BROAD-SPECTRUM MULTI-ANTIGEN PAN-CORONAVIRUS VACCINE
Waning immunity induced by first-generation Spike-alone-based COVID-19 has failed to prevent immune escape by many variants of concern (VOCs) that emerged from 2020 to 2024, resulting in a prolonged COVID-19 pandemic. Thus, a next-generation Coronavirus (CoV) vaccine incorporating highly conserved non-Spike SARS-COV-2 antigens is described herein. Conserved non-Spike T cell antigens in combination with a Spike antigen encapsulated in lipid nanoparticles: (i) Induced high frequencies of lung-resident antigen-specific CXCR5+CD4+ T follicular helper cells, GzmB+CD4+ and GzmB+CD8+ cytotoxic T cells, and CD69+IFN-γ+TNFα+CD4+ and CD69+IFN-γ+TNFα+CD8+ effector T cells; and (ii) Reduced viral load and COVID-19-like symptoms caused by various VOCs. The combined antigen/LNP-based pan-CoV vaccine could be rapidly adapted for clinical use to confer broader cross-protective immunity against emerging highly mutated and pathogenic VOCs.
1 .- 21 . (canceled)
22 . A composition comprising a lipid nanoparticle, wherein the lipid nanoparticle comprises a cationic lipid, a PEG-modified lipid, a sterol and a non-cationic lipid, wherein the lipid nanoparticle comprises a first messenger ribonucleic acid (mRNA) encapsulated therein and a second MRNA encapsulated therein, wherein:
i. the first mRNA comprises an open reading frame encoding a single Coronavirus NSP14 protein encoded by SEQ ID NO:4 or SEQ ID NO:5 and the second mRNA comprises an open reading frame encoding a single Coronavirus Nucleoprotein encoded by SEQ ID NO:7 or SEQ ID NO: 8;
ii. 2) the first mRNA comprises an open reading frame encoding a single Coronavirus NSP14 protein encoded by SEQ ID NO:4 or SEQ ID NO:5, and the second mRNA comprises an open reading frame encoding a single Coronavirus NSP2 protein encoded by SEQ ID NO:1 or SEQ ID NO: 2; or
iii. 3) the first mRNA comprises an open reading frame encoding a single Coronavirus NSP2 protein encoded by SEQ ID NO:1 or SEQ ID NO:2 and the second mRNA comprises an open reading frame encoding a single Coronavirus Nucleoprotein encoded by SEQ ID NO:7 or SEQ ID NO: 8.
23 . The composition of claim 22 , wherein each mRNA further comprises a 5′ untranslated region (UTR) and a 3′ UTR.
24 . The composition of claim 22 , wherein each mRNA further comprises a poly(A) tail and a 5′ cap.
25 . The composition of claim 22 , wherein the cationic lipid is an ionizable cationic lipid, the non-cationic lipid is a neutral lipid, and the sterol is a cholesterol.
26 . The composition of claim 25 , wherein the cationic lipid is selected from 2,2-dilinoleyl-4-dimethylaminoethyl-[1,3]-dioxolane, dilinoleyl-methyl-4-dimethylaminobutyrate, or di((Z)-non-2-en-1-yl) 9-((4-(dimethylamino)butanoyl)oxy)heptadecanedioate.
27 . The composition of claim 22 , wherein the lipid nanoparticle further comprises a third mRNA, wherein:
the first mRNA comprises an open reading frame encoding a single Coronavirus NSP14 protein encoded by SEQ ID NO:4 or SEQ ID NO:5;
the second mRNA comprises an open reading frame encoding a single Coronavirus Nucleoprotein encoded by SEQ ID NO:7 or SEQ ID NO: 8; and
the third mRNA comprises an open reading frame encoding a single Coronavirus NSP2 protein encoded by SEQ ID NO:1 or SEQ ID NO: 2.
28 . The composition of claim 27 , wherein each mRNA further comprises a 5′ untranslated region (UTR) and a 3′ UTR.
29 . The composition of claim 27 , wherein each mRNA further comprises a poly(A) tail and a 5′ cap.
30 . The composition of claim 27 , wherein the cationic lipid is an ionizable cationic lipid, the non-cationic lipid is a neutral lipid, and the sterol is a cholesterol.
31 . The composition of claim 30 , wherein the cationic lipid is selected from 2,2-dilinoleyl-4-dimethylaminoethyl-[1,3]-dioxolane, dilinoleyl-methyl-4-dimethylaminobutyrate, or di((Z)-non-2-en-1-yl) 9-((4-(dimethylamino)butanoyl)oxy)heptadecanedioate.
32 . A composition, comprising a first lipid nanoparticle and a second lipid nanoparticle, wherein:
i. the first lipid nanoparticle comprises a cationic lipid, a PEG-modified lipid, a sterol, and a non-cationic lipid, a first messenger ribonucleic acid (mRNA) encapsulated therein, and the first mRNA comprises an open reading frame encoding a single Coronavirus NSP14 protein encoded by SEQ ID NO:4 or SEQ ID NO:5, and the second lipid nanoparticle comprises a cationic lipid, a PEG-modified lipid, a sterol, and a non-cationic lipid, a second messenger ribonucleic acid (mRNA) encapsulated therein, and the second mRNA comprises an open reading frame encoding a single Coronavirus Nucleoprotein encoded by SEQ ID NO:7 or SEQ ID NO:8;
ii. the first lipid nanoparticle comprises a cationic lipid, a PEG-modified lipid, a sterol, and a non-cationic lipid, a first messenger ribonucleic acid (mRNA) encapsulated therein, and the first mRNA comprises an open reading frame encoding a single Coronavirus NSP14 protein encoded by SEQ ID NO:4 or SEQ ID NO:5, and the second lipid nanoparticle comprises a cationic lipid, a PEG-modified lipid, a sterol, and a non-cationic lipid, a second messenger ribonucleic acid (mRNA) encapsulated therein, and the second mRNA comprises an open reading frame encoding a single Coronavirus NSP2 protein encoded by SEQ ID NO: 1 or SEQ ID NO:2; or
iii. the first lipid nanoparticle comprises a cationic lipid, a PEG-modified lipid, a sterol, and a non-cationic lipid, a first messenger ribonucleic acid (mRNA) encapsulated therein, and the first mRNA comprises an open reading frame encoding a single Coronavirus NSP2 protein encoded by SEQ ID NO:1 or SEQ ID NO:2, and the second lipid nanoparticle comprises a cationic lipid, a PEG-modified lipid, a sterol, and a non-cationic lipid, a second messenger ribonucleic acid (mRNA) encapsulated therein, and the second mRNA comprises an open reading frame encoding a single Coronavirus Nucleoprotein encoded by SEQ ID NO:7 or SEQ ID NO:8.
33 . The composition of claim 32 , wherein each mRNA further comprises a 5′ untranslated region (UTR) and a 3′ UTR.
34 . The composition of claim 32 , wherein each mRNA further comprises a poly(A) tail and a 5′ cap.
35 . The Coronavirus vaccine composition of claim 32 , wherein the cationic lipid is an ionizable cationic lipid, the non-cationic lipid is a neutral lipid, and the sterol is a cholesterol.
36 . The Coronavirus vaccine composition of claim 35 , wherein the cationic lipid is selected from 2,2-dilinoleyl-4-dimethylaminoethyl-[1,3]-dioxolane, dilinoleyl-methyl-4-dimethylaminobutyrate, or di((Z)-non-2-en-1-yl) 9-((4-(dimethylamino)butanoyl)oxy)heptadecanedioate.
37 . The composition of claim 32 , wherein the composition further comprises a third lipid nanoparticle, wherein:
the first lipid nanoparticle comprises a cationic lipid, a PEG-modified lipid, a sterol, and a non-cationic lipid, a first messenger ribonucleic acid (mRNA) encapsulated therein, and the first mRNA comprises an open reading frame encoding a single Coronavirus NSP14 protein encoded by SEQ ID NO:4 or SEQ ID NO:5;
the second lipid nanoparticle comprises a cationic lipid, a PEG-modified lipid, a sterol, and a non-cationic lipid, a second messenger ribonucleic acid (mRNA) encapsulated therein, and the second mRNA comprises an open reading frame encoding a single Coronavirus Nucleoprotein encoded by SEQ ID NO:7 or SEQ ID NO:8; and
the third lipid nanoparticle comprises a cationic lipid, a PEG-modified lipid, a sterol, and a non-cationic lipid, a third messenger ribonucleic acid (mRNA) encapsulated therein, and the third mRNA comprises an open reading frame encoding a single Coronavirus NSP2 protein encoded by SEQ ID NO: 1 or SEQ ID NO:2.
38 . The composition of claim 37 , wherein each mRNA further comprises a 5′ untranslated region (UTR) and a 3′ UTR.
39 . The composition of claim 37 , wherein each mRNA further comprises a poly(A) tail and a 5′ cap.
40 . The Coronavirus vaccine composition of claim 37 , wherein the cationic lipid is an ionizable cationic lipid, the non-cationic lipid is a neutral lipid, and the sterol is a cholesterol.
41 . The Coronavirus vaccine composition of claim 40 , wherein the cationic lipid is selected from 2,2-dilinoleyl-4-dimethylaminoethyl-[1,3]-dioxolane, dilinoleyl-methyl-4-dimethylaminobutyrate, or di((Z)-non-2-en-1-yl)9-((4-(dimethylamino)butanoyl)oxy)heptadecanedioate.