RU2597764C2 - Use of compounds of 1,3,4-thiadiazine class as agent for correction of experimental alloxan diabetes - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: invention relates to medicine, in particular to experimental pharmacology, novel biologically active compounds of general formula I, representing 2-morpholino-5-phenyl-6H-1,3,4-thiadiazin, hydrobromide (L-17); 2-morpholino-5-(4′-fluorophenyl)-6H-1,3,4-thiadiazin, hydrobromide (L-31), or general formula II: 2-amino propyl morpholino-5-phenyl-6H-1,3,4-thiadiazin, dihydrobromide (L-14); 2-amino propyl morpholino-5-(4′-fluorophenyl)-6H-1,3,4-thiadiazin, dihydrobromide (L-91) as agent for correction of experimental alloxan diabetes mellitus.

EFFECT: technical outcome is above compounds at intramuscular introduction in dosage of 40 mg/kg have antidiabetics action in development of alloxan diabetes in rats.

1 cl, 2 dwg, 1 tbl

Description

The invention relates to medicine, in particular to experimental pharmacology (new biologically active compounds).

The widespread prevalence of diabetes mellitus (DM) necessitates the search for new means of pathogenetic therapy of this disease. Existing approaches to the treatment of diabetes are reduced to insulin replacement therapy in the case of absolute insulin deficiency in type 1 diabetes, or to the appointment of drugs that stimulate insulin secretion and increase the sensitivity of target cells to it, in type 2 diabetes [1, 2, 3]. Synthetic antidiabetic drugs are known mainly in the classes of nitrogen- and sulfur-containing organic compounds, such as sulfonylurea derivatives, biguanides, thiazolidinediones, etc. [1, 2, 3]. However, existing approaches to the treatment of diabetes cannot ensure the full normalization of indicators of impaired metabolism and the prevention of chronic complications of the disease - diabetic angiopathies and neuropathy. In the progression of these complications, a leading role is assigned to two interrelated biochemical disorders - activation of non-enzymatic protein glycosylation and oxidative stress [1, 4].

Non-enzymatic glycosylation of proteins is activated in diabetes due to prolonged and severe hyperglycemia. The glycosylation reaction proceeds in three stages: the formation of the initial products of the process - Schiff bases and fructosamine; destructive oxidation of initial products to carbonyl intermediates (glyoxal, methylglyoxal, etc.); the formation of various glycosylation end products that irreversibly damage the protein structure (see Fig. 1).

Oxidative stress in diabetes is associated with an increase in the formation of free radicals during hyperglycemia, inactivation of antioxidant enzymes during their glycosylation, depletion of stocks of low molecular weight antioxidants, including thiols, as well as vitamins E and C. However, practical medicine does not have the means of treating diabetes combining the properties of antioxidants and non-enzymatic protein glycosylation blockers.

In the treatment of diabetes mellitus (in the experiment and the clinic), the use of lipoic acid preparations is known [3-5]. In the mechanism of action of lipoic acid in diabetes, its ability to transform into a thiol form - dihydrolipoic acid as a result of enzymatic reduction in vivo of a disulfide bond in the dithiolane cycle is traced. Dihydrolipoic acid, when administered orally and intravenously, exhibits antioxidant and non-enzymatic glycosylation of protein blockers.

In the scientific search for new effective antidiabetic agents, we drew attention to the original synthetic compounds of the 1,3,4-thiadiazine class, which, depending on the nature of its substituents, have a distinctive property - the ability to open and narrow the hetero ring with the formation of their various thiol derivatives [6 ]. Over the past 15 years, new compounds with a unique complex of antioxidant, hypothermic, and hypometabolic action have been obtained in this class [7]. Compounds that affect hemostasis [8] and have anti-inflammatory effects [8] have also been found. However, the use of compounds of the class 1,3,4-thiadiazine for the correction of experimental diabetes has not been reported.

As a result of a wide screening of various groups of 1,3,4-thiadiazines, 4 compounds of the general formula I and II were identified in an in vitro experiment, containing residues of morpholine (I) and 1-aminopropylmorpholine (II) as substituents at position-2 of the thiadiazine ring, showing pronounced antidiabetic properties:

General formula I: 2-morpholino-5-phenyl-6H-1,3,4-thiadiazine, hydrobromide (L-17); 2-morpholino-5- (4′-fluorophenyl) -6H-1,3,4-thiadiazine, hydrobromide (L-31)

General formula II: 2-Aminopropylmorpholino-5-phenyl-6H-1,3,4-thiadiazine, dihydrobromide (L-14); 2-aminopropylmorpholino-5- (4′-fluorophenyl) -6H-1,3,4-thiadiazine, dihydrobromide (L-91)

For the claimed compounds of class 1,3,4-thiadiazine I and II, in experiments in vitro and then in vivo, their high ability to inhibit non-enzymatic glycosylation of proteins, as well as their dose-dependent ability to reduce the rate of oxidation of ascorbic acid by atmospheric oxygen ( see fig. 2).

We associate the inhibition of the reactions of non-enzymatic glycosylation of proteins and the oxidation of ascorbic acid to dehydroascorbic acid with oxygen by air with compounds of the class of 1,3,4-thiadiazines, with the probable formation of SH-substituted derivatives of 1,3,4-thiadiazines (scheme), similar to the way it is observed by the example of their chemical transformation in acidic and alkaline media [6].

Thiol derivatives of 1,3,4-thiadiazines can bind glucose and carbonyl intermediates of non-enzymatic glycosylation of proteins and reduce dehydroascorbic acid to ascorbic acid (scheme)

As a result of a search in the scientific literature for compounds that correct metabolic disorders in experimental alloxan diabetes mellitus, data on a complex compound, a derivative of 1,3,4-thiadiazole, bis {3-phenyl-1- [2- ( 5-ethyl-1,3,4-thiadiazolyl)] carboxamido-1,3-propanedionato} cadmium,

for which hypoglycemic activity was detected during testing. However, this publication does not contain information about the antioxidant activity of this compound and its ability to block non-enzymatic glycosylation of proteins.

Research Method and Results

The experiments were performed on white Wistar male rats weighing 200-300 g. To study the ability of 1,3,4-thiadiazine compounds to correct the course of experimental diabetes, alloxan diabetes was simulated by triple intraperitoneal administration of alloxan in a total dose of 300 mg / kg of animal weight.

Experiment:

Simultaneously with the induction of diabetes, a solution of one of the compounds in distilled water (L-14, L-17, L-31 and L-92) was injected intramuscularly in a pharmacological dose of 40 mg / kg of the animal’s body with a frequency of 3 times a week for 4 weeks .

At the end of the experiment, the concentration of glucose and total protein was determined in the blood of animals. To characterize the activity of the process of non-enzymatic glycosylation of proteins (OGB) in animals, the concentration of fructosamine (FA) plasma and glycosylated hemoglobin (HbA _1c ) blood, the concentration of FA in organ homogenates were determined. The activity of free radical lipid oxidation was evaluated by the concentration of malondialdehyde (MDA) in blood plasma.

According to the table, the introduction of alloxan led to persistent and severe hyperglycemia, the accumulation of glycosylated proteins in the blood and organs of animals. Hyperglycemia and activation of non-enzymatic glycosylation of proteins could trigger oxidative stress, leading to the accumulation of MDA in the blood. The studied compounds of 1,3,4-thiadiazine also had the ability to partially correct metabolic disturbances in the development of alloxan diabetes. Since 1,3,4-thiadiazine cause reduction of hyperglycemia and HbA _1c concentration in the blood, as well as FA in the kidneys and liver of animals.

The ability of 1,3,4-thiadiazine derivatives to attenuate metabolic disturbances in developing alloxan diabetes is associated primarily with the protection of β-cells of the islets of Langerhans from oxidative stress induced by alloxan, leading to the death of pancreatic endocrinocytes. As the biochemical mechanism of such an effect, the ability of 1,3,4-thiadiazine derivatives to transform into thiol derivatives (see diagram) that increase the efficiency of the antioxidant protection of β-cells can be considered.

The technical result of this invention is a previously unknown application for the correction of experimental alloxan diabetes mellitus compounds of class 1,3,4-thiadiazine.

This fact allows us to expand the methodology for the search for such preparations among synthetic chemical compounds capable of transformation into various thiol compounds and, thus, to bind glucose and carbonyl intermediates of non-enzymatic glycosylation of proteins.

The claimed compounds of the class 1,3,4-thiadiazine are promising for their in-depth study in order to create an effective drug for the correction of experimental alloxan diabetes.

Bibliography

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2. Spasov A.A., Petrov V.I., Cheplyaeva N.I., Lenskaya K.V. The fundamental basis of the search for drugs for the treatment of type 2 diabetes // Vestnik RAMS. - 2013 - No. 2. - S. 43-49.

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7. On.N. Chupakhin, L.P. Sidorova, E.A. Tarakhtiy A.P. Novikova, N.M. Perova et al., RF Patent No. 2152943 (2000).

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Claims (1)

Use of a compound of formula (I) or formula (II)

as a means of correcting experimental alloxan diabetes.

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