Chondroitinase, process for preparing the same, and pharmaceutical composition comprising the same

A pharmaceutical composition comprising

• (a) physiologically acceptable additives, and
• (b) a purified chondroitinase ABC derived from a Proteus vulgaris microorganism having the following characteristics:
• (i) a single polypeptide having a molecular weight of about 100,000 daltons, as measured by SDS-polyacrylamide gel electrophoresis (SDS-PAGE), in a reduced or unreduced state, and by gel permeation chromatography,
• (ii) isoelectric points of about 8.2 and about 8.5,
• (iii) an optimum pH of about 8.0 to 8.2,
• (iv) an optimum reaction temperature of about 30 to 42° C.,
• (v) a specific activity of at least 300 U/mg, and
• (vi) a single peak as determined by HPLC;
• wherein activity of the chondroitinase ABC is inhibited by a metal ion selected from the group consisting of Zn2+, Ni2+, Fe3+, and Cu2+, and
• wherein the purified chondroitinase ABC is essentially free of nucleic acid, endotoxin and protease activity, and has alanine as the N-terminal amino acid and proline as the C-terminal amino acid.

A method of treating disc herniation by intervertebral disc dissolution comprising the step of:

• administering to a patient in need thereof an effective amount of a purified chondroitinase ABC derived from a Proteus vulgaris microorganism having the following characteristics:
• (i) a single polypeptide having a molecular weight of about 100,000 daltons, as measured by SDS-polyacrylamide gel electrophoresis (SDS-PAGE), in a reduced or unreduced state, and by gel permeation chromatography,
• (ii) isoelectric points of about 8.2 and about 8.5,
• (iii) an optimum pH of about 8.0 to 8.2,
• (iv) an optimum reaction temperature of about 30 to 42° C.,
• (v) a specific activity of at least 300 U/mg, and
• (vi) a single peak as determined by HPLC;
• wherein activity of the chondroitinase ABC is inhibited by a metal ion selected from the group consisting of Zn2+, Ni2+, Fe3+, and Cu2+, and
• wherein the purified chondroitinase ABC is essentially free of nucleic acid, endotoxin and protease activity, and has alanine as the N-terminal amino acid and proline as the C-terminal amino acid.