Fusion polypeptide comprising polypeptide region that can be o-glycosylated

A fusion polypeptide and glycosylation technology, which is applied in fusion polypeptides, polypeptides containing positioning/targeting motifs, polypeptides containing affinity tags, etc., can solve problems such as protein denaturation

Pending Publication Date: 2021-04-13
LG CHEM LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

When using hydrophobic hyaluronic acid derivatives to prepare particles for drug release formulations, organic solvents must be used, therefore, protein drugs may be denatured by contact with organic solvents, and the possibility of protein denaturation is high due to the hydrophobicity of hyaluronic acid derivatives

Method used

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  • Fusion polypeptide comprising polypeptide region that can be o-glycosylated
  • Fusion polypeptide comprising polypeptide region that can be o-glycosylated
  • Fusion polypeptide comprising polypeptide region that can be o-glycosylated

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0124] Embodiment 1: the generation of fusion polypeptide

[0125] 1.1. Production of fusion polypeptides containing human growth hormone (hGH) as the target polypeptide

[0126] Fusion polypeptides IgD-hGH-His(DHDD-8His), IgD-hGH(DHDD), IgA-hGH(AHAA), IgD-hGH-IgA(DHAA) and IgA-hGH-IgD(AHDD) were generated (see figure 1 ; the underlined part of the IgD and IgA1 sequences is the part capable of performing O-glycosylation), wherein the IgD hinge IgA1 hinge Or a combination of IgD hinge and IgAl hinge fused to the target polypeptide (human growth hormone: hGH; SEQ ID NO: 3). The amino acid sequence of each moiety contained in the fusion polypeptide is summarized in Table 2 below.

[0127] 【Table 2】

[0128]

[0129] 1.1.1. IgD-hGH (DHDD)

[0130] Plasmid pAF-D1G1 (including Korean Patent No. 10-1868139B1 promoter)) into which is inserted the code '(N-terminus)-[BamHI restriction site-signal peptide (SEQ ID NO:4)-IgD hinge (IgDH1; SEQ ID NO:1)- Human growth hormone (hG...

Embodiment 2

[0186] Example 2: Pharmacokinetic properties (PK profile) test of fusion polypeptide (in vivo)

[0187] 2-1. Target protein: human growth hormone (hGH)

[0188] The fusion polypeptide IgD-hGH, IgD-hGH-His, IgA-hGH F3 (fraction 3 of Example 1.1.3), IgA-hGH F4 (fraction 4 of Example 1.1.3) prepared in Example 1.1 and IgA-hGH F5 (execution fraction 5 of Example 1.1.3) were subcutaneously administered to SD rats (Orientbio, 7 weeks old, about 300 g; n=3) at a dose of 2 mg / kg, and the pharmacokinetics were tested . Sampling was performed at 0, 0.5, 1, 2, 4, 6, 8, 24, 48 hours, and for comparison, hGH (Eutropin, LG Chem) was subcutaneously administered at a dose of 2 mg / kg in the same manner as above, and carry out testing.

[0189] After administration to SD rats as described above, the blood collected at the time point was centrifuged to obtain serum. ELISA was performed using the Human Growth Hormone Quantikine ELISA Kit (R&D Systems), and after confirmation of hGH and fusion...

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Abstract

Disclosed are: a fusion polypeptide comprising a target polypeptide and a hinge region of an immunoglobulin; a pharmaceutical composition containing the fusion polypeptide; and a method for increasing the in vivo period of a target polypeptide, comprising a step of fusing a hinge region of an immunoglobulin.

Description

technical field [0001] The present invention relates to a fusion polypeptide comprising a target polypeptide and an O-glycosylated polypeptide region, a pharmaceutical composition containing the fusion polypeptide; and a method of increasing the duration of a target polypeptide in vivo comprising fusing an O-glycosylatable polypeptide area steps. Background technique [0002] When administered by methods other than intravenous administration, most protein or peptide drugs have a shortened time to maintain activity in vivo and have low absorption rates. When long-term drug therapy is required, there is the inconvenience of having to repeat and continuously inject these drugs at short dosage intervals. In order to eliminate this inconvenience, it is necessary to develop a technique for continuously releasing the drug in a single administration. To meet these needs, sustained release formulations for continuous release are being developed. [0003] For example, research into...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/605C07K14/61C07K16/00A61K38/26A61K38/27A61K47/68
CPCA61K38/26A61K38/27A61K47/68C07K14/605C07K14/61C07K2319/30C07K2319/31C07K2319/21C07K2319/02C07K16/00A61K47/6811C07K2317/53A61K38/00A61K2039/505C07K19/00C07K2317/41C07K2317/94
Inventor 金渊哲李承柱高宗郁金奎用郑新孙荣德
Owner LG CHEM LTD
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