Polymer-cargo-complexes comprising cross-linked copolymers and cargo molecules
Polymer-cargo-complexes including cross-linked copolymers and cargo molecules bound to the copolymers by electrostatic interactions. The non-medical use thereof for transfection and to a kit including such polymer-cargo-complexes. The polymer-cargo-complexes for use in therapy, in particular for use in gene therapy or peptide/protein drug delivery. A method of preparing the polymer-cargo-complexes. A kit for preparing a polymer-cargo-complex of the invention.
1 . A polymer-cargo-complex, comprising:
a) a cross-linked copolymer, the copolymer comprising two alternating units A and B forming a repeat unit A-B such that the copolymer comprises a (A-B) n backbone with n being the number of repeat units of the backbone, wherein unit A is a derivative of an amino acid hydrazide and unit B is a derivate of a dialdehyde comprising a polyethylene glycol (PEG) group according to the following scheme:
wherein R 2 is selected from the group consisting of a carbazole, a pyrrole derivative, a benzyl group, and an alkyl group, and wherein m is an integer from 3 to 12,
wherein 40 to 100 mol % of unit A are derivatives of hydrazides of either cationic amino acids selected from the group consisting of lysine (Lys), arginine (Arg), histidine (His) and combinations of two or more thereof, or anionic amino acids selected from the group consisting of aspartic acid (Asp), glutamic acid (Glu) and combinations thereof,
wherein the copolymer comprises imine groups and acylhydrazone groups alternatingly linking together the alternating units A and B of the polymer backbone such that the units A and B of the backbone are each linked to one neighboring unit by an imine group and to the other neighboring unit by an acylhydrazone group, and
b) cargo molecules bound to the cross-linked copolymer by electrostatic interactions between the cargo molecules and the amino acid side chains of unit A,
wherein the copolymer comprises cross-linking groups linking together some distinct units A present such that one cross-linking group links together two units A.
2 . The polymer-cargo-complex according to claim 1 , wherein the ratio of the total number of cross-linking groups to the total number of units A of the polymer is in a range of 0.05:1 to 0.45:1.
3 . The polymer-cargo-complex according to claim 1 , wherein the cargo molecule is selected from the group consisting of nucleic acids and peptides.
4 . The polymer-cargo-complex according to claim 1 , wherein the dialdehyde comprising a PEG group is a carbazole dicarboxaldehyde comprising a PEG group.
5 . The polymer-cargo-complex according to claim 1 , wherein 0 to 60 mol % of unit A are derivatives of hydrazides of amino acids selected from the group consisting of serine (Ser), threonine (Thr), asparagine (Asn), glutamine (Gln), cysteine (Cys), glycine (Gly), proline (Pro), alanine (Ala), valine (Val), isoleucine (Ile), leucine (Leu), methionine (Met), phenylalanine (Phe), tyrosine (Tyr), tryptophan (Trp) and combinations of two or more thereof.
6 . The polymer-cargo-complex according to claim 1 , wherein the polymer-cargo-complex is a nanoparticle having a hydrodynamic diameter (D H ) in a range of from 50 nm to 350 nm.
7 . A non-therapeutic method, comprising the step of: performing transfection with the polymer-cargo-complex of claim 1 .
8 . A method, comprising the step of: performing therapy with the polymer-cargo-complex of claim 1 .
9 . A method, comprising the step of: performing gene therapy or peptide drug delivery with the polymer-cargo-complex according to claim 8 .
10 . A method, comprising the step of: performing treatment and/or prevention of viral diseases with the polymer-cargo-complex according to claim 8 .
11 . A kit comprising the polymer-cargo-complex of claim 1 , or amino acid hydrazides and dialdehydes comprising a PEG group for preparing said polymer-cargo-complex.
12 . A method of preparing a polymer-cargo-complex according to claim 1 comprising the following steps:
a) Providing an aqueous solution comprising (i) hydrazides of amino acids and (ii) dialdehydes comprising a PEG group, wherein the solution has a pH in the range of from 2 to 6, and wherein 40 to 100 mol % of the hydrazides are hydrazides of either cationic amino acids selected from the group consisting of lysine, arginine, histidine and combinations of two or more thereof, or anionic amino acids selected from the group consisting of aspartic acid, glutamic acid and combinations thereof,
b) Incubating the solution to allow copolymer formation,
c) Mixing the formed copolymer with a cargo molecule in an aqueous solution,
d) Optionally adding amino acid side chain-specific cross-linker to the solution,
e) Increase the pH of the solution to at least 7.
13 . The method according to claim 12 , wherein the cargo molecules of step c) are nucleotides, and wherein the nucleotides are added such that a molar ratio of amine groups to phosphate groups is from 1 to 50.
14 . The polymer-cargo-complex according to claim 1 , wherein the ratio of the total number of cross-linking groups to the total number of units A of the polymer is in a range of 0.05:1 to 0.5:1.