Nanozymes, methods of making nanozymes, and methods of using nanozymes

Patent metadata

Field	Value
Publication number	US-10538757-B2
Application number	US-201615042935-A
Country	US
Kind code	B2
Filing date	Feb 12, 2016
Priority date	Aug 14, 2013
Publication date	Jan 21, 2020
Grant date	Jan 21, 2020

Abstract

Embodiments of the present disclosure provide for nanozymes that can include a therapeutic agent, methods of making nanozymes, methods of using nanozymes, and the like. In some embodiments, the nanozymes can include a shell that can surround a hollow core that can be configured to receive a compound and the shell can include a recognition moiety and an enzyme.

First claim

We claim at least the following:

1. A nanozyme comprising:
a shell surrounding a hollow core configured to receive a compound, wherein the shell comprises:
a single stranded DNA (ssDNA) recognition moiety selected from the group consisting of: a polynucleotide set forth in SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 16, SEQ ID NO: 17, and combinations thereof,
wherein the ssDNA recognition moiety comprises an alkylthiol-terminated propargyl ether modified poly-thymine polynucleotide; wherein the alkylthiol-terminated, propargyl ether modified poly-thymine polynucleotide comprises one or more propargyl ether groups coupled via an amidohexylacrylamido linker to the 5′ position of one or more thymine bases, and wherein the ssDNA recognition moiety specifically binds, through reverse complementary binding, a polynucleotide selected from the group consisting of: a c-MET mRNA, a conserved Glycoprotein (cGP) region of a positive RNA strand of Ebola virus, a cGP region of a negative RNA strand of Ebola virus, a survivin mRNA, and a bcl 2 mRNA, and
an endoribonuclease-oligonucleotide complex comprising an endoribonuclease coupled to an alkylthiol-terminated, propargyl ether modified poly-thymine polynucleotide, wherein the alkylthiol-terminated, propargyl ether modified poly-thymine polynucleotide comprises one or more propargyl ether groups coupled via an amidohexylacrylamido linker to the 5′ position of one or more thymine bases, and
wherein at least some of the propargyl ether groups on the alkylthiol-terminated propargyl ether modified poly-thymine polynucleotide of the endoribonuclease-oligonucleotide complex, of the ssDNA recognition moiety, or of the endoribonuclease-oligonucleotide complex and the ssDNA recognition moiety are crosslinked.

2. The nanozyme of claim 1 , wherein the shell further comprises a block copolymer comprising polyethylene glycol and lysine.

3. The nanozyme of claim 2 , wherein the block copolymer is methoxy-PEG-block-poly(L-lysine) hydrobromide.

4. The nanozyme of claim 1 , wherein the shell is substantially solid.

5. The nanozyme of claim 1 , wherein the shell is porous.

6. The nanozyme of claim 1 , further comprising a protecting moiety, wherein the protecting moiety is attached to the shell of the nanozyme, and
wherein the protecting moiety is selected from the group consisting of: a DNA oligonucleotide, a locked nucleic acid, a peptide nucleic acid, a poly(ethylene glycol), a poly(vinyl alcohol), a poly(acrylic acid), a poly(propylene fumarate-co-ethylene glycol), a polyacrylamide, a polypeptide, a poly-N-substituted glycine oligomer, a hyaluronic acid, an alginate, a chitosan, an agarose, a collagen, a fibrin, a gelatin, a dextran, and any combination thereof.

7. The nanozyme of claim 1 , further comprising an intra- or inter-cellular traffic guiding moiety, wherein the intra- or inter-cellular traffic guiding moiety is attached to the shell of the nanozyme.

8. The nanozyme of claim 1 , further comprising an allosterically functional moiety, wherein the allosterically functional moiety is attached to the shell of the nanozyme.

9. The nanozyme of claim 1 , wherein the shell further comprises a polymer layer, wherein the polymer layer is attached to the recognition moiety, the enzyme or both the recognition moiety and the enzyme.

10. The nanozyme of claim 9 , wherein the polymer layer comprises a plurality of pores.

11. The nanozyme of claim 9 , wherein the polymer layer consists of a polymer selected from the group consisting of: of a polyalkyne, a polyolefin, a polyisoprene, a polyamide, a polyester, a polycarbonate, a silicone, co-polymers thereof, and combinations thereof.

12. The nanozyme of claim 1 , further comprising an active agent, wherein the agent is contained in the hollow core.

13. The nanozyme of claim 12 , wherein the agent is a therapeutic agent.

14. The nanozyme of claim 13 , wherein the agent is selected from the group consisting of: a small molecule, a peptide, a nucleic acid, a polymer, and combinations thereof.

15. The nanozyme of claim 1 , wherein the agent is an imaging agent.

16. The nanozyme of claim 15 , wherein the imaging agent is selected from the group consisting of: a dye, a metal oxide nanoparticle, a metal nanoparticle, a semiconductor ananoparticle quantum dots, and a diamond nanoparticle, and combinations thereof.

17. The nanozyme of claim 1 , further comprising an imaging agent, wherein the imaging agent is attached to the shell.

18. The nanozyme of claim 17 , wherein the imaging agent is selected from the group consisting of: a dye, a metal oxide nanoparticle, a metal nanoparticle, a semiconductor a nanoparticle quantum dots, and a diamond nanoparticle, and combinations thereof.

19. The nanozyme of claim 1 , wherein the endoribonuclease is selected from the group consisting of: RNA RNase A, RNase III, RNase H, RNase P, RNase T1, and combinations thereof.

20. A pharmaceutical formulation comprising:
a nanozyme, wherein the nanozyme comprises:
a shell surrounding a hollow core configured to receive a compound, wherein the shell comprises:
a single stranded DNA (ssDNA) recognition moiety selected from the group consisting of: a polynucleotide set forth in SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 16, SEQ ID NO: 17, and combinations thereof, wherein the ssDNA recognition moiety comprises an alkylthiol-terminated propargyl ether modified poly-thymine polynucleotide; wherein the alkylthiol-terminated, propargyl ether modified poly-thymine polynucleotide comprises one or more propargyl ether groups coupled via an amidohexylacrylamido linker to the 5′ position of one or more thymine bases, and wherein the ssDNA recognition moiety specifically binds, through reverse complementary binding, a polynucleotide selected from the group consisting of: a c-MET mRNA, a conserved Glycoprotein (cGP) region of a positive RNA strand of Ebola virus, a cGP region of a negative RNA strand of Ebola virus, a survivin mRNA, and a bcl 2 mRNA;

an endoribonuclease-oligonucleotide complex comprising an endoribonuclease coupled to an alkylthiol-terminated, propargyl ether modified poly-thymine polynucleotide, wherein the alkylthiol-terminated, propargyl ether modified poly-thymine polynucleotide comprises one or more propargyl ether groups coupled via an amidohexylacrylamido linker to the 5′ position of one or more thymine bases, and
wherein at least some of the propargyl ether groups on the alkylthiol-terminated propargyl ether modified poly-thymine polynucleotide of the endoribonuclease-oligonucleotide complex, of the ssDNA recognition moiety, or of the endoribonuclease-oligonucleotide complex and the ssDNA recognition moiety are crosslinked; and
a pharmaceutically acceptable carrier.

21. A method comprising:
administering a nanozyme to a subject, wherein the nanozyme comprises a shell surrounding a hollow core configured to receive a compound, wherein the shell comprises:
a single stranded DNA (ssDNA) recognition moiety selected from the group consisting of: a polynucleotide set forth in SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 16, SEQ ID NO: 17, and combinations thereof, wherein the ssDNA recognition moiety is functionalized with comprises an alkylthiol-terminated propargyl ether modified poly-thymine polynucleotide; wherein the alkylthiol-terminated, propargyl ether modified poly-thymine polynucleotide comprises one or more propargyl ether groups c

Assignees

• Univ Florida

Inventors

• Cao Yunwei Charles
• Liu Chen

Classifications

• A61K38/43

  Enzymes; Proenzymes; Derivatives thereof · CPC title

• A61K47/6923

  the form being an inorganic particle, e.g. ceramic particles, silica particles, ferrite or synsorb · CPC title

• C12N11/14 Primary

  Enzymes or microbial cells immobilised on or in an inorganic carrier · CPC title

• A61K47/62

  the modifying agent being a protein, peptide or polyamino acid · CPC title

• A61K9/5115

  Inorganic compounds · CPC title