Cell-targeting nanoparticles comprising polynucleotide agents and uses thereof

Patent metadata

Field	Value
Publication number	US-10179113-B2
Application number	US-201715402444-A
Country	US
Kind code	B2
Filing date	Jan 10, 2017
Priority date	Jul 31, 2009
Publication date	Jan 15, 2019
Grant date	Jan 15, 2019

Abstract

A method of generating a particle is disclosed, the particle being for delivery of a polynucleotide to a target cell. The method comprises (a) contacting the polynucleotide with a composition comprising cationic molecules, wherein the cationic molecules condense the polynucleotide by electrostatic interactions to generate a complex, wherein the cationic molecules are not comprised in a liposome; and (b) covalently binding the complex to a targeting moiety at a pH equal to or below about 4.5, thereby generating the particle for delivery of the polynucleotide agent to the target cell. Use of the particles and compositions comprising same are also disclosed.

First claim

What is claimed is:

1. A particle comprising a core and a shell, wherein the core comprises 1,2-Dilauroyl-sn-Glicero-3-Phosphorylglycerol (DLPG), a lipid having an amino group which is 1,2-Dilauroyl-sn-Glicero-3-Phosphoethanolamine (DLPE), a polynucleotide, and optionally a cationic lipid and/or cholesterol; wherein the shell comprises a glycosaminoglycan targeting moiety; and wherein the particle is not a liposome.

2. The particle of claim 1 , wherein the glycosaminoglycan targeting moiety is selected from the group consisting of hyaluronic acid (HA), keratan sulfate, chondroitin sulfate, heparin sulfate, heparan sulfate, dermatin sulfate, salts, and mixtures thereof.

3. The particle of claim 1 , wherein the polynucleotide comprises RNA or DNA.

4. The particle of claim 1 , wherein the polynucleotide is selected from a single stranded and a double stranded polynucleotide.

5. The particle of claim 1 , wherein the polynucleotide comprises an RNA silencing agent.

6. The particle of claim 5 , wherein the RNA silencing agent is selected from the group consisting of an siRNA, miRNA, an antisense oligonucleotide, and a ribozyme.

7. The particle of claim 1 , wherein the cationic lipid is selected from the group consisting of dioleoyl trimethylammonium-propane (DOTAP); N-[1-(2,3-dioleyloxy)propyl]-N , N, N-trimethlylammonium chloride (DOTMA); dimethyldioctadecylammonium (DDAB); 1,2-dilauroyl-sn-glycero-3-ethylphosphocholine; 1,2-di-(9Zoctadecenoyl)-3-dimethylammonium-propane; and 38-[N-(N′,N′-dimethylaminoethane)carbamoyl]cholesterol hydrochloride (DC-Cholesterol).

8. The particle of claim 1 , wherein the core comprises DOTAP, and cholesterol.

9. The particle of claim 1 , further comprising at least one additional targeting moiety, said additional targeting moiety being selected from the group consisting of an antibody, an antibody fragment, a receptor ligand and an aptamer.

10. The particle of claim 1 , wherein the particle is a nanoparticle of about 1 nm to about 500 nm in size.

11. A method for downregulating a gene of interest in a target cell, comprising administering the particle of claim 1 to the target cell.

12. The method of claim 11 , wherein the target cell expresses CD44.

13. The method of claim 11 , wherein the gene of interest is an oncogene and/or a gene associated with cell viability.

14. A method for treating cancer in a subject in need thereof, the method comprising administering to the subject a particle according to claim 1 .

Assignees

• Univ Ramot

Inventors

• Peer Dan

Classifications

• A61P35/00

  Antineoplastic agents · CPC title

• B82Y5/00

  Nanobiotechnology or nanomedicine, e.g. protein engineering or drug delivery · CPC title

• C12N15/88

  using microencapsulation, e.g. using {amphiphile} liposome vesicle · CPC title

• C12N2310/14

  interfering nucleic acids [NA] · CPC title

• A61K2121/00

  Preparations for use in therapy · CPC title