Biologically active synthetic nanoparticle constructs and methods of use thereof

1. A biologically active synthetic nanoparticle construct comprising a polypeptide-dendrimer nanoparticle, and a plurality of moieties attached to a surface of the polypeptide-dendrimer nanoparticle, wherein the moieties comprise:

i. a plurality of single copies of DNA binding domains,

ii. a plurality of single copies of nuclear localization signals, and

iii. a plurality of single copies of transcriptional domains,

wherein the DNA binding domains are each individually covalently attached to the surface of the polypeptide-dendrimer nanoparticle,

wherein the nuclear localization signals are each individually covalently attached to the surface of the polypeptide-dendrimer nanoparticle, and

wherein the transcriptional domains are each individually covalently attached to the surface of the polypeptide-dendrimer nanoparticle.

2. The biologically active synthetic nanoparticle construct of claim 1 wherein:

i. X % of the moieties on the polypeptide-dendrimer nanoparticle are DNA binding domains, wherein X % ranges from 10 to 70%;

ii. Y % of the moieties on the polypeptide-dendrimer nanoparticle are transcriptional repression domains, wherein Y % ranges from 10 to 70%;

iii. Z % of the moieties on the polypeptide-dendrimer nanoparticle are nuclear localization signals, wherein Z % ranges from 10 to 70%; and

iv. the sum of X, Y, and Z % is not greater than 100%.

3. The biologically active synthetic nanoparticle construct of claim 2 ,

wherein Z % is at least 50%.

4. The biologically active synthetic nanoparticle construct of claim 1 , wherein the moieties further comprise a plurality of at least one conjugant;

wherein W % of the moieties on the polypeptide-dendrimer nanoparticle are the conjugants, wherein W % ranges from 0 to 40%; and

wherein the sum of W, X, Y and Z % is not greater than 100%.

5. The biologically active synthetic nanoparticle construct of claim 1 wherein the polypeptide-dendrimer nanoparticle comprises:

i. a hydrophobic core comprising a dendrimer,

wherein the dendrimer comprises an amide linker and an amine functionalization suitable for N-carboxyanhydride polymerization;

ii. a hydrophobic inner shell comprising an inner shell polypeptide,

wherein the hydrophobic inner shell at least partially surrounds the hydrophobic core; and

iii. a hydrophilic outer shell comprising an outer shell polypeptide,

wherein the hydrophilic outer shell at least partially surrounds the hydrophobic inner shell.

6. The biologically active synthetic nanoparticle construct of claim 5 wherein the dendrimer comprises a polyamidoamine.

7. The biologically active synthetic nanoparticle construct of claim 5 wherein the inner shell polypeptide comprises polymerized hydrophobic amino acids selected from the group consisting of glycine, alanine, valine, leucine, isoleucine, phenylalanine, methionine, tryptophan, cysteine, and a combination thereof.

8. The biologically active synthetic nanoparticle construct of claim 5 wherein the outer shell polypeptide comprises polymerized alpha amino acids.

9. The biologically active synthetic nanoparticle construct of claim 8 wherein the alpha amino acids are selected from the group consisting of lysine, glutamic acid, aspartic acid, and a combination thereof.

10. The biologically active synthetic nanoparticle construct of claim 5 wherein the outer shell polypeptide has an isoelectric point of from at least 6.5 to no more than 8.

11. The biologically active synthetic nanoparticle construct of claim 5 , wherein the outer shell polypeptide comprises a functional group selected from the group consisting of an amine, a carboxyl, a phosphonate, a sulfonate, and a combination thereof.

12. The biologically active synthetic nanoparticle construct of claim 5 , wherein the outer shell polypeptide has a functional group consisting of sulfosuccinimidyl 4-(N-maleimido-methyl)cyclohexane-1-carboxylate.

13. The biologically active synthetic nanoparticle construct of claim 1 wherein the polypeptide-dendrimer nanoparticle is loaded with at least one of, hydrophobic small molecules, oligonucleotides, and ions.

14. The biologically active synthetic nanoparticle construct of claim 13 wherein the ion is selected from the group consisting of gadolinium, manganese, rhenium, and a combination thereof.

15. The biologically active synthetic nanoparticle construct of claim 1 wherein the polypeptide-dendrimer nanoparticle is prepared by the process of:

i. performing a Michael addition and amidation reaction between an amine and an acrylate to form a hydrophobic dendrimer core having an outer surface;

ii. polymerizing a plurality of hydrophobic amino acids, by N-carboxyanhydride polymerization, in the presence of the hydrophobic dendrimer core, to form a hydrophobic polypeptide inner shell having an outer surface,

wherein the hydrophobic polypeptide inner shell at least partially surrounds the outer surface of the hydrophobic core; and

iii. polymerizing a plurality of hydrophilic alpha amino acids, by N-carboxyanhydride polymerization, in the presence of the hydrophilic inner shell, to form a hydrophilic polypeptide outer shell having an outer surface,

wherein the hydrophilic polypeptide outer shell at least partially surrounds the outer surface of the hydrophobic inner shell.

16. The biologically active synthetic nanoparticle construct of claim 15 , wherein the amine comprises tri(2-aminoethyl)amine, and the acrylate comprises methyl acrylate.

17. The biologically active synthetic nanoparticle construct of claim 1 wherein the DNA binding domain is selected from the group consisting of hairpin polyamides, zinc finger domains, triplex-forming oligonucleotides (TFOs), transcription activator-like effectors, oligonucleotide analogs, locked-nucleic acids, peptide nucleic acids, and a combination thereof.

18. The biologically active synthetic nanoparticle construct of claim 1 ,

wherein the DNA binding domains are hairpin polyamides, and

wherein the hairpin polyamides comprise at least one N-methyl-imidazole moiety or at least one N-methyl pyrrole moiety, or combinations thereof, arranged sequentially on the polyamide to bind a target gene.

19. The biologically active synthetic nanoparticle construct according to claim 1 , wherein the transcriptional domains are transcriptional activation domains.

20. The biologically active synthetic nanoparticle construct according to claim 19 , wherein the transcriptional activation domains are selected from the group consisting of peptoids, amphipathic isoxasolidine, wrenchnolol, amphipathic helix peptides and combinations thereof.

21. The biologically active synthetic nanoparticle construct according to claim 1 , wherein the transcriptional domains consist of peptides.

22. The biologically active synthetic nanoparticle construct of claim 1 wherein the nuclear localization signals are derived from a SV-40 antigen, derived form an HIV-1 antigen, or derived from the group consisting of: TAT, Penetratin, MAP, Transportin/TP10, VP22, MPG, Pep1, pVEC, YTA2, YTA4, M918, CADY, and combinations thereof.

23. The biologically active synthetic nanoparticle construct according to claim 1 , wherein the transcriptional domains are transcriptional repression domains.