JPS61293926A - Interleukin-2 composition - Google Patents

Abstract

PURPOSE:To provide the titled stabilized composition administrable safely to living body and producible in large quantity, by compounding interleukin-2 with a surfactant. CONSTITUTION:A solution containing interleukin-2 (IL-2: e.g. human IL-2 pro duced by human lymphocyte, human leukemia cell, etc., mouse IL-2 produced by mouse T-lymphoma cell, etc., and recombinant IL-2 produced by E.coli,etc.) is added with >=0.0003wt% (especially 0.003-5.0% for injection) surfactant (e.g. polyoxyethylene alkyl ether, polyoxyethylene alkylphenyl ether, polyoxy ethylene sorbitan fatty acid ester, etc.) to obtain the objectiveIL-2 composition. EFFECT:The surfactant is available easily at a low cost, has high safety, exhibits the stabilizing effect at a small amount and can be administered to human body without causing too much influence on the body.

Description

[Detailed description of the invention] [Industrial application field] The present invention relates to compositions of interleukin-2.

[Prior Art] Interleukin-2 (hereinafter abbreviated as IL-2) is one of the lymphokines produced by T cells, and its main function is to proliferate cells, as well as to activate natural killer cells. This is necessary for proteins with multipurpose functions such as
Its application as a medicine is expected. However, IL
When applying IL-2 as a medicine, the problem is that IL-2 is extremely unstable. In particular, in IL-2 that has been highly purified to the point of clinical use, its activity can be significantly reduced by temperature, stirring, dilution, etc. Especially IL without sugar chains produced by genetic recombination technology.
-2 is said to be more unstable than natural IL-2, which has sugar chains. Therefore, it is much more difficult to maintain the activity of recombinant IL-2 stably than in the case of natural IL-2. Therefore, there is an urgent need to establish a stabilizer for IL-2 that is safe and can be administered to living organisms and can be supplied in large quantities.

[Problems to be Solved by the Invention] As described above, the present invention aims to establish a stabilized composition for IL-2 that can be administered to a living body, is safe, and can be supplied in large quantities.

[Means for Solving the Problems] The present inventors have discovered that the activity of IL-2 is significantly stabilized by blending IL-2 with a surfactant, and have achieved the present invention.

IL-2 here refers to human IL-2 produced by human lymphocytes, human leukemia cells, etc. (Gi 11is et al., Nature, 268° 154 (197
7); Farrar et al., Journal of Immunology (J, Immunol.

), 121.1353 (1978): Gi I l
iS et al., Journal of Immunology (J.

Immuno 1. >, 120, 2027 (1978
) HGi l l iS et al., Journal of Experimental Medicine (J, Exp, Med,),
152.1709 (1980)>, or mouse T
Mouse IL-2 produced by lymphoma cells etc. may be used, or Escherichia coli into which a human, mouse, or bovine IL-2 structural gene has been incorporated using genetic recombination technology,
Genetically recombinant IL-2 that can be produced by microorganisms such as yeast, or hamster or monkey animal cells may also be used.

As the surfactant used in the present invention, for example, as a polyoxyethylene alkyl ether type, "Br1j3
0", "13ri j35" (Atlas Corporation), "Nitsukol BC, "Nitsukol BL" (Nikko Chemical 1 Co., Ltd.), as polyoxyethylene alkyl phenyl ether type, "Tri tonX-100", "T r
i to n
20”, “TWeen40”, “TWeen60”,
“Tween80” (Atlas), “Nikkor TS”
”, “Nitsukor To” (Nikko Chemicals Co., Ltd.), “HCO-40” as a polyoxyethylene hydrogenated castor oil system
, "HCO-50", "HCO-60" (Nikko Chemicals Co., Ltd.), "Emarex HC-40", "Emarex HC-50", "'Emarex HC-60" (Nikko Emulsion Co., Ltd.) and other non-ionic Among these surfactants, polyoxyethylene sorbitan fatty acid ester type and polyoxyethylene hydrogenated castor oil type are suitable for administration into the body. It is particularly preferably used because it does not cause hemolysis.

The amount of surfactant added is 0.000 to the solution containing IL-2.
The content is preferably 0.0003% (weight %, the same hereinafter) or more, and particularly preferably 0.003 to 5.0% in the case of injections.

The composition of the present invention may be used by appropriately blending substances other than surfactants. For use in combination, IL-
Although not particularly limited as long as it does not affect the stability of No. 2, examples thereof include saccharides, hydroxyethyl starch, carboxymethyl cellulose, polyvinyl alcohol, and the like.

[Effects of the Invention] Although IL-2 has attracted attention as a pharmaceutical and its development is progressing, it is extremely unstable, so there is an urgent need to develop a stabilizer that is effective and can be administered to the human body.

In the present invention, it has been found that the addition of a surfactant is effective for stabilizing IL-2. Surfactants are very easy to obtain, inexpensive, highly safe, and exhibit a stabilizing effect with only a small amount added. Therefore, when administering Shishi-2 to the human body, it is important to avoid having little effect on the human body. It can be administered without giving.

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

Note that the measurement of IL-2 activity was performed using CTLL-2 cells (Gil
lis et al., Journal of Im/Long Yi (J, I
mmunol, >, 120°2027 (1978))
'T0MO8mann's method (Journal of Immunological Methods (J, Immuno
I OCI i CaIMethods>, 65.55
(983)).

Example 1 Crude purified sample of human recombinant IL-2 produced by the method described in Japanese Patent Application No. 1988-88896 (
Activity: 6. Qx 105 U/ml), 20mM
pH 7.0 containing phosphate buffer and 0.2M potassium chloride
"Tween20" as a surfactant in the solution of
Tween80”, “HCO60”, “Br1j3
5” and “TritonX-100” at 0.03 each.
After adding the solution containing % by weight and stirring for 30 seconds using a portex mixer, the mixture was left at 30'C overnight, and the residual IL-2 activity was measured.

As a control, carboxymethylcellulose 0°03%
, 0.6% hydroxyethyl starch, 1.0% polyvinyl alcohol, 1.0% mannitol, 1.0% maltose, 0.3% glutamic acid, and 0.3% glycine, and one without surfactant. The added one was used. The results are shown in Table 1.

Table 1

Claims (1)

[Claims] An interleukin-2 composition comprising interleukin-2 and a surfactant.