Antidiuretic preparation and process for preparation thereof

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

There are no Lithuanian medicines available for the treatment of this disease, or they are outdated and do not meet modern requirements.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 output.

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 document, "Approved by the former USSR Ministry of Medical Industry," Grupovoj promyšlenyj Reg. 7-11-45 na proizvodstvo pituitrina dlia injijj ", pp. 31-34). .

Disadvantages of the known preparation pituitrine:

1. The extract produced has a low biological activity of 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 U / mg) in increasing the mass 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 manufacture.

The active ingredient, arginine vasopressin, isolated from the buccal pituitary gland extract purified by flash chromatography (HPLC), is a cyclic nanopeptide of the general formula <RTI ID = 0.0> eis-Tyr-Phe-Gln-Asn-Cys-Pro-Arg-Gly-NH.₂It is an object of the present invention that the preparation consisting of an active ingredient isolated from bovine pituitary gland extract and a preservative comprises the following components:

<td colspan="2">Active substance:</td>

<td>Purified HPLC Vasopressin Preservative: Chlorobutanol hydrate</td><td>- 0.01 g</td>

<td>(Former USSR FS 42-1574-80) Stabilizer: Chemically pure sodium acetate</td><td>- 5.0 g</td>

<td>(Former USSR GOST 199-78) Chemically pure acetic acid 0,1 mol / l</td><td>- 2.2 g</td>

<td>(Former USSR GOST 61-75) Treasure Toner: Sodium chloride</td><td>836.60 ml</td>

<td>(Former USSR FS 42-2572-88) Water for injections</td><td>6.8 g</td>

<td>(ex-USSR FS 42-2520-91)</td><td>- up to 1000 ml of solution.</td>

<td>The essence of the preparation method is that</td><td>by dissolving the active substance,</td>

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 present invention is also directed to the treatment of 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 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. Stir well and add the remaining volume (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 are 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, non-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 of 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 non-diabetes mellitus, ie with anti-diuretic hormone deficiency. Osmolarity of the urine increased more than twice (63.82%) on average after injection of 5 W Bovipressin, thus 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 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 24 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 drug overdose.

The starting dose was minimal and gradually increased according to 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 a study of urinary osmolarity, the effect of bovipressin on intramuscular injection was found to be approximately 3 hours, with subsequent osmolarity approaching lower or lower levels (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, resulting in at least two-fold increase in antidiuresis and urinary osmolarity.

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

3. The sensitivity 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, potent 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-diabetic diabetes.

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

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

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

The claimed formulation is significantly superior to the known in that it is enhanced pharmacotherapeutic activity because it is prepared from a 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