JPS61137828A - Gamma-interferon preparation composition - Google Patents

Abstract

PURPOSE:To provide the titled composition containing human serum albumin and maltose, and useful for the prevention and remedy of diseases such as cancer. CONSTITUTION:A gamma-interferon preparation composition can be produced by dissolving a proper amount of gamma-interferon in a buffer solution, preferably a phosphate buffer solution together with human serum albumin (0.01-2w/v%, preferably 0.1-1w/v%), maltose (1.0-20.0w/v%, preferably 5-10w/v%) and optionally thiomalic acid or L-cysteine (0.005-0.1w/v%, preferably 0.1-0.05w.v%). The obtained product can be freeze-dried, and the freeze-dried product can be reconstituted with distilled water to obtain a solution. There occurs no lowering of the titer nor the clouding of the solution by the reconstitution of the freeze-dried preparation with a proper reconstitution agent such as distilled water, and the reconstituted solution can be left stably at room temperature for a long period without causing the clouding, etc.

Description

DETAILED DESCRIPTION OF THE INVENTION A.3 Objective of the Invention Industrial Application Field The present invention relates to a 7-interferon preparation composition used as a medicine, and the composition is used in humans as an antiviral drug, an anticancer drug, etc. It is used for the prevention or treatment of diseases.

Prior Art Bovine serum albumin is used to prevent a decrease in activity during freeze-drying of α-interferon (Cr
yobiology (1979), 16.

301-314), and a method of adding polyols such as glycerin, ethylene glycol, and saccharides is known as a method for stabilizing β-interferon that does not have sugars (Patent Application Publication No. 1982-25333).

On the other hand, a method using mercapto reagents containing reduced geltadeone, thioethanolamine, thiodiglycol, thioacetic acid, monothioglycol, and cysteine has been reported to stabilize interferon derived from fibroblasts (J, gan.

Virol, (1977), 36, 323-327).

However, the circular does not contain data regarding cysteine, and the specification of US Pat. No. 4,100,150, which is considered to correspond to this report, does not include cysteine. On the other hand, inactivation of interferon in chickens, mice, and rats by SH-containing compounds such as cysteine has not been reported. (nature (1964) 203
, 1048-1050. J,tiol.

Biol, (1965) ±3,679-691°Bi
ochim, Biophys, Acta, (1
966) Upper upper ball.

429-439).

Methods for adding amino acids to increase the amount of interferon include adding aromatic amino acids and freeze-drying (Patent Application Publication No. 102519/1982), and adding glycine or α-alanine together with human albumin if desired. Freeze-drying method (Patent application published in 1982-
146504), and a method of adding an amino acid or amino acid/acid and large serum albumin and freeze-drying at °C (Patent Application Publication No. 181224/1982).

In the prior art, it is not known how to stabilize 7-interferon with maltose, and it is not known that a more stable formulation can be obtained by adding thiomalic acid or cysteine.

Ming is trying to do it.

7-Since interferon is composed of proteins or glycoproteins, attempts to prepare preparations containing pharmaceutically effective units often result in failure to obtain clear solutions;
Furthermore, even if a clear solution is obtained, a decrease in titer and turbidity are observed over time due to cleavage of peptide bonds and intermolecular polymerization.

For example, when preparing a lyophilized preparation, a clear solution for lyophilization may not be obtained, a decrease in titer may be observed due to lyophilization, or a clear solution may not be obtained when a destabilizing agent is added to the lyophilized preparation. There are problems such as the reconstituted solution becoming cloudy or decreasing in activity if left undisturbed. The problem to be solved by the present invention is how to stably maintain 7-interferon in an aqueous solvent.

B6 Structure of the invention 1. As mentioned above, various methods for stabilizing interferon are already known, but there is currently no reliable method for 7-interferon. When human serum albumin and maltose were used in combination, 7-interferon showed unexpected stability and a clear liquid was obtained that dissolved enough units of 7-interferon to be used as a pharmaceutical. Ta. This solution is also suitable for freeze-drying, and even when the freeze-dried preparation is reconstituted by adding an appropriate restoring agent such as distilled water, no decrease in titer or turbidity of the solution is observed. Even when left at room temperature, no turbidity occurs and stability is maintained.

Stability is further enhanced by addition of thiomalic acid or L-cysteine or salts thereof.

The pharmaceutical composition according to the invention rBq combines an appropriate amount of 7-interferon with human serum albumin (approximately 0.01 to 2 k/V).
%, preferably about 0.1-1%), maltose (
About 1,0-20 Ow/v%, preferably about 5-20%
10 w/v%), optionally thiomalic acid or L-
It can be obtained by dissolving it in a buffer, preferably a phosphate buffer, together with cysteine (about 0,000 to 0.1%/V, preferably about 0.01 to 0.05%/V). The present invention also includes a lyophilized preparation obtained by lyophilizing the obtained solution, and a solution preparation prepared by adding distilled water, physiological saline, etc. to the lyophilized preparation to obtain an appropriate concentration.

An example of the formulation of the present invention is shown below.

7- Interferon Approximately I x 10' to 6 x 10'1. U.

Human serum albumin approximately 0.2-40mg maltose
Approximately 0.02-0.4gL-cysteine Approximately 0.1-2.0mg, 2.0 in total amount of 1M phosphate buffer
ml.

Note that the titer of 7-interferon in this specification is a value calculated by converting the activity corresponding to α-interferon 1 international level to 1 unit (I, U,).

The salt of L-cysteine that can be used is a pharmaceutically usable salt, such as hydrochloride. The buffer range of the buffer solution is preferably about pH 6.5 to 7.5.

If a lyophilized preparation is desired, the solution comprising the above composition is cooled to about -10 to -60°C, preferably about -2°C.
After rapidly freezing at 5 to -40°C for several minutes to 10-odd hours,
0.0 for about 5 to 72 hours, supplying sublimation heat if necessary.
It is preferable to use a conventional method such as keeping the water content at 0.005 to 1 mb, sublimating and removing water until a predetermined water content is reached, filling with an inert gas such as nitrogen or dry air if necessary, and sealing the container tightly. When using the dry preparation obtained by freeze-drying as an injection, it is sufficient to dissolve it in distilled water.
Approximately 1-2 ml of distilled water is used in the above formulation examples.

Examples and results of tests conducted on the stability of the 7-interferon preparation according to the present invention and its lyophilized preparation are shown below.

The test preparation consisted of the composition shown in Table 1 in 0.1M phosphate buffer (pH 6.8) containing 0.5% human serum albumin.
The filtrate obtained by dissolving the solution in 1 ml and filtering the same solution, and the preparation obtained by freeze-drying 2 ml of the same filtrate in a vial were added with distilled water after being left to stand under the time and temperature conditions shown in Table 1. This is a solution prepared by adding 1 ml. The state of the solution was observed according to 1. Insoluble Foreign Matter Testing Method for Injectables described in the Japanese Pharmacopoeia Law, - indicates that the solution is clear, and 10 to +++ indicates the degree of turbidity.

In this test, recombinant 7-interferon was used, and Test Examples 1 to 4 and 5 to 10 were conducted separately.

(Left below) Table 1; As shown, maltose is 7-interf;
By adding it to Ron's solution, better solubility improvement is observed compared to glucose and mannitol, which are already in use.

It has also been found that the combination of ma-retose and thiomalic acid or cysteine provides better solubility than the combination with mannitol and prevents a decrease in the activity of 7-interferon.

Example 1 0.1M Lyrin (dihydrogen) monosodium-phosphoric acid (1 water 1g) disodium buffer solution adjusted to pH 6.8 and human 7-interferon 3X10''1, U, equivalent amount Dissolve 1.25 g of human serum albumin (as a dry weight) and 12.5 g of pharmacopoeial maltose (as a dry weight), and make the total volume 500 ml with the above buffer.The obtained solution is filtered through an appropriate membrane filter. After injecting 2 ml of this sterile solution into a vial, freeze it at below 125°C, and freeze-dry according to a conventional method while maintaining the product temperature below 125°C. Obtain a lyophilized formulation.

Example 2 0.075 g of L-cysteine was added to the composition solution of Example 1.
The sterile solution obtained is lyophilized in the same manner as in Example 1 to obtain a lyophilized preparation.

Example 3 A lyophilized preparation is obtained using thiomalic acid in place of L-cysteine in Example 2.

Example 4-8 A lyophilized preparation is obtained using the following composition in the same manner as in Example 1.

(Margin below)

Claims (2)

[Claims] (1) A 7-interferon pharmaceutical composition containing human serum albumin and maltose. (2) The composition according to claim (1), which contains thiomalic acid or L-cysteine.