LV11727B - Pharmaceutical composition - Google Patents

Abstract

The invention relates to gamma -butyrobetaine-containing pharmaceutical compositions for oral, parenteral, subcutaneous or rectal administrations, that are providing for the treatment blood circulation disturbances of various genesis and localisation. This composition in the experiments on anaesthesized cats at a dose of 10 mg/kg, i.v. increases the total blood flow by 12 %, not considerably changing blood pressure and heart rhythm. The composition arrests adrenaline-induced isolated rabbit ear blood-vessel spasms. In a concentration of 2.0 mM it decreases reperfusion pressure by 18 %. NO-synthase blocking reverses the composition effect on adrenaline-caused blood-vessel spasms. Being infused the composition at a dose of 200 mg/kg significantly increases blood coagulation during phases I-II. In the comparative experiments the disclosed composition demonstrates more potent effect compared to known medicine [3-(2,2,2-trimethylhydrazinium)propionate]. The preparation is distinguished by a low toxicity and wide safety margin.

Description

A pharmaceutical composition

The invention relates to pharmaceutical compositions, in particular to pharmaceutical compositions associated with circulatory disorders of various genesis and localization.

The proposed therapeutic composition contains a known chemical, the use of which produces unexpected pharmacological effects. In particular, there is provided a pharmaceutical composition comprising γ-butyrobetaine as active ingredient in combination with pharmaceutically acceptable excipients and / or solvents.

γ-Butyrobetaine (actinin), from which carnitine is synthesized by the mammalian body, was initially characterized as a toxic substance that accelerates respiration, induces salivation and lacrimation, pupil expansion, vasoconstriction and cardiac arrest in diastole (1). At the same time, in subsequent works, other authors have found that γ-butyrobetaine is extremely low in toxicity (LD ₅₀ = 7000 mg / kg, subcutaneous).

Cardiovascular effects of unsubstituted γ-butyrobetaine have not been reported in the literature, although γ-butyrobetaine has been reported to be an acetylcholine-prolonged substance. However, the same author later reports that, by mistake, experiments have used the methyl ester, which has indeed cholinergic properties, instead of γ-butyrobetaine. In contrast, γ-butyrobetaine (unsterified) has been described as a pharmacologically inert substance (4, 5,6).

The closest structural analogue of γ-butyrobetaine used in the treatment of cardiovascular disease is the 3- (2,2,2-trimethylhydrazinium) propionate (Mildronate, Quaterin), an aza analogue of γ-butyrobetaine. Its mechanism of action is based on limiting the rate of carnitine biosynthesis and the consequent reduction of the transport of garlic fatty acids through mitochondrial membranes (7).

Acute toxicity was assessed in white mice (19-26 g) of both sexes in a group of 10 animals. The substances were administered as a 10% solution orally or intravenously at a rate of 0.004 ml / sec. It was found that the oral administration γbutirobetaīnam _LD50> 4500mg / kg, but at intravenous administration, LD _5u = 1860 (1430-2418) mg / kg, suggesting that γ-butyrobetaine is a practically nontoxic agent.

Special cat experiments have shown that a pharmaceutical composition containing purified γ-butyrobetaine, at a dose 186 times less toxic, has a stronger effect on vascular tone and blood flow than the known preparation and the closest structural analogue, Mildronate, and, unlike acetylcholine, decreases in heart rate and pulse rate while blood flow increases significantly (Table 1).

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Table 1

Effect of 3- (2,2,2-Trimethylhydrazmium) propionate (M), γ-butyrobetaine (GBB) and Acetylcholine (Ach) on Hemodynamics in Anesthetized Cats

Substance Deva, i.v., mg / kg % Change in blood pressure Pulse frequency changes % Change in blood flow rate M 5.0 ± 3 ± 3 + 5 10.0 ± 5 ± 3 + 8 * GBB 5.0 ± 4 ± 5 + 6 10.0 -7 - = - +3 ± 5 + 12 ' Ach 0.001 -35 ' -20 ' ± 8

*) p <0.05 versus baseline **) p <0.05 versus M dose

Experiments were performed on 2.9-3.8 kg narcotic cats / urethane (200 mg / kg) and chloralose (50 mg / kg), both i.p./. The experimental animals were exposed to the thoracic cavity, artificial respiration was performed, and blood pressure in the carotid artery and total blood flow to the aorta was measured with the help of the physiograph DMP-4B Narko Bio Systems, USA.

If the observed effect of γ-butyrobetaine on blood flow was due to a previously incorrectly attributed cholinergic component (mainly due to impurities in the γ-butyrobetaine esters (The Merck Index, Eleventh Edition, 1871) in poorly purified γ-butyrobetaine samples), then a significant decrease in blood pressure and pulse rate would be expected (see effect of acetylcholine, Table 1).

The observed cardiovascular effect indicates a positive inotropic effect of the proposed therapeutic composition, with a simultaneous reduction of peripheral resistance following a completely different mechanism that can be used in the treatment of heart failure and circulatory disorders of various genesis.

In experiments with isolated rabbit ear veins, a pharmaceutical composition containing γ-butyrobetaine is 2-3 times more potent than the closest structural analogue to the known preparation, 3- (2,2,2-trimethylhydrazinium) propionate (Mildronate), on adrenaline-induced vasospasm (Table 2). ).

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Table 2

Effect of 3- (2,2,2-Trimethylhydrazinium) propionate (M) and γ-butyrobetaine (GBB) on adrenaline-induced vascular spasms in isolated rabbit ear

Substances concentration (μΜ) Perfusion pressure (mm Hg) max / min Registered perfusion pressure downgrade (%) Original parameters Output data (after adding adrenaline 3.10 ' ⁷ M) max min max min M, 0.3 38 ± 5 8 ± 2 125 80 1 M, 1.0 38 ± 5 8 ± 2 123 77 4 M, 2.0 38 ± 5 8 ± 2 126 80 8 * GBB, 0.3 38 ± 5 8 ± 2 124 76 6th GBB, 1.0 38 ± 5 8 ± 2 125 80 15 * GBB, 2.0 38 ± 5 8 ± 2 125 78 18 **

* p <0.05 ** p <0.01

Unexpectedly, this vasodilating effect is found to be based on NO synthase activation, which is completely blocked by L-NO ₂ -arginine (Table 3).

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Table 3.

Effect of 3- (2,2,2-Trimethylhydrazine) propionate (M) and γ-butyrobetaine (GBB) on adrenaline-induced splenic rabbit vascular spasm in the presence of L-nitroarginine (L-NQ ₂ Arg) (10 mg / l)

Substances concentration (μΜ) Perfusion pressure (mm Hg) max / min Registered perfusion pressure downgrade (%) Original parameters Output data (after addition of adrenaline 3.10 ' ⁷ M and L-NO ₂ Arg) max min max min M, 0.3 36 ± 5 7 ± 2 165 102 0 M, 1.0 36 ± 5 7 ± 2 163,100 0 M, 2.0 36 ± 5 7 ± 2 165,100 2 GBB, 0.3 35 ± 5 8 ± 2 168 105 0 GBB, 1.0 35 ± 5 8 ± 2 165,100 0 GBB, 2.0 35 ± 5 8 ± 2 163,100 0

* p <0.05 γ-Butyrobetaine also has an effect on blood clotting time. We found it on the ICR-JCL line for men 24-28 g. heavy white mice per group of 10 mice using the Moravica method (TojiopoB ft. KjTHHHnecKHe jia6opaTopiiMe HCCJienoBaHHH b nejtHaTpHH, "MeaHu, η φκ3κ." . Blood was drawn from the jugular vein by pre-narcosis of the urethane (1000 mg / kg, i.p.). The solutions of the substances were infused immediately before the blood clotting time was determined.

As can be seen in Table 4, γ-butyrobetaine significantly prolongs the coagulation phase I-II - i.e. the time when fibrin strings occur. This means that pharmaceutical compositions based on γ-butyrobetaine can be used in the treatment of circulatory disorders associated with thrombus formation and thrombus embolism.

Table 4 Effect of γ-Butyrobetam (GBB) on Blood Clotting Time in Mice (by Moravica)

Substance, dose (mg / kg Aggregation Time (sec) GBB, 200 mg / kg, infused 46 ± 5.5 * Control (isotonic solution) 23.75 ± 3.4

*) p <0.05

Thus, we were able to discover that the γ-butyrobetaine-based pharmaceutical composition has a broad spectrum vascular effect that is related to vascular and myocardial tone and to NO synthase and is more potent than the known preparation of mildronate, which is a direct .

Thus, a pharmaceutical composition containing γ-butyrobetaine is a promising agent for the treatment of circulatory disorders of various genesis.

The preparation may be administered orally, parenterally, rectally or transcutaneously.

In the case of the active substances being administered by injection or orally in the form of drops, syrup or beverage, the pharmaceutical composition contains? -Butyrobetaine in a total amount of 0.5 to 40% by weight and as a pharmaceutically acceptable solvent such as distilled water, isotonic or glucose or buffer .

When the active substances are administered orally or sublingually in tablets, caplets, dragees, granules, powders or capsules, they contain and / or γ-butyrobetaine in a total amount of 0.01 to 0.5 g per tablet, caplet, dragee, capsule or per serving of powder or granules.

When the active substances are administered transcutaneously, their content in the ointment or patch is 0.5 to 40% by weight.

In the case of rectal administration, the content of the active ingredient in the suppository or microclimate is 0.5 - 40% by weight.

Claims (10)

References 1. Simkhovich B.Z., Shutenko Z.V., Meirena D.V. et al. 3- (2,2,2-trimethylhydrazinium) propionate (THP) - a novel γ-butylobetaine inhibitor with cardioprotective properties. Biochem. Pharmacol. 1988, 37, 195-202 2. W.Linneweh, Z.physiol.Chem., 42, 181, 1929 3. W.Rotzsch, I.Lorenz, E.Strack, Actabiol.med.ger.1959, 3, 28-36). 4. Hosein E.A., McLennan H. Pharmacological action of γ-butvrobetaine. Nature, 1959, 183, 328-329 5. E. A. Hosein, P. Proulx, Isolation and Probable Functions of Beta Esters in Brain Metabolism, Nature, 187, 321-322 (1960). 6. A.S. V.Burgen, F.Hobiger. Brit.Pharmacol. 4, 229 (1949). 7. E.Strack, K.Foesterling. Z.Physiol.Chem., 1953, 295,377). INVENTION FORMULA A pharmaceutical composition for the treatment of circulatory disorders, characterized in that it contains γ-butyrobetaine and pharmaceutically acceptable excipients as an active ingredient. Pharmaceutical composition according to claim 1, characterized in that the content of γ-butyrobetaine in the composition is 0.5-95% by weight. Pharmaceutical composition according to claim 1 or 2, characterized in that it is intended for oral or sublingual administration and it is in the form of tablets (coated or uncoated), capsules, capsules, dragees, granules, powder or solution containing 0.01 to 0.5 g of active ingredient per tablet, capsule, dragee, granule or powder dose, or as a 0.5 to 40% solution or syrup for oral administration. Pharmaceutical composition according to claim 3, characterized in that the pharmaceutically acceptable excipients are selected from the group consisting of stearic acid and its salts, lactose, glucose, sucrose, starch, talc, vegetable oils, polyethylene glycols, microcrystalline cellulose, aerosols. , flavorings, spices, colorings, ethyl alcohol and water, taken alone or in combination. Pharmaceutical composition according to claim 1 or 2, characterized in that it is intended for parenteral administration and is in the form of a solution for injection containing 0.5 - 40% by weight of the active ingredient and a pharmaceutically acceptable diluent. The pharmaceutical composition according to claim 5, wherein the pharmaceutically acceptable solvent is selected from the group consisting of distilled water, isotonic solution, buffer solution or glucose solution taken alone or in combination. Pharmaceutical composition according to claim 1 or 2, characterized in that it is intended for transcutaneous administration of the active ingredient and is in the form of an ointment, solution or patch containing 0.5 - 40% by weight of the active ingredient and a pharmaceutically acceptable excipient. . in the form of a solution or a patch containing 0.5 to 40% by weight of the active ingredient and a pharmaceutically acceptable excipient. 8. A pharmaceutical composition according to claim 7, wherein the pharmaceutically acceptable excipients are selected from the group consisting of water, polyethylene glycols 400, 1500, and 4000, vegetable oils, fats, glycerol, preservatives, emulsifiers, stabilizers, porous polymeric material, dimethylsulfoxide, alcohol and water, taken alone or in combination. Pharmaceutical composition according to claim 1 or 2, characterized in that it is intended for rectal administration of the active ingredient in the form of suppositories or micro-enemas, containing from 0.5 to 40% by weight of the active ingredients and pharmaceutically acceptable excipients. Pharmaceutical composition according to claim 9, characterized in that the pharmaceutically acceptable excipients are selected from the group consisting of water, polyethylene glycols 400, 1500 and 4000, vegetable oils, fats, glycerol, water, preservatives, emulsifiers and stabilizers taken alone or in combination.