Composition and formulation comprising recombinant human iduronate-2-sulfatase and preparation method thereof
A composition comprising recombinant iduronate-2-sulfatase (IDS) and a method for producing a purified recombinant IDS are provided. The glycosylation pattern and formylglycine content of the IDS composition are different from those of ELAPRASE® and have superior pharmaceutical efficacy and are safer than the conventional agent and thus can be effectively used for the therapy of Hunter Syndrome.
1. A method for treating Hunter syndrome in a subject in need thereof, the method comprising administering to said subject a composition comprising an effective amount of a purified recombinant iduronate-2-sulfatase (I2S) having the amino acid sequence of SEQ ID NO: 1and a carrier, wherein the purified recombinant I2S comprises at least 75% conversion of the cysteine residue corresponding to Cys59of SEQ ID NO:1to Ca-formylglycine (FGly), and wherein the purified recombinant I2S has a purity of at least 99.9% as measured using size exclusion high performance liquid chromatography (SE-HPLC).
2. The method of claim 1 , wherein the purified recombinant I2S comprises at least 80% conversion of the cysteine residue corresponding to Cys59of SEQ ID NO:1to Ca-formylglycine (FGly).
3. The method of claim 1 , wherein the composition is administered intravenously.
4. The method of claim 3 , wherein the composition is administered by intravenous injection.
5. The method of claim 3 , wherein the composition is administered at a dose of 0.5-1.0mg purified recombinant I2S/kg body weight.
6. The method of claim 5 , wherein the dose is 0.5mg purified recombinant I2S/kg body weight.
7. The method of claim 5 , wherein the dose is 1.0mg purified recombinant I2S/kg body weight.
8. The method of claim 1 , wherein administration of the composition results in a reduction of glycosaminoglycans within lysosomes in the subject.
9. The method of claim 1 , wherein the purified recombinant I2S is safe and efficacious.
10. The method of claim 1 , wherein the purified recombinant I2S is 99.9% pure or higher as characterized by silver stain SDS-PAGE.
11. The method of claim 1 , wherein the purified recombinant I2S is 100% pure as measured using size exclusion high performance liquid chromatography (SE-HPLC).
12. A method for treating Hunter syndrome in a subject in need thereof, the method comprising administering to said subject a composition comprising an effective amount of a purified recombinant iduronate-2-sulfatase (I2S) having the amino acid sequence of SEQ ID NO: 1and a carrier, wherein the purified recombinant I2S comprises at least 75% conversion of the cysteine residue corresponding to Cys59of SEQ ID NO:1to Ca-formylglycine (FGly), and wherein the purified recombinant I2S has a K uptake value of 18nM or less.
13. The method of claim 12 , wherein a Lineweaver-Burk plot of the purified recombinant I2S has an x-intercept of approximately −0.25, wherein the x-intercept is the negative reciprocal of the K uptake .
14. The method of claim 13 , wherein the purified recombinant I2S has approximately 3.0moles of mannose-6-phosphate (M6P) per mole of purified recombinant I2S.
15. The method of claim 12 , 13 , or 14 wherein the composition is administered intravenously.
16. The method of claim 15 , wherein the composition is administered by intravenous injection.
17. The method of claim 15 , wherein administration of the composition results in a reduction of glycosaminoglycans within lysosomes in the subject.
18. A method for treating Hunter syndrome in a subject in need thereof, the method comprising administering to said subject a composition comprising an effective amount of a purified recombinant iduronate-2-sulfatase (I2S) having the amino acid sequence of SEQ ID NO: 1and a carrier, wherein the purified recombinant I2S comprises at least 75% conversion of the cysteine residue corresponding to Cys59of SEQ ID NO:1 to Ca-formylglycine (FGly), and wherein the purified recombinant I2S has a specific activity of 19-55nmol/min/pg as determined by an in vitro fluorescent assay using 4-methylumbelliferyl-L-iduronide-2-sulfate Na2 (MU-IdoA-2S ) as a substrate.
19. A method for treating Hunter syndrome in a subject in need thereof, the method comprising administering to said subject a composition comprising an effective amount of a purified recombinant iduronate-2-sulfatase (I2S) having the amino acid sequence of SEQ ID NO: 1and a carrier, wherein the purified recombinant I2S comprises at least 75% conversion of the cysteine residue corresponding to Cys59of SEQ ID NO:1 to Ca-formylglycine (FGly), and wherein the purified recombinant I2S has a specific activity of 30.0 -70,0 nmol/min/μ g as determined by an in vitro fluorescent assay using 4-methylumbelliferyl-L-iduronide-2-sulfate Na2(MU-IdoA-2S) as a substrate.
20. A method for treating Hunter syndrome in a subject in need thereof, the method comprising administering to said subject a composition comprising an effective amount of a purified recombinant iduronate-2-sulfatase (I2S) having the amino acid sequence of SEQ ID NO: 1and a carrier, wherein the purified recombinant I2S comprises at least 75% conversion of the cysteine residue corresponding to Cys 59of SEQ ID NO:1 to Ca-formylglycine (FGly), and wherein the purified recombinant I2S has an isoelectric point of 3.5or less.
21. The method of claim 20 , wherein the purified recombinant I2S contains on average at least 16sialic acids per molecule.