RU2634272C1 - Composition with desamino-arginine vasotocin for parenteral introduction and method for its production - Google Patents

Abstract

FIELD: pharmacology.

SUBSTANCE: agent of the invention contains 10-25 g of 3-aqueous acetic acid sodium, 0.2-3 g of acetic acid, 0.005-0.500 g of dAVT in 1000 ml of water, and an acetate buffer solution providing pH 5-6 for parenteral administration in the form of aqueous solution for hypernatremia treatment, comprising a therapeutically effective amount of dAVT and an acetate buffer solution providing a solution pH of 5 to 6. The method of the invention involves agent preparation, for this purpose, water is poured into the reactor, a calculated amount of 3-aqueous acetic acid sodium is added, stirred until dissolved, adjusted to pH 5.0-6.0 with acetic acid. DAVT is added to the resulting solution, mixed, the resulting solution is subject to sterilizing filtration through membrane filters with a pore diameter preferably not more than 0.30 mcm in a stream of nitrogen and poured into dark glass ampoules.

EFFECT: spectrum of means used to regulate the urination process is expanding.

5 cl, 2 dwg, 4 tbl, 5 ex

Description

The invention relates to the field of pharmacy and relates to a technology for the preparation of a pharmaceutical agent in the form of a solution for injection based on desamine-arginine-vasotocin (dABT), intended for the prevention and treatment of hypernatremia. The composition for parenteral use is stable, convenient for use, safe for clinical use. The composition has the advantages of a simple production technology, simple and convenient quality control and lower production costs compared to solid dosage forms. Thus, the tool is economically advantageous for industrial production.

In clinical practice, associated with the treatment of edema and hypernatremia, the question of maintaining sodium balance has been and remains one of the most relevant and debated [Ellison DH Physiology and pathophysiology of diuretic action // Seldin and Geibisch's The Kidney. Physiology and Pathophysiology; eds. RJ Alpern [et al.]. - 5 ^th Edition. - New York: Elsevier, 2013 .-- Vol. 1. - P. 1353-1404]. Active therapeutic tactics are necessary with a rapid increase in the concentration of sodium in the blood of the patient or in the presence of clinical manifestations of hypernatremia. In contrast to the danger of a quick recovery of normal sodium concentration in the blood after hyponatremia, complications of the correction of hypernatremia in adults (with its duration from several minutes to 2 days) are practically undescribed [Bataille S. et al. Undercorrection of hypernatremia is frequent and associated with mortality // BMC Nephrol. - 2014 .-- Vol. 15. - P. 37]. In chronic hypernatremia (more than 2 days), the recommended rate of decrease in blood sodium level is 0.5 mM per hour to reduce the risk of cerebral edema and seizures [Sterns, RH Disorders of plasma sodium - causes, consequences, and correction // N. Engl . J. Med. - 2015. - Vol. 372. P. 55-65].

With an increase in sodium concentration in blood plasma, the restoration of normal osmolality requires increased excretion of sodium ions by the kidney and a simultaneous increase in the reabsorption of osmotically free water into the blood. Currently in clinical practice for the correction of dyselectrolyte disorders, mainly loop diuretics are used [Wargo K.A., Banta W.M. A comprehensive review of the loop diuretics: should furosemide be first line? // Ann. Pharmacother - 2009. - Vol. 43. - P. 1836-1847] and the introduction of hypotonic solutions. However, their effect may be different and, depending on the volemic status, lead to both hypo- and hypernatremia [Katzung B.G. Basic and clinical Pharmacology. - NY: McGraw-Hill, 2004]. Dehydration therapy with diuretics in patients with impaired consciousness (with strokes, traumatic brain injuries, etc.) is associated with a high risk of developing hypernatremia due to a lack of thirst and inadequate water intake. As a result, a fatal outcome is possible due to hyperosmolar coma. It is necessary to search for drugs to reduce the volume of extracellular fluid, to increase the excretion of sodium ions by the kidneys while increasing the reverse absorption of osmotically free water in the renal tubules.

Thus, the development of drugs to reduce the volume of extracellular fluid, enhance the excretion of sodium ions by the kidneys, but with an increase in the reverse absorption of osmotically free water in the renal tubules, is a very urgent task.

The arsenal of modern medicine has a number of effective means for normalizing the water-salt balance. But, despite this, the problem of correction of hypernatremia remains relevant, as the diuretic agents used simultaneously enhance the excretion of salts and water by the kidney.

It is known that deamino-Cys-Tyr-Ile-Gln-Asn-Cys-Pro-Arg-Gly-NH ₂ (desamino-arginine-vasotocin, dABT) is able to inhibit tubular reabsorption of sodium ions and increase their excretion by the kidney, which by virtue of the effect comparable with the action of loop diuretics [Patent RU 2342949, published January 10, 2009]. At the same time, davt is an analogue of vasopressin, an antidiuretic hormone. Due to the effect on vasopressin receptors, the drug increases the reverse absorption of water in the kidney, which, along with sodium excretion, contributes to a more rapid normalization of sodium levels in the blood.

Derivatives of vasotocin are known which differ from the natural hormone in the structure of vasotocin, which has modifications at position 1, 4, 8 and optionally 2. New derivatives of vasotocin are vasoactive most of all because of the specificity of increasing blood pressure and in some cases have a significant prolonged effect. The antidiuretic effect (decrease in urine output) is due to the removal of the source of the molecule [Patent RU 2067586, published 10.10.1996].

Known derivatives of vasotocin cause increased excretion of potassium by the kidney, but this also increases the excretion of sodium ions and water by the kidney, and the increase in sodium and water excretion is several times higher than the increase in potassium excretion.

A known method of maintaining and correcting the water-electrolyte balance in mammals, based on the introduction of a synthetic analogue of argenin-vasotocin. The main objective of the method was to develop a method for maintaining and correcting the water-electrolyte balance in mammals, which allows to increase the reabsorption of osmotically free water while maintaining the balance of potassium and sodium and not causing changes in the activity of the cardiovascular system [Patent RU 2342100, published December 27, 2008].

This patent discusses only the method of application of various analogues of vasotocin and does not disclose information about the technology for producing these analogues.

A known method of maintaining and correcting the water-electrolytic balance in primates, also based on the introduction of a synthetic analogue of argenin-vasotocin [Patent RU 2342101, published December 27, 2008].

This patent discusses only the method of application of various analogues of vasotocin and does not disclose information about the technology for producing these analogues.

It is known to use a potassium peptide peptide: 1-desamino-4-arginine-8-arginine-vasotocin (1-d-Arg-AVT) as a therapeutically active chemical compound that affects the urinary activity of the kidneys, causing a selective increase in the excretion of potassium ions by the kidney [RU Patent 2454426, published on June 27, 2012]. This invention relates to the field of production of peptides that cause a selective increase in the excretion of potassium ions by the kidney, which are useful for the treatment of hyperkalemia, and do not allow sodium salts to be excreted from the body as much as possible, for example, in case of hypernatremia, which is important for ensuring optimal blood osmolality.

It is known to use new exenatide analogs of the general formula HisGlyGluGlyThrPheThrSerAspLeuSerLysGlnNleGluGluGluAlaValArgLeuPheIleGluTrp-LeuLysAsnGlyGlyProSerSerGlyXPro and Ala for the treatment of diabetes and Ala diabetes and Ala can be used for The obtained analogues are more stable than exenatide, have high insulinotropic activity, and also have high activity in relation to the excretion of sodium ions by the kidney without loss of potassium ions, enhance the excretion of calcium and magnesium ions, the excretion of osmotically free water. These properties can be useful for restoring water-salt metabolism along with the normalization of glycemia [Patent RU 2422461, published 06.27.2011].

Presented exenatide relates to polypeptides containing more than 20 amino acids. The synthesis of this compound is complex and multi-stage, which entails huge economic costs for its production and will not allow its use in practice as an affordable means for the correction of hypernatremia.

Known pharmaceutical composition containing oxytocin or its analogues, pharmaceutically acceptable salts and at least one non-ionic cellulose ether [Patent application RU 2013147739, published 05/20/2015].

This patent application does not address the possibility of using oxytocin derivatives to correct hypernatremia.

Thus, direct analogues, i.e. ready-made drugs based on deamino-vasotocin to increase the excretion of sodium salts by the kidney and enhance reverse selective absorption from the renal tubules into the blood water were not found.

Closest to the proposed invention is the use of [desamino-1, isoleucine-3, arginine-8] -vazopressin as a means to increase the excretion of sodium salts by the kidney and enhance reverse selective absorption of water-solvent from the renal tubules into the blood [Patent 2342949, published 10.01 .2009]. The disadvantage of this invention is the lack of a description of the composition for parenteral administration and further use in the clinic.

The basis of the invention is the creation of a pharmaceutical preparation in the form of an aqueous solution for intravenous and intramuscular administration, which contains the necessary and sufficient amount of desamine-arginine-vasotocin and has storage stability.

The problem is solved in that the claimed composition for parenteral administration in the form of an aqueous solution for the treatment of hypernatremia according to the invention contains a therapeutically effective amount of desamine-arginine-vasotocin and an acetate buffer solution that provides a pH of the solution from 5 to 6.

The inventive composition may additionally contain 10.00-25.00 g of sodium acetic acid 3-aqueous and 0.200-3,000 g of acetic acid in 1000 ml of solution.

Preferably, an effective amount of desamino-arginine-vasotocin is 0.005-0.500 g / 1000 ml.

A preferred embodiment of the claimed composition contains 0.012 g of desamino-arginine-vasotocin, 19.5 g of sodium 3-aqueous acetic acid, 0.315 g of acetic acid and a sufficient amount of buffer to provide a pH of 5.5 in a volume of 1000 ml.

To obtain the dosage form, water was poured into the reactor, the calculated amount of sodium acetic acid 3-aqueous was added, stirred to dissolve, adjusted to pH 5.0-6.0 with acetic acid, desamine-arginine-vasotocin was added to the resulting solution, mixed, the resulting solution was subjected sterilizing filtration through membrane filters with a pore diameter of preferably not more than 0.30 μm, most preferably through a filter with a pore size of 0.22 μm, and poured 1 ml into dark glass ampoules.

All stages of the process were carried out at room temperature and relative humidity of 30-50%.

When studying the properties of the obtained solutions, it was also found that during cooling, freezing and thawing of solutions during the day, the properties of the drug do not change.

When studying stability at elevated temperatures (60 ± 0.2 ° C), the following indicators were selected for drug control: description, authenticity, transparency, color, pH, mechanical inclusions, impurities, quantification, bacterial endotoxins. It was found that over the entire shelf life of the solution for injection of desamine-arginine-vasotocin, it remained stable in all respects.

The practical application of the claimed invention to regulate the process of urine formation (to increase the total amount of sodium ions excreted in the urine and enhance reverse selective absorption from the renal tubules into the blood water) provides him with the criterion of "industrial applicability".

The present invention is illustrated by the following examples of the preparation of the composition and its use as a natriuretic agent that can be administered by injection.

Example 1. The preparation of a solution for injection means based on davt.

1000 ml of water for injection are loaded into the acetate buffer solution installation, 19.7947 g of sodium 3-aqueous sodium acetate are added, and they are mixed until complete dissolution using a PE-8100 type mixing device. Then add 0.3 ml of glacial acetic acid, mix and measure the pH of the resulting buffer solution. The pH of the resulting solution should be 5.0-6.0. Then add 12.2 mg of dABT, mix.

The resulting solution is subjected to sterilizing filtration (filtered through a membrane filter with a pore size of 0.22 μm) in a stream of nitrogen, then poured into aseptic conditions into brown glass ampoules. 1000 ampoules of 1 ml are obtained.

Example 2. Assessment of the stability of the funds based on davt.

The stability of the agent according to Example 1 was studied by the method of “accelerated aging” under conditions of elevated temperature (60.0 ± 0.2 ° C) to control the quality of the drug, the following indicators were selected: description, authenticity, transparency, color, pH, mechanical impurities, extraneous impurities, quantification, bacterial endotoxins.

It was found that the developed davt agent for parenteral administration is stable during storage. There is no change in the color of the solution, the appearance of any inclusions. Also, there is no significant change in the concentration of the active substance and the appearance of impurities. The data obtained indicate that the proposed tool meets the requirements of the State Pharmacopoeia of the Russian Federation XIII, Volume II, presented for solutions for injection (OFS.1.4.1.0007.15).

It was found that all indicators remain within normal limits.

The technical result of the invention lies in the possibility of obtaining storage stable and non-toxic when using an aqueous solution of dawt for parenteral administration with a pH of 5-6, obtained without the use of toxic organic solvents and high temperatures.

Example 3. Assessment of natriuretic activity.

The experiments were performed on non-narcotic outbred female rats. Urine collection was carried out within 4 hours after the administration of drugs and a control substance (saline).

Administration to rats of the preparation of Example 1 in doses of 0.1-1.5 μg / kg led to an increase in the excretion of osmotically active substances by the kidneys (table 1). The most pronounced effect was detected with a dose of 1.0 μg / kg (Figure 1).

The furosemide loop diuretic caused a more pronounced increase in diuresis, but an increase in excretion of osmotically active substances in rats, which is equal to that based on davt. Furosemide, in contrast to davt, did not affect the clearance of osmotically free water and reduced the glomerular filtration rate (table 1).

The diuretic activity of the drug was 59 ± 6% of the diuretic activity of furosemide. Less diuretic activity with greater natriuretic activity of the drug is due to the opposite effect of furosemide and the drug on the transport of water in the kidney. The clearance data of osmotically free water (table 1) indicate that, unlike furosemide, which prevented the concentration of urine, the drug sharply increased the reverse absorption of osmotically free water in the kidney.

In parallel with the increase in excretion of osmotically active substances, the reverse absorption of water in the kidney increased, as evidenced by a decrease in clearance of osmotically free water (table 1, figure 1). Urination increased significantly with the introduction of the drug in doses of 0.25-1.5 μg / kg (table 1), the maximum diuresis was observed after injection of the drug in a dose of 1.0 μg / kg (Figure 1). The average glomerular filtration rate estimated by the clearance of endogenous creatinine did not change. Thus, the dAWT-based agent in rats acted as a saluretic, increasing urination due to the excretion of osmotically active substances.

An analysis was made of the excretion of sodium ions by the kidney. The dAWT-based agent caused an increase in sodium excretion when administered in doses of 0.1-1.5 μg / kg (table 2). For the excretion of sodium and potassium ions, the effects of davt-based agents at a dose of 1 μg / kg and furosemide at a dose of 10 mg / kg did not differ from each other. Excretion of magnesium ions and especially calcium ions increased to a much greater extent after the introduction of furosemide.

Natriuresis increased with increasing dose, the maximum effect was observed after administration of 1.0 μg / kg dABT (Figure 2). The natriuretic effect of the drug lasted more than 4 hours from the moment of administration, the natriurez peaked in the range of 30-60 minutes and exceeded the basal value by more than 35-40 times (Figure 2). Thus, the dAWT-based agent had a pronounced natriuretic effect.

To analyze the selectivity of the drug on the transport of sodium in the kidney, the excretion of other cations — potassium, magnesium, and calcium — was analyzed. An increase in potassium excretion was detected under the action of a dAvT-based agent in doses of 0.05-1.5 μg / kg (table 3). Excretion of potassium increased to a lesser extent than excretion of sodium. The potassium uretic effect of the nonapeptide lasted more than 4 hours; potassium excretion significantly exceeded the basal level throughout the entire experiment. Calcium excretion significantly increased under the action of davt-based agents in doses of 0.25-1.5 μg / kg (table 3). The drug exerted the most pronounced effect on calcium excretion during the first hour after injection of the dAWT-based agent and after 1.5 hours the indicator returned to the basal level (Figure 2). The total excretion of magnesium ions for 2 hours of drug action has not changed (table 2). An analysis of the dynamics of the excretion of magnesium ions under the action of a dAWT-based agent in a dose of 1.0 μg / kg showed that in the first 30 minutes of the experiment, rats increased magnesium, then this indicator decreased to the initial level (after 1.5 h), and subsequently the excretion of magnesium became lower than in control (Figure 2).

Thus, the dAWT-based agent had a natriuretic effect and affected the excretion of potassium, magnesium and calcium ions.

Example 4. The study of diuretic activity.

Evaluation of the diuretic activity of the drug was carried out according to the method of Kau et al. (1984) as modified by Lahlou et al. (2007) with preliminary oral administration of 0.9% NaCl solution to animals in a volume of 5% of body weight (50 ml / kg) through a probe into the stomach to normalize the water-salt balance. The agent of Example 1 was diluted with saline and administered at a dose of 1.0 μg / kg. A reference preparation with diuretic activity (furosemide (Sanofi India Limited, India) 10 mg / kg) and saline was injected intramuscularly into rats.

Indicators of renal function (diuresis, clearance of osmotically active substances and osmotically free water, excretion of osmotically active substances, sodium, potassium, magnesium, calcium ions, glomerular filtration rate) are calculated according to standard formulas and normalized to kg of body weight.

Table 3 presents the indicators of renal function in rats by standardizing the conditions of water-salt metabolism by oral administration of saline (50 ml / kg 0.9% NaCl) through a tube into the stomach.

When administered against this background, a drug based on dAWT or loop diuretic furosemide in rats showed an increase in urination, excretion of osmotically active substances, sodium and potassium ions (table 3). The excretion of sodium and potassium ions under the action of the agent according to example 1 was significantly higher than under the action of the reference drug furosemide. The natriuretic and potassiumuretic activity of the agent according to example 1 was 126 ± 7% and 130 ± 7% of the corresponding activities of furosemide.

The diuretic activity of the agent according to example 1 was 61 ± 5% of the diuretic activity of furosemide. Less diuretic activity with greater natriuretic activity of the agent of Example 1 is explained by the opposite effect of furosemide and a dAWT-based agent on the transport of water in the kidney. The clearance data of osmotically free water (Table 3) indicate that, unlike furosemide, which prevented the concentration of urine, the dAWT-based drug dramatically increased the reverse absorption of osmotically free water in the kidney.

Example 5. The effect of davt-based agents on serum sodium in animals with hypernatremia.

The experiments were performed on outbred female rats. To simulate hypernatremia in an experiment, animals were injected with a 2.5% NaCl solution in a volume of 18 ml per kg of body weight. The control substance, test and reference preparations were administered intramuscularly. After 55 minutes of the experiment, the rats were anesthetized with zoletil (Virbach, France; 50 mg / kg intramuscularly). After 60 minutes of the experiment, blood was taken from the vessels of the neck in rats, after which the animals were euthanized by decapitation.

Under control conditions, administration of dAWT-based agents to rats in a natriuretic dose of 1 μg / kg led to a decrease in sodium concentration and blood osmolality (table 4). At the same time, these indicators remained within the normal range. Injection of the reference drug furosemide led to an increase in the level of sodium in the blood (table 4). Both with the action of the agent based on dAWT, and with the action of the reference drug furosemide, there was a decrease in the concentration of potassium in the blood serum (table 4).

It was shown that intraperitoneal administration of a hypertonic (2.5%) NaCl solution to rats led to the development of acute hypernatremia (the concentration of sodium in the blood for over an hour exceeded 150 mM) and increased serum osmolality.

The administration of dAWT-based agents simultaneously with a hypertonic NaCl solution prevented a significant increase in sodium level and blood osmolality. The introduction of furosemide loop diuretic, in contrast, exacerbated hyperosmia and hypernatremia (table 4).

Thus, a davt-based agent at a dose of 1 μg / kg effectively reduces the blood sodium level in rats with acute hypernatremia caused by the administration of a hypertonic NaCl solution.

Claims (5)

1. The agent for parenteral administration in the form of an aqueous solution for the treatment of hypernatremia, containing in 10 ml of water 10-25 g sodium 3-aqueous acetic acid, 0.2-3 g acetic acid, 0.005-0.500 g desamino-arginine-vasotocin and acetate buffer a solution providing a pH of 5-6. 2. The tool according to claim 1, characterized in that the amount of desamino-arginine-vasotocin is 0.012 g. 3. The tool according to p. 1, characterized in that the pH of the solution is 5.5. 4. The tool according to p. 1, characterized in that it can additionally contain pharmaceutically acceptable excipients. 5. The method of obtaining funds for PP. 1-4, characterized in that in order to obtain a dosage form, water is poured into the reactor, the calculated amount of sodium acetic acid 3-aqueous is added, stirred to dissolve, adjusted to pH 5.0-6.0 with acetic acid, desamine-arginine is added to the resulting solution β-vasotocin, stirred, the resulting solution is subjected to sterilizing filtration through membrane filters with a pore diameter of preferably not more than 0.30 μm in a stream of nitrogen and poured into dark glass ampoules.

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