RU2157698C1 - Method of preparing preparation "pensulin cc" of insulin suspension for cartridge - Google Patents

Abstract

FIELD: medicine, endocrinology. SUBSTANCE: invention relates to method of preparing suspension of porcine protamine-insulin in cartridge that is sustained-release preparation used in therapy of diabetes mellitus. Preparation that is able for prolonged storage is prepared by mixing four solutions in injection water in the definite sequence. Injection water has the following components: phenol and m-cresol, protamine sulfate and glycerol, substance of porcine monocomponent insulin and glycerol, phenol, m-cresol, sodium hydrogen phosphate dihydrate and 1 M hydrochloric acid (to pH 7.5-7.7). Then method involves filtration of mixture of solutions, addition of injection water to final insulin concentration 100 IU/ml, crystallization at 20-26 C and pouring the preparation into cartridges. Method provides safety of physical-chemical and biological properties of preparations. EFFECT: improved method of preparing. 3 cl, 1 tbl, 2 ex

Description

 The invention relates to medicine, in particular to the production of prolonged-acting insulin preparations used in the treatment of diabetes mellitus. The invention can be used in the medical industry for the manufacture of a suspension of porcine protamine-insulin in cartridges.

 A known method of obtaining a suspension of protamine-insulin in cattle with an insulin content of 100 PIECES / ml in vials (1). The disadvantages of this method include the need for freeze drying of a solution containing insulin and phenol in acidified water, after which a mixture of protamine sulfate, glycerol and phenol in water is added to the lyophilisate. In addition, this method of obtaining a suspension of protamine-insulin guarantees stability and preservation of the biological activity of the drug for only 3 months. According to modern pharmacopoeial requirements, the shelf life of insulin preparations should be at least 24 months.

 Biosynthetic human protamine-insulin preparations are known with concentrations of 40 and 100 IU / ml both in bottles and cartridges (2).

 As for the suspension of porcine protamine-insulin with an insulin content of 100 PIECES / ml, only such a preparation is known intended for filling into bottles, but not into cartridges (2).

 The objective of the invention is to develop a method of preparing a suspension of porcine protamine-insulin 100 IU / ml, intended for filling in cartridges.

The problem is solved by creating a method of preparing a suspension of insulin for a cartridge by using the substance of porcine insulin. The prepared preparation of a suspension of porcine protamine-insulin is poured into cartridges by mixing in a certain sequence four solutions in injection water, namely, a solution of preservatives phenol and m-cresol, a solution of protamine sulfate and glycerol, a solution of porcine insulin and glycerol, and a phosphate buffer solution, followed by crystallization at 20-25 ^o C and pouring a suspension of protamine-insulin into cartridges.

 The inventive method is as follows.

Solutions of the following components are preliminarily prepared (per 1 ml of injection water): 1.00-1.16 mg of phenol and 2.50-2.83 mg of m-cresol (solution A); 4-8 mg of protamine sulfate and 80-140 mg of glycerol (solution B); 34.62-42.31 mg of the substance of porcine monocomponent insulin and 95-105 mg of glycerol (solution B); 0.64-0.74 mg of phenol, 1.60-1.82 mg of m-cresol, 5.11 mg of sodium dihydrogen phosphate and 1 M hydrochloric acid (up to pH 7.5-7.7) (solution D) . Next, solution A is mixed with solution B and solution C at a stirring speed of 90-130 rpm, followed by sterilizing filtration. The resulting mixture at a stirring speed of 180-260 rpm is combined with a sterile solution G, injection water is added to a final insulin concentration of 100 U / ml and stirring is continued for 1 hour. Then crystallization is carried out at 20-25 ^° C for 72 hours and the resulting suspension of porcine protamine-insulin is poured into cartridges, in particular, with a volume of 1.5 ml, which are subsequently corked and rolled.

 The developed method allows to prepare a suspension of porcine protamine-insulin for cartridges (inserts), (which are usually used in conjunction with syringe pens), which retains its physicochemical and biological properties when stored in cartridges for a long time (see table). A draft provisional pharmacopoeial article (VFS) has been developed for the drug, providing for a shelf life of 2.5 years (30 months).

 The developed drug has successfully passed preclinical studies.

 The Pharmacological State Committee (FGC) of the Ministry of Health of Russia allowed clinical trials of the drug (protocol N 7 of June 25, 1998).

 The Commission on the instructions and nomenclature of the FGC recommended the developed drug the name "PENSULIN SS" ("PENSULIN SS") (protocol N 7 of September 28, 1998).

 The inventive drug PENSULIN SS allows you to provide healthcare with a new effective and inexpensive antidiabetic agent and reduce the penetration of expensive imported insulin preparations on the domestic market.

 The claimed method is illustrated by the following examples.

 Example 1

 Preparation of a solution of phenol and m-cresol. 3.0 g of phenol and 7.5 g of m-cresol are dissolved in 2 liters of injection water, and then the volume of the solution is adjusted with injection water to 3.0 liters.

Preparation of protamine sulfate solution. 2 g of protamine sulfate are dissolved in 100 ml of water for injection, heated to a temperature of 50 ^o C, 100 ml of injection water, cooled to a temperature of 5-10 ^o C, and 40.0 g of glycerol are added to the solution. The volume of the protamine sulfate solution was adjusted with injection water to 300 ml.

Preparation of the substance of insulin. 34.62 g of insulin substance with a biological activity of 26 PIECES / mg, with a zinc content of not more than 0.04 mg per 100 PIECES, is filled with 340 ml of injection water cooled to a temperature of 5-10 ^o C, mixed for 1-2 minutes. Then, with stirring, hydrochloric acid is added to the suspension to a pH of 3.0-3.4. Stirring is carried out until the crystals of insulin are completely dissolved, and then 100.0 g of glycerol are added to the solution and the volume of the solution is injected with injection water to 1000 ml.

 A solution of protamine sulfate is added to a solution of phenol and m-cresol with stirring at a speed of 100 rpm, the solution is stirred for 10 minutes, then a solution of the substance of insulin is added. Stirring is continued for 10 minutes, then sterilizing filtration is carried out using Millipore filters with 0.22 μm pore diameter and 0.45 μm prefilter.

 Preparation of a buffer solution with a pH of 7.5. 24 g of sodium phosphate disubstituted, 3.0 g of phenol and 7.7 g of m-cresol are dissolved in 3.5 l of injection water. After complete dissolution of the introduced components, 1M hydrochloric acid is added to the solution (up to pH 7.5). Next, sterilizing filtration is carried out using Millipore filters with a pore diameter of 0.22 μm and a prefilter of 0.45 μm.

 A mixture of sterile solutions of insulin, protamine sulfate and phenol with m-cresol is added to a sterile buffer solution with stirring at a speed of 200 rpm. Stirring is continued for 15 minutes, and then injection water is added to the suspension to obtain an insulin concentration of 100 U / ml and stirring is continued for 1 hour.

After the specified time, stirring is stopped and the product is left to crystallize for 72 hours at a temperature of 20-25 ^o C. Next, the drug is poured into 1.5 ml cartridges with their subsequent capping and running-in.

 The test results of the obtained drug are shown in the table.

 Example 2

 Preparation of a solution of phenol and m-cresol. 3.5 g of phenol and 8.5 g of m-cresol are dissolved in 2 l of injection water, and then the volume of the solution is adjusted with injection water to 3.0 l.

Preparation of protamine sulfate substance. 4 g of protamine sulfate are dissolved in 100 ml of water for injection, heated to a temperature of 50 ^o C. After dissolving the protamine sulfate, 100 ml of injection water, cooled to a temperature of 5-10 ^o C, and 70.0 g of glycerol are added to the solution.

 The volume of the protamine sulfate solution was adjusted with injection water to 300 ml.

Preparation of the substance of porcine insulin. 42.31 g of an insulin substance with a biological activity of 26 PIECES / mg, with a zinc content of not more than 0.04 mg per 100 PIECES, is filled with 340 ml of injection water cooled to a temperature of 5-10 ^o C and mixed for 1-2 minutes. Then, with stirring, hydrochloric acid is added to the suspension to a pH of 3.0-3.4. Stirring is carried out until the insulin is completely dissolved, and then 100.0 g of glycerol is added to the solution and the volume of the solution is injected with water to 1000 ml.

 A solution of protamine sulfate is added to a solution of phenol and m-cresol with stirring at a speed of 120 rpm, the resulting mixture is stirred for 15 minutes, then a solution of the substance of insulin is added. Stirring is continued for 15 minutes, after which sterilizing filtration is carried out using Millipore filters with a pore diameter of 0.22 μm and a prefilter of 0.45 μm.

 Preparation of buffer solution with a pH of 7.7. 24 g of sodium phosphate disubstituted, 3.5 g of phenol and 8.5 g of m-cresol are dissolved in 3.5 l of injection water. After complete dissolution of the introduced components, 1 M hydrochloric acid is added to the solution (up to pH 7.7). Next, sterilizing filtration is carried out using Millipore filters with a pore diameter of 0.22 μm and a prefilter of 0.45 μm.

 A mixture of sterile solutions of insulin, protamine sulfate and phenol with m-cresol is added to a sterile buffer solution with stirring at a speed of 250 rpm. Stirring is continued for 20 minutes, and then injection water is added to the suspension to obtain an insulin concentration of 100 U / ml and stirring is continued for 1 hour.

After the specified time, stirring is stopped and the product is left to crystallize for 72 hours at a temperature of 20-25 ^o C. Next, the drug is poured into 1.5 ml cartridges with their subsequent capping and running-in.

 The test results of the obtained drug are shown in the table.

Claims (3)

1. A method of preparing a suspension of insulin for a cartridge, comprising preparing a preparation of insulin in the form of a suspension at a concentration of 100 PIECES / ml, followed by adding it to the cartridge, characterized in that the preparation of the preparation uses the substance of porcine insulin sterilized using a prefilter with a pore diameter of 0, 45 microns and a filter with a pore diameter of 0.22 microns, followed by crystallization at 20 - 25 ^o C.  2. The method according to p. 1, characterized in that the preparation is prepared by mixing a solution containing 1.00 - 1.16 mg of phenol, 2.50 - 2.83 mg of m-cresol in 1 ml of injection water, with a solution of protamine sulfate with mixing speed of 90 - 130 rpm, containing 4 - 8 mg of protamine sulfate and 80 - 140 mg of glycerol in 1 ml of injection water, and a solution of porcine monocomponent insulin at the same mixing speed, containing 34.62 - 42.31 mg of insulin and 95 - 105 mg of glycerol 1 ml of acidified water, followed by sterilizing filtration of the resulting solution and combining it at a stirring speed of 180 - 260 rpm with a sterile buffer solution with a pH of 7.5 - 7.7, adding to the resulting injection water solution to a final insulin concentration of 100 PIECES / ml, mixing the prepared mixture, followed by crystallization and pouring the resulting preparation into cartridges with their further corking and running in.  3. The method according to PP.1 and 2, characterized in that the bottling of the drug suspension of porcine protamine-insulin is carried out in cartridges of 1.5 and 3 ml.

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