WO2002083165A1

WO2002083165A1 - Stable preparations for injection

Info

Publication number: WO2002083165A1
Application number: PCT/JP2001/003103
Authority: WO
Inventors: Katsumi Tanaka
Original assignee: Sumitomo Pharmaceuticals Co., Ltd.
Priority date: 2001-04-10
Filing date: 2001-04-10
Publication date: 2002-10-24

Abstract

Stable preparations for injection of human interferon α containing one or more subtypes which can be preserved over a long time. Stable preparations for injection which contain interferon a of one or more subtypes having an isoelectric point of 4.9 to 6.0 and polysorbate 80 and have a pH value controlled to 6.5 to 7.5.

Description

Description Stable injectable formulation Technical field

The present invention relates to a stable in-product feron formulation. More particularly, it relates to a stable injectable preparation containing interferon. Background art

Aqueous interferon solutions are sensitive to physical and chemical influences and contain many problems to be solved. Interferon, like other proteins, is chemically degraded in aqueous solutions, such as proteolysis, oxidation, disulphide exchange, oligomerization, deamidation and i3-elimination, as well as aggregation, precipitation and adsorption. Susceptible to physical effects. As a method for preventing these effects, it is known to maintain an interferon aqueous solution at a specific pH. In particular, a solution preparation containing interferon α whose pH is adjusted to 4.5 to 5.5 has the least degradation of covalent bond at pH 5 to 6 (JP-A-8-283176). . In addition, a method of adding a stabilizer is also known. For example, an aqueous solution of interferon containing a nonionic surfactant has been reported (JP-A-61-277633). However, depending on the subtype, stabilization cannot always be achieved by adjusting pH or adding a stabilizer. That is, the steric structure and the like are affected by slight differences in amino acid composition and addition of sugars, etc., and the stability differs among the subtypes. , pl-12, 1987). Therefore, it is not easy to stabilize an aqueous solution of an interface having a plurality of subtypes.

Inferon α with different subtypes has different physicochemical properties, and for example, interferon α has a plurality of different isoelectric points between ρ 〜 5.0 and 7.0. In particular, for interferon α derived from Namalba cells, 4.9-6 corrected paper (Fiber ¹ Exists between 0 (Bodo, G. And Adolf, GR The biology of Interferon

System, Elsevier _P 113, 1983). This difference in isoelectric point greatly affects the properties, adsorptivity and stability of injections. For example, since interferon α2a has an isoelectric point at pH 5.9, this PH has a problem of adsorption to the glass surface.

Many different subtypes of interferon α are produced simultaneously in the human body, and these subtypes play an important role in expressing the effects of interferon α. Therefore, development of more stable injectable preparations containing interferon α composed of one or more subtypes having different physicochemical properties is desired. Disclosure of the invention

An object of the present invention is to provide an injectable preparation which can be stored for a long time in human interferon containing one or more subtypes.

The present inventors have conducted intensive studies to solve the above-mentioned problems, and as a result, it has been found that, in human interferon α containing one or more subtypes, addition of polysorbate 80 and optimal ρ 維持 are maintained. As a result, they found that long-term storage was possible, and completed the present invention.

That is, the present invention

(1) It contains interferon α consisting of one or more subtypes having an isoelectric point of 4.9 to 6.0 and polysorbate 80, and the pH is adjusted to 6.5 to 7.5. Stable injection formulation,

(2) The injectable preparation according to (1), wherein the content of interferon α is 1 to 20 MU per 1 mL of the preparation.

(3) The stable injectable preparation according to (1) or (2), wherein the interferon α is derived from Namalva cells.

(4) Polysorbate 80 content formulation The stable injectable formulation according to (1) to (3), wherein the content is 0.01 to 1 mg / mL per 1 mL.

(5) A stable injectable preparation according to (1) to (4), containing sodium chloride. (6) The stable injectable preparation according to (1) to (5), which comprises trometamol.

(7) A stable injectable preparation according to (1) to (6), containing glycine. Interferon alpha refers to a family of highly homologous species-specific proteins that inhibit viral replication and cell growth and modulate immune responses, with partial amino acid substitutions, deletions, or sugar chains There are a plurality of types of subtypes due to such a combination. Typical interferon α subtypes include interferon a 2 a, a 2 b, and a 2 c. \ Other, interferon al, 4, 5, α6, α7 a , 7b, a7c, α8, α10, α13, α14, α17a, α17b, al7c, a21, 7

8, a88, λ2 and the like.

In the present invention, any type may be used as long as it includes a subtype whose isoelectric point selected from the above subtypes is 4.9 to 6.0. In particular, a purified mixture of natural interferon a derived from Namalva cells is suitable.

The sub-type isoelectric point refers to a specific iso-electric point of each of the above-mentioned sub-types, and refers to an iso-electric point measured according to an ordinary method. As a method for measuring the isoelectric point, there is, for example, a method described in Shinsei Kagaku Koza 1 Protein Separation 'Purification' Property 335-336 (1990).

In the case of interferon a containing a plurality of subtypes, the content of each subtype is not particularly limited, but a subtype mixture in which the ratio of each subtype is in the range of 2 to 50% is preferable.

There are no particular limitations on the plurality of subtypes used in the present invention as long as they are two or more.

Interferon injection is effective in treating various conditions, such as renal disease, multiple myeloma, leukemia, chronic and acute hepatitis B and c. Usually, 3 to 9 MU per day is administered subcutaneously or intramuscularly.

The polysorbate 80 used in the present invention can be used in the range of 0.001 to 10 mg, preferably in the range of 0.01 to 1 mg, per mL of the preparation.

The pH can be stabilized in the range of 6.5 to 7.5, but it is particularly preferable that the pH is 6.8 to 7.1. In the present invention, if necessary, an isotonic agent, a buffer and an additive can be appropriately added.

Shiojiri sodium is used as a tonicity agent and is added at a concentration that maintains the osmotic pressure ratio of the injection at 1-2.

Tromethamol and glycine are used as buffering agents and are not particularly limited as long as they can maintain the concentration, but are preferably l to 2 mg per lm L of tromethamolca S preparation, and glycine is 0.5 per ml of preparation. ~: 1 mg is preferable.

The injection according to the present invention can be produced, for example, by the following method. That is, a predetermined amount of polysorbate 80 is added to an aqueous solution containing one or more kinds of interferon α subtypes, sodium chloride, trometamol, daricin, and the like. Thereafter, it can be obtained by filtering the bacteria under sterile conditions and filling in a sealed container such as Noial. When an additive other than the above, for example, human serum albumin, is added, it can be added before sterilization filtration. Example

Next, the present invention will be described in detail with reference to Examples, Comparative Examples, and Test Examples, but the scope of the present invention is not limited thereto.

Example 1

Interferon a (NAMA LWA Sumitomo Pharma) 39.8 MU / mL, sodium chloride 8.8 mg / mL, trometamol 1.22 mg / mL, glycine 0.76 mg / mL, pH 7.1, aqueous solution of interferon alpha 25. IL The solution was dispensed into 4 mL glass vials, and an aqueous solution consisting of 80 lg g / mL of polysorbate was added. Thereafter, a buffer solution consisting of 8.8 mg / mL sodium salt, 1.22 mg / mL tromethamol, 0.76 mg / mL glycine, and pH 7.1 was added, and the whole amount was adjusted to l. OmL. Thereafter, the mixture was sealed with a rubber stopper to obtain a preparation for injection.

Comparative Example 1

A preparation for injection was obtained in the same manner as in Example 1 except that polysorbate 80 was not added. Was.

Example 2

Injection was performed in the same manner as in Example 1 except that 30 L of an aqueous solution containing 80 lmg / mL of polysorbate was added instead of the IOL of the aqueous solution containing 80 lmg / mL of the polysorbate of Example 1. Was obtained.

Example 3

In the same manner as in Example 1, except that an aqueous solution consisting of polysorbate 80 lmg / mL was added instead of the aqueous solution IOL consisting of 80 lmg / mL of polysorbate of Example 1 and lOO ^ L was added. A formulation for injection was obtained.

Example 4

Injection was carried out in the same manner as in Example 1, except that 30 L of an aqueous solution containing 80 lOmg / mL of polysorbate was added instead of the IOL of the aqueous solution containing 80 lmg / mL of the polysorbate of Example 1. A preparation was obtained.

Example 5

In the same manner as in Example 1 except that IOO L of an aqueous solution of polysorbate 800 _mg / mL was added instead of the aqueous solution IOL of 80 lmg / mL polysorbate of Example 1 A formulation for injection was obtained.

Test example 1

The preparations of Examples 1 to 5 and Comparative Example 1 were allowed to stand at 5 ° C. for 24 hours. Thereafter, the interferon α titer was measured by the radioimmunoassay (RIA) method. The results are shown in Table 1. Table 1 Intersorbing effect of polysorbate 80

*: Residual rate was calculated assuming that the charged amount was 1 MUZm L. Example 6

Interferon a (NAMA LWA Sumitomo Pharma) 39.8 MU / mL, sodium salt 8.8 mg / mL, trometamol 1.22 mg / mL, glycine 0.76 mg / mL, pH 7.1, consisting of interferon α 34.7 mL of the aqueous solution was dispensed into a 200 mL polystyrene bottle, and 200 tL of an aqueous solution containing 80 lOOmg / mL of polysorbate was added. Then, a buffer solution consisting of 8.8 mg / mL of sodium chloride, 1.22 mg / mL of tromethamol, 0.76 mg / mL of glycine, and pH 7.1 was added thereto, and the total amount was adjusted to 200 mL. The pH was adjusted to 7.1 using a 1M hydrochloric acid solution or a 1M sodium hydroxide solution. After filtration with a sterilization filter 1, l. OmL was filled into a 4 mL glass vial. The mixture was sealed with a rubber stopper to obtain an injectable preparation.

Example 7

A preparation for injection was obtained in the same manner as in Example 6, except that the pH was adjusted to 6.8 using a 1M hydrochloric acid solution.

Example 8

A preparation for injection was obtained in the same manner as in Example 6, except that the pH was adjusted to 6.5 using a 1 M hydrochloric acid solution.

Comparative Example 2

A preparation for injection was obtained in the same manner as in Example 6, except that the pH was adjusted to 6.0 using a 1 M hydrochloric acid solution.

Comparative Example 3

A preparation for injection was prepared in the same manner as in Example 6, except that the pH was adjusted to 5.0 using a 1M hydrochloric acid solution. Obtained.

Test example 2

The preparations of Examples 6, 7, and 8 and Comparative Examples 2 and 3 were stored at 10 ° C for 12 months. Thereafter, the dissolution of the drug product was confirmed by visual inspection. The results are shown in Table 2. Table 2.

Example 9

Interferon d (NAMA LWA Sumitomo Pharma) 21.5 MU / mL, sodium chloride 8.8 mg / mL, trometamol 1.22 mg / mL, glycine 0.76 mg / mL, pH 7.1 aqueous solution of interferon α 27 9 mL was dispensed into a 200 mL polystyrene bottle, and 600 L of an aqueous solution containing 80 lOOmg / mL of polysorbate was added. Then, a buffer solution consisting of 8.8 mg / mL of sodium chloride, 1.22 mg / mL of trometamol, 0.76 mg / mL of glycine, and pH 7.1 was added to make a total volume of 200 mL. The pH was adjusted to 7.1 with a 1M hydrochloric acid solution or a 1M 7J sodium oxide solution. After filtration with a sterilization filter 1, l. OmL was filled into a 4 mL glass vial. The mixture was sealed with a rubber stopper to obtain an injectable preparation.

Comparative Example 4

A formulation for injection was obtained in the same manner as in Example 9, except that polysorbate 80 was not added.

Test example 3

The preparations of Example 9 and Comparative Example 4 were stored at 10 ° C, and the titer of interferon α was measured by the radioimmunoassay (RIA) method immediately after production, and after storage for 3, 6, 9, and 12 months. did. The solution was visually inspected immediately after production and after storage for 12 months. The results are shown in Table 3 Table 3 Stability of Example 9 and Comparative Example 4 (stored at 10 ° C)

N.D .: No Data Example 1 o

Inter one Feron "2 b 8MU / mL, sodium chloride 8. 8 mg / mL, Torometamo Ichiru 1. 22 mg / mL, glycine 0. 76 mg / mL, inter one Feron alpha 2 b solution 37 consisting of _P H 7. 1 Add 300 L of an aqueous solution consisting of 80 lOOmg / mL of polysorbate to 5 mL, and add sodium chloride 8.8 mg / mL, tromethamol 1.22 mg / mL, glycine 0.76 mg / mL, and H 7.1. Adjust the pH to 7.1 using 1M hydrochloric acid solution or 1M * sodium chloride solution, and filter through a sterilizing filter. Fill the vial 1. Fill each OmL and seal with a rubber stopper to obtain an interferon a 2b injection product.

Industrial applicability

As shown in Tables 1, 2, and 3, the technology of the present invention has made it possible to obtain a long-term stable preparation for interface injection.

Claims

The scope of the claims

1. Contains interferon α consisting of one or more subtypes with isoelectric point of 4.9 to 6.0 and polysorbate 80, and has a pH of 6.5 to 7.5. Adjusted and stable injectable preparation.

2. The stable injectable preparation according to claim 1, wherein the content of interferon α is 1 to 20 MU per 1 mL of the preparation.

3. The stable injection preparation according to claim 1, wherein the interferon α is derived from Namalba cells.

4. The stable injectable preparation according to any one of claims 1 to 3, wherein the content of polysorbate 80 is 0.01 to: Img per mL of the preparation.

5. The stable injectable preparation according to any one of claims 1 to 4, which contains salt chick sodium.

6. The stable injectable preparation according to any one of claims 1 to 5, comprising trometamol.

7. The stable injectable preparation according to claim 1, which contains glycine.