JPS59181224A - Production of stabilized interferon preparation - Google Patents

Abstract

PURPOSE:An aminoacid or a combination thereof with human serum albumin is added to an interferon aqueous solution, then freeze drying is effected to produce the titled preparation having actions of inhibiting virus proliferation and resisting tumors. CONSTITUTION:An aminoacid or a combination thereof with human serum albumin is added to an interferon aqueous solution. Then, preferably the pH is adjusted to 7.1+ or -0.1 using hydrochloric acid or sodium hydroxide, in order to increase the stability of interferon. The resultant solution is freeze-dried to give the objective interferon preparation. The amount of the aminoacid added is preferably 0.05-0.3w/v%, while the amount of human serum albumin is preferably 0.01-1w/v%. When the resultant preparation is dissolved, the use of glucose aqueous solution or the addition of glucose to a solution of the interferon in a physiological salt solution enables the interferon to be kept stable, even after dissolving.

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to stable interferon formulations. More specifically, the present invention relates to a method for producing an interferon preparation, which comprises adding an amino acid or an amino acid and human serum albumin to an aqueous interferon solution, and freeze-drying the mixture.

Interferon is a type of glycoprotein produced by animal cells, including humans, when stimulated by viruses or other substances, and is a very useful substance that has viral growth-inhibiting and antitumor effects.

There are various types of interferon, which are broadly classified into α, β, and γ, and each type is expected to have slightly different effects in terms of medicinal efficacy, etc.

However, since all of these types of interferon are unstable, it was necessary to make some kind of effort to supply them as pharmaceuticals.

Up to now, attempts have been made to freeze-dry various formulations with the intention of improving stability, but none of them have yielded satisfactory results, making the studies extremely difficult.

The present inventors have conducted research on the purification of human interferon for many years and have also investigated methods for stabilizing the activity of this substance.As a result, they unexpectedly found that they added amino acids or amino acids and human serum albumin to an aqueous solution of interferon, and then froze it. They obtained the new finding that the activity of interferon in a lyophilized state is significantly stabilized by drying, and completed the present invention.

In the present invention, although any amino acid can be used to have a considerable effect, preferably, it is considered to be more effective to use a polar amino acid.

The polar amino acids referred to here are from ``Physics of Enzymes'' by Polken Schnain, translated by Yutaka Tanaka, published by Misuzu Shobo, 19
Among amino acids, it refers to amino acids whose properties are more hydrophilic as a whole, and more specifically refers to amino acids such as Arg, Asp (NO3
), GILI, Glu(NO3).

Examples include His, Lys, Ser, 1'hr, etc.

Furthermore, among these, glutamic acid is preferred.

In addition, salts of amino acids, such as sodium salts, can be used similarly.

The amount of amino acid to be added is not particularly limited, but it is more preferably added in an amount of 0.05 to Q, 3 W/V/%. Furthermore, although the addition of human serum albumin is not essential, a stabilizing effect is observed even in the presence of human serum albumin, and the amount added in this case is preferably 0.01 to IW/v.
% seems appropriate.

When carrying out the method of the present invention, it is better to add amino acids or amino acids and human serum albumin to the interferon aqueous solution, and then adjust the pH to about 7.1±0.1 using hydrochloric acid and sodium hydroxide. This is preferable in terms of increasing stability. Furthermore, the obtained solution is freeze-dried using a conventional method.

In carrying out the method of the present invention, tonicity agents, pH adjusters, soothing agents, excipients, etc. may be added according to conventional methods.

The present inventors also discovered that interferon can be stabilized by separately adding glucose to an aqueous interferon solution. Interferon can be stabilized even after dissolution by, for example, adding glucose after dissolving it in physiological saline.

The invention will be explained more clearly in the following experimental examples.

Example human serum albumin final concentration 0.15W/V%
Added human interferon solution (1.
Glutamic acid (added to a final concentration of 0.2 W/v %) to 6 x 106 international units of ink (including feron-α)
, Glucose (IW/V%) and Mannito (IW/V%)
After adding Q, lN-HCt and H0. The pH was adjusted to 7.1 ± 0.1 with IN-NaOH, filtered, and lyophilized. As a control, the human interferon α solution (pH 7.1 ± 0.1) was directly filtered and lyophilized. We compared the stability of the two.

The results of interferon titer measurement are shown in Table 1. When human serum albumin and glucose were added, coloration and a significant decrease in titer and pH were observed; A decrease in titer was observed in both samples and samples to which only human serum albumin was added, whereas the titer and pH of the sample to which human serum albumin and brutamic acid were added decreased.
No problems were observed in terms of properties or solubility.

Table 1 (Note) Each titer was expressed as a relative content percentage based on the titer at the start of storage as 100.

(Note) RIA using a monoclonal antibody for interferon α: Examples of the use of the interferon α kit manufactured by Dynabot R1 Laboratories are shown below, but these examples are not intended to limit the present invention.

Example 1 Human interferon solution (1.6 x 1 in lrrLt)
06 international units of interferon alpha) to a final concentration of 0.2 W/V%Q, lN -N
The aOHJgpH was adjusted to 7.1±0.1, filtered, and lyophilized to obtain a stable human interferon α preparation.

Example 2 Human interferon solution (1.6 xio in 1 mA)
human serum albumin and glutamic acid to a final concentration of 0.10% of human serum albumin and glutamic acid, respectively.
15 W/v% and 9.2 W/v%, adjusted pi-i to 7.1 ± 0.1 with Q, IN-HCl and 0, IN-NaOH, and filtered. By freeze-drying this, stable human interferon α is produced.
A formulation was obtained.

Claims (1)

[Scope of Claims] 1) A method for producing an interferon preparation, which comprises adding an amino acid or an amino acid and human serum albumin to an aqueous interferon solution, and freeze-drying the mixture. 2) The method for producing an interferon preparation according to claim 1, wherein the amino acid is a polar amino acid. 8) The method for producing a telEi interfero preparation according to claim 2, wherein the polar amino acid is glutamic acid.