Therapeutic nanoparticles and related compositions, methods and systems

Abstract

Disclosed are targeted sub-50 nanometer nanoparticles suitable for delivering bioactive agents of interest, and related compositions, methods, and systems, which improve the manufacturing, stability, efficacy and other aspects of therapeutic nanoparticles.

1 . A composition comprising nanoparticles, wherein the nanoparticles comprise: at least one bioactive agent, a surfactant having an HLB value of less than 6.0 units, a ligand, and Li+ and Cs+, wherein:
 i) the at least one bioactive agent and the surfactant form a surfactant micelle core;   ii) the ligand forms a shell; and   iii) the nanoparticles have an average diameter of less than about 50 nanometers.   
     
     
         2 . The composition of  claim 1 , wherein the nanoparticles are prepared using sterile water. 
     
     
         3 . The composition of  claim 1 , wherein the at least one bioactive agent is a polynucleotide or a plasmid DNA. 
     
     
         4 . The composition of  claim 1 , wherein:
 i) the at least one bioactive agent comprises a plurality of polynucleotides, each comprising a 3′ RNA portion and a 5′ primarily DNA portion, wherein the number 2 position from the 5′ end of each polynucleotide is a 2′-OMe modified RNA, wherein the sequence of more than about 40% and less than about 60% of the plurality of polynucleotides on average comprises SEQ ID NO: 8, and the sequence of the remainder of the plurality of polynucleotides on average comprises SEQ ID NO: 9, and   ii) the ligand is a protein targeting a tenascin receptor.   
     
     
         5 . The composition of  claim 4 , wherein the protein is tenfibgen. 
     
     
         6 . A method of administering at least one bioactive agent to a subject, the method comprising administering to the subject the composition of  claim 1 . 
     
     
         7 . A method for treating a patient with a solid tumor cancer, comprising administering to the patient a therapeutically effective amount of the composition of  claim 1 , wherein
 i) the at least one bioactive agent comprises a plurality of polynucleotides, each comprising a 3′ RNA portion and a 5′ primarily DNA portion, wherein the number 2 position from the 5′ end of each polynucleotide is a 2′-OMe modified RNA, wherein the sequence of more than about 40% and less than about 60% of the plurality of polynucleotides on average comprises SEQ ID NO: 8, and the sequence of the remainder of the plurality of polynucleotides on average comprises SEQ ID NO: 9, and   ii) the ligand is a protein targeting a tenascin receptor.   
     
     
         8 . A method for preparing the composition of  claim 1 , the method comprising:
 i) complexing at least one bioactive agent with a condensing agent to form a condensed bioactive agent;   ii) dispersing the condensed bioactive agent into a water-miscible solvent comprising a surfactant with an HLB of less than 6.0 to form a surfactant micelle;   iii) adsorbing a ligand to the exterior surface of the surfactant micelle to form a ligand particle; and   iv) mixing and incubating the ligand particle with (a) Li+ pre-treated with Cs+ and (b) sterile water to form the composition.   
     
     
         9 . The composition of  claim 1 , wherein the Li+ is pretreated with Cs+. 
     
     
         10 . The composition of  claim 1 , wherein the ligand comprises a protein, a peptide, a polypeptide, a carbohydrate, polyvinylpyrrolidone (PVP), an antibody, or a biocompatible polymer, or fragments thereof, or a small molecule. 
     
     
         11 . The method of  claim 8 , wherein the at least one bioactive agent is a polynucleotide or plasmid DNA.