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Pharmaceutical composition for prevention or treatment of inflammatory respiratory disease, with fusion protein of cell permeable peptide and ctctla4 peptide contained as effective ingredient therein

A technology for respiratory diseases and fusion proteins, applied in the field of pharmaceutical compositions for the prevention or treatment of inflammatory respiratory diseases comprising a fusion protein in which a cell-penetrating peptide is fused with a CTCTLA4 peptide as an active ingredient, capable of solving topical administration Difficulties, side effects, susceptibility to infection, etc., to achieve the effect of reducing airway hyperresponsiveness, inhibiting airway inflammation, rapid and effective treatment or prevention

Inactive Publication Date: 2019-10-22
IUCF HYU (IND UNIV COOP FOUND HANYANG UNIV)
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Among them, the problem with T cell-specific immunomodulators is that local administration (such as intranasal administration) for the treatment of asthma is difficult, and only systemic administration is possible
[0005] At the same time, monoclonal antibodies developed for the treatment of asthma, such as anti-IgE (omalizumab), anti-IL-5 (mepolizumab), anti-IL-13 (antrolizumab) and anti-TGF- β(daclizumab) also has limitations in that only systemic administration is possible and has problems associated with systemic administration such as side effects, susceptibility to infection and high cost
[0006] Also, the problem with antibodies is that only systemic administration is possible due to their tight binding in the respiratory system and biological size limitations

Method used

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  • Pharmaceutical composition for prevention or treatment of inflammatory respiratory disease, with fusion protein of cell permeable peptide and ctctla4 peptide contained as effective ingredient therein
  • Pharmaceutical composition for prevention or treatment of inflammatory respiratory disease, with fusion protein of cell permeable peptide and ctctla4 peptide contained as effective ingredient therein
  • Pharmaceutical composition for prevention or treatment of inflammatory respiratory disease, with fusion protein of cell permeable peptide and ctctla4 peptide contained as effective ingredient therein

Examples

Experimental program
Comparison scheme
Effect test

preparation Embodiment 1

[0091] Synthesis and purification of dNP2, ctCTLA-4 and Hph-1 peptides

[0092] Peptides having the amino acid sequences of SEQ ID NOs 1 to 3 were synthesized.

[0093] The cell penetrating peptide having the amino acid sequence of SEQ ID NO 1 was named 'dNP2', the peptide having the amino acid sequence of SEQ ID NO 2 was named 'ctCTLA-4' and the cell penetrating peptide having the amino acid sequence of SEQ ID NO 3 The transpeptide was named 'Hph-1'.

[0094] After synthesis of the sense and antisense oligodeoxynucleotides of the amino acid sequence, they were kept at 95° C. for 3 minutes to remove any secondary or tertiary structures formed (denaturation). Then, double-stranded DNA was prepared by changing the temperature to 50°C and then to 72°C. For insertion into the pRSET-b vector, in addition to sense and antisense oligodeoxynucleotides, restriction enzyme specific sequences were inserted at the 5' and 3' ends. Then, Escherichia coli BL21(DE3)Star pLysS was transfor...

Embodiment 1

[0101] Preparation of dNP2-ctCTLA-4 fusion protein

[0102] In order to fuse the cell penetrating peptide having the amino acid sequence of SEQ ID NO 1 prepared in Preparation Example 1 with the ctCTLA-4 peptide having the amino acid sequence of SEQ ID NO 2, a peptide that allows the cell penetrating peptide represented by SEQ ID NO 1 was prepared. Primer linked to the N-terminus of the ctCTLA-4 peptide. After generating the dNP2-ctCTLA-4 gene by PCR reaction, it was inserted into the vector (pRSET-b). After purification of proteins expressed in E. coli, intracellular delivery efficiency was investigated. Detailed experimental procedures are described below.

[0103] 1) Preparation of coding gene

[0104] By inserting the DNA base sequence encoding the cell penetrating peptide having the amino acid sequence of SEQ ID NO 1 prepared in Preparation Example 1 into a part of the N-terminus encoding the ctCTLA-4 peptide having the amino acid sequence of SEQ ID NO 2 The forward ...

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Abstract

The present invention relates to a pharmaceutical composition comprising a fusion protein as an effective ingredient for prevention or treatment of an inflammatory respiratory disease, wherein the fusion protein has a cell permeable peptide fused with the ctCTLA4 peptide. In an administration manner via a respiratory route through which an effective therapeutic or prophylactic effect has not beenconventionally achieved, the present invention has a remarkably outstanding permeability and efficiency to bronchial and pulmonary cells and remarkably improved activity of suppressing cytokine expression and airway inflammation, alleviating hypersensitive responses of the airway to allergens and Th2 inflammation, etc., compared to conventional therapeutic agents for inflammatory respiratory diseases, inter alia, allergic asthma. Therefore, the present invention can bring about a prompt, rapid, and efficient therapeutic or prophylactic effect.

Description

technical field [0001] The present disclosure relates to a pharmaceutical composition for preventing or treating inflammatory respiratory diseases, which comprises a fusion protein in which a cell-penetrating peptide and a ctCTLA4 peptide are fused as an active ingredient, and more particularly, the present disclosure relates to a pharmaceutical composition, since Improved cell permeability and enhanced targeting for inflammatory respiratory diseases by fusing the dNP2 peptide with the ctCTLA4 peptide can be shown to prevent or treat inflammatory respiratory diseases through the nasal mucosa without direct administration to the lung or Excellent effect in the bronchi. Background technique [0002] Inflammatory respiratory diseases are deadly chronic lung diseases caused by dust mites, pollen, cockroaches, etc., and cause serious health problems such as bad breath, difficulty breathing, coughing, wheezing, chest tightness, and pain. [0003] German cockroaches are one of the...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K38/16A61K9/00
CPCA61K9/00A61K38/16A61P11/00A61P11/02A61K9/007A61K9/0073A61P11/06A61K38/1774C07K2319/00C07K19/00A61K38/17
Inventor 崔济民林相昊
Owner IUCF HYU (IND UNIV COOP FOUND HANYANG UNIV)
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