Methods and compositions for needleless delivery of particles
Methods and compositions for needleless delivery of particles to the bloodstream of a subject are provided herein. In one aspect, the invention provides a delivery construct, comprising a receptor binding domain, a transcytosis domain, a particle to be delivered to a subject, and, optionally, a cleavable linker. In other aspects, the invention provides compositions comprising delivery constructs of the invention, kits comprising delivery constructs of the invention, and methods of using delivery constructs of the invention.
1 . A delivery construct, comprising:
a)—a receptor binding domain,
b)—a transcytosis domain, and
c)—a particle.
2 . The delivery construct of claim 1 , wherein the particle is a metal particle, a liposphere, a porous particle, a cell, a peptide or polypeptide aggregate, a peptide or polypeptide crystal, or a high-contrast.
3 . The delivery construct of claim 1 , wherein the particle is a platinum or gold particle.
4 . The delivery construct of claim 1 , wherein the particle is a liposphere.
5 . The delivery construct of claim 1 , wherein the particle is a porous particle.
6 . The delivery construct of claim 1 , wherein the particle is a cell.
7 . The delivery construct of claim 6 , wherein the cell is a mammalian cell.
8 . The delivery construct of claim 6 , wherein the cell is a human, rat, mouse, dog, hamster, chicken, or monkey cell.
9 . The delivery construct of claim 1 , wherein the particle is a high-contrast particle.
10 . The delivery construct of claim 1 , wherein the particle is a peptide or polypeptide aggregate.
11 . The delivery construct of claim 1 , wherein the particle is a peptide or polypeptide crystal.
12 . The delivery construct of claim 1 , further comprising a cleavable linker, wherein cleavage at the cleavable linker separates the particle from the remainder of the delivery construct.
13 . The delivery construct of claim 12 , further comprising a second cleavable linker.
14 . The delivery construct of claim 12 , wherein the 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).
15 . The delivery construct of claim 12 , wherein the cleavable linker is cleavable with an enzyme selected from the group consisting of Cathepsin GI, Chymotrypsin I, Elastase I, Subtilisin AI, Subtilisin AII, Thrombin I, and Urokinase I.
16 . The delivery construct of claim 1 , wherein the receptor binding domain is selected from the group consisting of receptor binding domains from Pseudomonas exotoxin A, cholera toxin, botulinum toxin, diphtheria toxin, shiga toxin, or shiga-like toxin; monoclonal antibodies; polyclonal antibodies; single-chain antibodies; 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.
17 . The delivery construct of claim 1 , wherein the receptor binding domain 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.
18 . The delivery construct of claim 16 , wherein the receptor binding domain of Pseudomonas exotoxin A is Domain Ia of Pseudomonas exotoxin A.
19 . The delivery construct of claim 18 , wherein the receptor binding domain of Pseudomonas exotoxin A has an amino acid sequence that is SEQ ID NO.:1.
20 . The delivery construct of claim 1 , wherein the transcytosis domain is selected from the group consisting of transcytosis domains from Pseudomonas exotoxin A, botulinum toxin, diphtheria toxin, pertussis toxin, cholera toxin, heat-labile E. coli enterotoxin, shiga toxin, and shiga-like toxin.
21 . The delivery construct of claim 20 , wherein the transcytosis domain is Pseudomonas exotoxin A transcytosis domain.
22 . The delivery construct of claim 21 , wherein the Pseudomonas exotoxin A transcytosis domain has an amino acid sequence that is SEQ ID NO.:2.
23 . A composition comprising a delivery construct, the delivery construct comprising:
a)—a receptor binding domain,
b)—a transcytosis domain, and
c)—a particle.
24 . The composition of claim 23 , wherein the composition further comprises a pharmaceutically acceptable diluent, excipient, vehicle, or carrier.
25 . The composition of claim 23 , wherein the composition is formulated for nasal or oral administration.
26 . A method for delivering a particle to a subject, comprising contacting an apical surface of a polarized epithelial cell of the subject with a delivery construct, wherein the delivery construct comprises a receptor binding domain, a transcytosis domain, and the particle, wherein the transcytosis domain transcytosis the macromolecule to and through the basal-lateral membrane of the epithelial cell.
27 . The method of claim 26 , wherein the particle is a metal particle, a liposphere, a porous particle, a cell, or a high-contrast particle.
28 . The method of claim 26 , wherein the particle is a platinum or gold particle.
29 . The method of claim 26 , wherein the particle is a liposphere.
30 . The method of claim 26 , wherein the particle is a porous particle.
31 . The method of claim 26 , wherein the particle is a cell.
32 . The method of claim 26 , wherein the cell is a mammalian cell.
33 . The method of claim 26 , wherein the cell is a human, rat, mouse, dog, hamster, chicken, or monkey cell.
34 . The method of claim 26 , wherein the particle is a high-contrast particle.
35 . The method of claim 26 , wherein the particle is a peptide or polypeptide aggregate.
36 . The method of claim 26 , wherein the particle is a peptide or polypeptide crystal.
37 . The method of claim 26 , further comprising a cleavable linker, wherein cleavage at the cleavable linker separates the particle from the remainder of the delivery construct.
38 . The method of claim 26 , further comprising a second cleavable linker.
39 . The method of claim 26 , wherein the 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).
40 . The method of claim 26 , wherein the cleavable linker is cleavable with an enzyme selected from the group consisting of Cathepsin GI, Chymotrypsin I, Elastase I, Subtilisin AI, Subtilisin AII, Thrombin I, and Urokinase I.
41 . The method of claim 26 , wherein the receptor binding domain is selected from the group consisting of receptor binding domains from Pseudomonas exotoxin A, cholera toxin, botulinum toxin, diphtheria toxin, shiga toxin, or shiga-like toxin; monoclonal antibodies; polyclonal antibodies; single-chain antibodies; 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.
42 . The method of claim 26 , wherein the receptor binding domain 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.
43 . The method of claim 26 , wherein the receptor binding domain of Pseudomonas exotoxin A is Domain Ia of Pseudomonas exotoxin A.
44 . The method of claim 26 , wherein the receptor binding domain of Pseudomonas exotoxin A has an amino acid sequence that is SEQ ID NO.:1.
45 . The method of claim 26 , wherein the transcytosis domain is selected from the group consisting of transcytosis domains from Pseudomonas exotoxin A, botulinum toxin, diphtheria toxin, pertussis toxin, cholera toxin, heat-labile E. coli enterotoxin, shiga toxin, and shiga-like toxin.
46 . The method of claim 26 , wherein the transcytosis domain is Pseudomonas exotoxin A transcytosis domain.
47 . The method of claim 26 , wherein the Pseudomonas exotoxin A transcytosis domain has an amino acid sequence that is SEQ ID NO.:2.
48 . The method of claim 26 , wherein the receptor binding domain is selected from the group consisting of receptor binding domains from Pseudomonas exotoxin A, cholera toxin, diphtheria toxin, shiga toxin, or shiga-like toxin; monoclonal antibodies; polyclonal antibodies; single-chain antibodies; 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.
49 . The method of claim 26 , wherein the receptor binding domain binds to a cell surface receptor selected from the group consisting of α2-macroglobulin receptor, EGFR, IGFR, transferrin receptor, chemokine receptor, CD25, CD11B, CD11C, CD80, CD86, TNFα receptor, TOLL receptor, M-CSF receptor, GM-CSF receptor, scavenger receptor, and VEGF receptor.
50 . The method of claim 26 , wherein the transcytosis domain is selected from the group consisting of transcytosis domains from Pseudomonas exotoxin A, botulinum toxin, diphtheria toxin, pertussis toxin, cholera toxin, heat-labile E. coli enterotoxin, shiga toxin, and shiga-like toxin.
51 . The method of claim 26 , wherein the macromolecule is selected from the group consisting of a peptide, a polypeptide, a protein, a nucleic acid, and a lipid.
52 . The method of claim 26 , wherein the 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.
53 . The method of claim 26 , wherein the 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).
54 . The method of claim 26 , 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.
55 . The method of claim 26 , wherein the mammal is a human.
56 . The method of claim 26 , wherein the delivery construct contacts the apical membrane of the epithelial cell.