Cross-linked polymer modified nanoparticles

Abstract

Disclosed herein are nanoconstructs comprising a nanoparticle, coated with additional agents such as cationic polymers, stabilizers, targeting molecules, labels, oligonucleotides and small molecules. These constructs may be used to deliver compounds to treat solid tumors and to diagnose cancer and other diseases. Further disclosed are methods of making such compounds and use of such compounds to treat or diagnose human disease.

We claim: 
     
         1 . A composition comprising:
 a plurality of multilayer nanoconstructs each comprising:
 a silica nanoparticle, a silicon nanoparticle, a silver nanoparticle, or a carbon nanotube nanoparticle; 
 polyethylenimine (PEI) bound to an exterior surface of the nanoparticle; and 
 polyethylene glycol (PEG) bound to the PEI and/or to the nanoparticle; 
   and   a protein or peptide loaded on the nanoconstructs,   
       wherein the nanoconstructs loaded with the protein or peptide have a hydrodynamic size Z-average diameter of about 200 nm or less. 
     
     
         2 . The composition of  claim 1 , wherein the nanoconstructs loaded with the protein or peptide have a polydispersity index (PDI) of no more than about 0.37. 
     
     
         3 . The composition of  claim 1 , wherein the PDI of the nanoconstructs loaded with the protein or peptide is about 0.2. 
     
     
         4 . The composition of  claim 1 , wherein the nanoparticle is a silica nanoparticle. 
     
     
         5 . The composition of  claim 4 , wherein the silica nanoparticle is a mesoporous silica nanoparticle (MSNP). 
     
     
         6 . The composition of  claim 1 , wherein the nanoparticle is a silicon nanoparticle. 
     
     
         7 . The composition of  claim 1 , wherein the nanoparticle is a silver nanoparticle. 
     
     
         8 . The composition of  claim 1 , wherein the nanoparticle is a carbon nanotube. 
     
     
         9 . The composition of  claim 1 , wherein the plurality of nanoparticles consists essentially of materials selected from the group consisting of: silica, silicon, silver, carbon nanotubes, and combinations of two or more thereof. 
     
     
         10 . The composition of  claim 1 , wherein the PEI of the nanoconstructs is from about 10% to about 30% by weight of the nanoconstructs. 
     
     
         11 . The composition of  claim 1 , wherein the PEI is cross-linked. 
     
     
         12 . The composition of  claim 11 , wherein the PEI is cross-linked using a cleavable cross-linker. 
     
     
         13 . The composition of  claim 1 , wherein the PEI has an average size of 1.8 kDa to 25 kDa. 
     
     
         14 . The composition of  claim 1 , wherein the protein or peptide is conjugated to the functional PEG. 
     
     
         15 . The composition of  claim 1 , wherein the protein or peptide is electrostatically bound to the PEI. 
     
     
         16 . The composition of  claim 1 , wherein the nanoparticles are from about 10 nm to about 90 nm in diameter. 
     
     
         17 . The composition of  claim 1 , wherein the hydrodynamic size Z-average diameter of the nanoconstructs is about 10 nm to about 200 nm. 
     
     
         18 . The composition of  claim 1 , wherein the hydrodynamic size Z-average diameter of the nanoconstructs is about 50 nm to about 200 nm. 
     
     
         19 . The composition of  claim 1 , wherein the hydrodynamic size Z-average diameter of the nanoconstructs is about 90 nm to about 120 nm. 
     
     
         20 . The composition of  claim 1 , wherein protein or peptide is a therapeutic agent. 
     
     
         21 . The composition of  claim 20 , wherein therapeutic agent comprises an antibody or an antigen-binding fragment thereof. 
     
     
         22 . The composition of  claim 21 , wherein therapeutic antibody comprises a monoclonal antibody. 
     
     
         23 . The composition of  claim 1 , wherein the protein or peptide comprises an antibody or an antigen-binding fragment thereof. 
     
     
         24 . The composition of  claim 1 , wherein the protein or peptide comprises a scFv antibody, a monoclonal antibody, an affibody, an aptamer, a peptide, a ligand, or small targeting molecule. 
     
     
         25 . The composition of  claim 1 , wherein the protein or peptide comprises a cytokine. 
     
     
         26 . The composition of  claim 1 , wherein the hydrodynamic diameter of the nanoconstructs loaded with the protein or peptide is about 50 nm to about 100 nm. 
     
     
         27 . The composition of  claim 1 , wherein the nanoconstructs further comprise a small molecule. 
     
     
         28 . The composition of  claim 27 , wherein the small molecule comprises:
 a chemotherapeutic agent,   an oligonucleotide,   a small molecule inhibitor, or   a label.   
     
     
         29 . The composition of  claim 1 , wherein the plurality of nanoconstructs comprises a targeting agent. 
     
     
         30 . The composition of  claim 1 , wherein the protein or peptide is a targeting agent. 
     
     
         31 . The composition of  claim 1 , further comprising trehalose. 
     
     
         32 . The composition of  claim 31 , wherein the trehalose is from about 1% to about 10% by weight of the plurality of nanoconstructs. 
     
     
         33 . The composition of  claim 1 , further comprising a label attached to the nanoconstructs. 
     
     
         34 . The composition of  claim 33 , wherein the label comprises at least one of a lanthanide, a fluorescent dye, a gold nanoparticle, a quantum dot, a PET tracer, or a MRI contrast agent. 
     
     
         35 . A method of labeling a target, comprising contacting the composition of  claim 37  with the target under conditions to bind the nanoconstruct to the target. 
     
     
         36 . The method of  claim 35 , wherein the contacting occurs ex vivo. 
     
     
         37 . The method of  claim 35 , wherein the contacting occurs in vivo in a subject. 
     
     
         38 . The method of  claim 35 , wherein the label comprises at least one of a lanthanide, a fluorescent dye, a gold nanoparticle, a quantum dot, a PET tracer, or a MRI contrast agent. 
     
     
         39 . The method of  claim 35 , further comprising:
 quantifying the amount of target by detecting the label after the nanoconstruct binds to the target.   
     
     
         40 . The method of  claim 35 , further comprising administering the labeled nanoconstruct to a subject and detecting the location of the labeled nanoconstruct after the administering. 
     
     
         41 . A method of delivering a protein or peptide to a site in a human or other mammalian subject, the method comprising administering an effective amount of the composition of  claim 1  to the human or other mammalian subject under conditions to deliver the nanoconstruct to the site. 
     
     
         42 . The method of  claim 41 , wherein the site is a cell. 
     
     
         43 . The method of  claim 42 , wherein the nanoconstruct is administered under conditions that the nanoconstruct is internalized by the cell. 
     
     
         44 . The method of  claim 41 , wherein the site is a tumor cell. 
     
     
         45 . The method of  claim 41 , wherein the protein or peptide is capable of modulating expression of a target protein. 
     
     
         46 . The method of  claim 41 , wherein the subject is diagnosed with cancer or diagnosed with or is at risk for fibrosis or inflammation, and the effective amount is a therapeutically effective amount. 
     
     
         47 . The method of  claim 41 , wherein the nanoconstruct further comprises a targeting agent. 
     
     
         48 . The method of  claim 41 , wherein administration of the composition reduces reactive oxygen species, bioavailable copper, and/or NOX expression level in the subject or modulates an adverse effect of one or more cytokines.