Novel class of therapeutic protein molecules

FIELD: medicine.

SUBSTANCE: invention relates to field of biotechnology. Claimed is protein, including catalytic fragment of sialidase and possessing sialidase activity, which is selected from protein, whose sequence includes amino acids 274-666, or 274-681, or 290-666, or 290-681 SEQ ID NO:12, or includes amino acid sequence SEQ ID NO:14, described herein. Described is fused protein, which includes described above protein and anchor domain. Also given is information about molecules of nucleic acids, which code said proteins, expression vectors, containing said nucleic acids, and pharmaceutical compositions, containing said protein or fusion protein. Claimed is method of treatment or prevention of viral infection, caused by virus of influenza or parainfluenza, including application of therapeutically efficient amount of said composition to epithelial cells of subject.

EFFECT: invention makes it possible to extend arsenal of means against infection, caused by influenza or parainfluenza vitus.

25 cl, 2 tbl, 13 dwg, 17 ex

 

The text descriptions are given in facsimile form.

1. Protein comprising the catalytic domain of sialidase and with sialidase activity, which is selected from:
(1) protein sequence which includes amino acid sequence shown in SEQ ID NO:14, and does not include amino acid residues extending from amino acid 1 to amino acid 273 amino acid sequence shown in SEQ ID NO:12;
(2) protein sequence which includes amino acid sequence starting from amino acid 274 amino acid sequence shown in SEQ ID NO:12, and ends with the amino acid 666 amino acid sequence shown in SEQ ID NO:12, and does not include amino acid residues 1 to 273 and to 667 901 amino acid sequence shown in SEQ D NO:12;
(3) protein sequence which includes amino acid sequence starting from amino acid 274 amino acid sequence shown in SEQ ID NO:12, and ends at amino acid 681 amino acid sequence shown in SEQ ID NO:12, and does not include amino acid residues 1 to 273 and 682 to 901 amino acid sequence shown in SEQ ID NO:12;
(4) protein sequence which includes amino acid sequence, which begins with amino acids 290 amino acid sequence shown in SEQ ID NO:12, and ends with the amino acid 666 amino acid sequence shown in SEQ ID NO:12, and does not include amino acid residues 1 to 289 and to 667 901 amino acid sequence shown in SEQ ID NO:12; and
(5) protein sequence which includes amino acid sequence, which begins with amino acids 290 amino acid sequence shown in SEQ ID NO:12, and ends at amino acid 681 amino acid sequence shown in SEQ ID NO:12, and does not include amino acid residues 1 to 289 and 682 to 901 amino acid sequence shown in SEQ ID NO:12.

2. The molecule is a nucleic acid represented by the nucleotide sequence that encodes a protein catalytic domain sialidase according to claim 1.

3. Expressionvisitor, comprising the nucleic acid molecule of claim 2.

4. Protein with sialidase activity and comprising a fragment of sialidase, including the catalytic domain sialidase, and the anchor domain, where this protein is selected from the following proteins:
(a) protein, where
the selection of sialidase includes the sequence of amino acids extending from amino acid 274 to amino acid 666 of SEQ ID NO:12, inclusive, and does not include a sequence that consists of the sequence extending from amino acid 1 to amino acids 273 and from 667 amino acids to amino acid 901 of SEQ ID NO:12; and
the specified anchor domain is glycosaminoglycan (GAG)binding domain of human amphiregulin comprising amino acid sequence SEQ ID NO:7;
b) protein, where
the selection of sialidase includes the sequence of amino acids extending from amino acid 274 to amino acids 681 of SEQ ID NO:12, inclusive, and does not include a sequence that consists of the sequence extending from amino acid 1 to amino acids 273 and from 682 amino acids to amino acid 901 of SEQ ID NO:12; and
the specified anchor domain is glycosaminoglycan (GAG)binding domain of human amphiregulin comprising amino acid sequence SEQ ID NO:7;
in proteins, where
the criminal code of the related fragment of sialidase includes an amino acid sequence that which extends from amino acid 290 to amino acid 666 of SEQ ID NO:12, inclusive, and does not include a sequence that consists of the sequence extending from amino acid 1 to amino acids 289 and from 667 amino acids to amino acid 901 of SEQ ID NO:12; and the specified anchor domain is glycosaminoglycan (GAG)binding domain of human amphiregulin comprising amino acid sequence SEQ ID NO:7;
g) protein, where
the selection of sialidase includes the sequence of amino acids extending from amino acids 290 to amino acids 681 of SEQ ID NO:12, inclusive, and does not include a sequence that consists of the sequence extending from amino acid 1 to amino acids 289 and from 682 amino acids to amino acid 901 of SEQ ID NO:12; and
the specified anchor domain is glycosaminoglycan (GAG)binding domain of human amphiregulin comprising amino acid sequence SEQ ID NO:7; and
e) protein, where
the selection of sialidase includes the sequence of amino acids extending from amino acid 274 to amino acid 667 of SEQ ID NO:12, inclusive, and does not include a sequence that consists of the sequence extending from amino acid 1 to amino acids 273 and from 668 amino acids to amino acid 901 of SEQ ID NO:12; and pointed to by the th anchor domain is glycosaminoglycan (GAG)binding domain of human amphiregulin, comprising the amino acid sequence of SEQ ID NO:7;
and advanced protein includes a peptide linker.

5. Fused protein according to claim 4, the sequence of which comprises the amino acid sequence shown in SEQ ID NO:21.

6. Fused protein according to claim 4, the sequence of which comprises the amino acid sequence shown in SEQ ID NO:23.

7. Fused protein according to claim 4, the sequence of which comprises the amino acid sequence shown in SEQ ID NO:25.

8. Fused protein according to claim 4, the sequence of which comprises the amino acid sequence shown in SEQ ID NO:27.

9. Fused protein according to claim 4, the sequence of which comprises the amino acid sequence shown in SEQ ID NO:35.

10. Fused protein according to claim 4, the sequence of which comprises the amino acid sequence shown in SEQ ID NO:37.

11. The nucleic acid molecule encoding a protein according to any one of claims 4 to 10.

12. Expressing a vector comprising the nucleic acid molecule according to item 11.

13. The nucleic acid molecule encoding a protein having sialidase activity and comprising a fragment of sialidase, including the catalytic domain sialidase, where the sequence of the nucleic acid molecule includes a nucleotide sequence selected from the sequences shown in SEQ ID NO:18,SEQ ID NO:20, SEQ ID NO:26, SEQ ID NO:28 and SEQ ID NO:36.

14. Pharmaceutical composition for treatment of influenza or parainfluenza and with sialidase activity containing protein according to claim 1 in an effective amount.

15. Pharmaceutical composition for treatment of influenza or parainfluenza and with sialidase activity containing the protein according to any one of claims 4 to 10 in an effective amount.

16. The pharmaceutical composition according to 14 or 15, prepared in the form of spray.

17. The pharmaceutical composition according to 14 or 15, prepared in the form of inhalation means.

18. The pharmaceutical composition according to 14 or 15, prepared in the form of a solution for injection.

19. The pharmaceutical composition according to 14 or 15, prepared in the form of a solution for eye drops.

20. The pharmaceutical composition according to 14 or 15, prepared in the form of a cream, balm, gel or ointment.

21. The pharmaceutical composition according to 14 or 15, prepared in the form of pills, tablets, lozenges, suspensions or solution for oral administration.

22. A method of treating or preventing a viral infection caused by influenza virus or parainfluenza, including:
the use of a therapeutically effective amount of a composition according to any one of p-21 to the epithelial cells of the subject.

23. The method according to item 22, where the specified application is carried out by using a nasal spray.

<> 24. The method according to item 22, where the specified application is carried out by use of the device.

25. The method according to item 22, where the specified application carry from one to four times per day.



 

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FIELD: medicine.

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4 tbl, 2 ex

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1 dwg, 4 ex

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1 tbl, 2 ex

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FIELD: medicine.

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1 tbl, 2 ex

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3 tbl, 3 ex

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