RU2467748C1 - 3-(2,2,2-trimethylhydrazinium) propionate derivative - 3-(2,2,2-trimethylhydrazinium) potassium propionate glycinate exhibiting endothelioprotective activity - Google Patents

Abstract

FIELD: medicine, pharmaceutics.

SUBSTANCE: what is presented is the new chemical compound - a 3-(2,2,2-trimethylhydrazinium)propionate derivative - 3-(2,2,2-trimethylhydrazinium)potassium propionate glycinate, (CH3)3N⁺NHCH2CH2COOKRCOO^-, wherein

exhibiting endothelioprotective activity. The presented derivative can find application in medicine in the integrated treatment for endothelial dysfunction correction in cardiovascular diseases.

EFFECT: enhanced endothelioprotective properties as compared with previously known mildronat due to the introduction of new functional group showing the most manifested antioxidant properties.

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Description

The invention relates to medicine, specifically to new chemical compounds, glycinate 3- (2,2,2-trimethylhydrazinium) potassium propionate having endothelioprotective activity.

Known pharmaceutical composition containing 3- (2,2,2-trimethylhydrazinium) propionate, D-glucuronic acid, isotonic solution to stimulate hematopoiesis (RU No. 2033152, publ. 20.04.1995).

Known combination, which includes oral hypoglycemic agents, angiotensin converting enzyme (ACE) inhibitors, antioxidants and 3- (2,2,2-trimethylhydrazinium) propionate, which reduces the severity of symptoms of perineural ischemia (RU No. 2402325, publ. 10.27.2010 )

The proposed pharmaceutical compositions are a combination comprising two or more compounds with 3- (2,2,2-trimethylhydrazinium) propionate acting unidirectionally to increase effectiveness in the treatment of diseases of the cardiovascular system. But the use of several components in treatment increases the possibility of side effects of a single drug, the inconvenience of use for the patient, and the likelihood of an unpredictable effect when interacting.

It is known to use pharmaceutical compositions containing 3- (2,2,2-trimethylhydrazinium) propionate dihydrate (mildronate) and a salt and / or salts of succinic acid, in solid and / or liquid form, used as anti-ischemic, antihypoxic, antioxidant when used in the acute and subacute stages of various hypoxic conditions, in particular in patients with acute myocardial infarction and heart failure (RU No. 2005125660, publ. 02.20.2007).

The endothelioprotective efficacy of 3- (2,2,2-trimethylhydrazinium) propionate (mildronate) itself (M.V. Korokina and E.V. Artyushkova et al. "Endothelioprotective properties of enalapril and mildronate" is known. Scientific reports of the Belarusian State University. Series of Medicine. Pharmacy. 2011 , No. 4, pp. 57-61, “Endothelio- and cardioprotective effects of the cytoprotector of the metabolic action of meldonium in experimental modeling of nitric oxide deficiency”, collection of inter-university scientific conferences dedicated to the memory of Prof. Vladislav Pichugin and 75- anniversary of KSMU, Kursk, 2009, p.95-97).

In experimental modeling of arterial hypertension in animals, an increase in reactive oxygen species leads to endothelial dysfunction, which is confirmed by an improvement in endothelium-dependent relaxation with the introduction of antioxidants. Currently, drugs with antioxidant properties are considered promising for the correction of endothelial dysfunction, since the main mechanism underlying endothelial dysfunction is a decrease in the production and bioavailability of NO while increasing the superoxide anion, which has the ability to inhibit the expression and activity of endothelial NO synthase ( eNOS), as well as bind and inactivate NO, reduce its content in the cell. A shift in the physiological equilibrium between NO and O ² leads to the formation of highly toxic pyroxynitrite (ONOO ^- ). 3- (2,2,2-trimethylhydrazinium) propionate itself does not have antioxidant properties, but it increases the concentration of gamma-butyrobetaine (GBB) in the body, since under the influence of mildronate it is more slowly oxidized to carnitine. In turn, GBB is able to induce the formation of nitric oxide (NO), which acts as one of the most active natural agents that bind free radicals in the body. The mechanism of endothelioprotective action of mildronate is indirect, i.e., by increasing the number of GBD, it is able to protect cells from the effects of free radicals, but this is done through the induction of NO biosynthesis.

Taking into account the importance of changing antioxidant status as one of the most important ways of realizing an increase in the endothelioprotective activity of mildronate, it is necessary to increase its antioxidant effect, which contributes to an increase in endothelial protective properties.

The objective of the invention is the creation of a new derivative of 3- (2,2,2-trimethylhydrazinium) propionate, which has a more pronounced endothelioprotective activity.

The problem is achieved in that a new derivative of 3- (2,2,2-trimethylhydrazinium) propionate is created, obtained by reacting 3- (2,2,2-trimethylhydrazinium) propionate with potassium hydroxide and glycine in the presence of a solvent when heated,

(глицинат), обладающее эндотелиопротекторной активностью.Where

(glycinate) with endothelioprotective activity.

(глицинат), что позволило усилить эндотелиопротекторное свойство нового химического соединения.Modification of 3- (2,2,2-trimethylhydrazinium) propionate - glycinate 3- (2,2,2-trimethylhydrazinium) potassium propionate is a new chemical compound and exhibits the most pronounced endothelioprotective effect before the reference drug mildronate due to the introduction of a new functional group

(glycinate), which allowed to strengthen the endothelial protective property of a new chemical compound.

Glycinate is the remainder of the glycine amino acid, which performs a number of essential functions in the body, in particular, it is the central mediator of the inhibitory type of action, improves metabolic processes in the brain tissue, has a sedative and antidepressant effect, reduces psycho-emotional stress, and has the property of an antioxidant, thereby it will provide an increase in the endothelial protective effect and enable wider use of the new derivative for the prevention and treatment of cardiovascular istyh diseases.

The proposed new derivative is not described in patent and scientific literature.

An example of obtaining glycinate 3- (2,2,2-trimethylhydrazinium) potassium propionate

To a solution of 1.46 g (0.024 mol) of potassium hydroxide in 40 ml of 2-propanol at 30-32 ° C with stirring, add 3.5 g (0.024 mol) of 3- (2,2,2-trimethylhydrazinium) propionate and incubated 20 -25 minutes. 1.80 g (0.024 mol) of glycine are loaded into the resulting solution. Obtain 3.57 glycinate 3- (2,2,2-trimethylhydrazinium) potassium propionate, (yield 50.5% of the calculated), a crystalline, very hygroscopic product, T _{decomposition} 195-197 ° C.

Found,%: C - 32.17; H, 7.23; N - 14.38; C ₈ H ₁₈ N ₃ O ₄ K ₁ • 2H ₂ O;

Calculated,%: C - 32.54; H 7.45; N - 14.23.

IR spectrum, cm ^-1 : 3448, 3416, 3376, 3180, 2976, 1572, 1494, 1048, 1324.

Pharmacological properties of the claimed compounds

The experiments were performed on white Wistar male rats weighing 250-300 g. L-NAME (N-nitro-L-arginine methyl ether) was administered intraperitoneally at a dose of 25 mg / kg / day. On the 8th day from the start of the experiment, under anesthesia (chloral hydrate 300 mg / kg), a catheter is inserted into the left carotid artery to record blood pressure (BP), bolus administration of pharmacological agents is carried out in the right femoral vein.

Hemodynamic parameters: systolic blood pressure (SBP), diastolic blood pressure (DBP) and heart rate (HR) are measured continuously using a sensor and the Biopac Systems, Inc. computer program. Functional tests: endothelium-dependent vasodilation (ESV) - intravenous administration of acetylcholine (AH) at a dose of 40 μg / kg, endothelium-independent vasodilation (ENZV) - intravenous administration of sodium nitroprusside (NP) at a dose of 30 μg / kg. When statistical data processing was calculated, the average value, the standard deviation. Differences were considered significant at p <0.05.

Concrete example

A bolus intravenous administration of AH for 3-5 s led to a sharp drop in blood pressure, reaching a peak in intact animals for systolic pressure (SBP) - 84.3 ± 4.4, for diastolic pressure (DBP) - 38.7 ± 2, 8, while during the first 2-3 s sharp bradycardia developed up to 130-150 beats per minute. ENZV was also characterized by a decrease in SBP to 83.0 ± 3.7, DBP to 42.1 ± 4.4 with subsequent recovery. Blockade of NO synthase with the help of a long, daily, intraperitoneal injection of L-NAME (at a dose of 25 mg / kg) for 7 days caused arterial hypertension (SBP - 190.3 ± 6.7, DBP - 145.0 ± 3.9 mmHg) and led to a smaller decrease in blood pressure after administration of AH (SBP to 110.6 ± 5.2, DBP to 82.8 ± 6.6 mmHg) and NP (SBP to 88.7 ± 4.7, DBP up to 50.8 ± 4.2 mm Hg) compared with intact animals.

The simultaneous administration of L-NAME and 3- (2,2,2-trimethylhydrazinium) propionate led to the following indicators of blood pressure (SBP - 181.8 ± 9.3, DBP - 128.8 ± 6.0 mm Hg), in experiments with glycinate 3- (2,2,2-trimethylhydrazinium) potassium propionate (at a dose of 159.6 mg / kg intraperitoneally) did not lead to a decrease in the initial BP values (SBP - 181.8 ± 8.8, DBP - 135 , 1 ± 4.2 mm Hg), therefore, both mildronate and the derivative under study did not prevent the development of severe hypertension in the experiments. The dose of the derivative was calculated by the formula:

,

,

Where

D ₁ - dose of the studied derivative of glycinate 3- (2,2,2-trimethylhydrazinium) potassium propionate;

D ₂ - the dose for the drug comparing mildronate;

M ₁ is the molecular weight of glycinate 3- (2,2,2-trimethylhydrazinium) potassium propionate;

M ₂ is the molecular weight of 3- (2,2,2-trimethylhydrazinium) propionate (mildronate),

When choosing a dose of mildronate, we proceeded from the minimum therapeutic dose of the drug using the dose conversion factor for animals in this case for rats (Freireich et.al., 1966, Ulanova I.P. et al., 1968).

The fundamental difference in EDV and ENZV in intact animals and animals with the introduction of an inhibitor of NO synthase L-NAME naturally led to the need to derive a special indicator characterizing the degree of endothelial dysfunction, which is the ratio of the area of the triangle over the trend of the restoration of blood pressure (BP) in response to the introduction NP (endothelium-independent vasodilation) to the area of the triangle above the trend of the restoration of blood pressure in response to the introduction of AH (endothelium-dependent vasodilation).

This ratio was calculated in each animal of the intact group and rats after modeling the blockade of NO synthase and obtained a difference of this indicator by 5 times — 1.1 ± 0.1 in intact and 5.4 ± 0.6 in animals treated with L-NAME, respectively .

Thus, to further evaluate the effect of AH and NP in ESV and ENZV, this ratio was used as an indicator of the correction of endothelial dysfunction. So, in the group where Mildronate was administered against the background of the administration of the L-NAME synthase inhibitor L-NAME, this ratio corresponded to a value of 2.9 ± 0.4, and in the group with the introduction of glycinate 3- (2,2,2-trimethylhydrazinium) potassium propionate - 2.1 ± 0.3 (table 1), which indicates the most pronounced endothelioprotective effect of the studied derivative.

Results of the effect of 3- (2,2,2-trimethylhydrazinium) propionate and glycinate 3- (2,2,2-trimethylhydrazinium) potassium propionate on the coefficient of endothelial dysfunction when modeling L-NAME-induced (25 mg / kg intraperitoneally, once per 7 days) nitric oxide deficiency are shown in table 1.

Table 1 Groups of animals Functional test GARDEN, mmHg DBP, mmHg S vascular reaction when conducting EDVD with AH and ENVD with NP, in conv. units QED, conv. units Intact Source 137.7 ± 3.7 101.9 ± 4.3 1.1 ± 0.1 OH 84.3 ± 4.5 38.7 ± 2.8 1268.0 ± 74.8 NP 83.0 ± 3.7 42.1 ± 4.4 1375.3 ± 93.7 Receiving L-NAME (25 mg / kg) Source 190.3 ± 6.7 * 145.0 ± 3.9 * 5.4 ± 0.6 * OH 110.6 ± 5.2 * 82.8 ± 6.6 * 695.3 ± 87.6 * NP 88.7 ± 4.7 50.8 ± 4.2 3322.7 ± 116.7 * L-NAME (25 mg / kg) + Mildronate (90 mg / kg) Source 181.8 ± 9.3 ** 128.8 ± 6, ** 2.9 ± 0.4 ** OH 84.6 ± 3.6 ** 52.2 ± 2.7 * 823.96 ± 111.4 ** NP 93.2 ± 4.9 ** 57.6 ± 6.1 2389.5 ± 179.6 **

L-NAME (25 mg / kg) +
potassium 3- (2,2,2-trimethylhydrazinium) propionate glycinate (159.6 mg / kg) Source 181.8 ± 8.8 ** 135.1 ± 4.2 2.1 ± 0.3 ** OH 98.5 ± 3.3 ** 64.9 ± 6.0 ** 1212.1 ± 169.3 ** NP 102.8 ± 6.5 ** 74.2 ± 5.3 ** 2545.6 ± 323.7 ** Note: * - p <0.05 in comparison with the intact group, ** - p <0.05 compared to L-NAME, GARDEN - systolic blood pressure, DBP - diastolic blood pressure, S is the area above the curve for the restoration of blood pressure during functional tests, QED - coefficient of endothelial dysfunction

Thus, the obtained results allow us to ascertain the activation of correction of endothelial dysfunction against the background of the administration of L-NAME 3- (2,2,2-trimethylhydrazinium) propionate and its derivative glycinate 3- (2,2,2-trimethylhydrazinium) potassium propionate, but the endothelioprotective effect the glycinate derivative of 3- (2,2,2-trimethylhydrazinium) potassium propionate is higher in comparison with the comparison drug mildronate due to the introduction of a new functional group with the most pronounced antioxidant properties. These properties of the proposed derivative can be used in medicine in complex treatment for the correction of endothelial dysfunction in cardiovascular diseases.

Claims (1)

Derivative of 3- (2,2,2-trimethylhydrazinium) propionate glycinate 3- (2,2,2-trimethylhydrazinium) propionate potassium, (CH3) 3H ⁺ NHCH2CH2COOKRCOO ^- ,
Where

possessing endothelioprotective activity.