Class of therapeutic protein based molecules
The present invention provides new compositions and methods for preventing and treating pathogen infection. In particular, the present invention provides compounds having an anchoring domain that anchors the compound to the surface of a target cell, and a therapeutic domain that can act extracellularly to prevent infection of a target cell by a pathogen, such as a virus. The present invention also comprises therapeutic compositions having sialidase activity, including protein-based compounds having sialidase catalytic domains. Compounds of the invention can be used for treating or preventing pathogen infection, and for treating and reducing allergic and inflammatory responses. The invention also provides compositions and methods for enhancing transduction of target cells by recombinant viruses. Such compositions and methods can be used in gene therapy.
1. A method for preparing a pharmaceutical composition, comprising
(a) providing a fusion protein comprising at least one catalytic domain of a sialidase and an anchoring domain, wherein the catalytic domain comprises:
(i) the sequence of amino acids extending from amino acid 274 to amino acid 681 of SEQ ID NO:12, inclusive and lacks the sequence that consists of the sequence extending from amino acid 1 to amino acid 273 and from amino acid 682 to amino acid 901 of SEQ ID NO:12;
(ii) the sequence of amino acids extending from amino acid 290 to amino acid 666 of SEQ ID NO:12, inclusive and lacks the sequence that consists of the sequence extending from amino acid 1 to amino acid 289 and from amino acid 667 to amino acid 901 of SEQ ID NO:12; or
(iii) the sequence of amino acids extending from amino acid 290 to amino acid 681 of SEQ ID NO:12, inclusive and lacks the sequence that consists of the sequence extending from amino acid 1 to amino acid 289 and from amino acid 682 to amino acid 901 of SEQ ID NO:12; and
the anchoring domain comprises (SEQ ID NO:7); and
(b) combining the fusion protein with a pharmaceutically acceptable carrier or diluent thereby creating a pharmaceutical composition.
2. The method of claim 1 wherein the pharmaceutical composition further comprises at least one of a preservative, a stabilizer, a dye and a flavoring agent.
3. The method of claim 1 wherein the pharmaceutical composition is a solution.
4. The method of claim 1 wherein the pharmaceutical composition is a suspension.
5. The method of claim 1 wherein the pharmaceutical composition is a powder.
6. The method of claim 1 wherein the catalytic domain comprises the sequence of amino acids extending from amino acid 274 to amino acid 681 of SEQ ID NO:12, inclusive and lacks the sequence that consists of the sequence extending from amino acid 1 to amino acid 273 and from amino acid 682 to amino acid 901 of SEQ ID NO:12.
7. The method of claim 1 wherein the catalytic domain comprises the sequence of amino acids extending from amino acid 290 to amino acid 666 of SEQ ID NO:12, inclusive and lacks the sequence that consists of the sequence extending from amino acid 1 to amino acid 289 and from amino acid 667 to amino acid 901 of SEQ ID NO:12.
8. The method of claim 1 wherein the catalytic domain comprises the sequence of amino acids extending from amino acid 290 to amino acid 681 of SEQ ID NO:12, inclusive and lacks the sequence that consists of the sequence extending from amino acid 1 to amino acid 289 and from amino acid 682 to amino acid 901 of SEQ ID NO:12.
9. The method of claim 1 wherein the fusion protein comprises an amino acid sequence selected from the group consisting of: SEQ ID NO:23, SEQ ID NO:25 and SEQ ID NO:27.
10. The method of claim 1 wherein the fusion protein comprises the amino acid sequence of SEQ ID NO:23.
11. The method of claim 1 wherein the fusion protein comprises the amino acid sequence of SEQ ID NO:25.
12. The method of claim 1 wherein the fusion protein comprises the amino acid sequence of SEQ ID NO:27.
13. The method of claim 1 wherein the step of providing a fusion protein comprises:
(a) providing a cell harboring an expression vector comprising a nucleotide sequence encoding the fusion protein;
(b) culturing the cell under conditions to express the polypeptide; and
(c) purifying the expressed fusion protein.
14. The method of claim 13 , wherein the cell is a eukaryotic cell.
15. The method of claim 13 , wherein the cell is a prokaryotic cell.
16. The method of claim 13 , wherein the prokaryotic cell is E. coli.