# US 12,691,166 B2 · Antiviral vaccines using spherical nucleic acids

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- Number: US 12,691,166 B2
- Document ID: US12691166B2
- Status: Granted
- Kind: B2
- Application number: 17/684,269
- Filed: 2022-03-01
- Published: 2022-09-15
- Granted: 2026-07-28
- Earliest claimed: 2021-03-12
- Assignees: Northwestern University
- Applicants: Northwestern University
- Inventors: Chad A. Mirkin; Michelle Hope Teplensky; Max Everett Distler; Caroline Danielle Kusmierz; Cassandra Elizabeth Callmann
- Web page: https://patentgrep.com/patent/US12691166B2

## CPC

- A61K 39/001192: {Glycoprotein 100 [Gp100]}
- A61K 39/12: Viral antigens
- A61K 39/39: characterised by the immunostimulating additives, e.g. chemical adjuvants
- A61P 35/00: Antineoplastic agents
- C12N 15/1138: {against receptors or cell surface proteins}
- A61K 2039/55555: {Liposomes; Vesicles, e.g. nanoparticles; Spheres, e.g. nanospheres; Polymers}
- A61K 2039/55561: {CpG containing adjuvants; Oligonucleotide containing adjuvants}
- C12N 2770/20034: Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein
- A61K: PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES (devices or methods specially adapted for bringing pharmaceutical products into particular physical or administering forms A61J3/00; chemical aspects of, or use of materials for deodorisation of air, for disinfection or sterilisation, or for bandages, dressings, absorbent pads or surgical articles A61L; soap compositions C11D)
- A61P: SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)

## Abstract

Spherical Nucleic Acids (SNAs) are chemically well-defined nanoscale architectures comprised of nucleic acids densely arranged around a spherical nanoparticle core in a highly oriented fashion. SNAs are highly modular structures produced by chemical synthesis and programmed assembly, allowing for their rapid modification to incorporate novel viral antigens. The present disclosure provides a SNA comprising (a) a nanoparticle core; (b) a shell of oligonucleotides attached to the external surface of the nanoparticle core, the shell of oligonucleotides comprising one or more immunostimulatory oligonucleotides; and (c) a viral antigen encapsulated in the nanoparticle core.

## Claims

### Claim 1

A spherical nucleic acid (SNA) comprising:
(a) a nanoparticle core, wherein the nanoparticle core is a liposomal core or a lipid nanoparticle core;
(b) a shell of oligonucleotides attached to the external surface of the nanoparticle core, the shell of oligonucleotides comprising one or more immunostimulatory oligonucleotides; and
(c) a coronavirus antigen or a nucleic acid that encodes a coronavirus antigen encapsulated in the nanoparticle core.

### Claim 2 (depends on 1)

The SNA of claim 1, wherein the coronavirus antigen is receptor binding domain (RBD) (SEQ ID NO: 4), S1 subunit of Spike (SEQ ID NO: 5), SARS-COV-2 Spike (SEQ ID NO: 2), a variant of any of the foregoing, or a combination thereof.

### Claim 3 (depends on 1)

The SNA of claim 1, wherein the coronavirus antigen comprises or consists of a sequence that is at least 80% identical to receptor binding domain (RBD) (SEQ ID NO: 4), S1 subunit of Spike (SEQ ID NO: 5), SARS-COV-2 Spike (SEQ ID NO: 2), SARS-COV-2 surface glycoprotein (SEQ ID NO: 14), SARS-COV-2 RBD (SEQ ID NO: 15), SARS-COV-2 Omicron variant RBD (SEQ ID NO: 16), RBD (SEQ ID NO: 17), or a combination thereof.

### Claim 4 (depends on 1)

The SNA of claim 1, wherein at least about 0.5 milligram (mg) of the coronavirus antigen is encapsulated in the nanoparticle core per micromole (μmol) of oligonucleotides in the shell of oligonucleotides.

### Claim 5 (depends on 4)

The SNA of claim 4, wherein about 0.5 mg to about 10 mg of the coronavirus antigen is encapsulated in the nanoparticle core per micromole (μmol) of oligonucleotides in the shell of oligonucleotides.

### Claim 6 (depends on 1)

The SNA of claim 1, wherein the nanoparticle core is a lipid nanoparticle core.

### Claim 7 (depends on 1)

The SNA of claim 1, wherein the shell of oligonucleotides comprises DNA, RNA, or a combination thereof.

### Claim 8 (depends on 1)

The SNA of claim 1, wherein the nanoparticle core comprises DNA, RNA, or a combination thereof encapsulated therein.

### Claim 9 (depends on 1)

The SNA of claim 1, wherein the ratio of oligonucleotides in the shell of oligonucleotides to the coronavirus antigen encapsulated in the nanoparticle core is about 10:1 to about 70:1 (number of oligonucleotides in the shell of oligonucleotides to the number of coronavirus antigen molecules encapsulated in the nanoparticle core).

### Claim 10 (depends on 9)

The SNA of claim 9, wherein the ratio of oligonucleotides in the shell of oligonucleotides to the coronavirus antigen encapsulated in the nanoparticle core is about 16:1.

### Claim 11 (depends on 9)

The SNA of claim 9, wherein the ratio of oligonucleotides in the shell of oligonucleotides to the coronavirus antigen encapsulated in the nanoparticle core is about 32:1.

### Claim 12 (depends on 9)

The SNA of claim 9, wherein the ratio of oligonucleotides in the shell of oligonucleotides to the coronavirus antigen encapsulated in the nanoparticle core is about 43:1.

### Claim 13 (depends on 1)

The SNA of claim 1, wherein the shell of oligonucleotides comprises a targeting oligonucleotide, an inhibitory oligonucleotide, or a combination thereof.

### Claim 14 (depends on 1)

The SNA of claim 1, wherein the immunostimulatory oligonucleotide is a CpG-motif containing oligonucleotide, a double-stranded DNA oligonucleotide, or a single-stranded RNA oligonucleotide.

### Claim 15 (depends on 6)

The SNA of claim 6, wherein each of the immunostimulatory oligonucleotides is a toll-like receptor (TLR) agonist.

### Claim 16 (depends on 1)

A pharmaceutical formulation comprising the SNA of claim 1 and a pharmaceutically acceptable carrier or diluent.

### Claim 17 (depends on 1)

An antigenic composition comprising the SNA of claim 1 in a pharmaceutically acceptable carrier, diluent, stabilizer, preservative, or adjuvant, wherein the antigenic composition is capable of generating an immune response including antibody generation or a protective immune response in a mammalian subject.

### Claim 18 (depends on 6)

The SNA of claim 6, wherein the nucleic acid is mRNA.

### Claim 19 (depends on 6)

The SNA of claim 6, wherein the lipid nanoparticle core comprises an ionizable lipid, a phospholipid, a sterol, and a lipid-polyethylene glycol (lipid-PEG) conjugate.

### Claim 20 (depends on 19)

The SNA of claim 19, wherein each oligonucleotide in the shell of oligonucleotides is covalently attached to the exterior of the lipid nanoparticle core through the lipid-PEG conjugate.

### Claim 21 (depends on 1)

The SNA of claim 1, wherein the nanoparticle core is a liposomal core.

### Claim 22 (depends on 21)

The SNA of claim 21, wherein the liposomal core comprises a plurality of lipid groups.

### Claim 23 (depends on 22)

The SNA of claim 22, wherein the plurality of lipid groups comprises a lipid selected from the group consisting of the phosphatidylcholine, phosphatidylglycerol, and phosphatidylethanolamine families of lipids.

### Claim 24 (depends on 1)

The SNA of claim 1, wherein at least one oligonucleotide in the shell of oligonucleotides is attached to the exterior of the liposomal or lipid nanoparticle core through a lipid anchor group.

### Claim 25 (depends on 24)

The SNA of claim 24, wherein the lipid anchor group is attached to the 5′ end or the 3′ end of the at least one oligonucleotide.

### Claim 26 (depends on 24)

The SNA of claim 24, wherein the lipid anchor group is tocopherol or cholesterol.

### Claim 27 (depends on 1)

The SNA of claim 1, wherein the shell of oligonucleotides comprises DNA oligonucleotides and RNA oligonucleotides.

### Claim 28 (depends on 1)

The SNA of claim 1, wherein the shell of oligonucleotides comprises single-stranded DNA, double-stranded DNA, single-stranded RNA, double-stranded RNA, or a combination thereof.

### Claim 29 (depends on 6)

The SNA of claim 6, wherein the nanoparticle core comprises single-stranded DNA, double-stranded DNA, single-stranded RNA, double-stranded RNA, or a combination thereof encapsulated therein.

### Claim 30 (depends on 1)

The SNA of claim 1, wherein the shell of oligonucleotides comprises at least 200 oligonucleotides.

### Claim 31 (depends on 1)

The SNA of claim 1, wherein the shell of oligonucleotides is attached to the external surface of the nanoparticle core at a surface density of about 2 pmol/cm2 to about 200 pmol/cm2.

### Claim 32 (depends on 6)

The SNA of claim 6, wherein each oligonucleotide in the shell of oligonucleotides is about 10 to about 50 nucleotides in length.

### Claim 33 (depends on 32)

The SNA of claim 32, wherein each oligonucleotide in the shell of oligonucleotides is about 20 to about 30 nucleotides in length.

### Claim 34 (depends on 6)

The SNA of claim 6, wherein diameter of the SNA is about 10 nanometer (nm) to about 150 nm.

### Claim 35 (depends on 1)

The SNA of claim 1, wherein diameter of the SNA is less than or equal to about 50 nanometers.

### Claim 36 (depends on 6)

The SNA of claim 6, wherein each oligonucleotide in the shell of oligonucleotides is an immunostimulatory oligonucleotide.

### Claim 37 (depends on 15)

The SNA of claim 15, wherein the TLR is toll-like receptor 9 (TLR9).

### Claim 38 (depends on 17)

A method of producing an immune response to a coronavirus antigen in a subject, comprising administering to the subject an effective amount of the antigenic composition of claim 17, thereby producing an immune response to the viral antigen in the subject.

### Claim 39 (depends on 1)

A method of treating a coronavirus infection in a subject in need thereof, comprising administering to the subject an effective amount of the SNA of claim 1, thereby treating the coronavirus infection in the subject.

### Claim 40 (depends on 38)

The method of claim 38, wherein the administering is intravenous, intraperitoneal, intranasal, subcutaneous, or intramuscular.

### Claim 41 (depends on 38)

The method of claim 38, wherein the administering comprises at least one dose of the antigenic composition.