Multi-Component Biological Transport Systems
Compositions and methods are provided that are useful for the delivery of therapeutic agents, including nucleic acids. The compositions can be prepared with components useful for targeting the delivery of the compositions as well as imaging components.
1 . A composition comprising a non-covalent association complex of:
a) a positively-charged backbone; and
b) at least two members selected from the group consisting of:
i) a first negatively-charged backbone having a plurality of attached imaging moieties;
ii) a second negatively-charged backbone having a plurality of attached targeting agents;
iii) at least one member selected from the group consisting of RNA, DNA, ribozymes, modified oligonucleotides and cDNA encoding a selected transgene;
iv) DNA encoding at least one persistence factor; and
v) a third negatively-charged backbone having a plurality of attached biological agents;
wherein said association complex carries a net positive charge and at least one of said two members from group b) is selected from groups i), iii) or v).
2 . A composition in accordance with claim 1 , wherein said biological agent is a therapeutic agent.
3 . A composition in accordance with claim 2 , wherein said therapeutic agent is selected from the group consisting of VEGF, botulinum toxin, a blocker of VEGF, and insulin.
4 . A composition in accordance with claim 1 , wherein said biological agent is a cosmeceutical agent.
5 . A composition in accordance with claim 4 , wherein said cosmeceutical agent is Epidermal growth factor.
6 . A composition in accordance with claim 1 , comprising at least three members selected from groups i) through v).
7 . A composition in accordance with claim 1 , comprising at least one member from each of groups i), ii), iii) and iv).
8 . A composition in accordance with claim 1 , comprising at least one member from each of groups i) and ii).
9 . A composition in accordance with claim 1 , comprising at least one member from each of groups ii), iii) and iv).
10 . A composition in accordance with claim 1 , wherein said positively-charged backbone has a length of from about 1 to 4 times the combined lengths of said members from group b).
11 . A composition in accordance with claim 1 , wherein said positively-charged backbone comprises a polymer having attached positively charged branching groups.
12 . A composition in accordance with claim 11 , wherein said polymer is a peptide and said positively charged branching groups are selected from the group consisting of -(gly) n -arg-arg-arg-arg-arg-arg-arg, HIV-TAT and fragments thereof, wherein the subscript n is an integer of from 0 to 20.
13 . A composition in accordance with claim 12 , wherein n is an integer of from 0 to 8.
14 . A composition in accordance with claim 12 , wherein n is an integer of from 2 to 5.
15 . A composition in accordance with claim 12 , wherein said HIV-TAT fragment has the formula (gly) p -RGRKKRRQRRR-(gly) q , wherein the subscripts p and q are each independently integers of from 0 to 20, and said HIV-TAT fragment is attached to said positively charged backbone via either the C-terminus or the N-terminus.
16 . A composition in accordance with claim 15 , wherein the subscripts p and q are each independently integers of from 0 to 8.
17 . A composition in accordance with claim 15 , wherein the subscripts p and q are each independently integers of from 2 to 5.
18 . A composition in accordance with claim 11 , wherein said polymer is a polylysine and said positively charged branching groups are attached to the lysine sidechain amino groups and are selected from the group consisting of -gly-gly-gly-arg-arg-arg-arg-arg-arg-arg and HIV-TAT.
19 . A composition comprising a non-covalent association complex of a positively-charged backbone having at least one attached efficiency group and at least one nucleic acid member selected from the group consisting of RNA, DNA, ribozymes, modified oligonucleotides and cDNA encoding a selected transgene.
20 . A composition in accordance with claim 19 , wherein said positively charged backbone is polylysine.
21 . A composition in accordance with claim 19 , wherein said efficiency group is selected from the group consisting of (Gly) n1 -(Arg) n2 , wherein the subscript n1 is an integer of from 3 to about 5, and the subscript n2 is an odd integer of from about 7 to about 17, and TAT domains.
22 . A composition in accordance with claim 19 , wherein said positively charged backbone having at least one attached efficiency group is a 150,000 to 300,000 polylysine backbone having a plurality of attached Gly 3 Arg 7 groups wherein the degree of lysine saturation is from about 5% to about 30%.
23 . A composition in accordance with claim 19 , wherein said nucleic acid member is cDNA encoding a selected transgene.
24 . A composition in accordance with claim 19 , wherein said nucleic acid member is part of a plasmid that expresses a detectable product.
25 . A composition in accordance with claim 24 , wherein said detectable product is a fluorescent protein.
26 . A composition in accordance with claim 24 , wherein said detectable product is a blue fluorescent protein.
27 . A composition in accordance with claim 24 , wherein said plasmid further comprises a CMV promoter.
28 . A method for delivery of a biological agent to a cell surface in a subject, said method comprising administering to said subject a composition comprising:
(a) a positively charged backbone;
(b) at least one biological agent selected from the group consisting of:
(i) a first negatively charged backbone having a plurality of attached imaging moieties;
(ii) at least one member selected from the group consisting of RNA, DNA, ribozymes, modified oligonucleotides and cDNA encoding a selected transgene; and
(ii) a third negatively charged backbone having a plurality of attached therapeutic agents; and
(c) a second negatively charged backbone having a plurality of attached targeting agents;
wherein said composition is a non-covalent association complex of said positively charged backbone, said biological agent and said second negatively charged backbone having a plurality of attached targeting agents, and carries a net positive charge.
29 . A method in accordance with claim 28 , wherein said biological agent is an oligonucleotide or a cDNA encoding a selected transgene, and said composition further comprises DNA encoding at least one persistence factor.
30 . A method in accordance with claim 28 , wherein said biological agent is a first negatively charged backbone having a plurality of attached imaging moieties.
31 . A method in accordance with claim 28 , wherein said biological agent is a third negatively charged backbone having a plurality of attached therapeutic agents.
32 . A method in accordance with claim 28 , wherein said administering is intravenous.
33 . A method in accordance with claim 28 , wherein said administering is transdermal.
34 . A method in accordance with claim 28 , wherein said administering is carried out using an angioplastic balloon.
35 . A method in accordance with claim 28 , wherein said administering is carried out using a catheter.
36 . A method in accordance with claim 28 , wherein said administering is intraperitoneal.
37 . A method in accordance with claim 28 , wherein said composition is in a gel formulation.
38 . A method for preparing a pharmaceutical composition, said method comprising combining a positively charged backbone component and at least two members selected from the group consisting of
i) a first negatively-charged backbone having a plurality of attached imaging moieties;
ii) a second negatively-charged backbone having a plurality of attached targeting agents;
iii) at least member selected from the group consisting of RNA, DNA, ribozymes, modified oligonucleotides and cDNA encoding a selected transgene;
iv) DNA encoding at least one persistence factor; and
v) a third negatively-charged backbone having a plurality of attached therapeutic agents;
with a pharmaceutically acceptable carrier to form a non-covalent association complex having a net positive charge, with the proviso that at least one of said two members from groups i) through v) is selected from groups i), iii) or v).
39 . A kit for formulating a pharmaceutical delivery composition, said kit comprising a positively charged backbone component and at least two members selected from the group consisting of
i) a first negatively-charged backbone having a plurality of attached imaging moieties;
ii) a second negatively-charged backbone having a plurality of attached targeting agents;
iii) at least one member selected from the group consisting of RNA, DNA, ribozymes, modified oligonucleotides and cDNA encoding a selected transgene;
iv) DNA encoding at least one persistence factor; and
v) a third negatively-charged backbone having a plurality of attached therapeutic agents;
and instructions for preparing said pharmaceutical delivery composition.