RU2677330C2 - Method of preparing injection form of chondroitin sulfate - Google Patents

Abstract

FIELD: medicine.SUBSTANCE: invention relates to medicine and pharmaceuticals and relates to the method for preparing an injectable form of the chondroitin sulfate preparation. Method comprises the sequential dissolution of sodium disulfite, chondroitin sodium sulfate, benzyl alcohol in injectable water, followed by adjusting, if necessary, the pH of the solution to 6.0–7.5, however, the entire process of dissolving the ingredients is carried out under nitrogen protection. Finished solution of the drug is filtered using sterilizing filtration, poured into the ampoules and the ampoules are sealed in the flow of nitrogen. Method provides for minimal risks of degradation of molecular structure and changes in the molecular weight range of chondroitin sulfate, as well as for minimal risks of initial contamination of the solution and the accumulation of backendotoxins. Method is quick and easy to implement. Method is also characterized by the elimination of the appearance of pink staining solution and the preservation of the required values of the quantitative content of chondroitin sulfate, color and pH of the solution for more than 2 years of storage.EFFECT: preparation is a stimulator of tissue repair.5 cl, 1 tbl, 1 ex

Description

The present invention relates to medicine and pharmaceuticals and relates to a method for the preparation of an injectable form of the drug chondroitin sulfate (hereinafter XC), which is a stimulator of tissue repair.

Chondroitin sulfate preparations (in particular, based on the Na salt of chondroitin sulfate, a mucopolysaccharide from animal tissues) are widely used in modern medicine to treat degenerative diseases of the joints and spine (osteoarthritis, osteochondrosis), and to accelerate the formation of bone marrow in fractures. Cholesterol preparations are produced in various dosage forms, including injection ones, in particular in the form of a solution for intramuscular administration.

From the patent RU No. 2021812, there is known an injection solution for the treatment of arthrological diseases, which is a mixture of chondroitin sulfates A and C in a ratio of 3: 2-1: 1, a preservative - benzyl alcohol and pyrogen-free water. This product is prepared by dissolving benzyl alcohol and cholesterol in pyrogen-free water, after which the pH of the sodium hydroxide solution is adjusted to 7.5, sterile filtered and poured into ampoules.

There is also known a method for producing an injection form of a chondroitin sulfate preparation, which includes dissolving benzyl alcohol in water, setting a pH of 7.2-7.4 with a 30% sodium hydroxide solution, then dissolving in the resulting cholesterol solution, heating the mixture to 50 ° C with further sterilizing filtration, filling into ampoules and their sealing (Regulation for the production of a solution of chondroitin sulfate for injection // All-Union Scientific Research Institute of the technology of blood substitutes and hormonal preparations. - M., 1994; cited from atentu RU №2200018).

The disadvantages of these two methods are described in patent RU No. 2200018 when characterizing the prior art: “Often in the manufacture of a preparation containing cholesterol (consisting of uronic acid and hexosamine) and benzyl alcohol, a pink color of the solution appears at a pH of 7.2-7.5, which is enhanced during storage. The appearance of a pink color indicates the appearance of some cholesterol derivatives in the solution, the presence of which excludes the use of the drug for therapeutic purposes. ”

For the prototype of the present invention, a method for preparing an injection form of a chondroitin sulfate preparation is selected, protected by RU patent No. 2200018. The prototype method involves dissolving benzyl alcohol, cholesterol and sodium hydroxide in injection water, using heating to 50 ° C, sterilizing filtration, filling in ampoules and seals, moreover, the solutions of the components of the drug are prepared separately, while first the cholesterol substance is dissolved in heated injection water followed by sterilization and cooling (solution A), then a solution of benzyl alcohol (solution B) and a solution of sodium hydroxide (solution C) are prepared, then solution A is added to solution A with stirring to pH 6.2-6.8 and solution B, the resulting mixture was diluted with water injection, dispensed and sealed into ampoules (RU №2200018. A method for preparing injectable forms of the drug chondroitin sulfate. 10.03.2003).

The prototype method avoids the appearance of pink staining, characteristic of the above two analogues. However, it is not without some drawbacks:

- the use of heating to 50 ° C and intermediate sterilization of a solution of a substance gives rise to risks of degradation of the structure of chondroitin sulfate molecules and a change in the interval of the declared molecular weight;

- the preparation of several solutions (solutions A, B, C) in different units of equipment increases the preparation time, complicates the method of preparation of the solution, increases the risks of the initial contamination of the solution, thereby not excluding the accumulation of backendotoxins.

The objective of the invention is to reduce the risks of degradation of the structure of cholesterol molecules and change the interval of the declared molecular weight of cholesterol, reduce the risks of initial contamination of the solution and the accumulation of bacendotoxins, simplify and speed up the method of preparation, provided all these results are achieved simultaneously with the exception of the appearance of pink staining of the solution.

This object is achieved in that a method for preparing an injection form of a chondroitin sulfate preparation is proposed, including sequential dissolution of sodium disulfite (synonyms: sodium metabisulfite; sodium pyrosulfite, sodium pyrosulphuric acid), chondroitin sulfate salt (INN chondroitin sulfate), benzyl alcohol, followed by adjusting, if necessary, the pH of the solution to 6.0-7.5, while the entire process of dissolving the ingredients is carried out under nitrogen protection, the finished solution of the drug is filtered, poured into ampoules and ampoules are sealed in a stream of nitrogen.

Some aspects of the proposed method are as follows:

- use injection water with a temperature of 15-30 ° C;

- the dissolution of each ingredient is carried out with constant stirring until completely dissolved;

- adjust the pH of the solution to 6.0-7.5 with a 1 M sodium hydroxide solution;

- the finished solution of the drug is successively filtered through a filter system of preliminary and sterilizing filtration;

- filling the ampoules is carried out in a stream of nitrogen.

A new technical result achieved by the claimed invention is as follows:

- the exclusion of heating to 50 ° C and intermediate sterilization of a solution of the substance allows you to save the structure of chondroitin molecules from degradation, to ensure the stated molecular weight range;

- the use of antioxidant and nitrogen protection allows you to stabilize the solution (to prevent changes in color, pH and quantitative content);

- elimination of the stages of separate preparation of several solutions (in several units of equipment) and preparation of the drug in one unit of equipment reduces the preparation time, simplifies the method of preparation of the solution, reduces the risks of initial contamination of the solution, thereby preventing the accumulation of backendotoxins (which reduces the risk of non-sterile filtrate).

When testing the technology of the drug, we studied the sequence of adding ingredients, the conditions for dissolving the active substance, the need to protect the drug with nitrogen during the preparation of the solution and when sealing ampoules, the possibility of using sterilizing filtration and heat sterilization, the need to add excipients, the light and frost resistance of the drug.

In laboratory conditions, samples of the drug were made:

- using full gas protection with nitrogen;

- with sealing in a stream of nitrogen;

- without nitrogen protection;

- with sterilizing filtration through a membrane with a pore diameter of 0.22 μm;

- with sterilization by autoclaving at 100 ° C for 30 minutes;

- with a different set of excipients (Trilon B, sodium metabisulfite, sodium sulfite anhydrous).

The use of thermal sterilization leads to a significant change in the color of the solution, a change in pH and the quantitative content of the active substance in the solution. Therefore, the method of sterilizing filtration of the solution was chosen.

The need to protect the drug with nitrogen during the preparation of the solution and when sealing ampoules was studied. In laboratory conditions, samples of the preparation were made using gas protection with nitrogen and without it. The samples were stored for stability studies by the method of “accelerated aging” at a temperature of 40 ° C for a period equivalent to 2 years of storage in vivo. During the shelf life, the samples were analyzed by OCC in appearance, pH, quantitative content of chondroitin and benzyl alcohol, and color. The use of nitrogen protection led to a decrease in the drop in the quantitative content of chondroitin sulfate and benzyl alcohol, to a slowdown in the processes of color change during storage of the solution. A change in the transparency of the solution was not recorded for any of the samples.

After operating the first industrial series, the stability of the preparation was studied during storage under natural conditions for 2 years and 3 months (temperature (25 ± 2) ° С). During the entire observation period, the values of the quantitative content, color and pH of the solution did not go beyond the limits regulated in the FSP. Change in other quality indicators was not recorded for any of the samples.

The proposed method is as follows.

Calculate the loading of the amount of ingredients:

- Amount of substance of chondroitin sodium sulfate:

, где

where

M _{subst ..} - the mass of the substance of chondroitin sodium sulfate required for loading, kg;

V _height. - regulated volume of loading, l;

With _subst - the quantitative content of chondroitin sodium sulfate in the substance, mass. % (if the quantitative content of the passport is more than 100%, when calculating it should be equal to 100%);

W _subsst - loss in mass when drying the substance of chondroitin sodium sulfate, mass. %

- Amount of benzyl alcohol:

where M _{b.s ..} is the mass of the benzyl alcohol solution required for loading, kg;

V _rast. - regulated volume of loading, l;

S _bs - quantitative content of benzyl alcohol in raw materials, mass. %;

W _b.s. - water content in raw materials, mass. %

- Amount of sodium disulfite:

where M _{nm ..} - the mass of sodium disulfite required for loading, kg;

V _rast. - regulated volume of loading, l;

With _N.M. - quantitative sodium disulfite, mass. %;

- The amount of water for injection for loading is calculated by the formula:

M _water = M _rast. - M _subst. - M _b.s. - M _N.M. - M _N.G. ,

where M _water is the mass of water for injection required for loading, kg;

M _rast. - mass of solution, kg;

M _N.G. - weight of 1 M sodium hydroxide solution, kg

Before starting the process of preparing the solution, a 1 M sodium hydroxide solution is prepared.

1 liter of the solution contains 0.04 g of sodium hydroxide.

About 500 ml of water for injection is poured into a 1-liter glass. In a separate glass, 40 g of dry sodium hydroxide is weighed, pour it into a glass of water and mix with a glass rod until completely dissolved. The solution obtained in a glass is carefully transferred to a 1 L volumetric flask (if necessary, use a glass funnel) and the solution volume in the volumetric flask is adjusted to 1 liter by adding water for injection with a temperature of 25 ± 2 ° С. To prepare a larger amount of the solution, the operation is repeated (or use a glass or flask of a larger volume, with an appropriate recount of the quantitative content of the ingredients).

The calculated amount of water for injection (with a temperature of 15-30 ° C) is loaded into a previously prepared reactor for the preparation of solutions.

The entire process of dissolving the components is protected under nitrogen, with a break for loading components

The calculated amount of sodium disulfite is loaded into the reactor for the preparation of solutions with a working stirrer (200-350 rpm) and stirred until complete dissolution, by visual inspection of the completeness of dissolution, through the inspection window of the reactor manhole.

Then, the calculated amount of chondroitin sulfate (sodium chondroitin sulfate) is portionwise loaded into the reactor for solution preparation and mixed until the substance is completely dissolved, by visual inspection of the completeness of dissolution, through the inspection window of the reactor manhole.

Next, the calculated amount of benzyl alcohol is loaded into the reactor for the preparation of solutions, stirred until complete dissolution, and a sample is taken. Measure the pH of the resulting solution, the value of which should be in the range from 6.0 to 7.5. If necessary, adjust the solution with a 1 M sodium hydroxide solution. After adding 1 M sodium hydroxide, the solution was stirred and the pH was re-measured (the procedure was repeated until a satisfactory result was obtained).

Upon receipt of satisfactory results of the analysis, water for injection is loaded into the reactor until the mass of the solution reaches the target and mix.

Conduct a control analysis of physico-chemical and microbiological indicators.

Then the solution is fed into a preliminary and sterilizing filtration system under a sterile nitrogen pressure of 0.1-4.0 bar, consisting of successively arranged filter holders for the pre-filter filter cartridge (pore size - from 1.00 to 0.22 μm,) and a filter holder for filter cartridge sterilizing filtration (pore size from 0.1 to 0.22 microns).

Then the product is bottled and sealed in ampoules and this is carried out with local product protection by a laminar flow of sterile air in a stream of nitrogen.

An example of a specific implementation of the method.

Using the proposed method, a preparation of chondroitin sulfate was prepared in the form of a solution for intramuscular injection of the following composition:

1 ampoule of 1 ml (or 2 ml) contains:

The active substance is chondroitin sodium sulfate - 100 or 200 mg.

Excipients:

benzyl alcohol - 9.0 mg (or 18.0 mg);

sodium disulfite -1.0 mg (or 2.0 mg);

sodium hydroxide - up to pH 6.0-7.5;

water for injection - up to 1 ml (or 2 ml).

We performed a load calculation and got the following indicators:

Before starting the preparation of the drug solution, a 1 M sodium hydroxide solution was prepared.

1 ml of the solution contains 0.04 g of sodium hydroxide.

About 500 ml of water for injection was poured into a 1 liter glass. In a separate glass, 40 g of dry sodium hydroxide was weighed, poured into a glass of water and mixed with a glass rod until completely dissolved. The solution obtained in a glass was carefully transferred to a 1 L volumetric flask and the volume of the solution in the volumetric flask was adjusted to 1 liter by adding water for injection with a temperature of 25 ± 2 ° С.

180 kg (360 kg to load 400 l) of water for injection with a temperature of (15-30) ° C were loaded into a previously prepared reactor for preparing solutions.

The entire process of dissolving the components was carried out under nitrogen protection, with a break for loading the components.

0.2 kg (0.4 kg to load 400 l) of sodium disulfite (in terms of 100% substance) was loaded into the reactor for the preparation of solutions with a working stirrer (200-350 rpm) and mixed until complete dissolution, by visual inspection for complete dissolution, through the inspection window of the reactor hatch.

Then, 20 kg (40 kg for loading 400 l) of sodium chondroitin sodium sulfate (in terms of 100% substance) were portioned into the reactor for the preparation of solutions and mixed until the substance was completely dissolved, by visual inspection of the completeness of dissolution, through the inspection window of the reactor hatch.

Next, 1.8 kg (3.6 kg to load 400 L) of benzyl alcohol (in terms of 10% substance) was loaded into the reactor for the preparation of solutions, stirred for at least 20 minutes and a sample was taken. The pH of the resulting solution was measured, the value of which should be in the range from 6.0 to 7.5. After adding 1 M sodium hydroxide, the solution was stirred for at least 10 min and the pH was re-measured (the procedure was repeated until a satisfactory result was obtained).

Conducted control on physico-chemical and microbiological indicators.

Then the solution was supplied to the preliminary and sterilizing filtration system under a pressure of sterile nitrogen of 0.1-4.0 bar, consisting of successively arranged filter holders for the pre-filtering filter cartridge (pore size 0.45 μm) and a filter holder for the sterilizing filter cartridge filtration (pore size - 0.22 μm,).

Pouring in a stream of nitrogen and sealing the drug into ampoules, this was done with local protection of the product with a laminar flow of sterile air.

The stability of the drug during storage under natural conditions for 2 years and 3 months (temperature (25 ± 2) ° C) was studied. During the entire observation period, the values of the quantitative content, color and pH of the solution did not go beyond the limits regulated in the FSP. Change in other quality indicators was not recorded for any of the samples.

Summing up everything stated in this application, it should be noted that the task has been achieved. The developed technology, in contrast to the prototype method, has minimal risks of degradation of the structure of cholesterol molecules and changes in the interval of the declared molecular weight of cholesterol (due to the exclusion of heating to 50 ° C), as well as minimal risks of the initial contamination of the solution and the accumulation of backendotoxins (due to the exclusion of stages separate preparation of several solutions). At the same time, a simplification and acceleration of the method was achieved (also due to the elimination of the stages of separate preparation of several solutions). All of the above results are ensured simultaneously with the exception of the appearance of pink staining of the solution and preservation of the required values of the quantitative content, color and pH of the solution for more than 2 years of storage.

Claims (5)

1. A method of preparing an injection form of a chondroitin sulfate preparation, comprising sequentially dissolving 1.0 mg / ml sodium disulfite, 100 mg / ml chondroitin sodium sulfate, 9.0 mg / ml benzyl alcohol in injection water, followed by adjusting the pH, if necessary solution up to 6.0-7.5, while the entire process of dissolving the ingredients is protected by nitrogen, the finished solution of the drug is filtered using sterilizing filtration, poured into ampoules and sealed ampoules in a stream of nitrogen. 2. The method according to p. 1, characterized in that they use injection water with a temperature of 15-30 ° C. 3. The method according to p. 1, characterized in that the dissolution of each ingredient is carried out with constant stirring until completely dissolved. 4. The method according to p. 1, characterized in that the pH of the solution is adjusted to 6.0-7.5 using 1M sodium hydroxide solution. 5. The method according to p. 1, characterized in that the filling of the ampoules is carried out under nitrogen protection.