LT4487B - Antidiuretic preparation and process for preparation thereof - Google Patents

Abstract

This invention belongs to the field of medicine. The aim of the invention is to increase an antidiuretic preparation pharmaceutical therapy activity and improve the production method thereof. An antidiuretic preparation contains active substances, secreted by the posterior pituitary gland, and a preservative. It consists of the following components:chromatographically purified vasopressin - 0.01 g,chlorobutanolhydrate - 5.0 g,sodium acetate - 2.2 g,ice acetic acid (0.1 mol/l concentration) - 836.6 ml,sodium chloride - 6.8 g,injection water up to 1,000 ml solution.The active substance, chromatographically purified vasopressin, is dissolved in an acetate buffer solution, isotonic by sodium chloride, the solution is preserved by chlorobutanolhydrate, it is filtered and poured into ampoules. The advantage of such a preparation is increased biological and pharmaceutical therapy activity, no present outside albumen admixtures.

Description

The present invention provides a pharmaceutical composition for use in the treatment of patients with antidiuretic hormone deficiency and a process for its preparation.

There are no Lithuanian medicines for the treatment of this disease, or they are outdated and do not meet modern requirements. In the case of anti-diuretic hormone deficiency, fluid circulation in the human body is impaired, with up to 20 liters of urine per day. Aitidiuretic drugs regulate the volume of urine excreted.

A known preparation with antidiuretic activity is pituitrine, which contains two posterior pituitary hormones (48% oxytocin, 39% vasopressin) and 13% foreign impurities. The preservative is phenol.

Pituitrine has a biological activity of 5 UU / ml. (See document approved by the former USSR Ministry of Medical Industry, "Grupovoj promyšlenyj Regulation No 7-11 -45 na proizvodstvo pituitrina dlia injection", pp. 2, 31, 32, 1975).

It is also known to prepare a formulation in which the active ingredient isolated from the bovine pituitary gland extract is dissolved in water, deprotected and preserved with a 0.3% phenol solution. The reconstituted solution is filtered and dispensed into 1.0 - 1.1 ml ampoules (See the document, "Approved by the former USSR Ministry of Medical Industry," Grupovoj promyšlenyj Regulation No 7-11-45 na proizvodstvo pituitrina dlia injectionj ", pp. 31-34). .

Disadvantages of the known preparation pituitrine:

1. Low bioactivity of the prepared extract in 3-5 UU / mg because the colloidal iron solution used for protein separation adsorbs the active ingredient.

2. Vasopressin adsorbed on Alt Bentonite was retained by the prior art and only partially isolated by elution with 0.1 N hydrochloric acid. Further purification was technologically impossible.

3. Low concentration of solutions and need for lyophilization.

4. By washing the absorbent colloidal iron with hydrochloric acid solution, the hormonal solutions become contaminated with minerals which must be subsequently removed and concentrated.

5. Low yields and high production losses due to the low activity of the resulting substance (no more than 5 UU / mg) to increase the weight of the active ingredient in the formulation.

The object of the invention is to increase the pharmacotherapeutic activity of the preparation and to improve the method of its preparation.

The active ingredient, arginine vasopressin, isolated from bovine pituitary glandular extract purified by flash chromatography (HPLC), is a cyclic nanopeptide of the general formula ε-Tyr-Phe-Gln-Asn-Cys-Pro-Arg-Gly-NH ₂ in that the preparation consisting of an active substance isolated from the bovine pituitary gland extract and a preservative comprises the following components:

Active substance: Purified HPLC Vasopressin Preservative: Chlorobutanol hydrate - 0.01 g (Former USSR FS 42-1574-80) Stabilizer: Chemically pure sodium acetate - 5.0 g (Former USSR GOST 199-78) Chemically pure acetic acid 0,1 mol / l - 2.2 g (Former USSR GOST 61-75) Treasure Toner: Sodium chloride - 836.60 ml (Former USSR FS 42-2572-88) Water for injections 6.8 g (ex-USSR FS 42-2520-91) - up to 1000 ml of solution. The essence of the preparation method is that by dissolving the active substance,

serving, filtering and dispensing into ampoules, - the active ingredient, HPLC-purified vasopressin, is dissolved in acetate buffer, isotonic with sodium chloride, and preserved with chlorobutanol hydrate.

The invention is also directed to the treatment of non-diabetes mellitus.

An example of a method of making an anti-diuretic called Bovipressin.

First, prepare a 0,1 mol / l acetic acid solution.

5 ml of glacial acetic acid GOST 61-75 (ex-USSR standard) is diluted with 11 water for injections.

A buffer is then formed. Dissolve 2.2 g of sodium acetate in 836.60 mol / l acetic acid and make up to one liter with water for injection.

The resulting buffer solution is dissolved in 6.8 g of sodium chloride solution for isotonization so that the osmotic pressure of the solution is equal to the osmotic pressure of the body fluids. Add the preservative to the solution thus prepared. Triturate 5 g of chlorobutanol hydrate in a mortar and make up to 300 ml with buffer acetate. Mix well and add the remaining solution (700 ml). Stir for 10-20 minutes. mixer. Leave in airtight container for 12 hours. After dissolving the chlorobutanol hydrate, check the pH, which should be between 3.80 and 3.95.

Then 0.01 g of HPLC-purified vasopressin powder with a biological activity of 450-500 UU / mg and a purity of at least 95% HPLC on oxytocin is dissolved in the prepared solution and mixed thoroughly. The prepared solution is filtered from mechanical impurities through membrane filters (through a membrane with a pore diameter of 0.22 µm). 860 ml of the product is obtained, which is dispensed in 1.1 ml into neutral glass ampoules in an inert gas atmosphere.

Clinical trials have been conducted in patients with non-diabetes mellitus with an antidiuretic agent called Bovipressin for injection.

The purpose of these studies is to determine the antidiuretic effect and tolerance of Bovipressin.

The studies involved 36 patients of both genders at various ages. They were divided into 4 groups: control - 7 patients, diabetic - 24 patients, psychological polydipsia - 4 patients, nephrogenic diabetes - 1 patient (Figure 1).

The course of investigations

The fluid retention-Bovipressin specimen was performed with a doubtful diagnosis of diabetes mellitus and administered 5 W Bovipressin intramuscularly and monitored for plasma and urine osmolarity every 1 hour. (4 hours total).

Fluid retention - vasopressin sample results are shown in Table 1 and Figure 1. The antidiuretic effect of bovipressin was observed in all patients. In one patient it was minimal and inadequate, as the osmolarity of the urine was below normal (in nephrogenic diabetes). In the control group, as in patients with psychogenic polydipsia, urinary osmolarity increased by less than 10% (9.46% and 4.99%, respectively). Meanwhile, fluid retention significantly increased urinary concentrations (71.25%), which helped to exclude them from antidiuretic hormone deficiency.

The most pronounced positive effect of bovipressin was observed in patients with diabetes mellitus, i.e. y. in the presence of antidiuretic hormone deficiency. On average, the osmolarity of urine increased more than twice (63.82%) after injection of 5 W Bovipressin, confirming the diagnosis of central neurohumoral diabetes mellitus.

Further study was conducted with Bovipressin only in patients whose diagnosis of neurohumoral diabetes mellitus was not in doubt.

Bovipressin was administered by intramuscular, subcutaneous and intravenous administration as small infusions. Administration of the drug has been shown to produce a rapid and shorter effect on the muscles. With subcutaneous injection, the effects last longer. Intravenous use in patients following neurosurgical surgery or in other indications for infusion therapy. The anti-diuretic effect of bovipressin was assessed by diuresis and fluid intake dynamics as well as changes in urinary osmolarity over a period of 3 hours (9-12-15-18-21-24-3-6 hours). The dose was adjusted to maintain the osmolarity of urethral within the normal range (600 ± 20Q mmol / kg) avoiding overdose.

The starting dose was minimal and gradually increased depending on the severity of the disease (Table 2). However, the effect of bovipressin was not only dependent on the severity of the disease, -1. the degree of antidiuretic hormone deficiency, but also according to the duration of the disease and the medication used, and was individual to each patient.

In the urinary osmolarity study, the effect of bovipressin on intramuscular injection was found to be approximately 3 hours, with subsequent urinary osmolarity approaching or lowering to the lower limit of normal (Table 3, Graph 1). With subcutaneous injection, the effect lasts for 4 to 6 hours, resulting in a more even effect at 4 times daily (schedule 2). During intravenous infusion, the drug stops working at the end of the infusion and is therefore suitable in cases where infusion therapy is required (the patient is unable to drink, is in an unconscious state, etc.) and should continue to administer Bovipine intramuscularly or subcutaneously.

conclusions

1. .Antiidiuretic agent Bovipressin is effective in neurohumoral non-diabetic diabetes, at least doubling antidiuresis and urinary osmolarity.

2. The dose of bovipressin to be administered to the patient should be individualized according to the degree of anti-diuretic hormone deficiency and individual sensitivity to the medicinal product.

3. Hypersensitivity to bovipressin depends on the duration of the disease and the vasopressin product used. Patients on long-term treatment with synthetic vasopressin have a worse drug effect.

4. Intramuscular injection of bovipressin has a rapid, strong and short duration of action, making it very suitable for the diagnosis of neurohumoral diabetes mellitus in a fluid retention - vasopressin test.

5. The infusion formulation is suitable for the treatment of patients with severe or unconscious conditions and following neurosurgery or head trauma, acute onset or iris-diabetes.

6. Bovipressin should be used when the product cannot be administered intranasally (runny nose, catarrh).

7. Due to the short duration of action, it is recommended that the drug be administered subcutaneously at least 3-4 times daily.

8. Adverse events of bovipressin did not occur at the optimal dose.

The claimed preparation is significantly superior to the known in that it is enhanced pharmacotherapeutic activity because it is produced from chromatographically pure drug substance (substance) with a biological activity (vasopressin purity of at least 95%) of at least 450 UU / mg (was 3-5 UU / mg). mg) and oxytocin not more than 0.1%.

DEFINITION OF INVENTION

Claims (2)

DEFINITION OF INVENTION Table 1 Fluid Retention - Vasopressin Sample Results Li pony groups Fluid retention sample Answer! Bovipressin PI is an osmotic. fluctuation limits (mmol / kg) In essence, fluctuation bounds (t miViol / kt) Osmoi of urine. in the oscillation bos ('mmoL'ks) Home During the sample Home During IvičiZinjo Checklist '^ UCC 285-283 N 2S9-295 M 537-905 N 739-1041 M / l ΓΓ c- ~ t s 1 5 C i J ~ i - J * A upper limit of nonos or t (9.46%) Sugar free diabetes 285 - 303 upper limit or min I 300-315 Φ not _β 'ϊΟζξ smaller than pi azrncs 144 - 331 slight f 593 - 720 Communication 77 / (63.82 / Psychological poipsips and 283 - 290 Normal or min 4 2S6 - 290 M 170 - 230 smaller than pi azmos 553 - 900 ryčkus1-IT 621 - Ib 'je further increased mini.'4 G'fr. ' Neurogenic diabetes 293 305 65 99 149 no placement or mm - f M - within normal range f - increased I - decreased (plasma osrnol. N285 - 295 mmol / kg, diaphoretic osmoi. N 400-900 mrnoi / kg) Table 2 Moderate changes in diuresis and urinary osmolarity during treatment with Bovipressin .ADH failure Bovipressin avg. daily dose (IU) Daily diuresis (1) In the urine osmcl. (mmo! / kg) before treatment in healing before treatment in healing Easy 10-15 <5 1,510.5 200-400 600-1100 Moderate 15-20 5-10 2.5 ₁ 1 100-200 500-100 Notable 25-30 > 10 4 11.5 <100 4001100 ADH - antidiuretic hormone Table 3 Urinary osmolarity dinar_____m _E v Bovipressin injections in absolute numbers and graphically \ Urine > osmol. Open Initial mmol / kg After 1 or. mmol / kg After 2 or. mmol / kg After 3 or. mmol / kg After 4 or. mmol / kg Notes 1 153 653 720 692 416 n 2S4 5 SS 741 535 257 -T J 128 467 671 497 323 4 276 309 680 542 513 5 283 460 623 523 414 6th 331 502 779 723 562 7th 458 415 612 418 S * 199 326 442 3S5 293 Consumed Adiurethin 9th 206 344 575 483 274 10th 298 635 786 841 462 11th 256 423 645 704 294 12 * 144 157 347 n-7 λ 190 Consumed Adiurethin * - The patient used synthetic vasopressin preparations for treatment before the sample Diagram I URINE OSMOLI. B0VIPRE3IN INJECTIONS -> A - control group B - Diabetes mellitus C - Psychogenic Polydipsia D - Nephrogenic Diabetes to ~ | - initial ”P ° Bovipressin 5VV injection f, | - urinary osmol.normal limits 1. An anti-diuretic preparation consisting of an active substance isolated from the buccal pituitary extract and a preservative, characterized in that it comprises the following components: Chromatographically purified Vasopressin - 0.01 g, Echlorobutanol hydrate, Titrate Acetic acid, glacial acetic acid (0.1 mol / l) Sodium chloride Water for injections 836.6 ml, 6.8 g, up to 1000 ml of solution. 2. A process for the preparation of a formulation according to claim 1, wherein the active ingredient is dissolved, preserved, filtered, and dispensed in ampoules, wherein the active ingredient - chromatographically purified vasopressin is dissolved in acetate buffer, isotonic with sodium chloride, and preserved with chic acid. A preparation according to claim 1 for the treatment of diabetes mellitus. Graph 2 DYNAMICS OF URINARY OS & IOLARITY DURING THE 4-DAY EOVIPRESSINE AFTER SKIN Becoming an osmol.