CS221172B1 - Stabilized solution of human gamaglobuline of low protein concentration - Google Patents

Abstract

It is used in immuno-biological production of medicinal products. The purpose of the invention is to improve the stability and appearance of the product as well as to increase the yield. The increased effect, especially the stability of the low gamma globulin solution of low biotin concentrations, is achieved by adding a 1-human albumin injectable solution to the diluted human gamma globulin solution containing 0.1-6.0% by weight of biotin. a stabilizer, in an amount of 0.05 - 0.15% by weight.

Description

It is used in immunological-biological production of medicinal products.

The purpose of the invention is to improve the stability and appearance of the product as well as to increase the yield.

The enhanced effect, in particular the stability of the human gamaglobulin solution with a low concentration of biotin, is achieved by adding to the dilute human gamaglobulin solution containing 0.1-6.0% by weight of the protein a solution of 1'human albumin as stabilizer in an amount of 0.05 - 0.15% w / w.

The invention relates to a stabilized solution of 1'-human gamaglobulin of low protein concentration using 1'-human albumin.

Ethanol fractionation of human plasma, respectively. For example, gamma globulin, which is ultimately prepared as a 0.5 to 16% solution for injection, is obtained from serum and other suitable technological operations. Gamaglobulin in solution is generally stabilized with amino acids or simple sugars. Of the amino acids, aminoacetic acid and s-aminocaproic acid are most commonly used in practice (J. T. Edsall: Advances in Protein Chem.

III. 1947 Strassle, R .: Expericalie, 9, 1953; Nikitina V. D .; Cholchev N.V., Kolesnikova L.I. and Korableva Z.I .: ZMEI, 1, 1975).

Of the simple sugars, it is mainly glucose (J.T. Edsall: Advences in Protein Chem. III. 1947).

In some workplaces, sodium thiosulphate is used as a stabilizer in the preparation of human low-protein gamaglobulin containing histamine as stabilizer (Naučno-issledovatelskij institut vaccin and syvorotok imeni I. I. Mečnikova: Adjustment after primeneniju suchoga Histaglobulin, 1972).

Practice has shown that aminoacetic acid and ε-aminocaproic acid reliably stabilize a 1'-human gamaglobulin solution when it contains a higher protein concentration (10-16%). Conversely, a low-protein gamaglobulin solution (0.5-5.0%), in particular after re-lyophilization, appears to opalescence to turbidity, suggesting that the stabilizer present does not adequately protect the protein against possible denaturing changes. Experimentally, it has been found that gamaglobulin with low protein concentration and high stabilizer concentration (aminoacetic acid, ε-aminocaproic acid and glucose) do not provide the necessary stabilizing effect of the formulation.

The literature describes the isolation of cholinesterase from fraction IV, used for intravenous administration, stabilized by 2.5% albumin (M. Steinbuch, Granier, C. Tavernier, A. Faure, Vox Sang 36: 142-149, 1979).

One of the low protein products of human gamaglobulin is prepared from fraction II obtained by ethanol fractionation of human plasma. Fraction II, prepared to the lyophilized state, is dissolved in a 0.01% mertiolate solution and 2.25% aminoacetic acid stabilizer to a 16% protein solution.

This was diluted with 0.45-0.50% sodium chloride solution to 3% protein and adjusted to pH 5.75-5.80 with 5% citric acid. Then 2% bentonite was added to the gamma globulin solution with stirring, and the mixture was allowed to stand for 1 hour. The gel formed is centrifuged and the supernatant is filtered through a pad.

The pH is adjusted to 6.8 to 6.9, followed by filtration through sterilizing pads, and the resulting gamaglobulin is diluted with isotonic sodium chloride solution to a 0.5 to 0.7% protein solution. The gamma globulin solution thus prepared is mixed with a sterile isotonic saline solution containing histamine so that the final histamine concentration is 0.05-0.10 µg / ml. After complete homogenization, the formulation is filled, frozen and lyophilized.

The essence of the novel method of stabilizing a low-protein gamma globulin solution of low protein concentration is that the gamma globulin obtained by fractionating human plasma is diluted with a suitable solution to 0.1 to 6.0% by weight protein, and this is stabilized by adding 0.05 to 0.15 % by weight of human albumin.

The advantage of said method of preparing low-protein gamma globulin over the prior art is that it improves the stability and appearance of the product, increases yield, reduces labor and shortens the technological cycle.

Example of preparation of 0,5 - 0,7% solution of human gamaglobulin containing histamine:

100 ml of a 16% solution of human gamma globulin is diluted to 0.55% protein by adding 2900 ml of isotonic sodium chloride solution. To this gamaglobulin solution was added 1.5 g of a 1'-human albumin solution, calculated on 100% protein. The solution is adjusted to pH

6.9 ± 0.5 and then sterile filtered and mixed with sterile histamine solution to give a final histamine concentration in the solution of 0.05-0.10 µg / ml. The solution is then dispensed in 2 ml portions into 5 ml penicillin vials, frozen and lyophilized.

100 ml of a 1% human gamaglobulin 16% solution for injection is diluted to 0.55% protein by adding 2900 ml of isotonic sodium chloride solution. To this gamaglobulin solution is added 3 g of a 1'-human albumin solution, calculated on 100% protein. The pH of the solution is adjusted to 6.9 ± 0.5 and then sterile filtered and mixed with a sterile solution of histamine so that the final concentration of histamine in the solution is 0.05-0.10 µg / ml. The solution is then dispensed in 2 ml portions into 5 ml penicillin vials, frozen and lyophilized.

yield

With the existing technology, about 500 to 550 pieces of a 0.5 to 0.7% gamaglobulin solution containing histamine are prepared from 100 ml of a 16% gamaglobulin solution.

With the new technology, about 1000 to 1100 pieces of a 0.5 to 0.7% solution of ga221172 maglobulin containing histamine are prepared from 100 ml of a 16% gamaglobulin solution, thereby increasing the yield by 100%.

Appraisal Appearance

In table no. 1 shows relative turbidity values in% transmittance versus

taboo

Type of product

Distilled water

Product prepared by existing technology Product prepared by new technology

Mean values of relative turbidity in Table no. 1 show that the appearance of products prepared by the new technology is substantially better than the products prepared by the existing technology.

It also dissipates relative haze values for products prepared by the new technology is substantially lower than for products prepared under the current technology.

This progress was achieved with perfectly distilled water, measured by a Pulfrich photometer (filter # 3, sensitivity 2) and dissipated relative haze values in% of a 0.5% histamine-containing gamma globulin solution prepared by existing and new technology.

1'ka č. 1

Average values Scatter relative haze values in% relative haze in%

100 0

42

12 by stabilizing a 0.5% solution of 1'-human gamma globulin containing histamine.

Analytical evaluation

In table no. 2 shows the analytical values determined for a commercial preparation and a preparation made according to the invention.

Table no. 2

Required Tests Commercial Formulation Formulation of the Invention

sterility suits suits harmlessness suits suits appearance suits suits Histamine content suits suits Protein content 0.61 g / 100 ml 0.74 g / 100 ml Sodium chloride content 0,96% 0,97% Moisture content 2,01% 2,25% Current acidity 6.70 6.84

Similarly, human albumin can be used to stabilize a 4.5-5.5% gamaglobulin solution for intravenous administration (Ivega) as well as 1.3-1.8% -

Claims (3)

* SUBJECT A stabilized low-protein solution of 1'-human gamaglobulin, characterized in that it comprises a diluted solution of 1'-human gamaglobulin in an amount of 0.1 to 6.0% by weight, and as a stabilizer it contains human albumin in an amount of 0.05 to 0 15 wt%. 2. A method according to claim 1, for stabilizing a human gamaglobulin solution having a low final gamaglobulin solution containing other classes of immunoglobulins, in particular IgA, IgM (IGAM). BACKGROUND OF THE INVENTION Human albumin is used in the form of a 4.5 to 22% solution for injection. 3. A process according to claim 1 or 2, characterized in that, as a stabilizer, a solution of 1 ' Protein is an injectable solution of 1'-human albumin with an electrophoretic purity of at least 90%.