Nano delivery systems for sirna

Abstract

The present invention makes use of a unique methodology of double nano-encapsulation for protecting and controlling the release of active agents, either hydrophobic or hydrophilic, from stable nanoparticles of opposite characteristics. The protection of the active agent was achieved by loading the agent to be protected, into nanocarriers, which were subsequently encapsulated into sub-micron nanoparticles. The sub-micron nanoparticles formation has been successfully achieved by the use of novel nanospray techniques.

1 . A polymeric nanoparticle encapsulating a plurality of nanocarriers, at least a portion of the nanocarriers containing at least one active agent, the nanoparticle having an averaged diameter of between 400 and 950 nm, wherein the plurality of nanocarriers being made of a material such that
 (i) where the active agent is hydrophobic, the nanocarrier material is hydrophobic and the nanoparticle material is hydrophilic;   (ii) where the active agent is hydrophilic, the nanocarrier material is hydrophilic and the nanoparticle material is hydrophobic;   (iii) where the active agent is hydrophobic, the nanocarrier material is hydrophobic and the nanoparticle material is hydrophobic; and   (iv) where the active agent is hydrophilic, the nanocarrier material is hydrophilic, and the nanoparticle material is hydrophilic.   
     
     
         2 . The nanoparticle of  claim 1 , wherein the nanoparticles and/or the nanocarriers are each obtainable by nanospraying. 
     
     
         3 . The nanoparticle of  claim 1 , wherein the nanoparticle and/or the nanocarrier are each cross-linked. 
     
     
         4 . The nanoparticle of  claim 1 , wherein the nanocarrier further comprises a polycationic lipid being 1,2-dioleoyl-3-trimethylammonium-propane (DOTAP). 
     
     
         5 . The nanoparticle of  claim 1 , wherein the average diameter of said nanocarrier is less about 300 nm. 
     
     
         6 . The nanoparticle of  claim 1 , wherein the nanoparticle and/or the nanocarrier being each in the form selected from a nanocapsule or a nanosphere. 
     
     
         7 . The nanoparticle of  claim 1 , wherein said hydrophobic material is selected from the group consisting of lactic acid, poly(D,L-lactic-co-glycolic acid) (PLGA), poly(D,L-lactic acid) (PLA), poly(ε-caprolactone), poly(2-dimethylamino-ethylmethacrylate) homopolymer, poly(2-dimethylamino-ethylmethacrylate)-b-poly(ethyleneglycop-α-methoxy-ω-metacrylate copolymers, polycyanoacrylates, polyanhydride polymers and combinations thereof. 
     
     
         8 . The nanoparticle of  claim 7 , wherein said PLGA has a molecular weight of between about 4,000 and 100,000 Da. 
     
     
         9 . The nanoparticle of  claim 1 , wherein said nanoparticle material is hydrophobic, an outer surface of said nanoparticle being associated with at least one polyethylene glycol (PEG) moiety. 
     
     
         10 . The nanoparticle of  claim 1 , wherein said hydrophilic material is selected from the group consisting of dextran, hylauronate, human serum albumin (HSA), being normal or cross-linked, bovine serum albumin (BSA) being normal or cross-linked, chitosan, shellac, collagen, gelatin, gum arabic, polyvinyl alcohol, cyclodextrin, and combinations thereof. 
     
     
         11 . The nanoparticle of  claim 1 , wherein said active agent is selected from a vitamin, a protein, an anti-oxidant, a nucleic acid, a short or long oligonucleotide, a siRNA and its chemical derivatives, a peptide, a polypeptide, a lipid, a carbohydrate, a hormone, an antibody, a monoclonal antibody, a vaccine, a prophylactic agent, a drug, a diagnostic agent, a contrasting agent, a nutraceutical agent, a small molecule, an electrolyte, an immunological agent and combination thereof. 
     
     
         12 . The nanoparticle of  claim 11 , wherein the nanocarrier comprises at least one hydrophilic active agent selected from the group consisting of exenatide, insulin, growth hormone, triptorelin acetate, buserelin, and nafarelin. 
     
     
         13 . The nanoparticle of  claim 11 , wherein the nanocarrier comprises at least one hydrophobic active agent selected from the group consisting of an analgesic or anti-inflammatory agent; an enthelmintic agent; an anti-arrhythmic agent; an anti-bacterial agent; an anti-coagulant; an anti-depressant; an antidiabetic; an anti-epileptic; an anti-fungal agent; an anti-gout agent; an anti-hypertensive agent; an anti-malarial agent; an anti-migraine agent; an anti-muscarinic agent; an anti-neuroplastic agent or immunosuppressant; an anti-protazoal agent; an anti-thyroid agent; an alixiolytic, sedative, hypnotic or neuroleptic agent; a beta-blocker; a cardiac inotropic agent; a corticosteroid; a diuretic agent; an anti-Parkinsonian agent; a gastro-intestinal agent; an histamine H1-receptor antagonist; a lipid regulating agent; a nitrate or anti-anginal agent; a nutritional agent; an HIV protease inhibitor; an opioid analgesic; a sex hormone; and a stimulant agent. 
     
     
         14 . The nanoparticle of  claim 1 , having an outer surface associated with at least one targeting agent, optionally selected from the group consisting of
 a monoclonal antibody; a small molecule; hyaluronic acid or hyaluronan; tumor penetrating peptides; epidermal growth factor (EGF); transferrin; ferritin; Arginine-Glycine-Aspartic acid (RGD) peptide; epithelial cell adhesion molecule (EpCAM); intercellular adhesion molecule 1 (ICAM-1); carcinoembrionic antigen (CEA); vasoactive intestinal peptide; CA 15-3 antigen; MUC1 protein; CD20; CD33; integrins; lymphatic targeting moieties (such as LyP-1); aptamers; and oligosaccharides.   
     
     
         15 . A composition comprising a plurality of nanoparticles of  claim 1 . 
     
     
         16 . A process for encapsulating a plurality of nanocarriers in a polymeric nanoparticle, the process comprising:
 (a) obtaining at least one nanocarrier, said nanocarrier comprising at least one active agent; and   (b) encapsulating a plurality of nanocarriers into said nanoparticle by nanospray drying.   
     
     
         17 . The process of  claim 16 , wherein the nanocarriers are obtained by:
 dissolving a hydrophobic polymer in organic solvent to form an organic phase;   contacting said organic phase with an aqueous phase, the aqueous phase optionally comprising a surfactant, to thereby obtain said nanocarriers; and   incubating said nanocarriers with a solution of said active agent to allow association of said active agent with at least a portion of the surface of said nanocarriers.   
     
     
         18 . The process of  claim 16 , wherein the nanocarriers are obtained by:
 dissolving a hydrophilic polymer in an aqueous solution of the active agent to form an aqueous phase; and   continuously adding an organic phase comprising a desolvating agent to the aqueous phase under a pH permitting the formation said nanocarriers, the active agent being distributed within the nanocarrier.   
     
     
         19 . The process of  claim 16 , further comprising lyophilizing the nanoparticle.