Nanocomplexes of polyanion-modified proteins

Abstract

A nanocomplex, 50 to 1000 nm in size, containing a lipid-like nanoparticle formed of a cationic lipid-based compound and a modified protein formed of a protein and an anionic polymer that includes a plurality of polar groups, the lipid-like nanoparticle and the modified protein being non-covalently bonded to each other. Also disclosed are a method of preparing the above-described nanocomplex and use thereof for treating a medical condition. Further disclosed is a pharmaceutical composition containing a nanocomplex.

1 - 22 . (canceled) 
     
     
         23 . A nanocomplex, comprising:
 i) a protein comprising an anionic polymer, and   ii) a lipid-like nanoparticle, comprising a cationic lipid-based compound formed from an amine headgroup, wherein the amine headgroup is selected from the group consisting of   
       
         
           
           
               
               
           
         
       
       wherein the protein is non-covalently assembled with the lipid-like nanoparticle; and 
       wherein the nanocomplex is characterized by a particle size of 50 nanometers (nm) to 1000 nm. 
     
     
         24 . The nanocomplex of  claim 23 , wherein the protein is a protein-based cytotoxin, an antibody, a transcription factor, or a genome-editing protein. 
     
     
         25 . The nanocomplex of  claim 23 , wherein the protein is a protein-based cytotoxin, and the protein-based cytotoxin is an RNase, saporin, gelonin, ricin chain A, shiga toxin chain A1, or botulinum neurotoxin. 
     
     
         26 . The nanocomplex of  claim 24 , wherein the protein is a genome-editing protein, and wherein the genome-editing protein is a Cas9 or Cpf1. 
     
     
         27 . The nanocomplex of  claim 23 , wherein the protein-based cytotoxin is an RNase. 
     
     
         28 . The nanocomplex of  claim 23 , wherein the anionic polymer comprises a plurality of polar groups selected from the group consisting of CO2H, CO2-, SO3H, SO3-, PO3H, and PO3-. 
     
     
         29 . The nanocomplex of  claim 23 , wherein the anionic polymer is selected from the group consisting of hyaluronic acid, heparin, DNA, RNA, polysialic acid, polyglutamic acid, pentosan polysulfate sodium, sulphated polysaccharide, negatively charged serum albumin, negatively charged milk protein, synthetic sulphated polymer, polymerized anionic surfactant, and polyphosphate. 
     
     
         30 . The nanocomplex of  claim 23 , wherein the anionic polymer is hyaluronic acid or heparin. 
     
     
         31 . The nanocomplex of  claim 23 , wherein the cationic lipid-based compound is formed from an electrophile and an amine headgroup. 
     
     
         32 . The nanocomplex of  claim 23  wherein the electrophile is an epoxide, an acrylate, or an acrylamide. 
     
     
         33 . The nanocomplex of  claim 23 , wherein the electrophile is an epoxide. 
     
     
         34 . The nanocomplex of  claim 33 , wherein the electrophile is an epoxide substituted with C1-C20 alkyl or C1-C20 heteroalkyl. 
     
     
         35 . The nanocomplex of  claim 23 , wherein the electrophile is 
       
         
           
           
               
               
           
         
       
     
     
         36 . A pharmaceutical composition, comprising a nanocomplex of claim  0 ; and a pharmaceutically acceptable carrier thereof. 
     
     
         37 . A method of treating a medical condition, comprising administering to a subject in need thereof an effective amount of a nanocomplex of  claim 23 . 
     
     
         38 . The method of  claim 23 , wherein the medical condition comprises a cancer. 
     
     
         39 . The method of  claim 38 , wherein the cancer is breast cancer, prostate cancer, ovarian cancer, or leukemia. 
     
     
         40 . The method of  claim 38 , wherein the cancer is associated with a cell comprising high levels of CD44 expression.