Protein ifnab-bpi, its precursor, dna molecule, expression vector, method of production, pharmaceutical composition for inhibiting ifn-α or ifn-β activity

FIELD: chemistry.

SUBSTANCE: invention relates to field of biotechnology, namely to genetic engineering. New interferon-binding proteins, which modulate activity of different interferon-α subtypes, as well as interferon-β activity, are obtained. Described is cloning of DNA fragment, which codes interferon-α/β, binding protein IFNAB-BPI, and expression of obtained DNA fragment in host-cells both with formation of respective polypeptide and in form of fused proteins. Practical application of obtained proteins as components of pharmaceutical compositions for inhibiting activity of IFN-α or IFN-β is suggested. Invention can be applied in medicine for inhibiting undesirable impact of IFN-α or IFN-β.

EFFECT: obtaining new interferon-binding proteins, which can find application in medicine for inhibiting undesirable impact of IFN-α or IFN-β.

13 cl, 10 dwg, 6 tbl, 17 ex

 

The text descriptions are given in facsimile form.

1. Protein IFNAB-BPI, possessing the ability to bind interferon-α/βcharacterized by the amino acid sequence

ISYDSPDYTDESCTFKISLRNFRSILSWELKNHSIVPTHYTLLYTIMSKPED LKVVKNCANTTRSFCDLTDEWRSTHEAYVTVLEGFSGNTTLFSCSHNF WLAIDMSFEPPEFEIVGFTNHINVMVKFPSIVEEELQFDLSLVIEEQSEGIV KKHKPEIKGNMSGNFTYIIDKLIPNTNYCVSVYLEHSDEQAVIKSPLKCT LLPPGQESESAESAKIGIITVFLIALVLTSTIVTLKWIGYICLRNSLPKVLR QGLTKGWNAVAIHRCSHNALQSETPELKQSSCLSFPSSWDYKRASLCPS D, or its functionally active mutein obtained by conservative substitutions in the sequence, or a functionally active fragment.

2. The predecessor of IFNAB-BPI, characterized by the amino acid sequence

MLLSQNAFIVRSLNLVLMVYISLVFGISYDSPDYTDESCTFKISLRNFRSIL

SWELKNHSIVPTHYTLLYTIMSKPEDLKVVKNCANTTRSFCDLTDEWRS

THEAYVTVLEGFSGNTTLFSCSHNFWLAIDMSFEPPEFEIVGFTNHINVM

VKFPSIVEEELQFDLSLVIEEQSEGIVKKHKPEIKGNMSGNFTYIIDKLIPN

TNYCVSVYLEHSDEQAVIKSPLKCTLLPPGQESESAESAKIGGIITVFLIAL

VLTSTIVTLKWIGYICLRNSLPKVLRQGLTKGWNAVAIHRCSHNALQSE

TPELKQSSCLSFPSSWDYKRASLCPSD, or functionally active mutein obtained by conservative substitutions in the sequence, or a functionally active fragment.

3. A protein having the ability to bind interferon-α/β and consisting of (a) amino acid sequence of the protein IFNAB-BPI or its functionally active mutein or fragment according to claim 1 and (b) the amino acid sequence of the immunoglobulin or its fragment.

4. A protein having the ability to bind interferon-α/β and consisting of (a) amino acid sequence of the protein IFNAB-BPI or its functionally active mutein or fragment according to claim 1 and (b) the amino acid sequence of the extracellular part of the receptor α-interferon (IFNAR) or its fragment.

5. A protein having the ability to bind interferon-α/β and Ostoji from (a) amino acid sequence of the protein IFNAB-BPI or its functionally active mutein or fragment according to claim 1 and (b) the amino acid sequence of the immunoglobulin, its fragment, or a combination of any of them with the extracellular part of the receptor α-interferon (IFNAR).

6. An isolated DNA molecule having a nucleotide sequence that encodes a protein IFNAB-BPI according to claim 1 or 2.

7. The DNA molecule according to claim 6, which is transcribed into an mRNA molecule size approximately 1.5 TPN

8. Hybrid DNA molecule encoding a protein according to claim 3 and comprising (a) a nucleotide sequence in accordance with the genetic code determines the amino acid sequence of the protein IFNAB-BPI or its functionally active mutein or fragment according to claim 1, and (b) a nucleotide sequence in accordance with the genetic code determines the amino acid sequence of the immunoglobulin or its fragment.

9. Hybrid DNA molecule encoding a protein according to claim 4 and comprising (a) a nucleotide sequence in accordance with the genetic code determines the amino acid sequence of the protein IFNAB-BPI or its functionally active mutein or fragment according to claim 1, and (b) a nucleotide sequence in accordance with the genetic code determines the amino acid sequence of the extracellular part of the receptor α-interferon (IFNAR) or its fragment.

10. Hybrid DNA molecule encoding a protein according to claim 5 and comprising (a) a nucleotide placenta is successive, in accordance with the genetic code determines the amino acid sequence of the protein IFNAB-BPI or its functionally active mutein or fragment according to claim 1, and (b) a nucleotide sequence in accordance with the genetic code determines the amino acid sequence of an antibody, fragment, or any combination of them with the extracellular part of the receptor α-interferon (IFNAR).

11. Replicated expressing a vector containing a DNA molecule according to any one of p-10 and regulatory elements operatively related to said DNA molecule and ensuring its expression in transformed cell of a mammal.

12. Pharmaceutical composition for inhibiting the activity of IFN-α or IFN-βcontaining protein IFNAB-BPI according to any one of claims 1 and 3-5, in combination with a pharmaceutically acceptable carrier.

13. Method of production of the protein IFNAB-BPI, involving the cultivation of a culture of mammalian cells, expressing transformed by the vector according to claim 11, and the allocation of the specified protein.

Priority items:

07.02.1994 according to claims 1-13.



 

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