JPS61277633A - Interferon composition - Google Patents

Abstract

PURPOSE:To obtain the titled composition having improved stability of interferon and useful as pharmaceuticals such as carcinostatic agent, etc., by adding a surfactant to interferon. CONSTITUTION:The objective composition is produced by compounding interferon with a surfactant (e.g. polyoxyethylene alkyl ether). Interferon (alpha-, beta- and gamma-interferon) has antiviral and carcinostatic activities and is expected as a medicinal drug, however, it has extremely poor stability and accordingly, the development of an effective stabilizer administrable to human body is highly requested. When the present surfactant is used in place of the conventional human serum albumin, sufficient stabilization effect can be attained at a low dose (>=0.0003wt% based on the interferon-containing solution) in high safety. The surfactant is a very useful stabilizer because it is available extremely easily at a low cost.

Description

[Detailed description of the invention] [Industrial application field] The present invention relates to interferon compositions.

"Prior Art" Interferon is a type of protein that has multipurpose effects including antiviral and anticancer effects, and its development as a drug has progressed rapidly in recent years.However, when applying interferon as a drug, One problem has been that interferon is extremely unstable. Particularly in interferon that has been highly purified to the point of clinical use, its activity can be significantly reduced by changes in temperature, stirring, dilution, etc. For this reason, interferon without sugar chains produced by genetic recombination technology is said to be more unstable than natural interferon with sugar chains.In particular, it is difficult to maintain the activity of genetically recombinant interferon stably. This is much more difficult than in the case of natural interferon. Currently, serum albumin protein is commonly used and is effective as a protective agent against such deactivation of interferon. When adding serum albumin, to avoid antigen-antibody reactions, those derived from animals other than humans such as cows cannot be used, and serum albumin of human origin (huma
n serum albumin; H3A) is required. However, since H3A is extracted from human blood, it is expensive and difficult to obtain. H3A derived from patients with acquired immunodeficiency syndrome (AIDS), which has been spreading worldwide in recent years.
is becoming a new problem, and the increase in AIDS patients is particularly serious in the United States, where Japan imports most of its blood products. It is very difficult to distinguish such H3A derived from AIDS patients from normal HSA, and moreover, when an interferon preparation using H3Δ derived from AIDS patients is administered to the human body, the administered patient may be infected with AI.
The risk of contracting DS is extremely high.

For the above reasons, there is an urgent need to establish an interferon stabilizer that is safe and can be supplied in large quantities as an alternative to H3A [Problems to be solved by the invention] As described above, the present invention is a safe alternative to H8A. The aim is to establish a stabilized interferon composition that can be supplied in large quantities.

[Means for Solving the Problems] The present inventors have discovered that the activity of interferon is significantly stabilized by blending interferon with a surfactant, and have achieved the present invention.

The interferon referred to here may be any of the α-type, β-type, or γ-type. In general, human interferon-α derived from human leukocytes, human interferon-β derived from human normal diploid cells, human interferon-γ derived from human lymphocytes, or mouse cells such as mouse L-929 cells and mouse C-243 cells. Natural interferon such as mouse interferon produced by humans may be used, or microorganisms such as Escherichia coli, yeast, etc. that have incorporated interferon structural genes from humans, mice, or cows using genetic recombination technology, or microorganisms such as hamsters, monkeys, etc. It may also be a recombinant interferon produced by animal cells. Among these, the present invention is particularly effective in producing recombinant interferons.

As the surfactant used in the present invention, for example, as a polyoxyethylene alkyl ether type, "Br1j3
0", "Br i j35" (Atlas Corporation), "Nitsuru BC", "Niracol BL" (Nikko Chemicals Co., Ltd.), as polyoxyethylene alkylphenyl ether type, ""rr 1tonX-100", "Tr i
tonX-405” (Sigma), “Niracol N
P” (Nikko Chemicals Co., Ltd.), “Twe en 20” as a polyoxyethylene sorbitan fatty acid ester type
, “Tween40”, “TWeen60”, “Tween
e n 80” (Atlas), “Niracol TS”
, "Niracol To" (Nikko Chemicals Co., Ltd.), "HCO-40" as a polyoxyethylene hydrogenated castor oil system,
"HCO-50", "'HCO-60" (Nikko Chemikaf 11 companies), "Emarex HC-40", "Emarex HC-50", "Emarex HC-60"
(Nikko Emulsion Co., Ltd.) and other nonionic surfactants. One or more of these may be selected as appropriate. Among the above-mentioned surfactants, polyoxyethylene sorbitan fatty acid ester type and polyoxyethylene hydrogenated castor oil type are particularly preferably used because they do not cause hemolysis when administered to the body.

The amount of the surfactant added is preferably 0.0003% (wt%, same hereinafter) or more based on the solution containing interferon. polyoxyethylene sorbitan fatty acid ester,
When using a polyoxyethylene hydrogenated castor oil-based surfactant, 0.0% of the solution containing interferon is used.
In the case of polyoxyethylene alkyl ether type and polyoxyethylene alkylphenyl ether type surfactants, 0.0003% to 0.05% is preferably used.

The composition of the present invention may be used by appropriately blending substances other than surfactants. There are no particular restrictions on what can be used in combination, as long as it does not affect the stability of interferon, but for example, sugars, hydroxyethyl starch, carboxymethyl cellulose, polyvinyl alcohol, etc. can significantly improve stability, so these should be used in combination as appropriate. The effect can be greatly increased by Sugars include monosaccharides such as glucose, galactose, and fructose, or maltose, lactose,
Oligosaccharides such as sucrose, cellobiose, maltotriose, cellotriose, mannitriose, and maltotetraose are preferred, and one or more of them are appropriately selected. The amount of sugar added is preferably 0.1 to 10.0% by weight based on the interferon-containing solution.

Effects of the Invention] Although interferon has attracted attention as a pharmaceutical and its development is progressing, it is extremely unstable, so there is an urgent need to develop an effective stabilizer that can be administered to the human body.

In the present invention, it has been found that addition of a surfactant as a substitute for the conventionally used human serum albumin is effective for stabilizing interferon. The surfactant is
Interferon is extremely easy to obtain, inexpensive, highly safe, and exhibits a stabilizing effect with only a small amount added, so interferon can be administered to the human body without having much of an effect on the human body. .

EXAMPLES Next, the present invention will be explained in detail with reference to Examples.

The interferon activity was measured using human FL cells and vesicular stomatitis virus (VSV) for human recombinant interferon-β, and between mouse LY cells and vesicular stomatitis virus (VSV) for mouse recombinant interferon-β. Genetically recombinant interferon
γ was measured by the cytopathic effect (CPE) inhibition method using human FL cells and Sindbis virus, respectively.

Example 1 A purified sample of human recombinant interferon-β (activity: 1.Ox 1070/ml) produced by the method described in JP-A-57-77654 was added to 10 m
1) H4°5 containing M acetate buffer and 0.15H NaCl
"Tween20" as a surfactant in the solution of
Tween40”, “Dingween80”
” , ”HCO60” , ”Br1j3
0'' and 0.03 each for “DoritonX-100”
The interferon activity of this mixed solution was measured by diluting it 400 times with a solution containing % by weight.

Theoretical dilution value (2.5x 104 U/ml) 10
The residual activity of interferon was measured at 0% concentration.

The value diluted with physiological saline is shown as a control. The results are shown in Table 1.

Table 1 Example 2 A purified sample of mouse recombinant interferon-β (activity: 6.0x1051/ml) produced by the method described in Japanese Patent Application No. 139399/1980
pH 4 containing 0mM acetate buffer and 0.15M NaCl
, 5 as a surfactant in the solution of ``TWeen20P,''
Tween40","TWeen80","HC
O60”, “Br1j30”, “TritonX-10”
Add a solution containing 0°03% by weight of each of
The mixture was stirred for 30 seconds using a portex mixer, and the residual interferon activity was measured. As a control, 0 and no surfactant were used. The results are shown in Table @2.

Table 2 Example 3 To a purified sample of human recombinant interferon-γ (activity 1, ox 10 SU/ml) produced by the method described in JP-A-58-90514, 10
m) DH with i acetate buffer and 10 mg/ml H3A
A solution containing O-0311t% of 'HCO60' and 003% by weight of maltose was added to the solution of 7.0.
Interferon residual activity was measured after being left for 17 days at 30'C. While the residual activity of the sample without additives, which was used as a control, was 12%, the residual activity of the sample to which "HCO60" and maltose were added increased to 59%.

Claims (1)

[Claims] An interferon composition comprising interferon and a surfactant.