate Lys-Ala-AMC at pH 5.5 (specific to DPP-2) in contrast to the hydrolysis of Gly-Pro-AMC, split with an optimum at pH 7.5.

To highlight the DPP-2 used a modified compared to that described for human placenta procedure. Partial purification DPP-2 of the homogenate was performed by fractionation with ammonium sulfate, chromatography on DEAE-cellulose (Wathman) and phenyl-sepharose (Amesham-Pharmacia). The resulting preparation of the enzyme was stored at -80°C.

In addition, as the enzyme preparation used the blood plasma of a person.

Polyangiitis.

Wipe polyangiitis from human blood erythrocytes was performed by the method of ion exchange chromatography on DEAE-Sephadex a-50 (Amersham-Phamacia), gel chromatography on Sephadex G-75 (Amersham-Phamacia) and high-performance chromatography on a MonoQ column (Amersham-Phamacia) in the FPLC system. The resulting preparation of the probes consisted of a monomer and was homogeneous when electrophorese in the gel Poliak is lamida dodecyl sulfate Na and had a value of molecular weight of 75 Kd.

The purified probe was concentrated by ultrafiltration and stored at

-80°C.

The determination of the activity of proteases

Per unit of enzyme activity took his number off 1 μmol of substrate in 1 min

Definition dipeptidylpeptidase-IV (EC 3.4.14.5, CD26).

The activity of DPP-4 was determined by hydrolysis of the substrate Gly-Pro-7-amino-4-methylcoumarin (Bachem) fluorometrically.

Measurement protocols:

Consistently mixing 20 μl of an enzyme solution (0.5-5 mg of enzyme) 760 μl M Tris-HCl buffer pH 8.0 and 20 μl of a solution of Gly-Pro-AMC (1 mg/ml in DMSO), vigorously mixed and incubated at 37°C for 20 minutes the Reaction was stopped with 200 μl of 20% acetic acid. The fluorescence of 4-methyl-coumaryl-7-amide was measured at a wavelength of excitation 380 nm and 460 nm fluorescence.

The specific activity was determined by the formula:

A=[(E-C)/(S-B)]*t-1*v-1,

where E is the fluorescence of the sample (380/460 nm); C - fluorescence of the mixture containing 20 μl of substrate and enzyme, 760 μl buffer (pH 6.0), and 200 μl of 20% acetic acid; B is the fluorescence of a mixture containing 20 μl of the substrate, 780 μl of buffer and 200 μl of acetic acid; S - fluorescence mixtures containing 20 μl of the substrate, 780 μl and 200 μl of acetic acid and 1 μl of a solution of 7-amino-4-methylcoumarin (1 nmol).

Definition dipeptidylpeptidase-II (EC 3.4.14.2, DPP-2)

The activity of DPP-2 was determined by the guide is Aliso substrate Lys-Ala-7-amino-4-methylcoumarin (Bachem) fluorometrically at pH 5.5 (for hydrolysis of the substrate Gly-Pro-AMC optimum was observed at pH 8).

Measurement protocols:

Consistently mixing 20 μl of an enzyme solution (0.5-5 mg of enzyme) 760 μl of 0.1M Na-msftlogo buffer pH 5.5 and 20 μl of a solution of Lys-Ala-AMC (1 mg/ml in DMSO), vigorously mixed and incubated at 37°C for 20 minutes the Reaction was stopped with 200 μl of 20% acetic acid. The fluorescence of 4-methyl-coumaryl-7-amide was measured at a wavelength of excitation 380 nm and 460 nm fluorescence.

The specific activity was determined by the formula:

A=[(E-C)/(S-B)]*t-1*v-1,

where E is the fluorescence of the sample (380/460 nm); C - fluorescence of the mixture containing 20 μl of substrate and enzyme, 760 μl buffer (pH 5.5), and 200 μl of 20% acetic acid; B is the fluorescence of a mixture containing 20 μl of the substrate, 780 μl of buffer and 200 μl of acetic acid; S - fluorescence mixtures containing 20 μl of the substrate, 780 μl and 200 μl of acetic acid and 1 μl of a solution of 7-amino-4-methylcoumarin (1 nmol).

Definition dipeptidylpeptidase-I (EC 3.4.14.1; cathepsin C, DPP-1)

The experiment used a commercial preparation of cathepsin C from the spleen of a bull (With-8511, Sigma-Aldrich) with specific activity of ≥5 units/mg protein.

The activity of DPP-1 was measured in two ways - with spectrophotometric substrate-His-Ser-p-nitroanilide (His-Ser-pNA, Bachem, Heidelberg, Germany) and fluorometrically with the substrate Gly-Arg-7-amino-4-methylcoumarine (Gly-Arg-AMC, Bachem).

Spectrophotometric method for the determination of the activity of DPP-1.

From 0.25 to 10 μg enzyme preparation DPP-1 were incubated for 5 min at 37°C in 500 ál of activation buffer (100 mM Na2HPO4with pH 6.0, containing 20 mM NaCl, 1 mM EDTA and 4 mM cysteine), was added 400 μl of substrate buffer (100 mM Na-phosphate buffer with pH 6, containing 20 mM NaCl, 1 mM EDTA, 125 mM His-Ser-pNA and 1% DMSO). The release of p-nitroaniline were recorded for 10 min in a spectrophotometer DU-50 (Beckman, USA) at 410 nm. The amount of p-nitroaniline was determined on the basis of the molar extinction coefficient of 8800 M-1l-1cm-1.

Fluorometrically method for determining the activity of DPP-1.

As the substrate in this definition used Gly-Arg-7-amino-4-methylcoumarine (Gly-Arg-AMC, Bachem). The release of 4-methyl-coumaryl-7-amine was registered on spectrofluorometer LS-5-B (Perkin-Elmer).

Measurement protocols:

Consistently mixing 20 μl of an enzyme solution (0.5-5 mg of enzyme) 760 μl of 0.1M Na-phosphate buffer pH 6.0 and 20 μl of a solution of Gly-Arg-AMC (1 mg/ml in DMSO), vigorously mixed and incubated at 37°C for 20 minutes the Reaction was stopped with 200 μl of 20% acetic acid. The fluorescence of 4-methyl-coumaryl-7-amide was measured at a wavelength of excitation 380 nm and 460 nm fluorescence.

The specific activity was determined by the formula:

A=[(E-C)/(S-B)]*t-1*v-1,

where E is the fluorescence of the sample (380/460 nm); C is the fluorescence of a mixture containing 20 µl of substrate to the enzyme, 760 μl buffer (pH 6.0), and 200 μl of 20% acetic acid; B is the fluorescence of a mixture containing 20 μl of the substrate, 780 μl of buffer and 200 μl of acetic acid; S - fluorescence mixtures containing 20 μl of the substrate, 780 μl and 200 μl of acetic acid and 1 μl of a solution of 7-amino-4-methylcoumarin (1 nmol).

The dipeptidyl peptidase 8/9 (DPP-8/9)

To determine the activity of dipeptidylpeptidase 8/9 used the so-called "gelatinases" method, which distinguishes it from other dipeptidylpeptidase that gelatino not hydrolized. The enzyme preparation was obtained from peripheral blood leukocytes of various chromatographic procedures.

Polyangiitis (EC 3.4.21.26, probe)

The activity of the probes was determined similarly to the definition of DPP-4 using fluorogenic substrate Z-Ala-Pro-AMC, where Z=benzyloxycarbonyl).

In the experiment used purified from human erythrocytes enzyme.

Prolidase (CFS).

In the experiment used purified from kidney pig prolidase (Sigma-Aldrich, R). The enzyme selectively hydrolyzes dipeptides type XAA-Pro (partially Xaa-Hyp) and not hydrolyzes dipeptide Pro-Pro.

The principle of determination of enzyme-based monitoring catalyzed by enzyme cleavage of substrate Gly-L-Pro at 40°C and pH 8.0 in the cuvette of a spectrophotometer at a wavelength of 242 nm in 1-cm quartz cuvette.

For 1 unit of activity was taken to the quantity of the enzyme, gidrolizuemye 1 µmol Gly-Pro for 1 minutes

In the definition used the following reagents company Sigma-Aldrich:

A. 50 mm Tris-HCl buffer, pH 8.0 (prepared 100 ml of Trizma base (T-1503), is brought 1H. HCl to pH 8.0 at 40°C);

B. 30 mm solution of glutathione (GSH) (prepared immediately before use 5 ml of deionized water of reduced glutathione (G-4251));

B. 20 mm solution of MnCl2(prepared 10 ml deionized water (MnCl2·4H2O, M-3634));

, 25 mm Gly-L-Pro (forthcoming 30 ml solution A, using Gly-L-Pro (G-3002). The pH value was adjusted to 8.0 at 40°C, using if necessary 1N HCl or NaOH);

D. Solution prolidase (prepared immediately before use, the solution containing the enzyme of about 5 mg/ml in cold solution A).

Measurement protocols:

Prepare the activated enzyme, mixing (in ml) of these solutions exactly in the order listed:

A solution (buffer)2.40
The solution In (MnCl2)0.40
Solution B (GSH)0.10
Solution D (prolidase)0.20

Mix and incubated at 40°C for 20, 25 and 30 minutes Immediately after the asset is tion is placed in a quartz cuvette of the spectrophotometer DU-50 (Beckman-Coulter):

ExperienceControl
The solution G (Gly-Pro ml)2.702.70
The solution In (MnCl2, ml)0.200.20

Stirred and heated to 40°C. Record the absorbance at 242 nm to stable values. Then add, ml:

The activation mixture with enzyme0.50
The activation mixture without enzyme0.50

Quickly mix and record the decrease in optical density of about 5 minutes

Calculations:

Ed.fepm./ml=(ΔA242nm/mandnAboutpst-(ΔAnd242nm/mandnKaboutntpaboutlb)(3.1 )(3.4)(0.0254)(0.5)(0.2)

3.1=volume (in ml) of the activation mixture
3.4=volume (in ml) of the reaction mixture
0.0254=millimolar extinction coefficient for Gly-Pro at 242 nm
0.5=volume (in ml) used in the determination of enzyme
0.2=volume (in ml) used for activation of the enzyme

Determination of the degree of inhibition (IC50) of peptidases by somigliana and Comparators (sitagliptin and vildagliptin).

In the experiment used drugs inhibitors at a concentration of 0.1, 1, 10 and 100 μm. For each concentration measurements were carried out in the presence of different concentrations of Gly-Pro-AMC (0.006, 0.024, 0.10, 0.39, 1.56, 1.25, 6.25, 25 and 100 nM. There were three separate experiments in three parallel measurements.

The parameters of inhibition was determined in the program Prism 4 (GraphPad Software Inc., San Diego, CA, USA).

The Protocol ISM is rhenium:

To 20 ml of an enzyme solution (approximately 5 mg/ml protein) or untreated plasma was added 740 μl of 0.02 M Tris-HCl pH 8.0, 20 μl of a solution of somigliana, sitagliptin or vildagliptin and preincubated at 37°C for 30 minutes Then to the mixture was added 20 μl of the substrate solution in DMSO and kept yourout in the same conditions for 20 minutes the Reaction was stopped by adding 200 μl of 20% solution of acetic acid. Fluorescence released during the enzymatic reaction of 4-methyl-coumaryl-7-amide was measured at a wavelength of excitation 380 nm and 460 nm fluorescence. A graphical representation of the results.

For statistical analysis and graphical presentation of results, we applied the software package Prizm 4 (GraphPad Software, Inc.). The results of studying the influence on the activity of DPP-4 in plasma samples of all studied compounds are shown in table 1.

Table 1
The effect of drugs on the activity of dipeptidylpeptidase IV in plasma of patients with diabetes (n=7) and healthy volunteers (n=6)
MedicationThe residual activity of dipeptidylpeptidase IV (%)
oncentrate connection nmol
0110100
Plasma 1100.0±8.2--
Plasma 1+Limpida100.0±8.272.5±6.927.8±3.44.9±0.5
Plasma 1+Sitagliptin100.0±8.243.8±5.712.3±3.51.5±0.4
Plasma 1+Vildagliptin100.0±8.229.5±4.88.4±1.60.6±0.2
Plasma 2100.0±5.9---
Plasma 2+Limpida100.0±5.981.4±9.535.9±5.75.7±2.3
Plasma 2+Sitagliptin100.0±5.934.6±6.823.8±6.7 3.1±1.8
Plasma 2+Vildagliptin100.0±5.935.8±7.219.5±7.22.1±1.2

As can be seen from the table, the activity of DPP-4 in plasma oppressed all studied drugs in the concentration range from 1 to 100 nmol.

You can say that the greatest inhibitory activity observed in the comparison drug Vildagliptin. Limpida and used by us as another drug compare Sitagliptin inferior to him slightly. For comparison, the inhibitory properties of Somigliana with drugs Sitagliptin and Vildagliptin conducted a study of the kinetics of inhibition purified from human blood plasma enzyme.

The study of the kinetics of inhibition of purified dipeptidylpeptidase IV Somigliana, Sitagliptin and Vildagliptin.

Inhibition by Somigliana activity purified. DPP-4 is presented in figure 1 (the influence of Diabenese activity dipeptidylpeptidase IV at various concentrations of substrate Gly-Pro-AMC).

As can be seen from figure 1, DPP-4 is inhibited by the drug Limpida effectively.

The following experiment was carried out a comparative study of the inhibitory activity of drugs Liegler, Sitagliptin and Weldgrip is in.

Obtained in one experiment, the IC50 values for comparative drugs Sitagliptin and Vildagliptin data corresponds to literature. The IC50 values for them, calculated from the equation of Cheng-Prussia, was in the range of 5.1±2.3 nmol/l (Vildagliptin), 24.7±9.4 nmol/l (Sitagliptin), and 150.5±17.9 nmol/l for Somigliana.

The interaction of Somigliana and drugs comparison with other related DPP-4 enzymes studied in purified commercial drugs prolidase and cathepsin S (PLT-1).

Polyangiitis, DPP-2 and DPP-4 were isolated and purified in the laboratory of psychopharmacology, Institute of pharmacology them. You RAMS.

The influence of Somigliana, Sitagliptin and Vildagliptin activity related DPP-4 enzymes.

It was also studied the effect of Somigliana and Comparators on the activity of DPP-1, DPP-2, prolidase, polyangiitis.

The results of this study are presented in table 2.

Table 2
The influence of Somigliana, Sitagliptin and Vildagliptin on the activity of dipeptidylpeptidase 1, 2, 8/9, polyangiitis and prolidase
MedicationIC50 (µmol/l)
The PP 8/9 DPP-2DPP-1ProbesProlidase
Limpida23±62400±3203200±2906500±870>800000
Sitagliptin9±23200±1609150±4901800±230>1000000
Vildagliptin6±21205±23021000±15001500±270>1000000

As can be seen from table 2, all studied drugs showed practically no selectivity with respect to the enzymes of similar specificity of DPP-4. Obtained in the experiment results showed that the drug Limpida has a high inhibitory activity against purified from human blood plasma the dipeptidyl peptidase IV. Comparators - Sitagliptin and Vildagliptin also showed in this study, the high inhibitory activity.

Limpida and Comparators has not shown sufficiently inhibiting properties is in relation to other proteinases dipeptidylpeptidase 1, 2, 8/9, Propylenediamine and prolidase. Thus, the drug Limpida has a high selectivity with respect to the dipeptidyl peptidase IV.

Diagnosed in Somigliana inhibiting DPP-4 activity and high selectivity in respect of the latter led to the following effects when using Somigliana. The reduction in the activity of DPP-4 and related enzymes leads to the stabilization of received from outside incretin, i.e. the use of Somigliana you can control the splitting of incretin in the blood than is achieved normalization of the level of incretins. This implies the possibility of applying Somigliana to prevent, slow the progression and/or treatment of conditions/diseases, mediated by the enzyme dipeptidyl peptidase IV, it is preferable to normalize the level of incretins in the blood and/or normalize the function of endothelial cells.

Thus, in accordance with the invention features the use of Somigliana to prevent, slow the progression and/or treatment of the following conditions or diseases, the pathogenesis of which involved the enzyme DPP-4: infertility, polycystic ovary syndrome, growth disturbances, weakness, arthritis, allograft rejection in transplantation, autoimmune diseases (such as scleroderma and Russian the th sclerosis), various immunomodulatory diseases (such as lupus or psoriasis), AIDS, intestinal diseases such as necrotic enteritis, a disease with inclusions of microvision or disease of the abdominal cavity), the syndrome of intestinal inflammation, atrophy or failure of the intestinal mucosa caused by chemotherapy, nervous anorexia, osteoporosis, diabetes, syndrome X, dysmetabolic syndrome, complications of diabetes, glucosuria, hyperlipidaemia, metabolic acidosi, hyperinsulinemia, obesity, atherosclerosis and related diseases, and inflammatory bowel disease (such as Crohn's disease and ulcerative colitis), while sick, the needy treatment prescribed for administration, a therapeutically effective amount of an inhibitor of DPP-4.

Sperm motility/contraception men: a pharmaceutical composition comprising a DPP-4 inhibitors, can be used to change sperm motility and for contraception men, as in the seminal fluid are important for mobility of sperm prostatotomy, organelles produced by the prostate, have high activity levels of DPP-4 (Eur. J. Clin. Chem. Clin. Biochem 30, 333-338 (1992)).

The growth hormone deficiency: pharmaceutical compositions can be used for the treatment of growth hormone deficiency, based on the hypothesis that the releasing vectorwoman growth (GRF), the peptide that stimulates the release of growth hormone from the anterior pituitary, is cleaved by the enzyme DPP-4 in vivo (WO 00/56297). These data indicate that the GRF is an endogenous substrate: (1) GRF efficiently cleaved in vitro by formation of inactive product GRF [3-44] (BBA 1122, 147-153 (1992)); (2) GRF is rapidly degraded in plasma to GRF [3-44]; these phenomena can be prevented by exposure to an inhibitor of DPP-4 diplotene A; and (3) GRF [3-44] found in the plasma of transgenic pigs with human GRF (J. Clin. Invest. 83, 1533-1540 (1989)). Therefore, DPP-4 inhibitors can be used for the same range of symptoms that are discussed for stimulators of growth hormone secretion.

Damage to the bowel: the potential use of pharmaceutical compositions containing the DPP-4 inhibitors for the treatment of damage to the bowel prompted by results of studies showing that like peptide-2 (GLP-2), likely endogenous substrate for DPP-4, can cause a trophic effect on the intestinal epithelium (Regulatory Peptides 90, 27-32 (2000)). The introduction of GLP-2 increases the mass of the small intestine in rodents and soothes the intestine damage in models of colitis and enteritis in rodents.

The immunosuppression: inhibition of DPP-4 by pharmaceutical compositions containing the DPP-4 inhibitors can be used to modulate the immune response, osnovy is citing studies of the involvement of the enzyme DPP-4 to activate T-cells and the processing of chemokines, and on the effectiveness of DPP-4 inhibitors in models of the disease in vivo.

HIV infection: a pharmaceutical composition comprising a DPP-4 inhibitors can be used to treat or prevent HIV infection or AIDS, because a number of chemokines that inhibit the penetration of HIV-cells, are potential substrates for DPP-4 (Immunology Today 20, 367-375 (1999)). In the case of alpha-SDF-l splitting reduces antiviral activity (PNAS 95, 6331-6 (1998)). Thus, when the inhibition of DPP-4 is likely to stabilize alpha-SDF-l to reduce the invasive ability of HIV.

Obesity: pharmaceutical composition containing an inhibitor of DPP-4, can be used to treat obesity. This treatment is based on the observed inhibitory effect of GLP-1 and GLP-2 on food intake and gastric emptying. Exogenous introduction of GLP-1 reduces food intake and slows gastric emptying in humans (AmJ. Physiol. 277, R910-R916, 1999). ICV-introduction of GLP-1 in rats and mice also significantly affect the absorption of food (Nature Medicine 2, 1254-1258 (1996)). In mice GLP-1R* this inhibition of the absorption of food eaten is not observed, which indicates that these effects are mediated by receptors GLP-1 in the brain. By analogy with GLP-1 is likely that GLP-2 is also regulated by DPP-4. ICV-introduction GLP-2 also inhibits the absorption of food, similar to the effects observed for GLP-1 (Nature Medicine 6, 80-807, 2000).

Haematopoiesis: a pharmaceutical composition comprising a DPP-4 inhibitors can be used to treat or prevent disorders, because DPP-4 may be involved in hematopoiesis. Inhibitor of DPP-4, Val-Boro-Pro, stimulate haematopoiesis in the model neutropenia mice induced by cyclophosphamide (WO 99/56753).

Neural disorders: a pharmaceutical composition comprising a DPP-4 inhibitors can be used to treat or prevent various neural or psychiatric disorders, because the number of peptides involved in a variety of neural processes, split DPP-4 in vitro. Therefore, the inhibitor of DPP-4 may have a therapeutic advantage in the treatment of neural disorders.

Invasion (spreading) tumors and metastases: a pharmaceutical composition comprising a DPP-4 inhibitors can be used to treat or prevent tumor invasion and metastasis, because during the transformation of normal cells into malignant phenotype observed increase or decrease in the expression of some octapeptides, including DPP-4 (J. Exp. Med. 190, 301 to 305 (1999)). The raising and lowering of the level of these proteins appears to be specific for the different types of tissues and cells. For example, there is increased expression of CD26/DPP-4 in T-cell lymphoma, T-cell acute lymphoblastic leukemia is, when thyroid carcinoma cell origin, basal cell carcinomas and carcinomas of the breast. Thus, pharmaceutical compositions containing the DPP-4 inhibitors can be used in the treatment of such carcinomas.

Benign prostatic hypertrophy (national Department of standardization): a pharmaceutical composition comprising a DPP-4 inhibitors can be used for the treatment of benign hypertrophy of the prostate, as in patients with BPH there is increased activity of DPP-4 in prostate tissue (Eur. J. Clin. Chem. Clin. Biochem 30, 333-338 (1992)).

Gingivitis: a pharmaceutical composition comprising a DPP-4 inhibitors can be used to treat gingivitis, as in some studies, establishes communication with the severity of periodontal disease, discovered the activity of DPP-4 in the gingival fluid, located in the gingival sulcus (Arch. Oral Biol. 37, 167-173 (1992)).

Osteoporosis: a pharmaceutical composition comprising a DPP-4 inhibitors can be used to treat or prevent osteoporosis, because osteoblasts are present GIP receptors.

Taking into account the effects of incretins on the secretion of glucagon, the use of Somigliana is appropriate for patients for whom it will be useful normalization of the levels of glucagon and its correlation with insulin, and the attainable due to this improved the control of glucose levels in the blood, for example, patients with diabetes or prediabetes, which shows the DPP-4 inhibitors, preferably for patients with diabetes mellitus type 2 with normal or overweight, diabetes with endocrinopathy, with slowly progressive diabetes mellitus type 1 in adults in the early stages and in combination with insulin, diabetes in the young (MODY diabetes)and diabetes related complications, such as retinopathy, neuropathy, nephropathy.

According to the invention it is also proposed pharmaceutical composition having inhibiting DPP-4 activity, which contains Limpida in a therapeutically effective amount and a pharmaceutically acceptable excipient.

The claimed composition is intended for a mammal, preferably human, male or female, in need of inhibition of enzyme activity of dipeptidylpeptidase-IV. Preferably the pharmaceutical composition is intended for normalization of the level of incretins in the blood. In another preferred embodiment, the pharmaceutical composition is designed to normalize the function of endothelial cells.

More specifically, the claimed composition is intended for treating such diseases or conditions, such as infertility, polycystic ovary syndrome, growth disturbances, weakness, arthritis, the allograft rejection in transplantation, autoimmune diseases (such as scleroderma and multiple sclerosis), various immunomodulatory diseases (such as lupus or psoriasis), AIDS, intestinal diseases such as necrotic enteritis, a disease with inclusions of microvision or disease of the abdominal cavity), the syndrome of intestinal inflammation, atrophy or failure of the intestinal mucosa caused by chemotherapy, nervous anorexia, osteoporosis, syndrome X, dysmetabolic syndrome, complications of diabetes, hyperinsulinemia, glucosuria, hyperlipidaemia, metabolic acidosi, obesity, atherosclerosis and related diseases, and inflammatory bowel disease (such as Crohn's disease and ulcerative colitis), as well as patients with diabetes, treatment which is effectively the use of DPP-4 inhibitors, and patients with diabetes related complications, such as retinopathy, neuropathy, nephropathy.

The proposed remedy perform in a variety of dosage forms, preferably of solid dosage forms - tablets, tablets, sustained/prolonged release capsules, granules, powders. As auxiliary substances may be used substances commonly used in the pharmaceutical industry the items for the production of solid dosage forms, for example, starch, sugar, cellulose and its derivatives, gelatin, polyvinylpyrrolidone, polyethylene oxide, calcium phosphate, lubricant, wetting agent, as nutriceuticals, esters of polyoxyethylenesorbitan and fatty acids (twins), esters sorbitan and fatty acids (spany), preferably starch, including modified, lactose, microcrystalline cellulose, nutritionstrategywales, polyvinylpyrrolidone, excipients inhibitors release lubricant. Examples of the latter include stearic acid and/or its salt is calcium stearate, magnesium stearate, zinc stearate, talc, colloidal silica, Aerosil, polyethylene glycol, hydrogensource vegetable oil, liquid paraffin. The new composition may also contain flavoring agents, colorants and/or flavorings.

Preferably, the preparation is made in the form of pills, which can be a shell. The presence of the latter improves the appearance and organoleptic properties of the dosage form, protects it from mechanical damage. Preferably the shell is performed based on, for example, ready-mix brand "Opadry".

Preferred quantities of the active start (Somigliana) in a unit dose from 50 mg to 250 mg, more preferably 150 mg In the framework of the present invented what I term Limpida also includes hydrates, the solvate and salt Somigliana, for example the dihydrochloride.

The inventive composition may additionally include one, two, three or more therapeutic agents of other types. Preferably the composition includes as an additional therapeutic agent thiazolidinedione, pioglitazone, Metformin or derived sulfonylureas, such as gliclazide and glimepiride, most preferably Metformin.

In another aspect of the invention features the use of Somigliana for the manufacture of a pharmaceutical composition having inhibiting DPP-4 activity. In a preferred variant the use of Somigliana for the manufacture of pharmaceutical compositions for the normalization of the level of incretins in the blood and/or normalize the function of endothelial cells.

Obtaining the inventive dosage forms can be carried out in accordance with known methods for the production of solid dosage forms. All methods include the stage of combining the active ingredient with the carrier which consists of one or more accessory ingredients. In the General case, the inventive pharmaceutical composition is produced by uniform and homogeneous Association of Somigliana with liquid or powdered solid carrier, or both, and then, if necessary, when the try to the product of the desired shape, for example, wet granulation, and then add to the dry granules with a lubricant, forming a final mixture of ingredients with the formation of the dosage forms of a given configuration and size, and, if necessary, by drawing the shell, or by direct pressing, followed by the application shell.

Example 1. The sifted powders somigliana in the amount of 150 g, 0.5 g hydroxypropylmethylcellulose and low-molecular polyvinylpyrrolidone in the amount of 14.0 g mix until smooth and moisturize 40% solution of polyvinylpyrrolidone low-molecular weight granularit, the granulate is dried at 45-55°C to a residual moisture of 2.0 to 3.0%. Dry matter again granularit, to granulate add 5.0 g of a mixture of calcium stearate and stearic acid and tabletirujut on a tablet press, getting pills, which contain 150 mg of Somigliana.

Composition for gastric-soluble coating is prepared as follows. 32,8 g hydroxypropylmethylcellulose type 2910 with stirring, pour in 295,2 g of water and leave to swell under stirring. Separately prepare a solution of polyethylene glycol of 4.0 g of dry matter and 30 g of water, add in 3.2 g of talc are mixed, the resulting suspension is added to a solution of hydroxypropylmethylcellulose. The mixture is stirred to homogeneity and used for applying the coating is on the pill-kernel to produce a uniform coating with an average weight gain of the tablets approximately 3.7 to 5.0%.

Example 2. Mix the sifted powders somigliana in the amount of 150 g and a modified polyvinylpyrrolidone (copovidone) in an amount of 7.5 g granularit in the fluidized bed in a stream of air at a temperature of 35-40°C, using as granulating liquid 100 g of the solution of copovidone made from 20,0 g dry matter is dried to a residual moisture 1,0-2,0%, the granulate is sieved, add 5.0 g of polyvinylpyrrolidone, 0.05 g dioxide roamnia and 2.5 g of magnesium stearate and tabletirujut on a tablet press, getting tablets for dosage of 0.15 g

1. A means of having inhibiting the enzyme dipeptidyl peptidase IV activity, which is Limpida.

2. Pharmaceutical composition having inhibiting the enzyme dipeptidyl peptidase IV activity, which contains Limpida in a therapeutically effective amount and a pharmaceutically acceptable excipient.

3. The pharmaceutical composition according to claim 2, made in the form of solid dosage forms

4. The pharmaceutical composition according to claim 3, made in the form of tablets.

5. The pharmaceutical composition according to claim 4, made in the form of coated tablets.

6. The pharmaceutical composition according to claim 4 or 5, which contains a single dose of 150 mg Somigliana

7. The use of Somigliana for the manufacture of pharmaceutical compositions having inhibiting the enzyme dipeptidyl peptidase IV activity.

8. The use according to claim 9, in which the composition is made in the form of coated tablets.

9. The use of claim 8, in which the composition contains a single dose of 150 mg.

10. The use of Somigliana to normalize the level of incretins in the blood.



 

Same patents:

FIELD: medicine.

SUBSTANCE: 20 minutes before radiation in the UV chamber, an infusion of nettle, birch, plantain and oak leaves, tansy blossom taken in 1:1:1:1:1 are introduced orally into laboratory animals daily at a rate of 5 ml/kg of body weight.

EFFECT: method enables providing a manifested actoprotective and antioxidant effect on the exposed animals that gives reason to recommend the above agent to increase the non-specific body resistance with underlying pro-oxidant factors.

5 tbl

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to pharmaceutical industry, namely an agent possessing general tonic, adaptogenic and immunity decrease preventing action. The composition having general tonic, adaptogenic and immunity decrease preventing action contains rose hips and Echinacea extracts, a sweetening agent, a flavour, carnitine, inositol, magnesium gluconate, magnesium citrate, ascorbic acid (vitamin C), vitamin E, zinc gluconate, vitamin A, fructose, a preserving agent and water taken in certain proportions. A method for preparing the composition possessing general tonic, adaptogenic and immunity decrease preventing action.

EFFECT: agent possesses effective general tonic, adaptogenic and immunity decrease preventing action.

2 cl, 5 ex

FIELD: medicine.

SUBSTANCE: invention refers to radiation biology and radiation hygiene, namely the use of the fine (4 mcm) natural zeolite mineral cremcep for the prevention and treatment of radiation pathologies caused by the ingestion of iodine, cesium, strontium radionuclides.

EFFECT: invention provides reducing the internal exposure of both organs and tissues, and a body.

3 ex

FIELD: medicine.

SUBSTANCE: invention refers to medicine, namely to efferent medicine and may be used in hemodiafiltration in acute hepatic failure in the cardiosurgical patients. For this purpose, the bilirubin values more than 80 mcmole/l and the urea values more than 15 mmol/l requires introducing a solution as a dialysate to increase the clearance of toxic metabolites associated with plasma proteins. This solution is a standard bicarbonate solution of the dialysate added by 10% poly-(O2-hydroxyethyl)-starch in ratio 1:1. That is followed by the prolonged venovenous hemodiafiltration at the length of 5-10 h.

EFFECT: method enables providing the high clinical effectiveness in the given pathology in the cardiosurgical patients by creating both a diffusion gradient, and an oncotic gradient ensuring a consistency of the oncotic pressure of the dialysate.

1 tbl, 2 ex

FIELD: medicine.

SUBSTANCE: invention refers to medicine and aims at the correction of the age-related cardiovascular changes. Functional food is introduced into a human diet. What is used is the functional food "Samarsky Zdorovyak" No.83 and the food "Samarsky Zdorovyak" No.61 in a daily dose of 210 g. In the morning and evening, the functional food "Samarsky Zdorovyak" No.61 is prescribed in equal doses of 60 g, at dinner-time the functional food "Samarsky Zdorovyak" No.83 is prescribed in equal doses of 90 g.

EFFECT: method enables increasing the clinical effectiveness, reducing body weight and correcting hyperlipidaemia.

5 tbl

FIELD: biotechnologies.

SUBSTANCE: protein complex is proposed with improved activity of long-term action and biostability, containing a physiologically active polypeptide, an Fc-domain of immunoglobulin and a non-peptidyl polymer, having three functional ends. The method is disclosed to produce a protein complex containing a physiologically active polypeptide, an Fc-domain of immunoglobulin and a non-peptidyl polymer, having three functional ends A pharmaceutical composition is proposed, which contains the effective quantity of the specified protein complex, having the improved resistance in-vivo of the physiologically active polypeptide.

EFFECT: invention makes it possible to produce a protein complex with improved activity of long-term action and biostability.

24 cl, 3 dwg, 4 tbl, 9 ex

Ergogenic agent // 2482860

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to medicine, in particular to creating herbal pharmacological preparations as an ergogenic agent in sports medicine for mental and physical fatigue. The ergogenic agent is an aqueous alcohol tincture of elevated parts of Dunce's Caps in 70% ethanol in ratio 1:7. The content of the extractants in the tincture is not less than 0.007 g per a dose.

EFFECT: agent extends the range of ergogenic products.

3 tbl

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to organic chemistry, more specifically to new biologically active peptides which can be used in pharmacology and medicine to create new drug preparations that have cytoprotective activity.

EFFECT: new biologically active peptides are presented.

12 ex, 28 dwg, 2 tbl

FIELD: medicine, pharmaceutics.

SUBSTANCE: present invention refers to organic chemistry, namely new 3,8-diaminotetrahydroquinoline derivatives of formula (1a) or to their pharmaceutically acceptable salts wherein X represents CH2, C=O or CH-OR; m is 1 or 2; Ar represents a phenyl group or a 5-merous or 6-merous aromatic heterocyclic group having one element specified in S and N, (wherein the phenyl group may be substituted by 1-2 halogen atoms); each R1 and R2 represents a hydrogen atom; R3 represents a C1-C6 alkyl group or indolyl-C1-4 alkyl group (the indolyl group is optionally substituted by a C1-C6 alkyl group or a halogen atom), n is 0; R4 and R5 which may be identical or different, each represents a hydrogen atom or a C1-C6 linear or branched alkyl group; each R6 and R7 represents a hydrogen atom; and R represents a hydrogen atom. Also, the present invention refers to a drug preparation and a pharmaceutical composition of the basis of the compound of formula (1a), to the compound of formula (F1), to a method for preparing an intermediate compound (e).

EFFECT: there are prepared new 3,8-diaminotetrahydroquinoline derivatives which possess high GHS-R antagonist activity.

10 cl, 1 tbl, 124 ex

FIELD: medicine.

SUBSTANCE: invention relates to experimental medicine, in particular to study of immunomodulator efficiency for prevention and treatment of cholera. Before infection rabbits weighing 1.5-2.0 kg are given 10 injections of imunofan in dose 0.2 mcg, diluted in 0.3 ml of novocaine, every second day. After that animals are kept without food for 24 hours. After that, infection is performed by introduction of virulent strain of cholera vibrio into isolated loop of small intestine. Loops of small intestine are taken out, ligatures are applied, equal 12 cm long parts of intestine are tied up with 4-5 cm intervals between them under ether anesthesia. 1 ml of 0.9% sodium chloride solution is introduced into one ligated loop for control, with 109 cells of 24-hour culture of Vibrio cholerae 5879 being introduced into another experimental loop. After that, abdominal wall is sutured and 18 hours later animals are killed for the further study and estimation of efficiency of imunofan application. Estimation is carried out by presence in experimental tied up loops of small intestine of enteropathogenic effect, whose expression is estimated in crosses, and presence of cholerogenic effect.The latter is calculated by formula: K=Vliq/Lloop, K is coefficient of loop stretching, Vliq is liquid volume, Lloop is loop length. If enteropathogenic effect is absent in experimental loops and K<1.0, application of imunofan is considered to be efficient.

EFFECT: method ensures objective assessment of imunofan efficiency in case of cholera.

2 ex, 2 tbl

FIELD: chemistry.

SUBSTANCE: invention relates to compositions enriched with phytosterols. The method of producing porous microparticles containing phytosterols involves preparation of a homogeneous melt of a composition containing a phytosterol component and a component selected from at least one C10-C26 fatty acid, monoglyceride of a C10-C26 fatty acid and/or a metal salt and a C10-C26 fatty acid; hardening the obtained melt to form a solid amorphous substance; treating the obtained solid amorphous substance in a fine powder; mixing the obtained powder in an aqueous phase; separating the porous microparticles from the aqueous phase and drying the porous microparticles. The obtained porous microparticles are used to produce a pharmaceutical composition for reducing blood cholesterol level in mammals and for making a food product rich in phytosterols. The porous microparticles are also used in granulate form.

EFFECT: invention enables to obtain a composition of phytosterols with high bioavailability and long storage life.

25 cl, 2 dwg, 8 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to a compound of formula (I) or to a salt thereof: The invention also refers to a method for preparing said compound, a pharmaceutical composition for treating the diseases caused by increased vascular contraction, proliferation or inflammation caused by endothelin on the basis of said compound.

EFFECT: what is prepared is the novel compound and the salts thereof to be used in medicine for treating hypertension, pulmonary hypertension, diabetic arteriopathy, cardiac failure, erectile dysfunction and angina pectoris.

14 cl, 4 tbl, 1 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: presented group of inventions refers to medicine. What is presented is a method for preventing and treating the diabetes mellitus complications associated with the developing degenerative processes of the nervous tissue, comprising administering a medicine containing a peptide of general formula Pro-Gly-Pro, or a pharmaceutically acceptable salt thereof in effective amounts. There are presented pharmaceutical composition comprising said peptide for the neuroprotective therapy of the complications of diabetes mellitus and the method for preparing it.

EFFECT: group of inventions enables the more effective prevention and treatment of the diabetes mellitus complications associated with the developing degenerative processes of the nervous tissue by the use of the peptide Pro-Gly-Pro, or a pharmaceutically acceptable salt thereof.

9 cl, 2 dwg, 4 tbl, 7 ex

FIELD: chemistry.

SUBSTANCE: invention relates to novel 5-halogen-substituted oxindole derivatives of formula I: , where: R1 denotes hydrogen, methoxy or ethoxy group; R2 denotes hydrogen or a methoxy group; R3 denotes hydrogen, methyl, ethyl, n-propyl or isopropyl; R4 denotes an ethoxy or isopropoxy group; R5 denotes H or methyl; R6 denotes Cl or F; X1 denotes O, NH or CH2; X2 and X3 denotes N or CH under the condition that X2 and X3 do not denote N at the same time; as well as pharmaceutically acceptable salts thereof. The invention also relates to pharmaceutical compositions for treating and/or preventing vasopressin-dependent diseases, which contain derivatives of formula I, and use thereof in treating vasopressin-dependent diseases.

EFFECT: high efficiency of using said compounds.

25 cl, 8 ex, 2 tbl

FIELD: medicine, pharmaceutics.

SUBSTANCE: present invention refers to medicine, namely to pharmaceutical compositions for topical application containing insulin and liposomes. What is described is a pharmaceutical composition for topical application, containing insulin and liposomes in the form of a biofilm having rapid cutaneous penetration, and an ability to reduce blood glucose.

EFFECT: what is presented is the composition having rapid cutaneous penetration.

4 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: present invention refers to medicine, namely to pharmaceutical compositions for topical application. What is described is a pharmaceutical composition for topical application, containing insulin and liposomes bound thereto containing hydrated lecithines in a combination with cholesterol in the form of a plaste having rapid cutaneous penetration, and an ability to reduce blood glucose.

EFFECT: usability and ease of dosing.

4 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention relates to pharmaceutical composition in form of solid dosage form for treatment or prevention of diabetes, which contains therapeutically efficient quantity of metformin, polyvinylpyrrolodone, stearic acid and/or its salt, starch, silicon dioxide, characterised by the fact that as polyvinylpyrrolidone it contains polyvinylpirrolidone with molecular weight from 1000000 to 1500000 and additionally glycerol and/or basic finely dispersed magnesium carbonate.

EFFECT: pharmaceutical composition possesses high strength and ensures high degree of active substance release.

10 cl, 1 tbl, 5 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to tetrahydroimidazo[1,5-a]pyrazine derivatives of formula I or to their pharmaceutically acceptable salts (I), wherein: Ar represents phenyl, wherein phenyl is additionally substituted by 1-3 substitutes independently specified in halogen; R1 represents trifluoromethyl; R2 is specified in a group consisting of hydrohyl, alkyl having 1 to 4 carbon atoms, alkoxyl having having 1 to 4 carbon atoms, cycloalkyl representing a 5-6-member monocyclic ring group consisting of carbon completely, and -NR4R5, wherein each alkoxyl is optionally substituted by one group specified in a group consisting of phenyl and -OC(O)OR8; R3 is specified in a group consisting of a hydrogen atom and alkyl having 1 to 4 carbon atoms; each of R4 and R5 is independently specified is a group consisting of a hydrogen atom, alkyl having 1 to 4 carbon atoms, cycloalkyl representing a 3-8-member monocyclic ring group consisting of carbon completely, phenyl and pyridinyl, wherein each alkyl or phenyl is optionally substituted by one or more group specified in a group consisting of halogen, a cyano group, -SO2R7, -NR4R5 and -C(=O)OCH3; or R4 and R5 together with an atom, whereto attached form a 5-6-member heterocycle wherein the 5-6-member heterocycle optionally contains one or more N, O or S atom, and each 5-6-member heterocycle is optionally substituted by one or more groups consisting of halogen, hydroxyl, an amino group, alkyl having 1 to 4 carbon atoms, hydroxyalkyl 1 to 4 carbon atoms, -SO2R7, -C(O)NR4R5, -C(O)R7, =O; R7 represents alkyl 1 to 4 carbon atoms; and R8 is specified in a group consisting of alkyl 1 to 4 carbon atoms, and cycloalkyl representing a 5-6-member monocyclic ring group consisting of carbon completely. The invention also refers to methods for preparing them, a pharmaceutical composition having dipeptidyl peptidase IV inhibitory activity and containing said derivatives.

EFFECT: there are produced new compounds and composition on their basis which can find application in medicine for treating type 2 diabetes mellitus or hyperglycemia.

17 cl

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to pharmaceutical composition and medicine, and represents a hypoglycemic, hypocholesteremic, hypolipidemic and (or) antioxidant combination of bis-(γ-L-glutamyl)-L-cysteinyl-glycine in the form of disodium salt and lipoic acid in the form of sodium salt, and coordination compounds formed by palladium, copper and γ-L-glutamyl-L-cysteinyl-glycine wherein the molar ratio of bis-(γ-L-glutamyl)-L-cysteinyl-glycine of disodium salt : sodium lipoate : palladium : copper found within the range of 100-10000:100-10000:1-10:1-10.

EFFECT: invention provides higher therapeutic effectiveness.

7 cl, 4 ex, 2 tbl

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention relates to field of medicine, namely, to antidiabetic composition. Method of obtaining antidiabetic pharmaceutical composition includes preparation of trituracio mixture of active substance repaglinide, taken in therapeutically efficient quantity, with complex-forming substance, solubiliser and colloid silicon dioxide, further addition of filling agent, disintegrant and lubricant, and tabletting by method of direct pressing.

EFFECT: pharmaceutical composition in form of tablet, obtained by claimed method, is characterised by high degree of active substance release, satisfactory strength and has storage term longer than 2 years.

10 cl, 1 tbl

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to pharmaceutical industry and represents a vitamin additive containing vitamin B1, vitamin B2, nicotinic acid, vitamin B6, biotin and pantothenic acid; and wherein the weight ratio of biotin to vitamin B1 makes 1:20 to 1:25; wherein the weight ratio of biotin to vitamin B2 makes 1:25 to 1:30; wherein the weight ratio of biotin to nicotinic acid makes 1:310 to 1:330; wherein the weight ratio of biotin to vitamin B6 makes 1:30 to 1:35; wherein the weight ratio of biotin to pantothenic acid makes 1:110 to 1:130.

EFFECT: invention provides increasing activity, reducing the environmental stress effects, improving immunity of an individual, reducing the cases, the length and severity of respiratory infections.

13 cl, 1 ex

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