Granted Patent B2

US 12,496,357 · App. 17/631,661 · Granted Dec 16, 2025

Inventors: Peter Glazer (Guilford, CT); Elias Quijano (Durham, CT); W. Mark Saltzman (New Haven, CT); Hee Won Suh (New Haven, CT)

Assignee: Yale University

A61K48/0066A61K9/5146A61K9/5153A61K48/0033C12N15/113C12N2310/15C12N2310/152C12N2310/3181

Abstract

Claims (26)

1 . A nanoparticle comprising

a core comprising poly(lactic acid) (PLA) comprising a number average molecular weight (Mn) of about 12 kDa;

a shell comprising a hyperbranched polyglycerol (HPG) comprising a number average molecular weight (Mn) of about 5 kDa; and

a plurality of a peptide nucleic acid (PNA) oligomer entrapped or encapsulated therein, the PNA oligomer comprising

a Hoogsteen binding peptide nucleic acid segment and a Watson-Crick binding PNA segment collectively totaling no more than 50 nucleobases in length,

wherein the two segments can bind or hybridize to a target region in genomic DNA and induce strand invasion, displacement, and formation of a triple-stranded molecule among the two PNA segments and the target region.

2 . The nanoparticle of claim 1 , wherein the Hoogsteen binding peptide nucleic acid segment comprises PNA residues comprising the nucleic acid sequence TTJJTJT and the Watson-Crick binding PNA segment comprises PNA residues comprising the nucleic acid sequence TCTCCTTAAACCTGT (SEQ ID NO: 1), TCTCCTTAAACCTGTCTT (SEQ ID NO:2), wherein “J” is pseudoisocytosine.

3 . The nanoparticle of claim 2 , wherein the PNA oligomer comprises the nucleic acid sequence TTJJTJT-linker-TCTCCTTAAACCTGT (SEQ ID NO: 3) or TTJJTJT-linker-TCTCCTTAAACCTGTCTT (SEQ ID NO:4), wherein “linker” is a flexible linker, linking the Hoogsteen binding segment to the Watson-Crick binding segment and “J” is pseudoisocytosine.

4 . The nanoparticle of claim 3 , wherein the PNA oligomer is free from gamma (γ) modifications in the PNA residues.

5 . The nanoparticle of claim 1 , wherein one or more of the PNA residues in the Hoogsteen binding segment, the Watson-Crick binding segment, or a combination thereof comprises a substitution at the gamma (γ) position of one or more PNA residues.

6 . The nanoparticle of claim 5 , wherein the PNA oligomer comprises the nucleic acid sequence lys-lys-lys-TTJJTJT-OOO- C C T A A C G C T-lys-lys-lys (SEQ ID NO:5), wherein the bolded and underlined residues are γ modified PNA and “J” is pseudoisocytosine.

7 . The nanoparticle of claim 6 , wherein the γ modification is serine.

8 . The nanoparticle of claim 1 , wherein core is poly (lactic acid) (PLA).

9 . The nanoparticle of claim 8 , wherein the shell comprises a low molecular weight HPG.

10 . The nanoparticle of claim 9 , wherein the number average molecular weight (Mn) of the PLA is 12.7 kDa, and the number average molecular weight (Mn) of the HPG is 5.4 kDa.

11 . The nanoparticle of claim 1 , wherein the conjugation ratio is about 1 and is optionally 1.3, the HPG (wt %) is about 40% and is optionally 38%, or a combination thereof.

12 . The nanoparticle of claim 11 , where the PNA oligomer comprises the nucleic acid sequence lys-lys-lys-TTJJTJT-OOO-TCTCCTTAAACCTGTCTT-lys-lys-lys (SEQ ID NO:5), wherein “J” is pseudoisocytosine.

13 . The nanoparticle of claim 12 , wherein the PNA oligomer comprises the nucleic acid sequence lys-lys-lys-TTJJTJT-OOO- C C T A A C G C T-lys-lys-lys (SEQ ID NO:5), wherein the bolded and underlined residues are serine γ modified PNA and “J” is pseudoisocytosine.

14 . The nanoparticle of claim 1 further comprising a donor oligonucleotide entrapped or encapsulated therein, the donor oligonucleotide comprising a sequence that can correct a mutation(s) in a cell's genome by recombination induced or enhanced by the peptide nucleic acid oligomer.

15 . A pharmaceutical composition comprising an effective amount of the nanoparticle of claim 1 .

16 . The pharmaceutical composition of claim 15 , further comprising a potentiating agent to increase the frequency of recombination of the donor oligonucleotide at a target site in the genome of a population of cells.

17 . The pharmaceutical composition of claim 16 , wherein the potentiating agent is selected from vorinostat, 2,4-Pyridinedicarboxylic Acid, Garcinol, Splitomycin, BML-210, Apicidin, Suberoyl bis Hydroxamic acid, Scriptaid, Nullscript, 5-Aza-2-deoxycitidine, Zebularine, ITSA1, Phenylbutyrate Na, Tranylcypromine, Valproic acid, EX-527, Resveratrol, M-344, Nicotinamide, BML-266, Piceatannol, AGK2, Salermide, MC-1293, Anacardic acid, B2, BIX-01294, Butyrolactone 3, CTPB, Oxamflatin, Sirtinol, NCH-51, CI-994, NSC-3852, Aminoresveratrol, BML-281, Triacetylresveratrol, Trichostatin A, Iso-Nicotinamide, Valproxam, BML-278, SAHA, Fluoro-SAHA, and Suramin Na.

18 . The pharmaceutical composition of claim 16 , wherein the PNA oligomer comprises the nucleic acid sequence lys-lys-lys-TTJJTJT-OOO-TCTCCTTAAACCTGTCTT-lys-lys-lys (SEQ ID NO:5), wherein “J” is pseudoisocytosine;

the number average molecular weight (Mn) of the PLA is about 12 kDa and optionally is 12.7 kDa, and the number average molecular weight (Mn) of the HPG is about 5 kDa, and optionally is 5.4 kDa; and

the potentiating agent is vorinostat, RI-1, B0-2, or 3E10 antibody or a fragment thereof.

19 . The pharmaceutical composition of claim 18 , wherein the PNA oligomer comprises the nucleic acid sequence lys-lys-lys-TTJJTJT-OOO- C C T A A C G C T-lys-lys-lys (SEQ ID NO:5), wherein the bolded and underlined residues are serine γ modified PNA and “J” is pseudoisocytosine.

Assignments (1)

Continuity (2)

References Cited (256)