KR880002037B1 - Interferon composition and its preparation method - Google Patents

Abstract

A composition of interferon is presented. Thus, more stable interferon was composed of 7.5 × 1010 I.U. α-interferon, 20g glycin, 1.0g human albumin, 0.55g NaH2PO4, and 2.27g Na2HPO4 in 1.0 litre of H2O.

Description

Interferon composition and preparation method thereof

The present invention relates to interferon compositions having improved safety and methods of making them.

The composition of the present invention is useful for the preparation of sterile solutions containing inteferon as active agents (particularly sterile solutions for use as injectables or nasal sprays, nasal solutions, or ophthalmic solutions) or for the preparation of ointments. Interferon has a powerful effect in the treatment of a wide range of disease states, and in particular in the treatment of various forms of viral infections.

Instability of well-known interferon solutions makes it difficult to formulate stable compositions for clinical or veterinary use. Therefore, it has been proposed to package the interferon in lyophilized form for re-formation with sterile water. Such compositions contain a buffer to maintain a pharmaceutically acceptable pH when the solution is reconstituted and also contain sufficient sodium chloride to make the reconstituted solution isotonic. However, even in such lyophilized compositions, interferon is very limited and exhibits insufficient stability.

The inventors have surprisingly found that the incorporation of a feature amino acid and a single derivative thereof in the lyophilized interferon composition results in a pharmaceutically acceptable lyophilized product with significantly improved stability. In addition, this method may provide another advantage, for example, the lyophilization step itself is easier, the re-formation of the lyophilized product is better and the appearance of the product is better (especially the degree of pigmentation). Low).

Therefore, the present invention provides a lyophilized pharmaceutical composition for preparation with sterile water, consisting of a stabilized amount of an amino acid selected from glycine, α-alanine and pharmaceutically acceptable salts thereof and an appropriate buffer system together with interfelon. to provide. These compositions advantageously contain about 1 × 10 ⁴ to 5 × 10 ⁸ , preferably 1 × 10 ⁶ to 1 × 10 ⁸ International Units (IU) of interferon per 5 to 150 mg of the amino acid or derivative thereof. Wherein the weight of the specific derivative of the amino acid is calculated on the basis of the free amino acid.

The interferon used in the composition according to the invention can be any interferon, but preferred is a human interferon or other interferon suitable for use in the human body, or a polypeptide exhibiting biological properties similar to the biological properties of such interferon. In particular, the human interferon may be leukocytes or fibroblast interferons, and such interferons or corresponding polys may be cultured by leukocytes or fibroblasts or by genetic engineering techniques, i.e. by culturing appropriately planned microorganisms by recombinant DNA methods. It can be prepared by producing a peptide.

Human interferon may be α-interferon, δ-interferon or γ-interferon. The number of α-interferon species is known and is generally numbered after the Greek letter. The scope of the present invention also includes the so-called hybrid interferon to which two or more natural interferon fragments are bound (see ePA 51873, for example, a hybrid of α-interferon). An example is a hybrid α-interferon in which the first 921 amino acids of α-1 interferon are bound to a fragment (carboxy terminus) consisting of 92 to 165 amino acids of α-2 interferon. Particularly preferred α-interferon types for use in the compositions of the present invention are α-2 interferons, in particular recombinant DNA techniques [Nagata et al. in "Nature", Vol. 284, pages 316-320 (1980)]. Another type of α-interferon preferred for use in the compositions of the present invention is α-1 interferon, in particular α-1 interferon prepared by recombinant DNA technology.

The intrinsic activity of α-2 interferon used in the composition of the present invention is preferably 5 × 10 ⁿ IU per mg protein (in the interferon component), and preferably 1 × 10 ⁸ IU per mg protein. This intrinsic activity can be assessed by measuring antiviral activity in comparison to NIH reference standards and by measuring total protein content using standard methods (eg Lowry method). Similarly, high intrinsic activity is desirable for clinical use of other interferons. Maintaining stability at high levels of intrinsic activity was a particularly difficult problem.

The amino acid is preferably used as amino acid itself, ie glycine and α-alanine. Under these conditions, about 5 to 150 mg, preferably 5 to 25 mg, especially 7 to 22 mg of α-alanine or especially glycine for interferon 1 × 10 ⁴ to 5 × 10 ⁸ IU.

Compositions containing this amount of components are suitable for reconstitution with 1 ml of sterile water.

The buffer system present in the composition according to the invention is physiologically suitable and is selected to maintain the re-formulated solution and the solution before lyophilization at the desired pH. The pH of the reconstituted composition of the present invention, especially the composition containing α-2 interferon, should be about 6.5 to 8.0 and preferably about 7.0 to 7.4. Particularly preferred buffer systems for the composition of α-2 interferon consist of dibasic sodium phosphate or monobasic sodium phosphate.

Whether the amino acid itself or a salt thereof is used, the buffer system is selected such that the pH of the composition (after reconstitution) is adjusted to a value within the desired range as described above.

Other ingredients may be present in the compositions according to the invention, provided they are physiologically compatible and do not harm any interferon. In particular, stabilizers may be further added. Preferred as further stabilizers are albumin, and human albumin is preferred, especially when the composition is intended for clinical use. It is possible to add up to about 10 mg of albumin, in particular about 1 mg, relative to the amount of the above-mentioned amino acids (or their derivatives calculated as amino acids), ie 5 to 150 mg (particularly preferably 5 to 25 mg of glycine).

If the amount of interferon (particularly α-interferon) in 1 ml of the reconstituted solution is less than about 5 × 10 ⁶ IU, the addition of albumin is very preferred.

Particularly preferred embodiments of the present invention contain the respective components in the following proportions. α-interferon (especially α-2 interferon) 1 × 10 ⁶ to 1 × 10 ⁸ IU: α-alanine or especially glycine about 5 to 25 mg, preferably 7 to 22 mg: pH of the re-formulated solution Suitable buffer system intended to be 7.0 to 7.4: and about 1 mg of albumin.

The present invention is also directed to dissolving an amino acid or derivative thereof selected from glycine, α-alanine and pharmaceutically acceptable salts thereof, a suitable buffer system, optionally human albumin, and interferon in sterile water, and then freezing and drying the resulting solution. It is characterized by consisting of steps to provide a method for producing a new composition described above.

The water used to prepare the compositions of the present invention may also contain suitable ingredients, especially preservatives such as methyl or isopropyl P -hydroxy benzoate. Such preservatives are advantageously used when the resulting solution is not a single dose but a solution for several applications, such as a rain or ophthalmic solution.

The following non-limiting examples specifically illustrate the preparation of sterile lyophilized powders for the preparation of interferon injectable solutions, with the preferred interferon being α-2 interferon.

Example 1

Lyophilized solution (1000 vials: 1 ml per vial).

Furtherance

α-interferon 7.5 × 10 ¹⁰ IU glycine, USP 20.0 g 1

Sodium Diphosphate Human Albumin USP 1.0 / g

Anhydrous, USP, reagent 2.27 g / 1 distilled water for injection. USP

Titanium Phosphate Monobasic, USP 0.55g / 1

Appropriate amount is made to 1.0ℓ.

Manufacturing method.

1: Place a portion of water in a suitable vessel equipped with a stirrer.

2: Add thorium phosphate and dissolve by stirring.

3: Glycine is added and stirred to dissolve.

4: Albumin is added and stirred to dissolve.

5: alpha -perferon is added and stirred to dissolve.

6: Add batch distilled water USP to make the batch final volume.

7: In the sterile zone, the solution is aseptically filtered through a sterile 0.2 Michaelon filter (washed and tested for stability) and placed into a sterile container.

8: The solution is aseptically filled into sterile vials.

9: Filled vials are aseptically loaded into the freeze dryer.

10: The solution is lyophilized.

11: Aseptically stop the vial stopper.

12: Seal and tighten the vial.

Freeze-drying method (using shelf freeze dryer)

1: The shelves are previously cooled to -30 deg.

2: A vial is put into the chamber.

3: Freeze the solution. Cool the condenser to -45 ° C or below. Make a vacuum The shelf is kept at a temperature of −30 ° C. for at least 12 hours.

4: The shelf temperature is increased to 25 ° C. or lower over a period of about 24 hours or more at a relatively constant speed.

5: When the temperature of the product reaches 25 degreeC, sterilized nitrogen is flowed in until a chamber reaches atmospheric pressure. Plug the vial into the chamber.

6: The vial is taken out of the chamber and sealed.

Example 2

Preferred compositions of the present invention which are interferon α-2 interferon are prepared according to the process of Example 1, using α-2 interferon as interferon and using α-alanine instead of glycine.

Example 3

Preferred compositions of the present invention wherein the interferon is α-1 interferon are prepared according to the process of Example 1, using α-1 interferon as the interferon.

Example 4

Another preferred composition of the present invention uses α-1 interferon as interferon and α-alanine instead of glycine. Prepared according to the process of Example 1.

Claims (26)

A method for producing an interferon composition, characterized by dissolving an amino acid selected from glycine, α-alanine and salts thereof, a buffer system, and interferon in sterile water and lyophilizing the resulting solution. A lyophilized pharmaceutical composition for preparation with sterile water, characterized in that it consists of a stabilizing effective amount of glycine, α-alanine, glycine salt or amino acid of α-alanine salt together with interferon and a buffer system. The method according to claim 2, wherein the amino acid or derivative thereof (the weight of the derivative of the amino acid is calculated based on the free amino acid) about 1 × 10 ⁴ to 5 × 10 ⁸ International Unit (IU) and human albumin 0 to 5 to 150 mg To about 10 mg. 4. A composition according to claim 2 or 3 containing about 1 mg of human albumin per 5 to 150 mg of an amino acid or derivative thereof. The composition of claim 2, wherein the buffer system is intended to maintain the pH at about 6.5 to 8.0 after re-formulation. The composition of claim 2, wherein the buffer system is intended to maintain the pH at about 7.0 to 7.4 after re-formulation. The composition according to claim 2, wherein the buffer system comprises disodium phosphate and sodium monophosphate. The composition of claim 2 wherein the interferon is human interferon. The composition of claim 8, wherein the interferon is α-interferon. The composition of claim 8, wherein the interferon is β-interferon. The composition of claim 9 wherein the interferon is α-2 interferon. The composition of claim 9, wherein the interferon is α-1 interferon. The composition of claim 2, wherein the interferon is an interferon prepared by recombinant DNA method. The composition according to claim 2, which contains 5 to 25 mg of glycine or α-alanine per 1 × 10 ⁴ to 5 × 10 ⁸ IU of interferon. The composition according to claim 2, which contains 7 to 22 mg of glycine or α-alanine per 1 × 10 ⁴ to 5 × 10 ⁸ IU of interferon. 3. Interferon 1 × 10 ⁶ to 5 × 10 ⁸ per 5 to 150 mg according to claim 2, wherein α-alanine, glycine or a salt thereof, wherein the weight of glycine or a derivative of α-alanine is calculated on the basis of free amino acids. A composition containing IU. 3. Interferon 1 × 10 ⁶ to 1 × 10 ⁸ IU per 5 to 25 mg according to claim 2, wherein α-alanine, glycine or a salt thereof, wherein the weight of glycine or a derivative of α-alanine is calculated on the basis of free amino acids. A composition containing a. 3. A composition according to claim 2, wherein glycine (in the case of derivatives of glycine, its weight is calculated on the basis of free amino acids). The composition contains 1 × 10 ⁶ to 1 × 10 ⁸ IU per 7 to 22 mg. The compound of claim 2, wherein the α-interferon 1 × 10 ⁶ to 1 × 10 ⁸ IU; about 5 to 25 mg of a-alanine or glycine; A buffer system intended to maintain the pH at about 7.0 to 7.4 in the reprepared solution; And about 1 mg of human albumin. The compound of claim 2, wherein the α-interferon 1 × 10 ⁶ to 1 × 10 ⁸ IU; About 7 to 22 mg glycine; A buffer system intended to maintain the pH at about 7.0 to 7.4 in the remanufactured solution; And about 1 mg of human albumin. The composition of claim 19 or 20, wherein the α-interferon is α-2 interferon. The composition of claim 19 or 20, wherein the α-interferon is α-1 interferon. 21. The composition of claim 19 or 20, wherein the buffer system is a mixture of dibasic sodium phosphate and dibasic sodium phosphate. The composition of claim 2 wherein the amino acid is glycine. The composition of claim 2 wherein the amino acid is α-alanine. The method of claim 1, further comprising human albumin.