Composition and formulation comprising recombinant human iduronate-2-sulfatase and preparation method thereof

a technology of iduronate and sulfatase, which is applied in the field of compounding for the treatment of hunter syndrome, can solve the problems of difficult to find a match of hla (human leukocyte antigen) for all patients, joint stiffness and limited motion, and inability to fully function, etc., to achieve the effect of superior pharmaceutical efficacy and safety, and effective us

Inactive Publication Date: 2016-05-26
THE GREEN CROSS CORP +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

[0057]The composition comprising the recombinant IDS and the formulation comprising the same in accordance with the present invention are superior in pharmaceutical efficacy and safety to the conventional agent ELAPRASE® and thus can be effectively used for the therapy of Hunter syndrome.

Problems solved by technology

Also, major joints may be affected by Hunter syndrome, leading to joint stiffness and limited motion.
In some cases of Hunter syndrome, central nervous system involvement leads to developmental delays and nervous system problems.
While bone marrow graft is able to stop most of the symptoms, it is difficult to find an HLA (human leukocyte antigen) match for all patients.
Further, a bone marrow graft is a major surgical operation accompanied by several adverse effects, including the patient's life being put under high risk if the HLA is mismatched.
However, gene therapy remains an experimental technique, and has not been clinically applied.
However, enzyme replacement must be continuously carried out, which incurs a high expense.
However, this drug is very expensive and suffers from the drawbacks of insufficient efficacy and safety.

Method used

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  • Composition and formulation comprising recombinant human iduronate-2-sulfatase and preparation method thereof
  • Composition and formulation comprising recombinant human iduronate-2-sulfatase and preparation method thereof
  • Composition and formulation comprising recombinant human iduronate-2-sulfatase and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

example 1

Preparation of IDS

[0080] Gene Acquisition

[0081]Peripheral blood mononuclear cells (PBMC) were isolated from human blood as described previously [S. Beckebaum et al., Immunology, 2003, 109:487-495]. Total RNA was extracted from the PBMC according to a protocol described previously [M. J. Holland et al., Clin. Exp. Immunol., 1996, 105:429-435]. In order to construct a cDNA library from the total RNA, single-stranded cDNA was synthesized using oligo-(dT) primer with the aid of a single-strand synthesis kit (Boehringer mannheim). In this regard, DEPC-treated distilled water was added to an eppendorf tube containing 1 μg of the total RNA so as to form a final volume of 12.5 μL. Then, 1 μL of a 20 pmol oligo(dT) primer was added to the tube, followed by incubation at 70° C. for 2 min and cooling. To this reaction mixture were added 4 μL of a reaction buffer, 1 μL of dNTP, 1 μL of an RNase inhibitor, and 1 μL of reverse transcriptase which were then reacted at 42° C. for one hour to synthe...

experimental example 1

Structural Analysis and Characterization of Inventive IDS

[0141] Amino Acid Sequencing—Internal Sequencing

[0142]Deglycosylated IDS was separated by SDS-PAGE, followed by gel slicing. Then, digests resulting from treatment with various endoproteinases (trypsin, chymotrypsin, AspN, chymotrypsin / trypsin, AspN / trypsin, GluC and GluC / trypsin) were analyzed using MALDI-MS / MS and LC-ESI-MS / MS (FIG. 5). As a result, a total of 525 amino acid sequences were identified. The amino acid sequences coincided with the theoretical sequence of human IDS (FIG. 6).

[0143] Disulfide Bond Analysis

[0144]In a polypeptide, a disulfide bond is a covalent linkage, usually derived by the coupling of two SH groups of cysteine residues, playing an important role in stabilizing the higher structure of proteins. Theoretically, the 525 amino acids of IDS contain six cysteine residues, four of which form disulfide bonds. In this example, the location of cysteine residues responsible for the disulfide bonds of IDS was...

experimental example 2

Clinical Analysis for Effect of IDS

[0219]Thirty one patients with Hunter syndrome were divided into three groups, administered with the IDS of the present invention and analyzed for parameters associated with Hunter syndrome. ELAPRASE®, a commercially available therapeutic agent for Hunter syndrome, was used as a positive control.

[0220] Change in Urine GAG Level (Primary Check Parameter for Validity Test)

[0221]The three groups of Hunter syndrome patients were administered for 24 weeks with ELAPRASE® (0.5 mg / kg) and the IDS of the present invention (0.5 mg / kg and 1.0 mg / kg), and urine GAG (Glycosaminoglycan) levels were measured as reported previously (Conn. Tissue Res. Vol. 28, pp 317-324, 1990.; Ann. Clin. Biochem. Vol. 31, pp 147-152, 1994). Measurements are summarized in Table 9, below.

TABLE 9Change in Urine GAG Level with IDS AdministrationELAPRASE ®Inventive IDSInventive IDSGroup(0.5 mg / kg)(0.5 mg / kg)(1.0 mg / kg)Change in urine−18.7−29.5−41.1GAG level (%)

[0222]In Hunter syndrome...

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Abstract

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.

Description

CROSS REFERENCE TO RELATED APPLICATIONS[0001]This application is continuation-in-part of application Ser. No. 14 / 809,856 filed Jul. 27, 2015 (allowed), which is a continuation of application Ser. No. 14 / 128,918 filed Dec. 23, 2013 (issued as U.S. Pat. No. 9,206,402), which is a National Stage of International Application No. PCT / KR2012 / 004734 filed Jun. 15, 2012, claiming priority based on Korean Patent Application No. 10-2012-0012718 filed Feb. 8, 2012 and U.S. Provisional Patent Application No. 61 / 500,994 filed Jun. 24, 2011, the contents of all of which are incorporated herein by reference in their entirety.TECHNICAL FIELD[0002]The present invention relates to a composition for the treatment of Hunter syndrome, comprising recombinant human iduronate-2-sulfatase (hereinafter referred to as “IDS”), a formulation comprising the same, and a method for preparing the same.[0003]More particularly, the composition for the treatment of Hunter syndrome in accordance with the present invent...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): C12N9/16
CPCC12N9/16A61K38/465C12Y301/06013A61K9/0019A61K47/02A61K47/26
Inventor JIN, THONG-GYUCHUNG, YO KYUNGPAIK, SANG HOONPARK, YOO CHANGSEO, JINWOOKCHOI, YONG WOONSON, JONG MUNKIM, YONG-CHUL
Owner THE GREEN CROSS CORP
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