Compositions and methods for intracellular delivery of biotinylated cargo

• US 7,534,819 B2
• Filed: 06/10/2005
• Issued: 05/19/2009
• Est. Priority Date: 06/10/2005
• Status: Expired due to Fees

• Current Assignee: University of Washington
• Original Assignee: University of Washington
• Inventors: To, Richard, Albarran, Brian, Stayton, Patrick S.
• Primary Examiner(s): Weber; Jon P
• Assistant Examiner(s): Liu; Samuel W
• Application Number:
  US11/150,588
• Publication Number:
  US 20060281677A1
• Time in Patent Office: 1,439 Days
• Field of Search: 435/7.5, 435/7.1, 530/367, 530/350, 514/2, 424/486
• US Class Current: 514/772
• CPC Class Codes: A61K 31/4188 condensed with other heterocyclic ring systems, e.g. biotin, sorbinil
• CPC Class Codes: A61K 47/645 Polycationic or polyanionic oligopeptides, polypeptides or polyamino acids, e.g. polylysine, polyarginine, polyglutamic acid or peptide TAT
• CPC Class Codes: A61K 47/665 the pre-targeting system, clearing therapy or rescue therapy involving biotin-(strept) avidin systems
• CPC Class Codes: B82Y 5/00 Nanobiotechnology or nanomedicine, e.g. protein engineering or drug delivery
• CPC Class Codes: Y10S 530/815 Carrier is a synthetic polymer

Abstract

The invention provides compositions for intracellular delivery of biotinylated cargo. The compositions comprise a complex formed between (a) a fusion protein comprising a protein transduction domain linked to streptavidin and (b) a biotinylated cargo for intracellular delivery. In some embodiments, the protein transduction domain comprises the protein transduction domain of the Human Immunodeficiency Virus type 1 (HIV-1) TAT protein. The complex may further comprise a biotinylated endosomal releasing polymer, such as a poly(propylacrylic acid) polymer. The invention also provides methods for obtaining intracellular delivery of biotinylated cargo and methods for obtaining cytoplasmic delivery of biotinylated cargo.

Claims

1. A composition, comprising a complex formed between (a) a fusion protein comprising a protein transduction domain linked to streptavidin (b) a biotinylated cargo for intracellular delivery and (c) a (propylacrylic acid)-containing polymer.

2. The composition of claim 1, wherein the protein transduction domain comprises the protein transduction domain of the Human Immunodeficiency Virus type 1 (HIV-1) TAT protein.

3. The method of claim 1, wherein the (propylacrylic acid)-containing polymer is poly(propylacrylic acid).

4. The composition of claim 1, wherein the (propylacrylic acid)-containing polymer is biotinylated.

5. A method for obtaining intracellular delivery of biotinylated cargo, comprising the step of exposing cells to a complex comprising (a) a fusion protein comprising a protein transduction domain linked to streptavidin to obtain intracellular delivery of the biotinylated cargo (b) a biotinylated cargo and (c) a (propylacrylic acid)-containing polymer.

6. The method of claim 5, wherein the protein transduction domain comprises the protein transduction domain of the HIV-1 TAT protein.

7. The method of claim 5, wherein the (propylacrylic acid)-containing polymer is biotinylated.

8. A method for obtaining cytoplasmic delivery of biotinylated cargo, comprising the step of exposing cells to a complex comprising (a) a fusion protein comprising a protein transduction domain linked to streptavidin to obtain cytoplasmic delivery of the biotinylated cargo, (b) a biotinylated cargo, and (c) a biotinylated (propylacrylic acid)-containing polymer.

9. The method of claim 8, wherein the (propylacrylic acid)-containing polymer is poly(propylacrylic acid).

10. The method of claim 9, wherein the protein transduction domain comprises the protein transduction domain of the HIV-1 TAT protein.