Highly stable biodegradable gene vector platforms for overcoming biological barriers
A major challenge in non-viral gene delivery remains finding a safe and effective delivery system. Colloidally stable non-viral gene vector delivery systems capable of overcoming various biological barriers, are disclosed. The gene vectors are biodegradable, non-toxic and highly tailorable for use in specific applications. The vectors include a mixture of biodegradable copolymers, such as PBAE, and biodegradable polymers conjugated with hydrophilic, neutrally charged polymer, such as PEG. The gene vectors demonstrate broad vector distribution and high transgene delivery in vivo, providing an efficient non-viral gene delivery system for localized therapeutic gene transfer. Methods of using the vectors to overcome biological barriers including mucus gel and extracellular matrix are provided. Methods of formulating the vectors are also provided.
1. A formulation of colloidally stable nanoparticles for delivery of nucleic acids across biological barriers, the nanoparticles comprising
(a) a polymer blend comprising poly (β-amino ester) (PBAE) and PBAE conjugated to a hydrophilic, neutrally charged polymer comprising polyethylene glycol or a copolymer thereof,
wherein the ratio of non-PEGylated PBAE to PBAE conjugated to polyethylene glycol or a copolymer thereof is between 25 and 1, and
and an effective amount of the polyethylene glycol is on the surface of the particle to create a near neutral surface charge, and
(b) nucleic acids complexed with the non-pegylated PBAE,
wherein the nucleic acids are encapsulated within the nanoparticle or are associated with the surface of the nanoparticle, and
the nanoparticles are less than 100 nm in size.
2. The formulation of claim 1 , wherein the mass ratio of PBAE to nucleic acid is up to 100, and wherein the concentration of the PBAE blend is up to 2,000 times the concentration of the nucleic acids.
3. The formulation of claim 2 , wherein the PBAE has a molecular weight between 4,000 and 7,000 Daltons.
4. The formulation of claim 1 , wherein the polyethylene glycol has a molecular weight between 1,000 Daltons and 10,000 Daltons.
5. The formulation of claim 4 , wherein the polyethylene glycol has a molecular weight of 5,000 Daltons.
6. The formulation of claim 1 , wherein the diameter, polydispersity and surface charge of the nanoparticle are stable for at least one week in aqueous solution.
7. The formulation of claim 1 , wherein the diameter, polydispersity index and surface charge of the nanoparticle are stable following lyophilization.
8. The formulation of claim 1 , wherein the molar ratio of the PEG to PBAE is between 0.5 and 50.
9. The formulation of- claim 8 ,
wherein the molar ratio of the PEG to PBAE is 1.2.
10. A dosage formulation for delivery of nucleic acids comprising
a. a therapeutically effective amount of the formulation of nanoparticles of claim 1 ; and
b. a pharmaceutically acceptable excipient for administration to an individual in need thereof.
11. The formulation of claim 10 , wherein the nanoparticles are formulated for administration to the brain.
12. The formulation of claim 10 , wherein the nanoparticles are formulated for administration to a mucosal layer.
13. The formulation of claim 10 , wherein the nanoparticles release an effective amount of the one or more nucleic acids over a period of at least 10 minutes, 20 minutes, 30 minutes, one hour, two hours, four hours, six hours, ten hours, one day, three days, seven days, ten days, two weeks, one month, or longer to treat a symptom of a disease or disorder.
14. A method for treating a disease or disorder, comprising administering to a subject in need thereof a formulation comprising
a therapeutically effective amount of the formulation of nanoparticles of claim 1 ; and
a pharmaceutically acceptable excipient for delivery.
15. The method of claim 14 , wherein the formulation is administered to the brain.
16. The method of claim 14 , wherein the formulation is administered to a mucosal layer.
17. The method of claim 15 , wherein the formulation of particles are administered in combination with one or more techniques to facilitate passage of the particles through the blood brain barrier.
18. The method of claim 17 , wherein the technique is selected from the group consisting of topical bolus administration, convection enhanced delivery, electron paramagnetic resonance, ultrasound, and ultrasound plus microbubbles.
19. The method of claim 15 , wherein the disease or disorder is selected from the group consisting of tumors, neurological disorders, and brain injury or trauma.
20. The method of claim 16 , wherein the disease or disorder is selected from the group consisting of tumors, lung injury or trauma, cystic fibrosis, diseases caused by infection with bacteria and diseases caused by infections with viruses.
21. The method of claim 14 , wherein the nanoparticles have been lyophilized or stored in an aqueous solution.