RU2252752C2 - Human insulin drug of short action - Google Patents

Abstract

FIELD: pharmaceutical industry, medicine.

SUBSTANCE: invention relates to human insulin drug with activity of 100 IU/ml, including cartridge forms. Drug contains active ingredient, glycerol as isotonic agent, conserving agent and water, wherein it contains human insulin substance of high purity with residual proteolysis activity not more than 0.005 adsorption units, sodium chloride as additional isotonic agent, m-cresol as conserving agent, and additionally sodium dihydrogenphosphate dihydrate or disodium hydrogenphosphate heptahydrate as substance with buffer capacity and pH 6.9-7.8.

EFFECT: human insulin drug of short action with increased physiological activity and physical and chemical storage stability.

6 ex, 1 tbl

Description

The invention relates to the pharmaceutical industry and can be used in the manufacture of finished dosage forms of human short-acting insulin with an activity of 100 units per ml (hereinafter - 100 IU / ml), including cartridge forms.

One of the biggest problems in the finished dosage forms of human insulin is their loss of stability during storage due to the formation of high molecular weight compounds, including dimers, that is, covalently cross-linked insulin molecules in two A-chains or through one A-chain and one B-chain - dimers AA and dimers of AB and polymers that increase the immunogenicity of the drug (Helbig HJ, Deutsche Wollporschungsinstitut, dissertation, 1976; Schlichtkrull et al., Horm. Metab. Res. Suppl. 5 (1974), pp. 134-143). Already during the first development of finished dosage forms of insulin, it was found that the acidic environment characteristic of the drugs of that time promoted the formation of desamidoinsulin (Sundby, F., L. Biol. Chem. 237 (1962), 3406-3411) and covalent dimers (Steiner et al., Diabetes 17 (1968), 725-736). Reducing their number is a very serious problem, which until now has no final solution. The ways in which they tried to solve it were different: they began to adjust the medium to neutral, change the composition of auxiliary components in the preparations and use a purer active substance, that is, monocomponent highly purified insulin substances. In recent years, analogues of human insulin substance have been created, obtained by recombinant technology, which were used as the active substance and which are characterized by a reduced ability to form insulin-related compounds that impair the quality of the drug during storage. So, it is known “Stable insulin preparation” (patent EP 0884053, 1998), which contains as an active substance an analog of insulin substance obtained by genetic engineering, as preservatives phenol and m-cresol, as an isotonic agent glycerin, as a substance with a buffer capacity TRIS or arginine and water. The use of an analogue of the insulin substance obtained by the recombinant, or, as it is also called, the genetic engineering method, provides greater chemical stability of the finished dosage form during storage, but this technical solution is mainly aimed at increasing the physical stability of the drug in relation to thermomechanical stress , that is, to create the possibility of its use in permanent infusion systems and in cartridge forms.

The closest in composition to the claimed dosage form are finished dosage forms according to patent EP No. 0280534, 1988, “New insulin analogs”, which contain, as active substance, an analogue of the substance of insulin GlyA ²¹ obtained in recombinant technology in an amount of 3.825-4.675 g / l, as the isotonic agent is glycerol in an amount of 16-20 g / l, as a preservative is phenol in an amount of 2.6-3.25 g / l and water. The pH of the drug is 3.5. The authors received a medicine with sufficiently high characteristics with regard to chemical and physical stability, but their own studies, carried out in accordance with the 1980 British Pharmacopoeia, showed that the biological activity of the GlyA ²¹ insulin substance analogue is approximately 85% of the activity of the insulin substance. That is, the use of this analogue of the substance of insulin as an active substance necessitates an increase in its quantity in order to obtain a medicine of a certain biological activity. In addition, its acidic pH can be attributed to the disadvantages of this medicine, since according to the current level of knowledge in the field of insulin, drugs with a neutral pH are considered to be more physiological for the human body. But the authors of patent EP No. 0884053 by their experiments show that with this composition of the finished dosage form, which they claim, with increasing pH, the number of harmful macromolecular compounds during storage of the drug increases, that is, the preparation of the claimed composition can be stable only under an acidic pH of 3.5 .

The objective of the invention is the creation of a finished dosage form of human insulin short-acting, which ensures the economical use of the active substance and increase the physiology of the drug for the human body with high physical and chemical stability of the drug during storage.

The problem is solved in that the finished dosage form of human short-acting insulin, which contains the active substance, glycerin as an isotonic agent, preservative and water, according to the invention is characterized in that it contains a highly purified substance of human insulin having residual proteolytic activity not more than 0.005 absorption units, as an additional isotonic agent - sodium chloride, as a preservative m-cresol, and additionally contains as a substance with a buffer capacity, sodium dihydrogen phosphate dihydrate or disodium hydrogen phosphate heptahydrate at a composition pH of 6.9-7.8 and the following ratio of components, g / l: highly purified human insulin substance 3.214-3.928; glycerin 14-18; m-cresol 2.5-3.0; sodium dihydrogen phosphate dihydrate 2.4-2.8 or disodium hydrogen phosphate heptahydrate 2.0-2.3; sodium chloride 0.15-0.3; water is the rest.

In the claimed solution, as an active substance, that is, a highly purified substance of human insulin, a semisynthetic human insulin substance obtained by the transpeptidation of porcine insulin with accompanying related impurities, which is carried out in the presence of trypsin with a molar excess of threonine di-tert-butyl ester and postad purified by high pressure chromatography, or a substance of human insulin, obtained by recombinant technology, both substances of human insulin ka should be characterized by a residual proteolytic activity of 0.005 units of absorption. As an isotonic agent in the claimed solution, glycerin and sodium chloride are used, a preservative is m-cresol, substances with a buffer capacity are sodium dihydrogen phosphate dihydrate or disodium hydrogen phosphate heptahydrate. Dissolution of human insulin and adjusting the pH of the mixture to 6.9-7.8 is carried out by known methods with using Hcl and NaOH, respectively.

It is known that the chemical stability of a medicinal product during its storage is affected by the quality of the used insulin substance, the type and concentration of preservatives, isotonic agents and substances with a buffer capacity (Acta Pharm. Nord. 4 (3), p. 149, 1992). The use of a highly purified substance of human insulin, which is claimed as an ingredient in a medicinal product, allows not only to achieve savings in the expenditure of the precious active substance in the preparation of a dosage form of a certain biological activity, but also to increase the physiology of the medicinal product for the human body due to its neutral pH, while providing a small content dimers and polymers in the finished dosage form. In turn, as shown by studies of the authors of the proposed solution, the use of m-cresol as a preservative in combination with isotonic agents - glycerin and sodium chloride - and with substances with a buffer capacity - sodium dihydrogen phosphate dihydrate or disodium hydrogen phosphate heptahydrate - within the claimed limits provides a stabilizing effect on the chemical composition and the physical state of the dosage form during its storage, that is, prevents the increase in the number of dimers and polymers in it, as well as a change in the physical about the state of the solution.

The solution of the problem is proved by examples of specific performance.

Example 1

3.571 g of human insulin substance was added to 200 ml of water and mixed until a fine suspension was obtained, the pH of the suspension was adjusted to 3.2 with 1 M Hcl. Then 2.5 g of sodium dihydrogen phosphate dihydrate was dissolved in 700 ml of water, 0.2 g of sodium chloride, 16 g of glycerol and 2.7 g of m-cresol were added and the pH of the solution was adjusted to 7.6 with 1 M NaOH solution. Then both solutions were mixed and the pH of the resulting solution was adjusted to 7.3. The volume was supplemented with water up to 1 L, it was sterilized by filtration, followed by aseptic filling into 1.5-3.0 ml cartridges. Studies of the finished short-form dosage form using high-performance liquid chromatography (HPLC) showed that the medicinal product has an activity of 95.8 IU / ml, the content of high molecular weight compounds is 0.11%, A21-insulin desamide is 0.3%.

Example 2

The method of preparation of the finished dosage form, as in example 1, but disodium hydrogen phosphate heptahydrate in the amount of 2.1 g was taken as a substance with a buffer capacity. Analysis of the medicinal product using HPLC showed that its activity was 98.1 IU / ml, the content of high molecular weight compounds of 0.1%, A21-insulin desamide - 0.15%.

Example 3

The method of preparing the finished dosage form, as in example 1, but sodium dihydrogen phosphate dihydrate was taken 2.0 g. Analysis of the drug using HPLC showed that its activity is 94.3 IU / ml, the content of high molecular weight compounds is 0.3%, A21- insulin desamide - 0.6%.

Example 4

The method of preparing the finished dosage form, as in example 2, but disodium hydrogen phosphate heptahydrate was taken 2.5 g. Analysis of the drug using HPLC showed that its activity is 95 IU / ml, the content of high molecular weight compounds is 0.25%, A21-insulin desamide - 0.4%.

Example 5

The method of preparing the finished dosage form, as in example 1, but sodium chloride was taken 0.08 g. Analysis of the drug using HPLC showed that its activity is 94.2 IU / ml, the content of high molecular weight compounds is 0.4%, A21- insulin desamide - 0.4%.

Example 6

The method of preparing the finished dosage form, as in example 1, but sodium chloride was taken 0.4 g. Analysis of the medicinal product using HPLC showed that its activity is 95.1 IU / ml, the content of high molecular weight compounds is 0.45%, A21- insulin desamide - 0.45%.

Thus, the analysis of human insulin preparations at the time of preparation showed that all obtained finished dosage forms meet the requirements for them. But then, in order to study the indicators of the chemical and physical stability of medicines during their storage, the obtained samples were subjected to accelerated aging at a temperature of 45 ° C for two weeks. After that, the samples were analyzed by high performance liquid chromatography (HPLC) for the content of high molecular weight compounds (IUDs) and the optical density of the samples was determined spectrophotometrically at 450 nm. The data obtained are summarized in the table below.

Sample number according to example The content of high molecular weight compounds (%) The optical density of the sample at 450 nm 1 1,0 0.005 2 0.99 0.003 3 1.25 0.05 4 1.37 0,03 5 1.48 0.04 6 1.34 0.02 prototype 1,0 - * - data on the prototype are not available.

As can be seen from the above examples and the table, in the finished dosage form of human short-acting insulin, which is claimed, the content of macromolecular compounds during storage is at the level of the prototype (examples 1 and 2), a pH value of 7.3 indicates a higher physiology of the drug, the optical value density at 450 nm indicates a high physical stability of the drug, that is, the possibility of its use in cartridge form, and the saving of the active substance due to the use in the inventive solution a highly purified substance of human insulin obtained by recombinant or chemical means is 0.628 g / l.

Claims (1)

The finished dosage form of human short-acting insulin containing the active substance, glycerin as an isotonic agent, preservative and water, characterized in that it contains a highly purified human insulin substance having a residual proteolytic activity of not more than 0.005 adsorption units, as an additional isotonic sodium chloride agent, m-cresol as a preservative and additionally contains sodium dihydrogen phosphate dihydrate or disodium hydrogen phosphate as a substance with a buffer capacity heptahydrate at a pH of the composition of 6.9-7.8 and the following ratio of components, g / l: highly purified human insulin substance 3.214-3.928 glycerin 14-18 m-cresol 2.5-3.0 sodium dihydrogen phosphate dihydrate 2.4-2.8 or disodium hydrogen phosphate heptahydrate 2.0-2.3 sodium chloride 0.15-0.3 water rest