RU2020954C1 - Method of preparing of insulin aqueous solution - Google Patents

Abstract

FIELD: medicinal industry. SUBSTANCE: method involves successive performing of the following procedures: preparing solution 1 - 4.1-5.1 mg insulin is dissolved in 1 ml 0.05N acetic acid aqueous solution; preparing solution 2 - nipagin is dissolved in 0.1N sodium hydroxide solution (4.6-6.8 mg in solution). Then both solutions are mixed, glycerol and water are added up to required volume. Preparation is sterilized by filtration, kept at temperature 1-5 C for 14-30 days, and after repeated sterilizing filtration is poured into small bottles. EFFECT: increased stability of insulin aqueous solution to low temperature. 1 tbl

Description

 The invention relates to medicine, in particular to a method for producing dosage forms of insulin for injection, and can be used in the medical industry in the manufacture of an aqueous solution of insulin for injection.

 Various methods are known for preparing aqueous insulin solutions containing 40 IU / ml, which include the following stages: dissolving the insulin substance in acidified distilled water, adding a preservative (phenol, cresol or mixtures thereof, methyl paraoxybenzoic acid - nipagin), if necessary, mixing with buffer component and sodium hydroxide solution, adjusting the pH to a neutral value. The solution is subjected to sterilizing filtration and poured into vials under aseptic conditions.

 By these methods, insulin preparations are obtained that have a quick but short action: actrapid (Novo Denmark), Iletin (Eli-Lilly USA), etc.

 Closest to the proposed is a method of preparing a drug company "Eli Lilly" USA. The composition of the drug includes the following components: insulin substance 40 PIECES; glycerin 0.014-0.016 g; m-cresol 0.0025 g; water for injection - up to 1 ml.

 The method of preparation of the drug is as follows.

The zinc insulin substance is dissolved in distilled water acidified with hydrochloric acid. Separately, aqueous solutions of glycerol and m-cresol are prepared. The resulting solutions are combined, sodium hydroxide adjust the pH to a neutral value and the volume of the solution to the desired value. The prepared aqueous insulin solution is subjected to sterilizing filtration using membrane filters and poured into vials. The drug has a neutral pH, transparent, colorless and stored at 4 - 5 ° ^C. To this drug it is possible freezing, since at low temperatures (between ^-5 and -21 ° ^C) of protein aggregation is observed in the solution (in the solution opalescence ), which leads to the impossibility of its further use. It should be noted that this drawback is inherent in other fast-acting injectable forms of insulin.

 The need to increase the stability of an aqueous solution of insulin exposed to low temperatures is caused by the fact that in winter conditions the storage of the drug may be violated, for example, during storage, transportation, etc.

 The aim of the invention is to obtain aqueous insulin solutions that retain their physicochemical and pharmacological properties when exposed to low temperatures, i.e. more stable during storage.

The goal is achieved by sequentially conducting the following operations:
preparation of solution 1: the substance of insulin is dissolved at the rate of 4.1 - 5.1 mg in 1 ml of 0.05 N. an aqueous solution of acetic acid;
preparation of solution 2: dissolve nipagin in 0.1 N. an aqueous solution of sodium hydroxide based on its content in 1 ml of a solution of 4.6 - 6.8 mg;
with stirring at a speed of 50-60 rpm, solution 2 is added to solution 1;
after mixing the solutions add glycerin and water to the desired volume of the drug;
the resulting aqueous solution is subjected to sterilizing filtration is maintained at 1 - 5 ^{° C} for 14-30 days and after repeated sterilizing filtration bottled.

The following component composition of the preparation is proposed: Insulin substance 40 PIECES Acetic acid 0.9-1.1 mg Glycerin 14-17 mg Nipagin 1.0-1.3 mg Water for injection Up to 1 ml
The proposed method allows to obtain an insulin preparation that retains its physicochemical and pharmacological properties when exposed to extreme temperature conditions.

 The results of the analysis of drugs are summarized in the table.

 The claimed method has been tested at enterprises producing insulin preparations: Kiev Darnitsa Production Chemical and Pharmaceutical Association, Lviv Meat Processing Plant and Production Association Belmedpreparaty. The described preparations underwent a preclinical, clinical study and by the decision of the Pharmacological Committee No. 13 dated 09/12/91 were allowed for production and release into the pharmacy network.

Using the proposed method in an industrial environment will provide an opportunity to provide healthcare with a stable, transparent, neutral fast-acting insulin preparation, which can be exposed to temperature effects from + 40 ^o to -21 ^o C.

PRI me R 1. 1.54 g (total activity of 40,000 PIECES) of the substance of porcine insulin with a biological activity of 26 PIECES / mg, with a zinc content of 0.42% dissolved in 340 ml of 0.05 N. acetic acid solution (1020 mg of CH ₃ COOH) (solution 1). Next, prepare solution 2, adding to 200 ml of 0.1 N. sodium hydroxide (800 mg NaOH) 350 ml of water and 1.2 g of nipagin and stirring until the nipagin dissolves. To solution 1, with stirring at a speed of 50 rpm, a solution of 2.15 ml of glycerol and water for injection up to 1 liter are added. The obtained insulin aqueous solution was subjected to sterilizing filtration using filters "Sartorius" (Sweden) with a 0.22 micron, 0.45 micron pre-filter and kept for 21 days at a temperature of 1 - 5 ^° C sterile insulin solution was aseptically dispensed into vials .

 The test results of the resulting solution are shown in the table.

PRI me R 2. 1.54 g (40,000 IU) of a substance of porcine insulin with an activity of 26 IU / ml with a zinc content of 0.42% is dissolved in 367 ml (1101 mg of CH ₃ COOH) 0.05 N. acetic acid solution (solution 1). Then prepare solution 2, adding to 250 ml of 0.1 N. a solution of sodium hydroxide (1000 mg NaOH) 350 ml of water and 1.2 g of nipagin. The resulting solution 1 is mixed with solution 2 with stirring at a speed of 60 rpm and then the experiment is carried out as described in example 1.

Sterile filtration of the obtained aqueous insulin solution is carried out using sterile Pall filters (Germany) on a filter unit consisting of 2 Silklin filter holders installed in series with SLK7002NBP filter elements placed in them, pore size of filtering membranes 0.45 , and filter elements SLК7002NFZP, pore size 0.2 microns. After holding the solution for 14 days at a temperature of 1 - 5 ^about With the process is carried out as described in example 1.

 The test results of the resulting solution are shown in the table.

PRI me R 3. 1.54 g (total activity of 40,000 PIECES) of the substance of porcine insulin with a biological activity of 26 PIECES / mg, with a zinc content of 0.4% is dissolved in 300 ml (900 mg of CH ₃ COOH) 0.05 n acetic acid solution (solution 1). Then, solution 2 was prepared by adding 0.1 N to 175 ml (700 mg of NaOH). a solution of sodium hydroxide 350 ml of water and 1.2 g of nipagin. The resulting solution 1 is mixed with solution 2 and then after a 30-day exposure, the process is carried out as described in example 1.

 The test results of the resulting solution are shown in the table.

PRI me R 4. 1.53 g (40,000 PIECES) of the substance of pig insulin company "Novo" (Denmark) with a biological activity of 26.1 PIECES / mg, with a zinc content of 0.43% dissolved in 340 ml of 0.05 n acetic acid solution (1020 mg of CH ₃ COOH) (solution 1). Further, the process is conducted as indicated in example 1. The test results of the resulting solution are shown in the table.

As seen from the table, the aqueous solution of insulin in Examples 1 - 4 at the time of preparation and after freezing in cryothermostats at -21 ^{° C} for 48 hours and subsequent thawing to a temperature of 5 - 10 ^C is a colorless transparent liquid, and in similar preparations of Novo (Denmark) and Eli Lilly (USA) form opalescence.

 The analysis showed that the preparations actrapid "Novo" (Denmark) and letin 1 "Eli-Lilly" (USA) have a reduced stability to the effects of extreme temperature conditions in comparison with the proposed drugs.

Claims (1)

METHOD FOR PRODUCING AN INSULIN AQUEOUS SOLUTION containing 40 IU / ml, dissolving the crystalline insulin substance in distilled water, acidified with hydrochloric acid, adding sodium hydroxide, glycerin, a preservative, sterilized filtration, characterized in that the preparation is prepared by mixing solution 1 containing 4.1 - 5.1 mg of insulin in 1 ml of 0.05 N. acetic acid solution, with a solution of 2 containing 4.8 - 6.8 mg of nipagin in 0.1 N. sodium hydroxide solution, with a stirring speed of 50-60 rpm, 14-17 mg of glycerol is added to the resulting solution, and the sterile distilled water is adjusted to an insulin concentration of 40 U / ml, sterilizing filtration is carried out, the preparation is kept at 1-5 ^o C for 14-30 days and then the process is conducted in a known manner.

Images