Compositions for needleless delivery of antibodies
The present invention relates, in part, to methods and compositions for needleless delivery of antibodies to a subject. The present invention also relates, in part, to methods for needleless delivery of fusion proteins comprising a bioactive molecule and an antibody fragment to subject. In one aspect, the methods and compositions involve administering to the subject a delivery construct comprising a carrier construct non-covalently bound to the antibody or fusion protein to be delivered, wherein the carrier construct comprises a receptor-binding domain, a transcytosis domain, and an antibody-binding domain to which the antibody or the antibody fragment of the fusion protein non-covalently binds.
1. A delivery construct, comprising an isolated carrier construct non-covalently bound to an isolated antibody, wherein said carrier construct comprises:
a) a receptor-binding domain, wherein said receptor-binding domain (i) binds to a cell-surface receptor that is selected from the group consisting of α2-macroglobulin receptor, epidermal growth factor receptor, transferrin receptor, chemokine receptor, CD25, CD11B, CD11C, CD80, CD86, TNFα receptor, TOLL receptor, M-CSF receptor, GM-CSF receptor, scavenger receptor, and VEGF receptor, or (ii) is selected from the group consisting of a receptor-binding domain from Pseudomonas exotoxin A; cholera toxin; botulinum toxin; diptheria toxin; shiga toxin; shiga-like toxin; TGF α; EGF; IGF-I; IGF-II; IGF-III; IL-1; IL-2; IL-3; IL-6; MIP-1a; MIP-1b; MCAF; and IL-8,
b) a transcytosis domain, and
c) an antibody-binding domain to which the antibody non-covalently binds, wherein said antibody-binding domain is an Fc binding domain.
2. The delivery construct of claim 1 , wherein said Fc binding domain is a Protein G, Protein A, an Fc receptor, or an antibody-binding fragment thereof.
3. The delivery construct of claim 2 , wherein said Fc receptor is Fc receptor γ or neonatal Fc receptor.
4. The delivery construct of claim 1 , wherein the antibody is selected from the group consisting of a monoclonal antibody, a human antibody, a humanized antibody, a chimeric antibody, a single chain antibody, a single domain antibody, an antigen-binding antibody fragment, a Fab fragment and a F(ab′)2 fragment.
5. The delivery construct of claim 1 , wherein the antibody is selected from rituximab, bevacizumab, trastuzumab, cetuximab, natalizumab, infliximab, etanercept, adalimumab, omalizumab, and efalizumab.
6. The delivery construct of claim 1 , wherein the carrier construct further comprises a cleavable linker, wherein cleavage at said cleavable linker separates said antibody-binding domain from the remainder of said carrier construct, and wherein said cleavable linker is cleavable by an enzyme that i) exhibits greater activity at a basal-lateral membrane of a polarized epithelial cell than at an apical membrane of the polarized epithelial cell, or ii) exhibits greater activity in the plasma of a subject than at an apical membrane of the polarized epithelial cell of the subject.
7. The delivery construct of claim 6 , wherein said cleavable linker comprises an amino acid sequence that is selected from the group consisting of Ala-Ala-Pro-Phe (SEQ ID NO.:4), Gly-Gly-Phe (SEQ ID NO.:5), Ala-Ala-Pro-Val (SEQ ID NO.:6), Gly-Gly-Leu (SEQ ID NO.:7), Ala-Ala-Leu (SEQ ID NO.:8), Phe-Val-Arg (SEQ ID NO.:9), Val-Gly-Arg (SEQ ID NO.:10).
8. The delivery construct of claim 6 , wherein the epithelial cell is selected from the group consisting of nasal epithelial cells, oral epithelial cells, intestinal epithelial cells, rectal epithelial cells, vaginal epithelial cells, and pulmonary epithelial cells.
9. The delivery construct of claim 6 , wherein said enzyme that is present at a basal-lateral membrane of a polarized epithelial cell is selected from the group consisting of Cathepsin GI, Chymotrypsin I, Elastase I, Subtilisin AI, Subtilisin AII, Thrombin I, and Urokinase I.
10. The delivery construct of claim 1 , wherein said receptor-binding domain is selected from the group consisting of a receptor-binding domain from Pseudomonas exotoxin A; cholera toxin; botulinum toxin; diptheria toxin; shiga toxin; shiga-like toxin; TGF α; EGF; IGF-I; IGF-II; IGF-III; IL-1; IL-2; IL-3; IL-6; MIP-1a; MIP-1b; MCAF; and IL-8.
11. The delivery construct of claim 1 , wherein said receptor-binding domain binds to an α2-macroglobulin receptor.
12. The delivery construct of claim 10 , wherein said receptor-binding domain of Pseudomonas exotoxin A is Domain Ia of Pseudomonas exotoxin A.
13. The delivery construct of claim 10 , wherein said receptor-binding domain of Pseudomonas exotoxin A has an amino acid sequence that is SEQ ID NO.:1.
14. The delivery construct of claim 1 , wherein said transcytosis domain is selected from the group consisting of a transcytosis domain from Pseudomonas exotoxin A, botulinum toxin, diptheria toxin, pertussis toxin, cholera toxin, heat-labile E. coli enterotoxin, shiga toxin, and shiga-like toxin.
15. The delivery construct of claim 14 , wherein said transcytosis domain is a Pseudomonas exotoxin A transcytosis domain.
16. The delivery construct of claim 15 , wherein said Pseudomonas exotoxin A transcytosis domain has an amino acid sequence that is SEQ ID NO.:2.
17. A composition comprising a delivery construct of claim 1 .
18. The composition of claim 17 , wherein said composition further comprises a pharmaceutically acceptable diluent, excipient, vehicle, or carrier.
19. The composition of claim 17 , wherein said composition is formulated for nasal or oral administration.
20. The delivery construct of claim 12 , wherein said transcytosis domain is a Pseudomonas exotoxin A transcytosis domain.
21. The delivery construct of claim 20 , wherein said Pseudomonas exotoxin A transcytosis domain has an amino acid sequence that is SEQ ID NO.:2.
22. The delivery construct of claim 13 , wherein said Pseudomonas exotoxin A transcytosis domain has an amino acid sequence that is SEQ ID NO.:2.