초록

This invention relates to a stable liquid formulation comprising hyaluronidase and a stabilizer for hyaluronidase and more particularly, to a stable liquid formulation by the addition of the stabilizer to the hyaluronidase.

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1. A method of stabilizing a hyaluronidase in liquid formulation, comprising contacting a stabilizer consisting of (i) a buffering agent to provide pH 4.0 to 6.0, (ii) 0.01 to 0.5 v/v % of a non-ionic surfactant and (iii) 0.1 to 5 mM of a chelating agent or MgCl2. 2. The method of claim 1, wherein t

1. A method of stabilizing a hyaluronidase in liquid formulation, comprising contacting a stabilizer consisting of (i) a buffering agent to provide pH 4.0 to 6.0, (ii) 0.01 to 0.5 v/v % of a non-ionic surfactant and (iii) 0.1 to 5 mM of a chelating agent or MgCl2. 2. The method of claim 1, wherein the hyaluronidase has a purity of 95% or higher. 3. The method of claim 1, wherein the hyaluronidase has a specific activity of 70,000 IU/mg or higher. 4. The method of claim 1, wherein the liquid formulation is an injectable formulation. 5. The method of claim 1, wherein the hyaluronidase has a stability of maintaining the activity up to 90% or more with regard to its initial enzymatic activity 100 at a temperature condition of 2 to 8° C. 6. The method of claim 1, wherein the hyaluronidase is an active component of the formulation. 7. The method of claim 1, wherein the chelating agent is EDTA. 8. The method of claim 1, wherein the non-ionic surfactant is selected from the group consisting of polyoxyethylene-sorbitan fatty acid ester and Triton X-100. 9. The method of claim 8, wherein the non-ionic surfactant is selected from the group consisting of polysorbate 20, polysorbate 80, and Triton X-100. 10. The method of claim 1, wherein the buffering agent is selected from the group consisting of succinate buffer, acetate buffer, phosphate buffer, citrate buffer, malonate buffer, 2-(N-Morpholino)ethanesulphonic acid(MES) buffer, Tris buffer and glycine buffer. 11. The method of claim 1, wherein the hyaluronidase is derived from sheep, cows, pigs, or humans. 12. The method of claim 1, wherein the hyaluronidase is a recombinant hyaluronidase produced by transducing a mammalian hyaluronidase gene into microbes, animal cells or plant cells, or is an extract derived from sheep, cows, pigs, or humans. 13. The method of claim 1, further comprising purifying the hyaluronidase from a hyaluronidase-containing material. 14. The method of claim 13, further comprising purifying the hyaluronidase from a hyaluronidase-containing material by one or more methods selected from the group consisting of affinity chromatography, ion exchange chromatography, and gel filtration. 15. The method of claim 14, wherein the affinity chromatography is an affinity chromatography in which a matrix is composed of cross-linked agarose beads modified with a modified triazine dye, or an affinity chromatography in which a matrix is composed of ross-linked agarose beads modified with heparin. 16. The method of claim 15, wherein the purifying hyaluronidase from a hyaluronidase-containing material is performed sequentially by using an affinity chromatography in which a matrix is composed of cross-linked agarose beads modified with a modified triazine dye, a cation exchange chromatography,an anion exchange chromatography,an affinity chromatography in which a matrix is composed of ross-linked agarose beads modified with heparin, anda gel filtration.