Antiviral vaccines using spherical nucleic acids
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.
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.
2 . 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.
3 . 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.
4 . 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.
5 . 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.
6 . The SNA of claim 1 , wherein the nanoparticle core is a lipid nanoparticle core.
7 . The SNA of claim 1 , wherein the shell of oligonucleotides comprises DNA, RNA, or a combination thereof.
8 . The SNA of claim 1 , wherein the nanoparticle core comprises DNA, RNA, or a combination thereof encapsulated therein.
9 . 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).
10 . 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.
11 . 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.
12 . 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.
13 . The SNA of claim 1 , wherein the shell of oligonucleotides comprises a targeting oligonucleotide, an inhibitory oligonucleotide, or a combination thereof.
14 . 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.
15 . The SNA of claim 6 , wherein each of the immunostimulatory oligonucleotides is a toll-like receptor (TLR) agonist.
16 . A pharmaceutical formulation comprising the SNA of claim 1 and a pharmaceutically acceptable carrier or diluent.
17 . 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.
18 . The SNA of claim 6 , wherein the nucleic acid is mRNA.
19 . 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.
20 . 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.
21 . The SNA of claim 1 , wherein the nanoparticle core is a liposomal core.
22 . The SNA of claim 21 , wherein the liposomal core comprises a plurality of lipid groups.
23 . 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.
24 . 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.
25 . 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.
26 . The SNA of claim 24 , wherein the lipid anchor group is tocopherol or cholesterol.
27 . The SNA of claim 1 , wherein the shell of oligonucleotides comprises DNA oligonucleotides and RNA oligonucleotides.
28 . 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.
29 . 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.
30 . The SNA of claim 1 , wherein the shell of oligonucleotides comprises at least 200 oligonucleotides.
31 . 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/cm 2 to about 200 pmol/cm 2 .
32 . The SNA of claim 6 , wherein each oligonucleotide in the shell of oligonucleotides is about 10 to about 50 nucleotides in length.
33 . The SNA of claim 32 , wherein each oligonucleotide in the shell of oligonucleotides is about 20 to about 30 nucleotides in length.
34 . The SNA of claim 6 , wherein diameter of the SNA is about 10 nanometer (nm) to about 150 nm.
35 . The SNA of claim 1 , wherein diameter of the SNA is less than or equal to about 50 nanometers.
36 . The SNA of claim 6 , wherein each oligonucleotide in the shell of oligonucleotides is an immunostimulatory oligonucleotide.
37 . The SNA of claim 15 , wherein the TLR is toll-like receptor 9 (TLR9).
38 . 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.
39 . 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.
40 . The method of claim 38 , wherein the administering is intravenous, intraperitoneal, intranasal, subcutaneous, or intramuscular.
41 . The method of claim 38 , wherein the administering comprises at least one dose of the antigenic composition.