RU2061481C1 - Agent showing antiaggregation activity - Google Patents

Abstract

FIELD: medicine, diabetology. SUBSTANCE: proposed agent is 9-diethylaminoethyl-2,3-dihydroimidazo[1,2-a] benzimidazole dihydrochloride showing hypoglycaemic activity. At the dose single dose 25 mg/kg compound decreases glucose content in animal with alloxane diabetes by 38% and after 10 day administration (25 mg/kg, every day) hypoglycaemic effect is increased by 1.5 fold. Hypoglycaemic potency of compound is equal with gliclazide, it exceeds that of maninil by 1.5 time and that of adebite by 2.4 times that are the known sugar-decreasing drugs. Duration of sugar-decreasing effect of proposed preparation corresponds with that of hypoglycaemic action of sulfonylurea derivatives, - gliclazide and maninil and exceeds this index of butylbiguanide derivative - adebite - by 3 times. Sugar-decreasing effect of substance is due to its stimulating effect on insulin secretion from β-cells of pancreas and potentiating (by 2.7 times) and prolonged (by 1.5 times) influence on hypoglycaemic action of this hormone. Compound is an active inhibitor of platelet aggregation and exceeds this effect of gliclazide and acetylsalicylic acid by 37 and 16.4 times, respectively, in vitro experiments. Antiaggregation effect of compound on intravessel platelet aggregation exceeds that of acetylsalicylic acid by 23.3 times. Proposed compound can be used for diabetes mellitus treatment, in part, at complicated states and used orally. EFFECT: increased effectiveness of new proposed antidiabetic agent. 3 tbl

Description

 The invention relates to medicine, in particular, to the use of a compound for which hypoglycemic activity is known, as an agent having anti-aggregation action, due to which, after conducting clinical trials, it can be proposed for the treatment of diabetes and diabetic angiopathies.

 Diabetes is one of the most common diseases of the century. According to experts of the World Health Organization, by 2000 there will be 120 million patients with diabetes in the world. The focus of modern diabetology is the fight against vascular complications of diabetes. The main thing in the treatment and prevention of diabetic angiopathies is the stable compensation of diabetes mellitus regardless of its clinical type. However, the currently used antidiabetic drugs do not allow to achieve constant normoglycemia, and therefore the risk factors for the occurrence and progression of diabetic angiopathies are not completely eliminated. In the case of developed angiopathies, symptomatic therapy of vascular complications is used, including the appointment of antiaggregatory drugs - trental and acetylsalicylic acid. Side effects of the latter limit its widespread and long-term use.

 Particularly promising for the treatment of diabetic complications and are of justifiable interest are drugs that simultaneously affect the pathogenetic links of both the main (diabetes mellitus) disease and its vascular complications, i.e. combining hypoglycemic effect with microvascular action.

 These drugs include midaglizole. Its hypoglycemic effect is observed after 0.5-1 hours after administration and lasts up to 5 hours, the maximum effect is observed after 1-1.5 hours. When administered weekly, it inhibits platelet aggregation. This drug is rated as a short-acting sugar-lowering drug.

 Another oral hypoglycemic agent that has an additional antiplatelet property is glyclazide (diamicron), which, along with a pronounced hypoglycemic effect, affects the hemostatic system: it reduces the ability of platelets to aggregate, normalizes the impaired fibrinolytic potential of the vascular wall and the fibrinolysis process slowed down in diabetes mellitus. Gliclazide, as a hypoglycemic, begins to act 1 hour after administration, the maximum effect is observed between the 4th and 8th hour, the duration of action is slightly less than 20 hours. Angioprotective effect appears only after 3 months of treatment and even more distinctly after 6-12 months.

 Thus, the hypoglycemic drugs currently used in clinical practice, combining the hypoglycemic effect with the microvascular effect, are not perfect, since in some of them the angioprotective effect appears only after 3-12 months after the start of administration (gliclazide), while others a short hypoglycemic effect can be attributed to short-acting drugs (midaglizole). Therefore, the need for drugs that combine a hypoglycemic effect with an angioprotective effect, but more effective than those already used in clinical practice, is very high, and the creation of drugs with a similar original mechanism of action is relevant.

 The aim of the present invention was the search among compounds with a hypoglycemic effect, substances with antiplatelet properties, which can become the basis for creating a new peroral means for the treatment of diabetes mellitus complicated by angiopathies, more effective than those used in clinical practice.

выраженного антиагрегационного эффекта, превосходящего таковой ацетилсалициловой кислоты, широко применяемой в медицинской практике в качестве антиагреганта, и гликлазида, наиболее эффективного перорального гипогликемического средства для лечения диабета, осложненного ангиопатиями.This goal is achieved by the detection of the known derivative 9H-2,3-dihydroimidazo [1,2-a] benzimidazole with hypoglycemic activity, namely the dihydrochloride of 9-diethylaminoethyl-2,3-dihydroimidazo [1,2-a] benzimidazole of the formula I :

a pronounced anti-aggregation effect, superior to that of acetylsalicylic acid, widely used in medical practice as an antiplatelet agent, and gliclazide, the most effective oral hypoglycemic agent for the treatment of diabetes complicated by angiopathies.

The method of preparation and the properties of dihydrochloride I are described in [4]. This compound is a white crystalline powder, readily soluble in water. Its melting point is 257-258 ^o C.

In the IR spectrum of the base of compound I, namely 9-diethylaminoethyl-2,3-dihydroimidazo [1,2-a] benzimidazole, taken in a solution of chloroform, an absorption band at 1665 cm ^-1 is observed, which refers to the absorption of the C = N 2,3-dihydroimidazo [1,2-a] benzimidazole core.

и CH₂N), 3,78 (4Н, м, циклич. (СН₂)₂), 4,2 (2Н, т, CH₂), 6,82 (4Н, м, ароматич. Н).PMR spectrum of the base (CDCl ₃ ), ppm 0.96 (6H, t, 2CH ₃ ), 2.55 and 2.73 (6H, partially overlapping quartets and triplets,

and CH ₂ N), 3.78 (4H, m, cyclic (CH ₂ ) ₂ ), 4.2 (2H, t, CH ₂ ), 6.82 (4H, m, aromatic H).

), 3,68 (2Н, т, CH₂), 4,41 (4Н, с, циклич. (CН₂)₂), 4,6 (2Н, т, сливающийся с сигналом D₂O, CH₂), 7,41 (4Н, м, ароматич. Н).PMR spectrum of dihydrochloride I (D ₂ O), ppm 1.29 (6H, t, 2CH ₃ ), 3.34 (4H, q,

), 3.68 (2H, t, CH ₂ ), 4.41 (4H, s, cyclic (CH ₂ ) ₂ ), 4.6 (2H, t, merging with the signal D ₂ O, CH ₂ ), 7.41 (4H, m aromatic H).

 UV spectrum of a 0.0025 alcohol solution of compound I, nm: 217 ± 2, 279 ± 2 and shoulder at (282-286) ± 2.

 pH of a 2.5% aqueous solution of compound I (pH metric): from 3.7 to 4.2.

 The following are the results of pharmacological studies of compound I.

 The effect of dihydrochloride I and comparison preparations on platelet aggregation was carried out in experiments in vitro and in vivo.

In vitro platelet aggregation was studied by the method of detecting changes in the optical density of rabbit plasma platelet-rich plasma (from 2.5 x 10 ^-8 to 3.0 x 10 ^-8 blood platelets in 1 ml) when platelet aggregation-inducing substances were added to the latter under constant stirring. An ADF disodium salt (Reanal, Hungary) at a final concentration of 2 μg / ml (3.6 x 10-6 M) was used as an aggregation inducer.

 To evaluate the anti-aggregation effect of the substance, 0.02 ml of the substance was added to 0.5 ml of platelet-rich blood plasma of Chinchilla rabbits, after a 5-minute incubation with the substance, an aggregation inducer was introduced. In control experiments, instead of a substance solution, a 0.9% sodium chloride solution was added.

The anti-aggregation effect of the substance was evaluated by EC ₅₀ (concentration in mole, in which the substance inhibited the adhesion of blood platelets by 50%). The value of the EC _{50 was} determined by the calculation method according to the schedule "dose effect".

 In vitro experiments have established the presence of a pronounced anti-aggregation effect in the studied compounds. As follows from the data in table 1, compound I in its absolute antiaggregation activity exceeds acetylsalicylic acid by 16.4 times and glycazide by 37 times.

For in vivo studies, intravascular platelet aggregation was simulated. The experiments were performed on white non-linear rats of both sexes weighing 250,350 g, anesthetized with Nembutal (50 mg / kg intraperitoneally). 30 seconds after the administration of ADP (1 mg / kg) into the femoral vein, two portions of blood were taken from the abdominal aorta, one of them was stabilized with a formalin fixative to prevent the breakdown of platelet aggregates, and the other was fixed without formalin. In the supernatant, the platelet count was calculated using a phase-contrast method. The level of intravascular platelet aggregation was determined by the decrease in the number of platelets in the first sample compared to the second. To assess the effect of the test compound on intravascular aggregation of blood platelets, the substance was administered intragastrically 30 minutes before anesthesia of animals. The indicator of anti-aggregation activity was chosen as the value of ED ₅₀ dose, in which the substance inhibits platelet aggregation by 50%
As a result of studies, it was found that compound I exerts a dose-dependent antiplatelet effect, which excels acetylsalicylic acid 23.3 times more pronounced (Table 2).

As is known [4], compound I has a hypoglycemic effect. Additional experiments were conducted comparing this effect with that of gliclazide. As a result of the experiments, the ED ₃₀ values were established (the dose at which the substance lowers the blood glucose content by 30%) for compound I and glycazide, as well as for hypoglycemic preparations of mannyl and adebite (Table 3). From the data given in table. 3, it follows that compound I in terms of ED ₃₀ corresponds to the effect of gliclazide on blood glucose, 1.5 times more active than mannyl and 2.4 times adebite. It has a long hypoglycemic effect: when administered intragastrically at a dose of ED ₃₀ (25 mg / kg) it reduces blood glucose by 26-41% for more than 44 hours.

On the model of alloxan diabetes, the antidiabetic effect of compound I was studied and it was shown that with a single injection (intragastrically) of a substance at a dose of 25 mg / kg, a maximum glucose decrease of 38% is observed at the 6th hour of the experiment. With prolonged intragastric administration of substance I (10 days, administration once a day at a dose of 25 mg / kg), there is a tendency to increase its hypoglycemic effect. Under his influence, the decrease in blood glucose in experimental animals averaged 47%
To establish the mechanism of the hypoglycemic effect of compound I, its effect on the secretion and sugar-lowering effect of insulin was studied. As a result of the experiments, it was found that the test substance increases insulin secretion, potentiates and prolongs the sugar-lowering effect of this hormone.

The average lethal dose for substance I, determined when it was once administered into the stomach to white rats of both sexes weighing 160-200 g, was
LD ₅₀ female 2321.25 mg / kg
LD ₅₀ males 2740.31 mg / kg,
which allows us to attribute this compound to low-toxic substances.

 Thus, compound I has a pronounced antiplatelet effect. In its absolute anti-aggregation activity (in vitro experiments), it is 16.4 times higher than acetylsalicylic acid and 37 times higher than glyclazide. In experiments conducted to study the effect on intravascular platelet aggregation, compound I exerts a dose-dependent antiplatelet effect that is 23.3 times greater in severity of effect than acetylsalicylic acid.

 In addition, compound I also exhibits a hypoglycemic effect, which, according to the severity of the hypoglycemic effect, corresponds to the effect of gliclazide, is 1.5 times more active than mannyl, and 2.4 times more than adebite. A distinct antidiabetic effect of this substance on animals with experimental alloxan diabetes was noted with a single and prolonged (within 10 days) administration of it. It was established that under the influence of this substance there is an increase in insulin secretion, potentiation and prolongation of its sugar-lowering effect.

 The combination of the pronounced antiplatelet activity of compound I with a hypoglycemic effect, the increase under its influence of insulin secretion, potentiation and prolongation of the sugar-lowering effect of this hormone, low toxicity of the compound justify its use for the treatment of diabetes mellitus complicated by angiopathies. TTT1 TTT2

Claims (1)

 An anti-aggregation agent, characterized in that it is 9-diethylaminoethyl-2,3-dihydroimidazo (1,2-a) benzimidazole dihydrochloride.

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