Methods for promoting entry of an agent into a cell

Inventors

Good, Liam • Chindera, Kantaraja • Gburcik, Valentina

Assignees

Tecrea Ltd

Abstract

A method for promoting entry of an agent (introduced agent) into a cell, the method comprising the step of complexing the introduced agent in the presence of an entry-promoting agent and then exposing to cells, wherein the entry-promoting agent comprises a linear and/or branched or cyclic polymonoguanide/polyguanidine, polybiguanide, analogue or derivative thereof according to the following Formula 1a & b. The method also provides a means for formation of nanoparticles formed between the entry promoting agent and the introduced agent. wherein: “n”, refers to number of repeating units in the polymer, and n can vary from 2 to 1000, for example from 2 or 5 to 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250, 300, 350, 400, 450, 500, 600, 700, 800 or 900; G1 and G2 independently represent a cationic group comprising biguanide or guanidine, wherein L1, and L2 are directly joined to a Nitrogen atom of the guanide; L1 and L2 are linking groups between the G1 and G2 cationic groups in the polymer and independently represent an aliphatic group containing C1-C40 carbon atoms, for example an alkyl group such as methylene, ethylene, propylene, C4, C5, C6, C7, C8, C9 or C10; C1-C10, -C20, -C30, -C40, -C50 -C60 -C70, -C80, -C90, -C-100, -C110, -C120, -C130 or -C140, alkyl; or a C1-C140 (for example C1, C2, C3, C4, C5, C6, C7, C8, C9 or C10; C1-C10, -C20, -C30, -C40, -C50 -C60, - C70, -C80, -C90, -C100, -C110, -C120, -C130o or -C140), cycloaliphatic, heterocyclic, aromatic, aryl, alkylaryl, arylalkyl, or oxyalkylene radical; or a polyalkylene radical optionally interrupted by one or more, preferably one, oxygen, nitrogen or sulphur atoms, functional groups or saturated or unsaturated cyclic moiety; N and G3 are optional end groups; X can be either present or absent; L3, L4 and X are linking groups between the G4 and G5 cationic groups in the polymer and independently represent an aliphatic group containing C1-C140 carbon atoms, for example an alkyl group such as methylene, ethylene, propylene, C4, C5, C6, C7, C8, C9 or C10; C1-C10, -C20, -C30, -C40, -C50 -C60, -C70, -C80, -C90, -C100, -C110, -C120, -C130 or -C140, alkyl; or L3 and L4 and X can independently be C1-C140 (for example C1, C2, C3, C4, C5, C6, C7, C8, C9 or C10; C1-C10, -C20, -C30, -C40, -C50 -C60, -C70, -C80, -C90, -C100, -C110, -C120, -C130 or -C140), cycloaliphatic, heterocyclic, aromatic, aryl, alkylaryl, arylalkyl, oxyalkylene radicals; or a polyalkylene radical optionally interrupted by one or more, preferably one, oxygen, nitrogen or sulphur atoms, functional groups as well as saturated or unsaturated cyclic moiety; “G4” and “G5” are cationic moieties and can be same or different, and at least one of them is a biguanidine moiety or carbamoylguanidine, and the other moiety may be biguanidine or carbamoylguanidine or amine; and cationic moieties G4 and G5 do not contain single guanidine groups. The entry-promoting agent may comprise homogeneous or heterogeneous mixture of one or more of agents arising from formula 1 a and b, for example polyhexamethylene biguanide (PHMB), polyhexamethylene monoguanide (PHMG), polyethylene biguanide (PEB), polytetramethylene biguanide (PTMB), polyethylene hexamethylene biguanide (PEHMB), polymethylene biguanides (PMB), poly(allylbiguanidnio-co-allylamine), poly(N-vinylbiguanide), polyallybiguanide.

Core Innovation

The invention provides a method for promoting entry of an introduced agent into a cell by exposing the cell to non-covalently complexed nanoparticles consisting of the introduced agent and polyhexamethlene biguanide (PHMB). The introduced agent is a nucleic acid or a nucleic acid analogue.

The disclosed compositions and polymers include polyguanidine and polymonoguanide materials that form noncovalent nanoparticles with an introduced agent. The polymers are described as linear, branched, or cyclic, and include biguanidine/guanidine/carbamoylguanidine cationic groups and linking groups as defined by Formula 1a and Formula 1b, including repeat units and end groups.

The invention further describes utility in delivering nucleic acids and nucleic acid analogues to functional locations within cells, including interaction with nucleic acids and delivery to the nucleus. It also describes forming nanoparticles with diverse cargo, including nucleic acids, proteins/peptides, and small molecules, together with broad cell-type uptake (bacteria, fungi, and mammalian).

Claims Coverage

The independent claim defines one inventive feature: promoting cellular entry using non-covalently complexed nanoparticles comprising the introduced agent and PHMB, where the introduced agent is limited to nucleic acids or nucleic acid analogues. Dependent claims refine target cell type, example nucleic-acid cargo, and performance contexts.

The claims are centered on promoting entry of nucleic acids or nucleic acid analogues by exposing cells to non-covalently complexed PHMB nanoparticles containing the introduced agent, with dependent claim limitations covering particular nucleic-acid types, target cell categories, and experimental performance contexts.

Stated Advantages

Documented Applications