Hydrophobic nanotubes and nanoparticles as transporters for the delivery of drugs into cells
Methods and materials for delivering biologically active molecules to cells in vitro or in vivo are provided. The methods and materials use carbon nanotubes or other hydrophobic particles, tubes and wires, functionalized with a linking group that is covalently bound to the nanotubes, or, alternatively, to the biologically active molecule, such as a protein. The biologically active molecule is preferably released from the nanotube when the complex has been taken up in an endosome.
1. A method for preparing an active agent for delivery to a living cell, comprising the steps of:
(a) providing a hydrophilic polymer linked to the active agent and also linked to a hydrophobic polymer;
(b) adsorbing the hydrophobic polymer onto a nanoparticle, thereby forming a complex with said nanoparticle, wherein the complex is noncovalently adsorbed onto the nanoparticle by hydrophobic interaction with the hydrophobic polymer, and wherein the nanoparticle is a carbon nanotube; and
preparing a stable suspension comprising suspended nanoparticles wherein the suspended nanoparticles are comprised in the complex formed in step (b).
2. The method of claim 1 further comprising the step of sonicating the nanoparticles.
3. The method of claim 1 wherein the active agent is selected from the group consisting of polynucleic acids and polypeptides.
4. The method of claim 3 wherein said polypeptide is selected from the group consisting of enzymes, antibodies, peptides, and transcription factors.
5. The method of claim 3 wherein said polypeptide has a molecular weight between 20 kD and 150 kD.
6. The method of claim 3 wherein said polynucleic acid is selected from the group consisting of mRNA, dsRNA, siRNA, synthetic DNA encoding a polypeptide, synthetic DNA complementary to a regulatory region of the cell, and cDNA encoding a polypeptide.
7. The method of claim 1 wherein the hydrophilic polymer is linked to the agent with a cleavable linkage.
8. The method of claim 7 wherein the cleavable linkage is cleavable by lowering the pH in the cell after the complex is delivered in order to release the active agent.
9. The method of claim 1 wherein the active agent induces apoptosis in the cell.
10. The method of claim 1 wherein the active agent is siRNA.
11. The method of claim 10 wherein the siRNA silences an oncogene.
12. The method of claim 10 wherein the siRNA silences a receptor.
13. The method of claim 1 wherein the hydrophilic polymer is polyethylene glycol.
14. The method of claim 1 wherein the hydrophobic polymer is comprised in a phospholipid.
15. The method of claim 1 wherein the linking agent comprises polyethylene glycol-phospholipid.
16. The method of claim 15 wherein said phospholipid is terminated with an amine group for linkage of the hydrophilic polymer.
17. The method of claim 7 wherein the cleavable linkage is selected from the group consisting of disulfide and hydrazone.
18. The method of claim 1 wherein the carbon nanotube is a single walled carbon nanotube.